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Commentaries on this Article from A Novel Combinatorial Nanotechnology-Based Oral Chemopreventive Regimen Demonstrates Significant Suppression of Pancreatic Cancer Neoplastic Lesions

Pancreatic cancer is a deadly disease killing 37,000 Americans each year. Despite two decades of research on treatment options, the chances of survival are still less than 5% upon diagnosis. Recently, chemopreventive strategies have gained considerable attention as an alternative to treatment. We have previously shown significant in vitro chemopreventive effects with low-dose combinations of aspirin, curcumin, and sulforaphane (ACS) on pancreatic cancer cell lines. Here, we report the results of 24-week chemopreventive study with the oral administration of ACS combinations on the N-nitrosobis (2-oxopropyl) amine (BOP)-treated Syrian golden hamster model to suppress the progression of pancreatic intraepithelial neoplasms (PanIN) using unmodified (free drug) combinations of ACS, and nanoencapsulated (solid lipid nanoparticles; SLN) combinations of aspirin, curcumin, and free sulforaphane. The use of three different doses (low, medium, and high) of unmodified ACS combinations exhibited reduction in tumor incidence by 18%, 50%, and 68.7% respectively; whereas the modified nanoencapsulated ACS regimens reduced tumor incidence by 33%, 67%, and 75%, respectively, at 10 times lower dose compared with the free drug combinations. Similarly, although the unmodified free ACS showed a notable reduction in cell proliferation, the SLN encapsulated ACS regimens showed significant reduction in cell proliferation at 6.3%, 58.6%, and 72.8% as evidenced by proliferating cell nuclear antigen expression. Cell apoptotic indices were also upregulated by 1.5, 2.8, and 3.2 times, respectively, compared with BOP control. These studies provide a proof-of-concept for the use of an oral, low-dose, nanotechnology-based combinatorial regimen for the long-term chemoprevention of pancreatic cancer. Cancer Prev Res; 6(10); 1015–25. ©2013 AACR.

Colorectal cancer (CRC) is the third most common cause of cancer death in the United States, accounting for approximately 51,000 deaths each year. We have previously shown in vitro chemopreventive effects of mixtures of aspirin, folic acid, and calcium (AFAC) on colon cancer cell lines. The objective of the present study was to evaluate the in vivo effects of orally administered, colon targeted chemopreventive combination regimens on the inhibition of aberrant crypt foci (ACF) in a rat model of colon carcinogenesis using (i) unmodified (free drug) combinations of AFAC and (ii) nanoparticle-encapsulated combinations of the same agents. A 14-week animal study was conducted in three phases to determine an optimal effective dose from AFAC combinations and evaluate the efficacy of nanotechnology-based chemopreventive regimens administered in combined (mixtures) and individual (single entity) forms. ACF inhibition when compared with azoxymethane-treated rat control group was significant in both the unmodified and the modified nanoparticle-mediated chemopreventive regimens, showing a range of 31% to 38% (P < 0.05) and 50% to 75% (P < 0.001) reduction, respectively, in the number of ACFs. In addition, the nanoparticulate combination regimens of AFAC showed a 2-fold increase in suppression of ACF compared with free drug mixtures. Individual administration of nanoparticle-encapsulated drugs showed no significant effect on the reduction of ACF. Histochemical analysis provided further confirmation of chemopreventive effects, showing a significant reduction in cell nuclear proliferation. Overall, our results provide a strong proof of concept using nanoparticle-mediated combination treatment in the chemoprevention of colon cancer. Cancer Prev Res; 4(10); 1655–64. ©2011 AACR.

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Pancreatic cancer (PC), currently the third leading cause of cancer-related deaths in the USA, is projected to become the second leading cause, behind lung cancer, by 2020. The increasing incidence, low survival rate, and limited treatment opportunities necessitate the use of alternative approaches such as chemoprevention, to tackle PC. In this study, we report significant synergistic chemoprevention efficacy for the first time from a low-dose combination of a classical antihistaminic drug, Loratadine (LOR) and a neutraceutical compound, Sulforaphane (SFN) using a self-microemulsifying drug delivery system (SMEDDS) formulation. The formulation was developed using Quality by Design approach (globule size, 95.13 ± 7.9 nm; PDI, 0.17 ± 0.04) and revealed significant (p < 0.05) enhancement in the in vitro dissolution profile confirming the enhanced solubility of BCS class II drug LOR with SMEDDS formulation. The LOR-SFN combination revealed ~ 40-fold reduction in IC50 concentration compared to LOR alone in MIA PaCa-2 and Panc-1 cell lines respectively, confirming the synergistic enhancement in chemoprevention. Further, the nanoformulation resulted in ~ 7-fold and ~ 11-fold reduction in IC50 values compared to LOR-SFN combination. Hence, our studies successfully demonstrate that a unique low-dose combination of LOR encapsulated within SMEDDs with SFN shows significantly enhanced chemopreventive efficacy of PC.

Celastrol (also called tripterine) is a quinone methide triterpene isolated from the root extract of Tripterygium wilfordii (thunder god vine in traditional Chinese medicine). Over the past two decades, celastrol has gained wide attention as a potent anti-inflammatory, anti-autoimmune, anti-cancer, anti-oxidant, and neuroprotective agent. However, its clinical translation is very challenging due to its lower aqueous solubility, poor oral bioavailability, and high organ toxicity. To deal with these issues, various formulation strategies have been investigated to augment the overall celastrol efficacy in vivo by attempting to increase the bioavailability and/or reduce the toxicity. Among these, nanotechnology-based celastrol formulations are most widely explored by pharmaceutical scientists worldwide. Based on the survey of literature over the past 15 years, this mini-review is aimed at summarizing a multitude of celastrol nanoformulations that have been developed and tested for various therapeutic applications. In addition, the review highlights the unmet need in the clinical translation of celastrol nanoformulations and the path forward.

Tumor Treating Fields (TTFields) are used in cancer treatment as they offer increased efficacy, safety, and low adverse effects. They have shown significant potential in pancreatic cancer (PC) clinical trials when combined with chemotherapy. It is pertinent to know that although TTFields-chemotherapy combination definitely enhances the treatment efficacy, it also results in chemotherapy induced severe adverse effects. To resolve this, we earlier reported development of Gemcitabine loaded self-assembling cationic-anionic polymer nanoparticles (S-CAP NPs) capable of achieving TTFields-triggered targeted drug release at the PC tumor site. Initially, four S-CAP NPs [C4 and C7 from chitosan- bovine serum albumin (Chitosan-BSA) S-CAP NPs; P5 and P8 from polyethylenimine- bovine serum albumin (PEI-BSA) S-CAP NPs] were shortlisted [Table 1 - particle size: ~ 200 nm; % encapsulation efficiency (EE): 55-65%]. For proof-of-concept studies, effect of TTFields on the % EE and % cumulative drug release of the S-CAP-NPs was investigated. The studies revealed (Table 1) significant reduction (p value < 0.05) in the % EE of S-CAP NPs within 48h TTFields treatment, confirming that the TTFields destabilize the NPs triggering targeted drug release. Similarly, In vitro drug release studies showed significant increase (p< 0.01) in rate of drug release from S-Cap NPs in presence of TTFields. The cell inhibition assay confirmed the synergism in cell apoptosis wherein PEI-BSA S-CAP NPs showed higher efficacy compared to chitosan-BSA S-CAP NPs. Further, additional stability studies continued over a period of 18 months confirmed stable formulations. In vivo acute and chronic toxicity studies are underway to identify the most optimum dose of S-CAP NPs for further in vivo efficacy studies. To summarize, S-CAP-NPs present a promising avenue to achieve targeted chemotherapy in conjunction with TTFields. This work is supported by AACR-Novocure tumor treating fields research fellowship. Table 1. Effect of TTFields on % encapsulation efficiency (n=3). Data represented as Mean ± SD. Batch Particle size (nm) Encapsulation efficiency (%) - 0h Encapsulation efficiency (%) - 24h - 150 KHZ Encapsulation efficiency (%) - 48h - 150 KHZ Chitosan-BSA S-CAP NPs C4 210.54 ± 38.96 61.26 ± 5.11 36.21 ± 3.53 24.86 ± 3.19 C7 215.67 ± 32.55 65.31 ± 5.84 32.19 ± 2.68 21.86 ± 4.64 PEI-BSA S-CAP NPs P5 198.29 ± 41.05 58.83 ± 3.33 27.66 ± 3.31 18.96 ± 3.96 P8 209.92 ± 31.33 64.31 ± 5.13 24.83 ± 4.29 19.42 ± 2.73 Citation Format: Preshita Prafulla Desai, Sunil Prabhu. Self-assembling nanoparticles: A novel approach for targeted cancer treatment using tumor treating fields [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 299.

