Mandip Sachdeva

The Nuclear Receptor TR3 Regulates mTORC1 Signaling in Lung Cancer Cells Expressing Wild-type p53

The orphan nuclear receptor TR3 (NR41A and Nur77) is overexpressed in most lung cancer patients and is a negative prognostic factor for patient survival. The function of TR3 was investigated in non-small-cell lung cancer A549 and H460 cells, and knockdown of TR3 by RNA interference (siTR3) inhibited cancer cell growth and induced apoptosis. The prosurvival activity of TR3 was due, in part, to formation of a p300/TR3/ specificity protein 1 complex bound to GC-rich promoter regions of survivin and other Sp-regulated genes (mechanism 1). However, in p53 wild-type A549 and H460 cells, siTR3 inhibited the mTORC1 pathway, and this was due to activation of p53 and induction of the p53-responsive gene sestrin 2, which subsequently activated the mTORC1 inhibitor AMP-activated protein kinase α (AMPKα) (mechanism 2). This demonstrates that the pro-oncogenic activity of TR3 in lung cancer cells was due to inhibition of p53 and activation of mTORC1. 1,1-Bis(3’-indolyl)-1-(p-hydroxyphenyl)methane (DIM-C-pPhOH) is a recently discovered inhibitor of TR3, which mimics the effects of siTR3. DIM-C-pPhOH inhibited growth and induced apoptosis in lung cancer cells and lung tumors in murine orthotopic and metastatic models, and this was accompanied by decreased expression of survivin and inhibition of mTORC1 signaling, demonstrating that inactivators of TR3 represent a novel class of mTORC1 inhibitors.

Drug targeting systems for cancer chemotherapy.

Tumour specific drug targeting has been a very actively investigated area for over 2 decades. Various approaches have involved the use of drug delivery systems that can localise the anticancer agent at the tumour site without damaging the normal cells. For this purpose, various delivery systems that have been utilised are liposomes, microspheres and recently, nanoparticles. Two liposome formulations containing anticancer drugs for example, adriamycin and daunomycin are already on the market in the USA and Europe. Microspheres are also being investigated for delivering various anticancer drugs and protein/peptides for anticancer treatment, and several formulations are in Phase I/II clinical trials. Antitumour drugs have also been linked to tumour specific monoclonal antibodies via various chemical linkages. Doxorubicin was linked to a chimeric monoclonal antibody that was targeted to the Lewis Y antigen. Though this conjugate initially showed potential, it was recently dropped from Phase II clinical trials. Another approach with monoclonal antibodies has been the use of immunotoxins. Immunotoxins initially showed promise as potential anticancer agents at picomolar concentrations but several clinical and preclinical studies have not shown much promise in this regard. Drug containing liposomes and microspheres have been further linked to tumour specific monoclonal antibodies to enhance their tumour specificity. Most of the studies with immunoliposomes or targeted microspheres have not gone beyond the preclinical studies. New therapeutic approaches are presently emerging based on natural products like cytokines, peptide growth factor antagonists, antisense oligonucleotides and specific genes. These approaches need the help of delivery systems to deliver these complex molecules to tumour cells. One of the current pursued approaches is the use of cationic liposomes. Several clinical studies are undergoing with various cationic liposomes and the next few years will demonstrate the usefulness of this approach. In recent years, the problems in cancer treatment have been complicated with the emergence of resistance strains leading to resistant and cross-resistant tumour cells. Several agents have been used to overcome or reverse drug-resistance in solid tumours and it remains a highly pursued area in cancer treatment.

Abstract 3738: Knockdown of Annexin A2 by liposomal formulation of Annexin A2 small hairpin RNA induces apoptosis and inhibits angiogenesis in lung cancer stem cells.