Celastrol (CL), a compound isolated from Tripterygium wilfordii, possesses various bioactivities such as antitumor, anti-inflammatory and anti-obesity effects. In previous studies, we developed CL-encapsulated silk fibroin nanoparticles (CL-SFNP) with satisfactory formulation properties and in vitro cancer cytotoxicity effect. For further in vivo oral bioavailability evaluation, in this study, a simple and reliable LC-MS/MS method was optimized and validated to determine CL concentration in rat plasma. The separation of CL was performed on a C18 column (150 by 2 mm, 5 &micro, m) following sample preparation using liquid&ndash, liquid extraction with the optimized extraction solvent of tert-butyl methylether. The assay exhibited a good linearity in the concentration range of 0.5&ndash, 500 ng/mL with the lower limit of quantification (LLOQ) of 0.5 ng/mL. The method was validated to meet the requirements for bioassay with accuracy of 91.1&ndash, 110.0%, precision (RSD%) less than 9.1%, extraction recovery of 63.5&ndash, 74.7% and matrix effect of 87.3&ndash, 101.2%. The developed method was successfully applied to the oral bioavailability evaluation of CL-SFNP. The pharmacokinetic results indicated the AUC0-&infin, values of CL were both significantly (P &lt, 0.05) higher than those for pure CL after intravenous (IV) or oral (PO) administration of equivalent CL in rats. The oral absolute bioavailability (F, %) of CL significantly (P &lt, 0.05) increased from 3.14% for pure CL to 7.56% for CL-SFNP after dosage normalization. This study provides valuable information for future CL product development.

Tumor Treating Fields (TTFields) have been clinically proven as safe, effective, and non-invasive approach for cancer treatment. Specifically, TTFields in conjunction with Gemcitabine/nab-Paclitaxel have shown promising results in Phase II pancreatic cancer (PC) PANOVA study. It is imperative to understand here that while TTFields are very safe, concurrent use of anticancer drugs will continue to elicit non-site-specific adverse effects resulting in overall low patient compliance. To overcome this drawback, we have developed an innovative strategy called ‘Tumor Treating Fields Triggered Targeting of Nanoparticles in Cancer (TTFields-TTONIC)'. For this, self-assembling cationic-anionic polymer nanoparticles (S-CAP NPs) encapsulating Gemcitabine as a model anticancer drug were developed. The hypothesis involves combination of NPs and TTFields wherein the developed NPs will be preferentially taken up by the tumor owing to leaky vasculature. Further, only under the applied TTFields, the NPs will be destabilized due to high charge density of cationic and anionic polymers leading to targeted release of encapsulated drug at the tumor site (reduction in non-site-specific side effects). For this, multiple batches of two types of S-CAP NPs [chitosan- bovine serum albumin (Chitosan-BSA) and polyethylenimine- bovine serum albumin (PEI-BSA)] were developed. The formulations were optimized using mathematical modelling and Design Expert® software to achieve low particle size and optimum encapsulation efficiency. Based on the results, 2 formulations from each type i.e., chitosan- BSA S-CAP NPs [Batch C4 - particle size: 210.54 ± 38.96 nm, PDI: 0.194, encapsulation efficiency: 61.26 ± 5.11%, zeta potential: (+) 7.38 ± 3.11; Batch C7 - particle size: 215.67 ± 32.55 nm, PDI: 0.201, encapsulation efficiency: 65.31 ± 5.84 %, zeta potential: (+) 3.22 ± 1.28] and PEI-BSA S-CAP NPs [Batch P5 - particle size: 198.29 ± 41.05 nm, PDI: 0.227, encapsulation efficiency: 58.83 ± 3.33%, zeta potential: (+) 8.17 ± 2.63; Batch P8 - particle size: 209.92 ± 31.33 nm, PDI: 0.196, encapsulation efficiency: 64.31 ± 5.13%, zeta potential: (+) 11.49 ± 2.99] were shortlisted that exhibited particle size ~ 200 nm and encapsulation efficiency in range of 55-65 %. All the formulations exhibited sustained drug release profile over a period of 60 h wherein chitosan- BSA S-CAP NPs showed slower drug release compared to PEI-BSA S-CAP NPs (P Citation Format: Preshita Prafulla Desai, Sunil Prabhu. Drug loaded nanoparticle targeting of pancreatic cancer using tumor treating fields (TTFields) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr LB023.

Psoriasis is considered to be a systemic disease of immune dysfunction. It is still unclear what triggers the inflammatory cascade associated with psoriasis but recent evidences suggest the vital role of IL-23/IL-17A cytokine axis in etiology of psoriasis. Several studies have been conducted in psoriatic-like animal models but ethical issues and complexity surrounding it halts the screening of new anti-psoriatic drug candidates. Hence, in this study, we developed a new in-vitro model for psoriasis using imiquimod (IMQ) induced differentiated HaCaT cells which could be used for screening of new anti-psoriatic drug candidates. The differentiated HaCaT cells were treated with IMQ (100μM) to induce psoriatic like inflammation and its effect was investigated using a natural anti-psoriatic compound, curcumin. The proliferation of psoriatic-like cells was inhibited by curcumin at 25 and 50µM concentrations. The psoriatic-like cells decreased in number with increase in apoptotic and dead cells upon curcumin treatment. Curcumin inhibited the proliferation of IMQ-induced differentiated HaCaT cells (Psoriatic-like cells) by down-regulation of pro-inflammatory cytokines, interleukin-17, tumor necrosis factor-α, interferon-γ, and interleukin-6. Apart from this, curcumin significantly enhanced the skin-barrier function by up-regulation of involucrin (iNV) and filaggrin (FLG), the regulators of epidermal skin barrier. The IMQ-induced differentiated HaCaT in vitro model recapitulated some aspects of the psoriasis pathogenesis similar to murine model. Henceforth, we conclude that this model may be used for rapid screening of anti-psoriatic drug candidates and warrant further mechanistic studies.

In gas chromatographic separation of hydrogen isotopes iron-alumina as a stationary phase is conventionally used. Further, Chromium-alumina as an alternative stationary phase has also been mentioned in the literature. In this present study, a detail comparative study of both the stationary phases in terms of their hydrogen isotope separation ability has been carried out. This study shows that chromium-alumina is better alternative than iron-alumina which might be attributed to its improved resolution, higher column efficiency, easy mode of preparation and shorter retention time along with equally simple regeneration procedure for regaining the performance of column material. To understand the basis behind their difference in ability of hydrogen isotope separation, hydrogen adsorption/desorption phenomenon at experimentally condition along with different surface related properties of both stationary phases have been evaluated. These experimental findings have been correlated with hydrogen isotope separation ability in terms of its retention time and resolution obtained during hydrogen isotopic separation and it illustrates the fact that hydrogen adsorption/desorption phenomenon is one of the governing factor in deciding the performance of the column materials.

Cancer research in pursuit of better diagnostic and treatment modalities has seen great advances in recent years. However, the incidence rate of cancer is still very high. Almost 40% of women and men are diagnosed with cancer during their lifetime. Such high incidence has not only resulted in high mortality but also severely compromised patient lifestyles, and added a great socioeconomic burden. In view of this, chemoprevention has gained wide attention as a method to reduce cancer incidence and its relapse after treatment. Among various stems of chemoprevention research, nanotechnology-based chemoprevention approaches have established their potential to offer better efficacy and safety. This review summarizes recent advances in nanotechnology-based chemoprevention strategies for various cancers with emphasis on lung and bronchial cancer, colorectal, pancreatic, and breast cancer and highlights the unmet needs in this developing field towards successful clinical translation.

Celastrol (CL), a bioactive compound isolated from Tripterygium wilfordii, has demonstrated bioactivities against a variety of diseases including cancer and obesity. However, its poor water solubility and rapid in vivo clearance limit its clinical applications. To overcome these limitations, nanotechnology has been employed to improve its pharmacokinetic properties. Nanoparticles made of biological materials offer minimal adverse effects while maintaining the efficacy of encapsulated therapeutics. Silk fibroin (SF) solution was prepared successfully by extraction from the cocoons of silkworms, and a final concentration of 2 mg/mL SF solution was used for the preparation of CL-loaded SF nanoparticles (CL-SFNP) by the desolvation method. A stirring speed of 750 rpm and storage time of 20 h at &minus, 20 &deg, C resulted in optimized product yield. A high-performance liquid chromatography (HPLC) method was developed and validated for the analysis of CL in rat plasma in terms of selectivity, linearity, intra-/inter-day precision and accuracy, and recovery. No interference was observed in rat plasma. Linearity in the concentration range of 0.05&ndash, 5 &micro, g/mL was observed with R2 of 0.999. Precision and accuracy values were below the limit of acceptance criteria, i.e., 15% for quality control (QC) samples and 20% for lower limit of quantification (LLOQ) samples. Rats were given intravenous (IV) administration of 1 mg/kg of pure CL in PEG 300 solution or CL-SFNP. The pharmacokinetic profile was improved with CL-SFNP compared to pure CL. Pure CL resulted in a maximum concentration (Cmax) value of 0.17 &micro, g mL&minus, 1 at 5 min following administration, whereas that for CL-SFNP was 0.87 &micro, 1 and the extrapolated initial concentrations (C0) were 0.25 and 1.09 &micro, 1, respectively, for pure CL and CL-SFNP. A 2.4-fold increase in total area under the curve (AUC0-inf) (&micro, g h mL&minus, 1) was observed with CL-SFNP when compared with pure CL. CL-SFNP demonstrated longer mean residence time (MRT, 0.67 h) than pure CL (0.26 h). In conclusion, the preparation of CL-SFNP was optimized and the formulation demonstrated improved pharmacokinetic properties compared to CL in solution following IV administration.

Pancreatic cancer is the fourth leading cause of death in the United States and has a 5-year life expectancy of ~8%. Currently, only a few drugs have been approved by the United States Food and Drug Administration for pancreatic cancer treatment. Despite available drug therapy and ongoing clinical investigations, the high prevalence and mortality associated with pancreatic cancer mean that there is an unmet chemopreventive and therapeutic need. From ongoing studies with various novel formulations, it is evident that the development of smart drug delivery systems will improve delivery of drug cargo to the pancreatic target site to ensure and enhance the therapeutic/chemoprevention efficacy of existing drugs and newly designed drugs in the future. With this in view, nanotechnology is emerging as a promising avenue to enhance drug delivery to the pancreas via both passive and active targeting mechanisms. Research in this field has grown extensively over the past decade, as is evident from available scientific literature. This review summarizes the recent advances that have brought nanotechnology-based formulations to the forefront of pancreatic cancer treatment.

Arvind Thakkar,1 Sushma Chenreddy,1 Astrid Thio,1 Wael Khamas,2 Jeffrey Wang,1 Sunil Prabhu1 1Department of Pharmaceutical Sciences, College of Pharmacy, 2College of Veterinary Medicine, Western University of Health Sciences, Pomona, CA, USA Abstract: Our previous studies have established the efficacy of chemopreventive regimens of aspirin and curcumin (CUR) encapsulated within solid lipid nanoparticles (SLNs) in combination with free sulforaphane (ACS combination) to prevent or delay the initiation and progression of pancreatic cancer, classified as one of the deadliest diseases with very low chances of survival upon diagnosis. Although toxicity of individual drugs and SLN has been studied previously, there are no studies in current literature that evaluate the potential toxicity of a combined regimen of ACS, especially when encapsulated within chitosan-SLNs (c-SLNs). Hence, objective of the current study was to investigate the potential toxic effects of ACS c-SLN combined chemopreventive regimens following acute (3days), subacute (28days), and subchronic (90days) administrations by oral gavage in BALB/c mice. Mice were administered the following regimens: saline, blank c-SLN, low-dose ACS c-SLN (2+4.5+0.16mg/kg), medium-dose ACS c-SLN (20+45+1.6mg/kg), and high-dose ACS c-SLN (60+135+4.8mg/kg). The potential toxicity was evaluated based on animal survival, body weight, hematology, blood chemistry, and organ histopathology. During 3-day, 28-day, and 90-day study periods, no animal deaths were observed. Treatment with ACS c-SLNs did not cause alteration in complete blood counts and blood chemistry data. Histopathological examination of various organ sections (pancreas, heart, liver, kidney, and brain) appeared normal. Based on the results of this study, no signs of toxicity in acute, subacute, and subchronic studies following oral administration of ACS c-SLNs were found indicating that the oral dosing regimens were safe at the levels tested for long-term administration to prevent the onset of pancreatic cancer. Keywords: pancreatic cancer, toxicity, chitosan-solid lipid nanoparticle, aspirin, curcumin, sulforaphane

Celastrol (CL), a compound isolated from Tripterygium wilfordii, possesses various bioactivities such as anti-tumor, anti-inflammatory and anti-obesity effects. Many new drug delivery systems of CL have been designed to overcome its poor water solubility and low bioavailability. However, few studies are available concerning its in vivo pharmacokinetics. In this study, a simple and sensitive LC-MS/MS method was developed and validated to determine CL in rat plasma using 18α-glycyrrhetinic acid as the internal standard. The separation of CL was performed on a Phenomenex Gemini C18 column (150 mm × 2 mm, 5 µm) following sample preparation using liquid-liquid extraction with the optimized extraction solvent of tert-butyl methyl ether and a small plasma sample volume of 50 μL. The analyte was quantitated under positive electrospray ionization and multiple reactions monitoring mode. The assay exhibited a good linearity in the concentration range of 0.5-500 ng/mL. The lower limit of quantification (LLOQ) was 0.5 ng/mL. The method was fully validated to meet the requirements for bioassay with accuracy of 91.1-110.0%, precision (RSD %) of less than 9.1 %, extraction recovery of 63.5-74.7%, and matrix effect of 87.3-101.2%. The developed method was successfully applied to the pharmacokinetic evaluation of CL-encapsulated silk fibroin nanoparticles (CL-SFNP). The results show, after intravenous (IV) and oral (PO) administration of equivalent CL (1 mg/kg and 3 mg/kg, respectively) in rats, the AUC values of CL were 8646.1±1998.9 ng/mL·h after IV dosing and 1065.5±494.6 ng/mL·h after PO dosing for CL-SFNP, which were significantly (P Citation Format: Shuyu Zhan, Amy Paik, Felicia Onyeabor, Baoyue Ding, Sunil Prabhu, Jeffrey Wang. Development and validation of an LC/MS/MS method for the determination of celastrol in rat plasma and its application to the pharmacokinetic evaluation of celastrol-encapsulated silk fibroin nanoparticles [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 6252.

Pancreatic cancer is a lethal disease with a dreadful 5-year survival rate of only 5%. In spite of several treatment options, the prognosis still remains extremely poor. Therefore, novel therapy strategies with combinations of drugs are urgently required to combat this fatal disease. Triptolide (TPL) and celastrol (CL), two main compounds in traditional Chinese medicine isolated from Thunder God Vine, have a broad range of bioactivities including anticancer activity. Silk fibroin (SF), a naturally occurring protein with several unique properties, is an ideal carrier material. In this study, we prepared TPL and CL loaded silk fibroin nanoparticles (TPL-SFNPs and CL-SFNPs) by a modified desolvation method and evaluated their synergistic effects against human pancreatic cancer cells. Both SFNPs were characterized for particle size and zeta potential. The entrapment efficiency, drug loading, and drug release profiles were evaluated by HPLC. The cytotoxicity and synergistic effect of SFNPs were investigated in MIA PaCa-2 and PANC-1 human pancreatic cells. The results showed that the particle sizes of TPL-SFNPs and CL-SFNPs were 166.4 ± 4.6 nm and 170.4 ± 2.3 nm, with a mean zeta potential -27.2 ± 2.0 mV and -25.5 ± 2.57 mV, respectively. TPL-SFNPs and CL-SFNPs have a drug loading of 57.0 ± 4.7 μg mg-1 and 63.5 ± 3.8 μg mg-1 along with an encapsulation efficiency of 81.8 ± 2.8% and 87.0 ± 5.1%, respectively. Drug release studies revealed that a rapid release of the drugs from SFNPs was observed at pH 4.5 (lysosomal pH) and a delayed release was observed at pH 7.4 (plasma pH). TPL-SFNPs (IC50 3.80 and 4.75 nM) and CL-SFNPs (IC50 0.38 and 0.64 μM) were 2-3 fold more potent against MIA PaCa-2 and PANC-1 cells than free TPL (IC50 11.25 and 11.58 nM) and CL (IC50 0.84 and 1.23 μM). Furthermore, co-treatment with TPL-SFNPs and CL-SFNPs increased the growth inhibition of the same cells significantly in comparison with TPL-SFNPs or CL-SFNPs alone. Almost all combination index (CI) values, calculated using the CompuSyn software, were