Proceedings: AACR 104th Annual Meeting 2013; Apr 6-10, 2013; Washington, DC Recent studies support the existence of small populations of pluripotent stem-like cancer cells capable of self-renewal. These cancer stem cells (CSCs) are attributed to the resistance phenotype occurring in many tumors such as invasion and metastasis. Annexin A2 (Anxa2) overexpression is associated with invasion, metastasis and drug resistance in many cancers. We investigated the effect of Anxa2 knockdown by a liposomal (nanosome) formulation of Anxa2 shRNA (shAnxa2) on growth and metastasis in H1650 CSCs. H1650 CSCs were maintained on basement membrane-coated supports in DMEM:F12 media enriched with growth factors. Nanosomes were prepared by solvent evaporation and bath sonication. H1650 CSCs were transfected with lipoplexes of shAnxa2 and varying amounts of nanosome in serum-free media. After 4 hrs of incubation, the medium was replaced with regular media and knockdown of Anxa2 analyzed after 48 hrs by Western blot. Inhibition of H1650 CSC sphere formation over 7 days was investigated in lipidure® plates following treatment with lipoplexes for 4 hrs. CSCs were assessed for induction of apoptosis after treatment with lipoplexes (4 hrs) by acridine orange-ethidium bromide staining. The effect of Anxa2 lipoplexes on proliferation, migration and capillary-like tube formation was investigated in human primary pulmonary artery endothelial cells (HPAEC). The effect of Anxa2 lipoplexes on the expression of Anxa2, EGFR, VEGF, and NFkβ was evaluated by Western blot. Efficient loading of shAnxa2 in nanosomes was achieved with high transfection efficiency compared to lipofectamine. Anxa2 shRNA lipoplex was cytotoxic to H1650 CSC spheroids, inhibiting spheroid growth by 30-60% between day 3 and 7. Early and late apoptosis induction was observed after treatment with shAnxa2 with a 2-fold increase of cells undergoing apoptosis compared to shAnxa2-lipofectamine 2000 complex. Anxa2 knockdown (90%) was observed in H1650 CSCs parallel to decreases in EGFR, VEGF, and NFkβ expression as shown by Western blot. High loading and enhanced transfection of shAnxa2 was achieved with the nanosome formulation. Anxa2 knockdown was achieved in parallel with inhibition of H1650 CSC spheroid growth, angiogenesis and induction of apoptosis. These results altogether support evidence that Anxa2 could be a therapeutic target for treatment of non-small cell lung cancer and that nanosome formulation enhances delivery of shAnxa2 as an anticancer agent. Citation Format: Terrick A. Andey, Srujan Marepally, Pomila Singh, Mandip S. Sachdeva. Knockdown of Annexin A2 by liposomal formulation of Annexin A2 small hairpin RNA induces apoptosis and inhibits angiogenesis in lung cancer stem cells. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3738. doi:10.1158/1538-7445.AM2013-3738

Abstract 2039: The use of self nano-emulsifying drug delivery systems to improve bioavailability of combination therapy and overcome erlotinib resistance in non-small cell lung cancer