Pancreatic cancer (PC) is the third leading cause of deaths in the United States claiming approximately 45,000 lives every year and is predicted to be the second cause of cancer related deaths by 2020. Late diagnosis, poor prognosis, limited success and significant side effects with existing anticancer drugs has resulted in such high mortality rates (5 year survival rate is less than 8%). These limitations have encouraged scientists to develop chemopreventive strategies and combination drug alternatives that can serve as better therapeutics. In our previous studies, we have reported the synergistic chemoprevention potential (formulation patent pending) of lipid nanocarriers (SMEDDS) of Loratadine (LOR), H1-receptor antagonist drug; with free Sulforaphane (SFN), a neutraceutical compound from broccoli when administered at low doses in human PC cell lines (MIA PaCa-2 and Panc-1). To confirm the hypothesis that enhanced efficacy using the SMEDDS formulation results from bioavailability enhancement, preclinical in vivo pharmacokinetic studies were performed in Sprague Dawley rats. The results confirmed that the bioavailability and Cmax of LOR increased 5-fold (2203.2 ± 293.7 ng.h/mL for plain LOR and 10266.1 ± 250.0 ng.h/mL for LOR SMEDDS) and 4-fold (120.9 ± 10.5ng/mL for plain LOR and 503.2 ± 5.8ng/mL for LOR SMEDDS) when administered as SMEDDS compared to plain drug. This corroborated the significant enhancement in bioavailbility with SMEDDS formulation. This promising results in bioenhancement and chemoprevention activity led to an hypothesis to assess the potential of LOR SMEDDS-SFN combination as an adjuvant therapy with classical anticancer drugs like Gemcitabine (GEM) to give synergistic efficacy enhancement in PC treatment. For this, IC50 concentrations were calculated for various treatment groups [GEM alone or in addition to the LOR free drug or as SMEDDS with or without SFN] using MTS viability assay on human PC cell lines in accordance with published protocol over an incubation time of 72 h. The LOR SMEDDS-SFN-GEM combination revealed 2-fold and 2.5-fold enhancement in anticancer activity compared to free drug combination and GEM alone respectively. This confirmed the synergistic anticancer potential of the free drug combination followed by further activity potentiation using SMEDDS formulation. In conclusion, a novel SMEDDS of LOR and free SFN was invented (provisional patent applied) to successfully elicit PC chemoprevention and adjuvant treatment in combination with GEM. We anticipate that, this adjuvant therapy will lead to relatively low doses of anticancer drugs and thereby can reduce the side effects with enhanced efficacy. Citation Format: Sunil Prabhu, Preshita Desai, Naga Jyothi Thumma, Jeffrey Wang. Self-microemulsifying drug delivery systems (SMEDDS) containing novel compounds in combination with gemcitabine: Bioavailability enhancement and synergistic anticancer efficacy studies for pancreatic cancer management [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2165.

Pancreatic cancer is a deadly disease killing 37,000 Americans each year. Despite two decades of research on treatment options, the chances of survival are still less than 5% upon diagnosis. Recently, chemopreventive strategies have gained considerable attention as an alternative to treatment. We have previously shown significant in vitro chemopreventive effects with low-dose combinations of aspirin, curcumin, and sulforaphane (ACS) on pancreatic cancer cell lines. Here, we report the results of 24-week chemopreventive study with the oral administration of ACS combinations on the N-nitrosobis (2-oxopropyl) amine (BOP)-treated Syrian golden hamster model to suppress the progression of pancreatic intraepithelial neoplasms (PanIN) using unmodified (free drug) combinations of ACS, and nanoencapsulated (solid lipid nanoparticles; SLN) combinations of aspirin, curcumin, and free sulforaphane. The use of three different doses (low, medium, and high) of unmodified ACS combinations exhibited reduction in tumor incidence by 18%, 50%, and 68.7% respectively; whereas the modified nanoencapsulated ACS regimens reduced tumor incidence by 33%, 67%, and 75%, respectively, at 10 times lower dose compared with the free drug combinations. Similarly, although the unmodified free ACS showed a notable reduction in cell proliferation, the SLN encapsulated ACS regimens showed significant reduction in cell proliferation at 6.3%, 58.6%, and 72.8% as evidenced by proliferating cell nuclear antigen expression. Cell apoptotic indices were also upregulated by 1.5, 2.8, and 3.2 times, respectively, compared with BOP control. These studies provide a proof-of-concept for the use of an oral, low-dose, nanotechnology-based combinatorial regimen for the long-term chemoprevention of pancreatic cancer. Cancer Prev Res; 6(10); 1015–25. ©2013 AACR.

Human skin is body's vital organ constantly exposed to abiotic oxidative stress. This can have deleterious effects on skin such as darkening, skin damage, and aging. Plant-derived products having skin-protective effects are well-known traditionally. Triphala, a formulation of three fruit products, is one of the most important rasayana drugs used in Ayurveda. Several skin care products based on Triphala are available that claim its protective effects on facial skin. However, the skin protective effects of Triphala extract (TE) and its mechanistic action on skin cells have not been elucidated in vitro. Gallic acid, ellagic acid, and chebulinic acid were deduced by LC-MS as the major constituents of TE. The identified key compounds were docked with skin-related proteins to predict their binding affinity. The IC50 values for TE on human dermal fibroblasts (HDF) and human keratinocytes (HaCaT) were 204.90 ± 7.6 and 239.13 ± 4.3 μg/mL respectively. The antioxidant capacity of TE was 481.33 ± 1.5 mM Trolox equivalents in HaCaT cells. Triphala extract inhibited hydrogen peroxide (H2O2) induced RBC haemolysis (IC50 64.95 μg/mL), nitric oxide production by 48.62 ± 2.2%, and showed high reducing power activity. TE also rescued HDF from H2O2-induced damage; inhibited H2O2 induced cellular senescence and protected HDF from DNA damage. TE increased collagen-I, involucrin and filaggrin synthesis by 70.72 ± 2.3%, 67.61 ± 2.1% and 51.91 ± 3.5% in HDF or HaCaT cells respectively. TE also exhibited anti-tyrosinase and melanin inhibition properties in a dose-dependent manner. TE increased the mRNA expression of collagen-I, elastin, superoxide dismutase (SOD-2), aquaporin-3 (AQP-3), filaggrin, involucrin, transglutaminase in HDF or HaCaT cells, and decreased the mRNA levels of tyrosinase in B16F10 cells. Thus, Triphala exhibits protective benefits on skin cells in vitro and can be used as a potential ingredient in skin care formulations.

Pancreatic cancer is the fourth largest cause of cancer deaths in the Unites States and the prognosis is grim with

Colorectal cancer (CRC) is the third most common cause of cancer death in the United States, accounting for approximately 51,000 deaths each year. We have previously shown in vitro chemopreventive effects of mixtures of aspirin, folic acid, and calcium (AFAC) on colon cancer cell lines. The objective of the present study was to evaluate the in vivo effects of orally administered, colon targeted chemopreventive combination regimens on the inhibition of aberrant crypt foci (ACF) in a rat model of colon carcinogenesis using (i) unmodified (free drug) combinations of AFAC and (ii) nanoparticle-encapsulated combinations of the same agents. A 14-week animal study was conducted in three phases to determine an optimal effective dose from AFAC combinations and evaluate the efficacy of nanotechnology-based chemopreventive regimens administered in combined (mixtures) and individual (single entity) forms. ACF inhibition when compared with azoxymethane-treated rat control group was significant in both the unmodified and the modified nanoparticle-mediated chemopreventive regimens, showing a range of 31% to 38% (P < 0.05) and 50% to 75% (P < 0.001) reduction, respectively, in the number of ACFs. In addition, the nanoparticulate combination regimens of AFAC showed a 2-fold increase in suppression of ACF compared with free drug mixtures. Individual administration of nanoparticle-encapsulated drugs showed no significant effect on the reduction of ACF. Histochemical analysis provided further confirmation of chemopreventive effects, showing a significant reduction in cell nuclear proliferation. Overall, our results provide a strong proof of concept using nanoparticle-mediated combination treatment in the chemoprevention of colon cancer. Cancer Prev Res; 4(10); 1655–64. ©2011 AACR.

The purpose of this research was to prepare and characterize a novel lyophilized chitosan-based hydrogel complex (termed CS-M) for controlled drug delivery applications using a highly water soluble model drug, niacinamide. Characterization studies were undertaken to evaluate the physical-chemical properties, polymer swelling, in vitro controlled release kinetics, tissue bioadhesion, intestinal permeability and stability of the novel chitosan complex. Additionally, a comparative analysis was conducted with commercial polymers namely, Carbopol 974P-NF® and hydroxypropyl methylcellulose (HPMC K4M). FT–IR and 1H NMR studies confirmed that despite alteration in physical structure and morphology of the chitosan complex the chemical properties remained unchanged, when compared to the parent chitosan compound. Polymer swelling studies showed consistency in the structural integrity and water uptake of CS-M compared to other polymers which showed inconsistent swelling and disintegration behavior over a 5 h period. In vitro controlled release profiles of CS-M showed a slower, more controlled release rate of niacinamide than other polymers indicating the influence of polymer swelling capacity on water uptake and subsequent drug release. CS-M demonstrated an overall increase in bioadhesion to intestinal tissue when compared to commercial polymers at same concentrations. Similarly, drug transport through everted sac intestinal tissue showed enhanced absorption properties of CS-M when compared to other polymers. Finally, a 3 month accelerated stability study showed all polymers including CS-M to be stable when formulated with niacinamide. Overall, the modified chitosan-based hydrogel polymer, CS-M, demonstrated enhanced characteristics indicating its potential to be used as a controlled release excipient in oral drug formulations. Keywords: Chitosan hydrogel complex, controlled drug delivery, niacinamide, lyophilization, Carbopol 974P-NF, HPMC K4M

The topic of adapting one’s leadership style is controversial and can generate great debate. When one considers adapting one’s leadership style, the question of authenticity often bubbles to the surface. This is even more compelling in the context of generational differences in our society and workforce. How can faculty members be effective if they refuse to adapt their leadership style to students’ learning styles and generational characteristics? Leadership development is an ongoing focus for the American Association of Colleges of Pharmacy (AACP) to prepare faculty for roles as future leaders in higher education while a major redistribution of generations is occurring within the workforce. These generations are molded by many factors, including the economy, significant events, technology, and education. As experienced leaders, we affirm that one’s leadership styles must be honed over time and adapted to reflect generational differences in academic pharmacy among colleagues, students, alumni, and stakeholders, including patients. Current and future leaders need further education and development so that their awareness of generational characteristics prepares them to adapt their leadership styles to more effectively engage and lead people of any generation.

Pancreatic cancer (PC) is the fourth leading cause of deaths in the United States, claiming approximately 43,000 lives every year. Existing trends suggest that it will be the second cause of death after lung cancer within the next 20 years. With mortality rates remaining high even after decades of research, scientists are now turning to develop chemopreventive strategies that may stop or delay the initiation and progression of the disease. Our study reports the chemoprevention potential of (1) a free drug combination of H1-receptor antagonist drug with Sulforaphane (SFN), a neutraceutical compound derived from broccoli and (2) lipid nanocarriers of H1-receptor antagonist drug (SP1 SMEDDS; formulation patent pending) with free SFN when administered at low doses in human pancreatic cancer cell lines (MIA PaCa-2 and Panc-1). IC50 concentrations were calculated for various treatment groups using MTS viability assay in accordance with published protocol, using 96-well microplates and incubation time of 72 h. The free drug combination revealed almost 70-fold and 20-fold enhancement in chemoprevention efficacy (based decrease in cancer cell viability) as compared to controls. Interestingly, the free drug combination (at 5µM) showed ~55 % (p Citation Format: Preshita Desai, Arvind Thakkar, Jeffrey Wang, Sunil Prabhu. Self-microemulsifying drug delivery systems (SMEDDS) containing novel compounds for the chemoprevention of pancreatic cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2238.

Objectives There is a significant shortage of pharmacy faculty. During the next decade, academia will need at least 1200 new faculty members to staff the various pharmacy programs around the country. This survey identifies factors that influence pharmacy students in considering academia as a career option. Primary objectives were to evaluate whether students have considered academia as a career pathway, how they were exposed, their perspectives on this career path, and to compare students' and faculty's perspectives on why students choose or do not choose academia as a career path. Methods Online surveys were developed, one for students and one for faculty, and administered at six institutions. This exploratory study used descriptive analysis and logistic regression models. Results Almost half of the students surveyed were unaware of the shortage in pharmacy faculty members. Low patient contact was the top reason students did not intend to pursue academia. The top reasons students were interested in academia were teaching responsibilities and work-life balance. Salary was not as significant a factor for students as faculty predicted. Current school initiatives did not significantly impact a student's career decision. Conclusion Student and faculty perspectives were significantly different in many areas. Future methods to educate students on academia as a career path need to be more effective in reaching students' interests and addressing misconceptions regarding the activities of a faculty member. Patient contact, teaching responsibilities, and a healthy work-life balance are important to our students and should be highlighted within our schools to encourage academic careers.

Our objective was to investigate the controlled release and transport of leuprolide acetate polypeptide in the colon using novel combinations of rate-controlling polymers. Polymer swelling, disintegration, drug release, and transport characteristics were measured using different polymers, carbopol, chitosan and polyox, alone and in combination. Studies demonstrated Carbopol® -containing combination formulations had maximum swelling and the slowest disintegration properties. A decrease in dissolution rate was observed from all combination formulations when compared with their individual counterparts. Carbopol® combinations showed the slowest overall release. Drug transport studies using the everted sac technique demonstrated good correlation to the swelling, disintegration, and dissolution studies. Thus, novel polymer combinations can be used to deliver polypeptide drug to the colon effectively compared with individual polymers.

The objective of this study was to design an in situ biodegradable polymer implant controlled-release drug delivery system, using novel combinations of co-solvents and a model polypeptide, calcitonin (CT), and to assess the release of drug as a function of these co-solvents. Formulations were prepared by dissolving/ suspending CT polypeptide in poly-(lactic acid) (PLA) polymer solutions/suspensions containing combinations of a hydrophobic (benzyl benzoate, BB) and a hydrophilic (benzyl alcohol, BA) solvent. The CT-PLA mixtures were each injected into test tubes containing phosphate buffered saline solution to form the in situ implant and sampling was conducted over a 28-day period. The samples were analyzed for drug content using a modified Lowry protein assay procedure. Cumulative drug release demonstrated a rank-order correlation depending on the amount of the hydrophobic (BB) and hydrophilic (BA) solvents within each system. Increasing the amounts of the hydrophobic solvent, BB, in formulations demonstrated a 1.2-4.4-fold increase in CT release. Stability studies of all formulations over a 4-month period showed progressive increase in degradation of the CT polypeptide, especially at 37 degrees C, but a slower degradation pattern prevailed at 4 degrees and 20 degrees C. Differential scanning calorimetric studies revealed a homogenous mixture of drug in the polymer matrix. Overall, these studies demonstrated the feasibility of designing controlled release systems capable of releasing a polypeptide drug as a function of influence of different co-solvent combinations.

In vitro dissolution of zinc insulin suspensions can be promoted by the complexation of zinc with an ionic species for which the zinc ion has a greater affinity. Studies conducted by our group have previously shown that the rate-limiting steps that govern the dissolution of zinc-complexed insulin suspension may be (1) chemical complexation (surface reaction) and (2) subsequent drug mass transport (diffusion and solubility). The purpose of this work was to use a computer simulation model to predict the dissolution behavior of zinc-complexed insulin suspensions and determine the influence of the above rate-limiting steps on the overall process of dissolution. A quasi-steady-state model was chosen which included the effects of a shrinking particle radius, the drug's solubility, and a convective mass transfer term. Based on this model, the computer simulation program evaluated dissolution behaviors of various model drugs, including zinc insulin suspensions. The experimental data obtained from actual dissolution experiments were superimposed on computer-generated profiles that incorporated quantitative values to key terms, namely the alpha (diffusion resistance) and beta (surface reaction resistance) values. Results demonstrated that the computer simulations could be used to predict the dissolution behavior of zinc-complexed protein suspensions by manipulating the alpha and beta values. Overall, the computer simulations indicated the involvement of both the surface reaction and the diffusion rate-limiting steps in zinc insulin dissolution, which was consistent with the results obtained from actual experimental studies.