Background:Erlotinib, an EGFR selective reversible inhibitor, improved response of patients diagnosed with non-small cell lung cancer (NSCLC) with 70% of patients showing significant tumor regression upon treatment. Unfortunately,most patients eventually relapse due to development of resistance. Our research goal is to quantify the effects of combination treatment of erlotinib with a glycyrrhetinic acid analogs and to improve the oral bioavailability of this treatment using self nano-emulsifying drug delivery systems. Methods: NSCLC cell lines HCC827, HCC827 (4µM erlotinib resistant), HCC827 Cl4 (second site EGFR mutated), HCC827BEAS2B, and H1975 (4µMerlotinib resistant) were treated in combination with CDODA-Me, CF3CDODA-Me and Erlotinib. The cell viability assays were performed and combination index (CI) values were calculated by isobolographic analysis. Self nano-emulsifying drug delivery systems (SNEDDs) were formulated and characterized through in vitro and in vivo studies. Results: CF3CDODA and improved efficacy for erlotinib in all cell lines with IC50 values of 6.0µM, 7.8µM, 4.6µM and 4.2±1.56µM for HCC827(RESISTANT), H1975(RESISTANT), HCC827CL4 and HCC827 respectively compared to IC50values of erlotinib alone (25µM, 21µM, 23µM, and 8µM). This was superior when compared to IC50 values of CDODA-Me combination treatment (6.66µM, 7.32µM, 12.06µM, and 5.45µM respectively). Combination treatment showed strong synergism with a constant concentration of 0.5µM CF3CDODA and CI values of 0.461, 0.548, 0.389, 0.550 for HCC827 4µM, H1975µM, HCC827CL4, and HCC827 respectively and antagonism in HCC827BEAS2B (1.508) cells. This is comparable to the synergism observed at a minimum concentration of 2µM CDODA-Me with CI values of 0.337, 0.67, 0.64, and 0.657 respectively and antagonism CI of 1.175 for HCC827BEAS2B. CF3CDODA Combination therapy showed a 2.5-fold decrease in colony formation in resistant cells making them comparable to wild type. This was comparable to CDODA-Me combination treatment with a 1.46-fold decrease in colony formation. Both combination treatments showed little effect on HCC827BEAS2B. Western Blot analysis showed decrease MET expression and decreased apoptosis inhibiting markers BCL2 and Survivin. SNEDD formulations increased maximum drug plasma concentrations for Erlotinib (281ng/ml to 442ng/ml) and circulating time as well (12hrs - 21hrs). Conclusions: Based on the results of these studies, CDODA-Me and CF3CDODA have shown to overcome erlotinib resistance in non-small cell lung cancer and when combining these treatments with the use of SNEDDS, they have shown to be a promising treatment regimen. Citation Format: Ebony Nottingham, Vasanth Sekar, Arindam Mondal, Mandip Sachdeva. The use of self nano-emulsifying drug delivery systems to improve bioavailability of combination therapy and overcome erlotinib resistance in non-small cell lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2039. doi:10.1158/1538-7445.AM2017-2039

Abstract 2086: Noscapine chemosensitization enhances docetaxel anticancer activity and tumor stroma disruption against triple negative breast cancer

Purpose: The use of Noscapine (Nos) as a chemosensitizer followed by docetaxel (DTX) treatment therapy could be a novel approach for the treatment for breast cancer and possibly reduce the adverse side effects associated with DTX based chemotherapy. The goal of this study was to examine the chemo-sensitizing effect of Nos to DTX and also tumor stromal disruption effect of Nos in mice bearing xenograft TNBC tumors. Methods: Effect of Nos chemosensitization on DTX cytotoxicity was evaluated in MDA-MB-231 cells by trypan blue dye method. Apoptosis was measured by AnnexinV/FITC method using flow cytometer. Expression of different proteins like phospho-p38, pJNK, bcl-2, α-tubulin, Akt, pAkt, survivin was evaluated by immunoblot. Alpha-tubulin binding assay was done by fluorescent microscopy. In vivo antifibrotic efficacy of Nos and uptake of coumarin-6 loaded fluorescent liposomes was evaluated by picro-sirius red staining and fluorescent microscopy respectively in xenograft breast tumors. Results: MDA-MB-231 TNBC cells were exposed at sub-therapeutic dose of Nos (4 μM) which increased the cytotoxicity of DTX by 3.0-fold. Flow-cytometric analysis showed significant increase (30 percent) of late apoptotic cells in Nos chemosensitized, DTX-treated MDA-MB-231 cells compared with DTX alone treatment. Further, chemosensitization of TNBC cells with Nos at different time intervals (6 h, 12 h and 24 h), the effect on stress transducer p38 stress activator protein kinase was significantly activated (p Conclusion: In conclusion, chemosensitization with sub-therapeutic dose of oral noscapine could be a promising approach to increase anticancer activity of DTX and can further enhance uptake of liposomes suggesting that this approach may have potential in TNBC treatment. Citation Format: Ravi Doddapaneni, Ketan Patel, Nusrat Chowdhury, Mandip Sachdeva. Noscapine chemosensitization enhances docetaxel anticancer activity and tumor stroma disruption against triple negative breast cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 2086.