The objective of the present study was to establish the individual and combined chemopreventive potential of a widely used non-steroidal anti-inflammatory drug, ibuprofen (IBU), encapsulated in solid lipid nanoparticles (SLNs) for the chemoprevention of pancreatic cancer. The IBU SLNs were optimized using various lipids (Stearic acid, Compritol 888 ATO and Tripalmitin) and surfactants (Poloxamer 188, Tween-80). The synergistic effect of combination of IBU with sulforaphane (SFN) was also evaluated. Cell viability studies were conducted followed by colony formation and NF-κB DNA binding assays. The IC50 concentration of free IBU in human pancreatic cancer Panc-1 and MIA PaCa-2 cells were 1.25 and 1.26 mM, respectively. SLN optimization study of IBU revealed stearic acid (1:2 drug to lipid ratio) formulated with Poloxamer 188 to be the most efficacious in cell viability study. Upon encapsulation in SLNs, IC50 concentration of IBU-SLN was 113.8 and 122.6 µM for Panc-1 and MIA PACa-2 cells, respectively, reflecting a 10-fold reduction compared to free IBU. Combinations of low doses of free IBU (250 µM) and SFN (5 µM) reduced cell viability by ~55% (P

The purpose of this study was to improve the dissolution properties of a poorly water soluble and bioavailable drug, ethopropazine HCl (ET), by incorporating the drug in three different types of solid dispersion systems. Solid dispersions of ET were prepared using 1:1 (w/w) ratios of (1) phospholipid (1,2 dimyristoyl-sn-glycerophosphocholine) (DMPC), (2) polyethylene glycol 8000 (PEG8000), and (3) a novel combination of both DMPC and PEG8000. Using the solvent method of preparation, ET and DMPC and/or PEG were dissolved in chloroform, and solvent subsequently was evaporated using nitrogen gas. The resulting solid dispersion(s) was passed through a 60-mesh sieve. Characterization of ET/DMPC solid dispersion was performed by differential scanning calorimetry (DSC) and X-ray diffractometry studies. Dissolution studies conducted in phosphate buffered saline (PBS) (pH 7.4, 37 degrees C +/- 0.5 degrees C) using the USP type II (paddle) dissolution apparatus showed significant increases in the dissolution rate of ET with all the solid dispersions in this study. Specifically, within the first 5 min (D5), solid dispersions containing ET/DMPC (1:1) showed an eightfold increase in dissolution; in combination with DMPC and PEG8000 (1:1), there was an approximately sixfold increase; and a fourfold increase was observed with PEG8000 (1:1). Complete dissolution of all solid dispersions occurred within 60 min (D60) of the run. Storage of the ET/DMPC sample for over 4.5 months revealed a decrease in the dissolution rate when compared to freshly prepared sample. Overall, it was concluded that the dissolution rate of ET significantly improved when dispersed in all the selected carrier systems. However, the solid dispersion of ET/DMPC was observed to be superior to the other combinations used.

Cancer is the second leading cause of death in the world, and current therapeutic outcomes are still not satisfactory. Cancer chemoprevention is an important strategy to reduce cancer morbidity by using non- or low-toxic natural or synthetic commodities to prevent and/or reverse the processes of neoplastic initiation and subsequent cancer development. Substantial evidence from human, animal and cell line studies has demonstrated that many herbal products used in traditional Chinese medicine (TCM) and Ayurveda medicine or traditional Indian medicine (TIM) can exert cancer chemopreventive effects. The underlying theories or philosophies for TCM and TIM to prevent cancer are very similar. The common central theme is to bring the patient back to a healthy state by modifying multiple abnormalities in the patient’s body in a holistic manner. Since carcinogenesis involves multiple abnormal gene expressions and signal transduction pathways, using TCM and TIM in cancer chemoprevention may not only be possible, but can be superior to agents aiming at single molecular target. However, before TCM and TIM can be accepted widely as complementary and alternative medicines for cancer prevention, it is crucial to evaluate their clinical efficacy and understand the molecular basis of their effects based on scientific evidence. This chapter highlights major molecular mechanisms in chemoprevention and summarizes the research on ten promising cancer preventive herbal products, including nine from TCM (Danshen, Danggui, Qianghuo, Huangqin, Zicao, Dasuan, Lingzhi, Yunzhi, and Xianggu) and five from TIM (Triphala, Turmeric, Neem, Guggulu, and Sapthaparna). The challenges in developing high quality TCM and TIM cancer preventive products are discussed and an integrated strategy is proposed to facilitate the process.

The purpose of this work was to assess synergistic inhibitory responses of a novel chemopreventive combination regimen of drugs namely, aspirin in combination with calcium and folic acid on two human colon cancer cell lines, HT-29 and SW-480. Subsequently, based on positive responses, nanotechnology-based formulations were developed for the targeted delivery of these combinatorial regimens to the colon for the chemoprevention of colon cancer. Additionally, conventional drug formulations using controlled release polymers chitosan, pectin and hydroxypropyl methylcellulose (HPMC) were tested for release of the drugs, for comparison purposes. Chemopreventive combination regimens demonstrated significant synergistic efficacy in both cell lines from XTT assay studies, when compared to the effects of individual agents. Approximately 45% decrease in cell viability for aspirin (15 mM) and calcium (30 mM) mixtures was observed in HT-29 cell lines, compared to approximately 55% decrease by the same combination in SW-480 cell lines. With combinations of aspirin (5 mM) and folic acid (1.5 mM), HT-29 cells demonstrated a 30% decrease in cell viability compared to approximately 38% decrease in the SW-480 cell line. Overall, all drug combinations demonstrated significant synergistic responses in the cell lines tested with the SW-480 cell line being more significantly affected by the drug regimens than the HT-29 cell line. Drug encapsulated nanoparticles demonstrated a spherical morphology,125 nm average particle size (aspirin and folic acid) of nanoparticles and encapsulation efficiencies in the range of 80-91%. Drug release from nanoparticles was controlled with approximately 60% of the original amount released over a 96 h period. Conventional formulations exhibited faster kinetics of drug release when compared to the PLGA nanoparticles. Overall, the cell line studies demonstrate, for the first time, the ability of novel chemopreventive combinations to inhibit the growth of colon cancer cells whereas the nanotechnology-based drug delivery system provides valuable evidence for targeted therapy towards colon cancer chemoprevention.

Cancer is the second leading cause of death, for which current therapeutic approaches are still very limited. Chemoprevention is an important approach to decreasing cancer morbidity and mortality by the use of non-toxic natural or synthetic substances to reverse the processes of initiation and subsequent progression of cancer. A substantial amount of evidence from human, animal and cell line studies has shown that many herbal products used for traditional Chinese medicine (TCM) can exert chemopreventive effects. The underlying theory for TCM to treat or prevent cancer is to bring the patient back to a healthy state by modifying multiple cancer-causing events. Since carcinogenesis involves multiple abnormal genes/pathways, using TCM in cancer chemoprevention may be superior to the agents targeting a single molecular target alone. However, before TCM can be accepted universally as complementary and alternative medicine for cancer treatment and prevention, it is crucial to understand the molecular basis for their effects. This review highlights several known molecular mechanisms of selected TCM in chemoprevention. Many TCM products or single active components have been reported to inhibit a variety of processes in cancer cell growth, invasion and metastasis by modulating a wide range of molecular targets, including cyclooxygenase-2 (COX-2), nuclear factor-Kappa B (NF-kappaB) and nuclear factor erythroid 2 -related factor 2 (Nrf2)-mediated antioxidant signaling pathways. The TCM and their active components with potent chemopreventive effects can be considered as promising lead agents for the design of more effective and less toxic agents for cancer chemoprevention.

Attention has shifted from the treatment of colorectal cancer (CRC) to chemoprevention using aspirin and folic acid as agents capable of preventing the onset of colon cancer. However, no sensitive analytical method exists to simultaneously quantify the two drugs when released from polymer-based nanoparticles. Thus, a rapid, highly sensitive method of high-performance liquid chromatography analysis to simultaneously detect low quantities of aspirin (hydrolyzed to salicylic acid, the active moiety) and folic acid released from biodegradable polylactide-co-glycolide (PLGA) copolymer nanoparticles was developed. Analysis was done on a reversed-phase C(18) column using a photodiode array detector at wavelengths of 233 nm (salicylic acid) and 277 nm (folic acid). The mobile phase consisted of acetonitrile-0.1% trifluoroacetic acid mixture programmed for a 30 min gradient elution analysis. In the range of 0.1-100 microg/mL, the assay showed good linearity for salicylic acid (R(2) = 0.9996) and folic acid (R(2) = 0.9998). The method demonstrated good reproducibility, intra- and inter-day precision and accuracy (99.67, 100.1%) and low values of detection (0.03, 0.01 microg/mL) and quantitation (0.1 and 0.05 microg/mL) for salicylic acid and folic acid, respectively. The suitability of the method was demonstrated by simultaneously determining salicylic acid and folic acid released from PLGA nanoparticles.

Established in 2003, the fully accredited international postbaccalaureate doctor of pharmacy (IPBP) program has attracted internationally trained pharmacists from approximately 25 countries and 6 continents, mostly residents of the United States, to attain the doctor of pharmacy (PharmD) degree at the Western University of Health Sciences. While recent trends in the IPBP applicant pool have shown a decline from its peak numbers in 2009 (222 applicants) for the 20 available seats each year, the quality of students remains high. Benchmark measures assessed for this group of students include the internal assessment entrance examination, admissions scores, academic assessments from didactic blocks, and scores on the North American Pharmacy Licensure Examination (NAPLEX), all of which indicate this quality. Moreover, graduates from the program not only consistently demonstrate excellence in the pharmacy curriculum and board examinations, but also go on to establish themselves as competent practitioners and educators. While the long-term future of the program is unknown, the status of the program and its graduates provides ample evidence of its value and ensures its continued success going forward.

Colorectal cancer (CRC) is the third most common cause of cancer death in the US, accounting for ∼51,000 deaths each year. Although the prognosis of this disease is improving due to better treatment options, attention has focused on the chemoprevention of this disease rather than on diagnosis and treatment. The objective of the present study was to evaluate the mechanisms of action of orally administered, colon targeted chemopreventive regimens aspirin (ASP), folic acid (FA) and calcium (Ca) alone and in combination AFAC (ASP+FA+Ca) in in-vitro and in-vivo rat model. We used colon cancer cells, HCT-116 and HT-29 as the in-vitro model. For colon targeting, ASP and FA was encapsulated in polylactide-co-glycolide (PLGA 50:50) polymer nanoparticles. Our previous studies demonstrated that combination of ASP (1mM), FA (1.5mM) and Ca (1.5mM) showed synergistic effect on inhibition of colon cancer cell viability. Follow-up study on azoxymethane (AOM) induced colon cancer in rats showed that the low (40+3.2+65 mg/kg), medium (80+6.4+130 mg/kg) and high dose (133+11+216 mg/kg) of modified AFAC-PLGA nanoparticle chemopreventive regimens demonstrated 1.7 to 2.0 fold reduction in the number of aberrant crypt foci (ACF). In continuation, a mechanistic approach was evaluated whereby colon cancer cells HCT-116 and HT-29 were treated with unmodified AFAC for 6h to determine gene expression using RT-qPCR analysis, and treated for 24h for protein expression study using western blot. The results obtained from the in-vitro study were further validated on rat colon tissue treated AFAC-PLGA nanoparticles, using immunohistochemistry. The RT-qPCR analysis showed that AFAC promoted reduction in cell viability was associated with down-regulation of proliferation genes (PCNA, and MKI67), cell cycle genes (CCND1, RB1, CDK4, E2F1, and JUN), and up-regulation of anti-apoptotic p21 gene. The protein expression studies revealed that AFAC treatment inhibited phospho-extracellular signal-regulated kinase 1/2 (p-ERK1/2), and phospho-p38 MAPK protein levels. The AFAC treatment also inhibited protein expression of PCNA, MKI67 and p-RB1 compared to control and individual drug treatment. Immunohistochemistry study using rat colon tissues revealed that the low, medium and high dose of AFAC-PLGA nanoparticles showed significant reduction in cell proliferation at 10.6%, 26.5%, and 48.5% as evidenced by PCNA protein expression. The AFAC treatment significantly increased apoptosis as evaluated by Tunnel assay, as well as decreased p-ERK1/2 and p-RB1 index. Overall, our results illustrate the mechanisms of action which confirms anti-proliferative activity of AFAC-PLGA nanoparticles, thus confirming the importance of novel combination chemoprevention strategies against colon cancer. Citation Format: Arvind Thakkar, Sushma Chenreddy, Jeffrey Wang, Sunil Prabhu. Mechanistic study of aspirin, folic acid and calcium (AFAC) combinations delivered using targeted polymeric nanoparticles in colon cancer prevention. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1900. doi:10.1158/1538-7445.AM2015-1900

The present study aims to evaluate the preclinical systemic toxicity profile of chitosan-solid lipid nanoparticles (c-SLNs) encapsulated aspirin (ASP), curcumin (CUR), and free sulforaphane (SFN) combination (ACS) in BALB/c mice. Recently, we demonstrated pancreatic cancer chemopreventive efficacy using ACS-SLNs delivery system in the BOP carcinogen induced pancreatic cancer hamster model. In order to develop safe ACS c-SLNs for clinical applications, it is vital to investigate their toxicity. Thus, we performed acute 3-day, sub-acute 28-day and sub-chronic 90-day toxicity studies of ACS c-SLNs in the BALB/c mice model. The mice were orally gavaged once every 24h with 200 μl ACS c-SLNs at three concentrations: low dose ACS (2+4.5+0.16 mg/kg; 1/30th of high dose), medium dose ACS (20+45+1.6 mg/kg; 1/3rd of high dose) and high dose ACS (60+135+4.8 mg/kg). Total of 60 mice were used for this study (4 mice each group X 3 timepoints X 5 groups). A standard evaluation was performed, which included daily clinical signs, weekly body weight, end-of-study blood hematology, blood clinical chemistry, gross necropsy on all animals and hematoxylin and eosin-stained examination of range of tissues from each animal. The clinical chemistry tests include blood urea nitrogen (BUN) and creatine (CRE), which indicate renal toxicity. Alanine transaminase (ALT) and alkaline phosphatase (ALP) levels indicate hepatic toxicity, while glucose (GLU) and total protein are measures of healthy growth. At the end-of-study, data revealed that mean body weight gain was consistent with low-, medium- and high-ACS c-SLN dose during acute, sub-acute and sub-chronic treatment period. The blood count and blood chemistry profiles revealed lack of systemic toxicity as the numbers were within normal ranges. Extensive histopathological examination of liver, kidney, heart, brain and pancreatic tissues showed no increase in treatment-related microscopic lesions in any of the three ACS c-SLN dose groups. Collectively, these findings demonstrate that repetitive oral administration of ACS c-SLN even at high concentrations was not associated with local or systemic toxicity and did not adversely affect the blood clinical chemistry of mice. Hence, long-term ACS c-SLN oral therapy is deemed safe for prevention of pancreatic cancer. Citation Format: Arvind Thakkar, Sushma Chenreddy, Jeffrey Wang, Sunil Prabhu. Preclinical systemic toxicity evaluation of chitosan-solid lipid nanoparticles encapsulated aspirin, curcumin and free sulforaphane (ACS) combinations in BALB/c mice. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2621. doi:10.1158/1538-7445.AM2015-2621