Abstract 4833: CDODA-Me augments the efficacy of chemotherapeutic agents and overcomes chemo-resistance in breast and lung cancer cells

Purpose: The purpose of this research is to determine the effects of the combination of (CDODA-Me), a derivative of glycyrrhetinic acid with chemotherapeutic agents like docetaxel (DTX) and Erlotinib (a TKI inhibitor) to treat triple negative breast cancer (TNBC) and TKI resistant lung cancer. Methods: TNBC lines MDA-MB-231 MDA-MB-468 and DTX resistant (MDA231) cells were treated in combination with CDODA-Me (nontoxic dose of 2 μM) and DTX. Wildtype (HCC827) and resistant (Erlotinib resistant) lung cancer cells were treated with Erlotinib and CDODA-Me. The cell viability of MDA-MB-231, MDA-MB-468, Docetaxel resistant (MDA231) cells and HCC 827 Erlotinib resistant (4 μM) cells in each treatment group was determined by crystal violet assay. Combination index values were calculated by isobolographic analysis. Western blot annalysis was used to investigate the influence of CDODA-Me combinations on drug resistance and key apoptotic proteins such as bcl2, survivin, SP1, SP3, and SP4. 2’,7’ -dichlorofluorescin diacetate (DCFDA) was used to measure reactive oxygen species (ROS) levels in all cell lines and examined by flowcytometry. Results: Breast Cancer cells (MDA-MB-468 and MDA-MB-231) showed increased cytotoxicity with a ten fold decrease in IC50 concentration (0.3 μM DTX and 0.03 μM DTX in combination with 2 μM CDODA-Me). DTX resistant (MDA231) cells showed IC50 values comparable to wildtype cells at 0.29 μM DTX in combination with CDODA-Me. HCC827 (Erlotinib resistant) showed a three fold decrease in cytotoxicity (IC 50 values of 4.9 μM and 16.9 μM Erlotinib for the combination and Erlotinib alone respectively). The combination treatment also showed higher response in resistant cells with IC 50 values comparable to wildtypes. The expression of bcl2, survivin, specific transcription factors (like SP1, SP3 and SP4) was downregulated in cells treated with CDODA-ME and DTX combination compared to CDODA-ME alone, DTX alone and control cells. Similar results were observed with CDODA-Me and Erlotinib combination in HCC827 erlotonib resistant (4 μM) cells. CDODA-Me alone treatment showed ROS levels in HCC 827 (4 μM) resistant cell lines were increased (8-fold) significantly as compared to other breast cancer cells. Conclusion: In conclusion, CDODA-Me inhibits growth of TNBC cells and Erlotinib resistant lung cancer cells and downregulates the SP proteins and anti-apoptotic proteins. Therefore, these results indicate that CDODA-Me is a promising anticancer agent and can overcome resistance of chemotherapeutic agents against breast and lung cancer. Citation Format: Ebony Nottingham, Ravi Doddapaneni, Mandip Sachdeva. CDODA-Me augments the efficacy of chemotherapeutic agents and overcomes chemo-resistance in breast and lung cancer cells. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4833.

Abstract 3139: Nurr1, a novel target of 1,1-bis(3’-indolyl)-1-(p-chlorophenyl) methane for inhibition of the initiation and progression of skin cancer tumorigenesis