Lipid-based delivery systems are becoming increasingly popular as carriers of drugs due to their ability to overcome barriers to oral absorption. The purpose of this study was to prepare novel lipid-based formulations of a model drug, piroxicam (PXCM), a poorly water-soluble non-steroidal anti-inflammatory drug (NSAID) using 1,2-dimyristoyl-sn-glycero-3-phosphatidylcholine (DMPC) phospholipid alone, and in combination with polyethylene glycol (PEG 4600). Lipid-based drug delivery systems were prepared using conventional methods of preparation and the following aspects were evaluated (1) in vitro dissolution behavior, (2) absorption via Caco-2 cell monolayers and (3) stability of formulations over a 12-month period. In addition, physical characterization studies using differential scanning calorimetry (DSC) were also performed. Formulations of PXCM were prepared using DMPC in the following combinations (A) 1:1 and (B) 2:1 and a mixture of DMPC and PEG 4600 (C) 2:1:1, respectively. Dissolution studies conducted in phosphate buffered saline (PBS, pH 7.4, 37+/-0.5 degrees C) using the USP type II (paddle) dissolution apparatus showed an increase in dissolution rate and extent of the PXCM from all solid dispersion formulations when compared to the control. As such, the rate of drug release was observed to be fastest with formulation (C) showing the greatest increase of over two-fold compared to the control. Release of PXCM from formulations (A) and (B) was intermediate with the latter showing superior dissolution behavior despite containing lower amounts of the carrier lipid than the former. This observation indicates a possible existence of threshold levels for phospholipids carriers beyond which dissolution could be adversely affected. DSC studies further confirmed the dissolution behavior of these formulations demonstrating different levels of amorphous to crystalline nature. Results of HPLC analysis from Caco-2 cell culture studies showed increase in transport of PXCM from all formulations, with formulation (C) showing the maximum increase followed by formulations (B) and (A), when compared to control. The apparent permeability coefficients (Papp) were calculated to be 7.92x10(-6), 9.48x10(-6), 9.2x10(-6) and 5.6x10(-6)cm/s for formulations (A)-(C) and control, respectively. Overall, permeation appeared to improve for all formulations over the control. Stability studies at various temperatures showed all formulations to have good stability for the first 6 months; then a decline in dissolution rates was observed, especially for PEG-based lipid carrier systems, attributed to the increase in crystalline content of the solid dispersions upon storage.

A common approach utilized in validating the cleaning of multi-product equipment is to challenge the cleaning cycle with the hardest-to-clean, or "worst-case", soil. The worst-case soil is often determined through bench-scale experiments. These experiments can be tricky and time-consuming, especially when the dissolution dynamics of the soils are characterized by crossovers beyond the limit of visual detection. This paper describes a simple in situ method for monitoring soil dissolution dynamics and rapidly determining worst-case soils for cleaning validation. The method utilizes the real-time response of electrical conductivity to identify crossovers and to determine the optimal time for endpoint analysis. The results address the sensitivity and reproducibility of the method and the identification of critical experimental parameters. The dissolution dynamics of several parenteral products are compared.

Pancreatic cancer ranks as the fourth most deadly form of cancer in the United States with about 37,000 deaths each year. This study evaluated the chemopreventive potential of combinations of free and nano-encapsulated mixtures of ferulic acid (FA) and aspirin (ASP) in low doses to human pancreatic cancer cells, MIA PaCa-2 and Panc-1. Cell lines were tested according to published protocols wherein cells were seeded in 96-well microplates and incubated with the unmodified and modified combinations for 72 h and analyzed using MTS cell viability assay method. Apoptosis was measured by flow cytometry. Results demonstrated that free FA (200 µM) and ASP (1 mM) combinations on average reduced cell viability by 60% (P Citation Format: Arvind Thakkar, Jeffrey Wang, Sunil Prabhu. Novel combinations of nano-formulated ferulic acid and aspirin show high potential for chemoprevention of pancreatic cancer. [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 2155. doi:10.1158/1538-7445.AM2014-2155

The presence of bound and unbound zinc in the crystal matrix of protein suspensions helps physically stabilize the crystal and limits the dissolution of the drug. In case of zinc insulin suspensions, dissolution can be promoted by complexation of zinc with an ionic species for which the zinc has a greater affinity, these complexing ions having either formulation and/or physiological relevance. The purpose of this work was to use ligand-complexed formulations of insulin suspensions to gain an understanding of these products' dissolution performance and to establish a fundamental understanding of the rate-limiting steps in zinc insulin dissolution. Our group has suggested that the critical factors to zinc insulin dissolution are: (1) chemical complexation (zinc with ionic species); and (2) drug transport (insulin diffusion and solubility). Dissolution studies conducted using different ionic species (acetate, phosphate, citrate and EDTA) in a spin-filter device demonstrated a rank order correlation for different sources of zinc insulin; it was observed that human zinc insulin dissolved faster than bovine insulin, these differences attributed to their binding properties and the respective affinities to the various ionic species used. Also, as the amount of crystallinity increased in a formulation, a rank order increase in dissolution times was observed. The project also identified a sensitive and reproducible dissolution testing methodology. Overall, this study demonstrated that: (1) the complexation rate-limiting step was more significant in the dissolution of zinc insulin than the diffusion rate-limiting step; and (2) that dissolution kinetics depended primarily on the source and solid state differences and the binding affinities of the zinc insulin.

Pancreatic cancer ranks as the fourth most deadly form of cancer in the United States with approximately 37,000 deaths each year. The present study aims to evaluate the combinatorial chemopreventive effects of low doses of chloroquine (CQ) and aspirin (ASP) alone and in combination CQA (CQ+ASP) on three pancreatic cancer cell lines, MIA PaCa-2, Panc-1 and BxPC-3. Results demonstrated that low doses of CQ (6.25μM) and ASP (1mM) alone did not reduce cell viability, whereas combination CQA reduced cell viability by ∼80% (P Citation Format: Arvind Thakkar, B. Karthik Grandhi, Jeffrey Wang, Sunil Prabhu. Inhibition of PI3K-mTOR pathway using combination of chloroquine and aspirin regimen leads to apoptosis in pancreatic cancer 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 2596. doi:10.1158/1538-7445.AM2013-2596

Pancreatic cancer ranks as the fourth most deadly form of cancer in the United States with approximately 37,000 deaths each year. Due to poor prognosis, attention has recently focused on the chemoprevention of this disease rather than on diagnosis and treatment. The present study aims to evaluate the combinatorial chemopreventive effects of low doses of aspirin (ASP), curcumin (CUR), sulforaphane (SFN) alone and in combination ACS (ASP+CUR+SFN) on two pancreatic cancer cell lines, MIA PaCa-2 and Panc-1. Results demonstrated that low doses of ASP (1mM), CUR (10μM) and SFN (5μM) alone did not reduce cell viability, whereas combination ACS reduced cell viability by ∼70% (P Citation Format: Arvind Thakkar, Dhruvitkumar Sutaria, Karthik Grandhi, Ying Huang, Jeffrey Wang, Sunil Prabhu. Activation of ERK1/2 using combination of aspirin, curcumin, and sulforaphane regimen leads to NF-κB inhibition and apoptosis in pancreatic cancer cells. [abstract]. In: Proceedings of the Eleventh Annual AACR International Conference on Frontiers in Cancer Prevention Research; 2012 Oct 16-19; Anaheim, CA. Philadelphia (PA): AACR; Cancer Prev Res 2012;5(11 Suppl):Abstract nr A65.

Pancreatic cancer caused 40,000 deaths in 2010 and is currently ranked as the fourth leading cause of deaths in the United States. Due to poor prognosis of this disease, it is imperative that chemopreventive strategies be developed to control the formation of the initial neoplastic lesions. We have previously reported the effectiveness of a low dose combination regimen of aspirin, curcumin and sulforaphane (ACS) on MIA-PaCa-2 and Panc-1 pancreatic cancer cell lines. The objective of the current study was to use the ACS combination regimens in vivo to demonstrate the molecular targets responsible for prevention of pancreatic cancer in N-nitrosobis(2-oxopropyl) amine (BOP) carcinogen treated Syrian golden hamsters. BOP treated Syrian golden hamsters (n=6/group) were sacrificed after 24 weeks of treatment with a low (20+45+1.6), medium (67+150+5) and high (200+450+16) doses (in mg/kg) of ACS, respectively. Saline and BOP controls were included in the study. The pancreatic tissues were processed for isolation of nuclear and cytoplasmic proteins, and 20 mg of tissue was used for isolation of total RNA. Results from the Western blot show that low dose of ACS combined regimen down-regulated active NF-κB by inhibiting phosphorylation of IαBβ and p65. Low dose ACS also inhibited phosphorylation of Akt, needed for IKK activation. Consequently at the transcription level, low dose ACS significant down-regulated NF-κB regulated gene products TNFα, IL1α and IL6 compared to BOP controls (p2 fold down-regulation in low dose regimen compared to BOP controls. With the high and medium doses, a trend similar to the low dose ACS was evident, whereby the effects were more significant at higher doses (p Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 2589. doi:1538-7445.AM2012-2589