Proceedings: AACR Annual Meeting 2014; April 5-9, 2014; San Diego, CA The objective of this study was to demonstrate the chemopreventive potential of 1,1-bis(3′-indolyl)-1-(p-chlorophenyl methane) (DIM-D) in skin cancer using an in vitro and in vivo models. In vitro cell cytotoxicity and viability assays were carried out in A431 human epidermoid carcinoma cell line (A431) and normal human epidermal keratinocytes (NHEK) respectively. Apoptosis induction and accumulation of ROS following UVB exposure in DIM-D pretreated NHEK cells (2 hr prior) was also evaluated. Immunocytochemistry and western blot analysis were performed to determine proapoptotic and proinflammatory markers expressions in DIM-D treated A431 cells and in UVB irradiated NHEK cells. For in vivo experiments, DIM-D was encapsulated within nanocarriers by hot melt homogenization using Nano DeBEE. Nanocarriers were surface modified with oleic acid and incorporated into a gel using established procedures (G-NOD) and characterized for particle size, zeta potential, entrapment efficiency, rheology, skin permeation and drug release. The chemo-preventive efficiency of G-NOD was evaluated using UVB-induced skin cancer model in SKH hairless mice and skin samples were collected and investigated for lipid peroxidation, protein carbonyls, and molecular markers by western blot. The IC50 values of DIM-D were 24.5±2.6, 17.2±3.6and 4.1±1.1 whilst for EGCG were 192.1±3.8, 85.3±2.4 and 26.0 ±1.4 for 24, 48 and 72 hr treatments respectively. DIM-D induced higher expression in A431 cells compared to EGCG of cleaved caspase 3 (3.0-fold vs. 2.4-fold changes), Nurr1 (2.7-fold vs. 1.7-fold changes) and NFκB (1.3-fold vs. 1.1-fold changes). Nanocarriers containing DIM-D were 188.00±8.00 nm in size with polydispersity of 0.59±0.01. The zeta potential and entrapment efficiency were 37.17±0.90mV and 89±0.50%, respectively. There was significant (p<0.05) reduction in tumor size/number for G-NOD pretreated group (2 tumors/mouse, average diameter, 2±0.50 mm) compared to DIM-PG (4 tumors/mouse, average diameter, 4±0.45mm) and EGCG PEG-gel (EGCG-PG; 6 tumors/mouse, average diameter, 5±0.25mm). IHC and western blot results indicated significant increase (p<0.05) in expression of Nurr1 in G-NOD pretreated group compared to EGCG-PG. There was however pronounced reduction in expression of STAT3, NFkB, cleaved Caspase-3, BCl2 and 8-OHdG for G-NOD pretreated group compared to DIM-PG and EGCG-PG. Lipid peroxidation assay revealed decrease in amount of Malondialehyde (MDA) produced in G-NOD pretreated group prior to UV exposure by 2 fold compared to DIM-PG and 1.3 fold compared to EGCG-PG treatments. Therefore, findings in both in vitro and in vivo studies suggest that the enhanced percutaneous delivery of DIM-D reduces UVB-induced damage to skin lipids and protein and inhibits initiation and progression of skin photocarcinogenesis in SKH mice via the transactivation of Nurr1. Citation Format: Ravi Doddapaneni, Cedar Boakye, Punit Shah, Apurva R. Patel, Chandraiah Godugu, Stephen Safe, Santosh Katiyar, Mandip Sachdeva. Nurr1, a novel target of 1,1-bis(3’-indolyl)-1-(p-chlorophenyl) methane for inhibition of the initiation and progression of skin cancer tumorigenesis. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3139. doi:10.1158/1538-7445.AM2014-3139

Abstract 3361: Pharmacokinetic and pharmacodynamic evaluation of novel dual channel spray dried enteric coated self-emulsifying capsules for anti-cancer agents in dogs.

Purpose: Oral administration of anti-cancer agents presents a series of advantages for patients. However, most of the anticancer agents are hydrophobic (poor water solubility) thus associated with low bioavailability. Self-emulsifying drug delivery systems (SEDDS), known to improve the oral absorption of highly lipophilic compounds, are normally prepared in a liquid form. However, solid-SEDDS has several advantages over conventional SEDDS. Thus we have incorporated spray drying for this purpose using customized spray gun system, which allows simultaneously/pulsatile flow of two different liquid systems through single nozzle. This modification allowed us to formulate Enteric coated SEDDS by using SEDDS as one liquid system and Enteric coating solution as second liquid system. Objective of this study was to design and evaluate this formulation in-vivo using DIM-C-pPhC6H5 (DIM-P), a novel anti-cancer agent, in dogs. Methods: SEDDS formulations were prepared as capsules and optimized based on pseudo-ternary phase diagram analysis as well as characteristics such as turbidity or phase separation prior to self-emulsification, emulsion droplet size and self-emulsification efficiency before/after spray drying. Evaluation for enteric coating was done by drug release of spray dried E-SEDDS. Pharmacokinetic and pharmacodynamic properties in dogs were determined following intravenous and oral administration of formulations. Pharmacokinetic parameters were determined using non-compartmental and compartmental techniques with WinNonlin® 5.0 software. Efficiency of anti-tumor activity was carried out using metastatic H1650 tumor model in nude mice using ‘M’ size capsules. Results: Three formulations of SEDDS were selected based on pseudo-ternary phase diagram and physicochemical properties of SEDDS. Droplet size of emulsion was in range of 5-10 μm. In-vitro drug release study showed 5 suggesting coating of SEDDS particles. Pharmacokinetic evaluation in dogs showed improved absorption of SEDDS formulations compare to solution; increased Cmax (39.18 ± 7.34 vs 21.68 ± 6.3 μg dL-1), higher AUC0-t (34481.34 ± 1125.46 vs 14159.53 ± 702.20 μg min dL-1). The relative oral bioavailability of SEDDS calculated on the basis of AUC0-t was about 43% as compared to solution. Also, plasma concentration vs time profile following intravenous administration and tissue distribution analysis confirmed three compartment distribution of DIM-P. SEDDS formulation showed ∼20-25% tumor volume and weight reduction in-vivo H1650 tumor models. Conclusion: The results emanating from these studies demonstrate a potential use of dual channel spray dried enteric coated self-emulsifying capsules for enhanced oral absorption and thus increased anti-cancer activity for treatment of Lung cancer. Citation Format: Apurva R. Patel, Chandraiah Godugu, Wilson R. Heather, Stephen Safe, Mandip S. Sachdeva. Pharmacokinetic and pharmacodynamic evaluation of novel dual channel spray dried enteric coated self-emulsifying capsules for anti-cancer agents in dogs. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3361. doi:10.1158/1538-7445.AM2013-3361

Abstract 2139: Effect of telmisartan on triple negative breast cancer (TNBC) and lung cancer tumor progression and intratumoral distribution of nanoparticles.

Proceedings: AACR 104th Annual Meeting 2013; Apr 6-10, 2013; Washington, DC Introduction: The intratumoral distribution of tumor targeted nanoparticles (NP) is hindered by dense collagen network and highly fibrous interstitium. Use of antifibrotic agents can decrease tumor interstitial fibrosis and promote NP intratumoral distribution. The objective of this study was to evaluate the effect of Telmisartan (TEL, AT1 blocker/PPAR-γ agonist) and Losartan (LOS, AT1 blocker) on NPs intratumoral distribution and anticancer effects in TNBC and lung cancer. Methods: Anticancer effect of TEL (5 mg/kg) and LOS (20 mg/kg) by inhalation and oral route were evaluated using Non-small cell lung cancer (A549 orthotopic and metastatic model) and triple negative breast cancer (TNBC, MDA-MB-231 orthotopic model) in Nu/nu mice. Also, to study intratumoral distribution, fluorescent polystyrene NP (FPNP) were administered intravenously followed by fluorescent microscopy. Tumor fibrous nature was characterized by estimating collagen-1, transforming growth factor beta1 (TGF-β;1) expression by IHC and ELISA. Collagen deposition in the tumor interstitium was studied by Massons Trichome staining. To correlate the AT1 receptor blockage to anticancer effects, expression of VEGF, cleaved caspase-3, MMP-9 and microvessel densities (CD31) were quantified by IHC and western blotting. Epithelial Mesenchymal transition (EMT) mediated tumor metastasis was characterized by expressions of E-Cadherin and Vimentin. Results: TEL showed significant antitumor effects in both lung and TNBC tumor models. TEL at 1/4th dose produced superior anticancer effects than LOS. Survival rate was extended with TEL compare to LOS & untreated control. Further, Paclitaxel-NP produced better tumor regression in inhalation with TEL compared to single agent treatment and control. The fibrous nature of lung tumors and TNBC was decreased significantly (p<0.05) with TEL and LOS treatment based on expression of collagen-1 and TGF-β;1. This allowed better FPNP distribution into the tumors compared to control. Further, compared to control tumors, LOS and TEL treated group resulted in 5.33 and 14.33 fold increase in NP distribution. Similarly, Paclitaxel-NP distribution was superior in combination with TEL & LOS treatment, which in turn exhibited synergistic anticancer effects. TEL treatment resulted in the decreased metastatic tumor nodules and tumor volumes. The metastatic markers MMP-9 & vimentin levels were significantly decreased (p<0.001) in TEL group compared to LOS & control, suggesting the promising effects of TEL in tumor anti-metastasis. Further, our study also demonstrated the safety of TEL upon inhalation route. Conclusion: Telmisartan showed significant anticancer effects in Lung cancer and TNBC. Telmisartan could be an ideal candidate for combination therapy to improve the NPs intratumoral distribution and anticancer effects in various cancer types. Citation Format: Chandraiah Godugu, Apurva R. Patel, Srujan Marepally, Ravi Doddapaneni, Mandip Singh Sachdeva, Mahavir B. Chougule. Effect of telmisartan on triple negative breast cancer (TNBC) and lung cancer tumor progression and intratumoral distribution of nanoparticles. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2139. doi:10.1158/1538-7445.AM2013-2139

Abstract 5511: Multifunctional CREKA peptide conjugated lipid nanocarriers of synergistically acting Noscapine and Doxorubicin for breast cancer therapy

Proceedings: AACR 101st Annual Meeting 2010‐‐ Apr 17‐21, 2010; Washington, DC Estrogen receptor negative (ER−) breast cancers (∼40%) are clinically aggressive with poor clinical outcome. Combination of different mechanism based antimicrotubular Noscapine and DNA intercalating Doxorubicin may lead to additive/synergistic activity against ER− breast cancer. Clinical utility of safer oral Noscapine (poor bioavailability and short half life) and Doxorubicin (cardiotoxicity and myelosuppression) has been limited. Encapsulation of Noscapine and Doxorubicin in nanocarriers whose surface is modified with pegylated CREKA peptide (MNCs) will significantly deliver nanocarriers to tumors by homing to tumor stroma, vessel wall and thereby releasing both drugs in controlled manner to exert anticancer activity. The purpose of this study was to encapsulate synergistically acting Noscapine and Doxorubicin in nanocarrier and modify the nanocarrier surface with CREKA and evaluate for anticancer activity in MDA-MB 231 ER− breast cancer cells. Isobolographic method and TUNEL assay were used to study Noscapine (10, 20 and 30 µM) and Doxorubicin interaction in MDA-MB 231 cells. For preparation of MNCs, Noscapine and Doxorubicin (molar ratio of 1:400) were dissolved in lipophilic phase composed of Miglyol (6% w/v), Compritol (3% w/v) and DOGS-NTA-Ni (0.2 % w/v). Lipophilic phase was poured to aqueous phase containing Polaxamer 188 (1.2 % w/v in water) and subjected to high-pressure homogenization to yield DOGS-NTA-Ni engrafted nanocarriers. Six-Histidine tagged PEG (1K)-CREKA (0.01-0.04 % w/v) was incubated with nanocarriers for 30 min for conjugation of DOGS-NTA-Ni engrafted nanocarrier with Histidine tagged peptide to yield MNCs. MNCs were characterized for size, drug release, antiproliferative and clot binding efficiency. Noscapine and Doxorubicin alone showed IC50 of 42 ± 4 µM and 0.25 ± 0.02 µM against MDA-MB cells respectively. In presence of Noscapine solution (20 µM), the IC50 of Doxorubicin solution was reduced to 0.05 µM (5-fold). Further, the combination Index values (< 0.6) and higher apoptotic cells (P 96 % of encapsulation and controlled release of both drugs (8 hr∼15 % and 48 hr∼ 60 %). A significantly (P < 0.01) higher binding of MNCs (CREKA concentration 0.045 % w/v) to the clotted plasma proteins showed the targeting ability of nanocarriers. MNCs showed similar IC50 values (20 µM Noscapine + 0.05 µM Doxorubicin) to that of solution combination. In conclusion, Noscapine acts synergistically with Doxorubicin and combination delivery of Noscapine and Doxorubicin using nanocarriers conjugated with CREKA showed significant increase in cytotoxicity with controlled drug release and significant binding efficiency. Multifunctional nanocarriers can effectively inhibit breast cancer and may reduce limitations associated with chemotherapy. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5511.