Nanocurcumin Formulation Research Articles

Is oral nano-curcumin formulation a safe and effective measure for preventing cisplatin-induced nephrotoxicity in cancer patients?

Nephrotoxicity is one of the most important complications in cancer patients under treatment with cisplatin-containing regimens. Curcumin, as the most important active component of Curcuma longa, is an antioxidant and anti-inflammatory compound. In this clinical trial, we assessed the preventive effect of nano-curcumin oral formulation against cisplatin-induced nephrotoxicity in cancer patients. In this triple-blind clinical trial 30 cancer patients on cisplatin were randomly included in the treatment group, receiving nano-curcumin 40 mg capsules ( n = 15) or the placebo group ( n = 15) twice a day during four chemotherapy courses. Kidney function was measured at the beginning of the study and then at the end of each course of chemotherapy. There was no significant difference in acute kidney injury occurrence rate and creatinine and blood urine nitrogen serum levels between the treatment and placebo groups at the end of each chemotherapy course ( P value >0.05). Just at the end of the first course, the difference was close to significant ( P = 0.055). We also found no difference in mortality and recurrence rate in an average 30-month follow-up. Nano-curcumin in the prescribed dose and duration was not effective in preventing cisplatin-induced nephrotoxicity in cancer patients in comparison with the placebo. Further studies with larger sample size using different doses and duration of nano-curcumin are recommended.

Microbial fermentation-based synthesis of nano-curcumin suggesting the role of pullulan in nano-formulation

Curcumin is a multitargeting nutraceutical with numerous health benefits, however, its efficacy is limited due to poor aqueous solubility and reduced bioavailability. While nano-formulation has emerged as an alternative to encounter such issues, it often involves use of toxic solvents. Microbial synthesis may be an innovative solution to address this lacuna. Present study, for the first time, reports exploitation of Aureobasidium pullulans RBF4A3 for production of nano-curcumin. For this purpose, Aureobasidium pullulans RBF4A3 was inoculated in YPD media along with curcumin (0.1 mg/mL) and incubated for 24 h, 48 h, and 72 h. Subsequently, residual sugar, biomass, EPS concentration, curcumin concentration, and curcumin nanoparticle size were measured. As a result, nano-curcumin with an average particle size of 31.63 nm and enhanced aqueous solubility was obtained after 72 h. Further, investigations suggested that pullulan, a reducing polysaccharide, played a significant role in curcumin nano-formulation. Pullulan-mediated nano-curcumin formulation, with an average particle size of 24 nm was achieved with conversion rate of around 59.19 %, suggesting improved aqueous solubility. Additionally, the anti-oxidant assay of the resulting nano-curcumin was around 53.7 % per μg. Moreover, kinetics and thermodynamic studies of pullulan-based nano-curcumin revealed that it followed first-order kinetics and was favored by elevated temperature for efficient bio-conversion. Also, various physico-chemical investigations like FT-IR, NMR, and XRD reveal that pullulan backbone remains intact while forming curcumin nanoparticle. This study may open up new avenues for synthesizing nano-polyphenols through a completely green and solvent free process with plausible diverse applications.

Preparation and evaluation of transdermal hydrogel of chitosan coated nanocurcumin for enhanced stability and skin permeability

Curcumin (CR) is a natural product that has great interest in biopharmaceutical applications. It has several biological effects that antagonizes inflammation, oxidative stress, and cancer. On the other hand, low aqueous solubility, stability, and dissolution rate are the main challenges to using curcumin. In the present investigation, curcumin was formulated as a nanosystem known as nanocurcumin using a combination of polyvinyl pyrrolidone (PVP) with different polymers as stabilizing agents. The nanocurcumin formulations were prepared and evaluated by measuring the particle size, surface charge, drug content, and stability. The selected nanocurcumin formulation prepared using PVPK30-Poloxamer 188 demonstrated a significantly smaller particle size (272.1 ± 15.32 nm) and a higher settlement volume ration as compared with other formulations. Also, the selected formulation showed higher solubility and dissolution rates as compared with other stabilizer combinations. Chitosan was added to the selected nanocurcumin formulation as a coating polymer to enhance short-term storage colloidal stability. The result of stability proved that chitosan (0.05%)-coated nanocurcumin dispersion showed good stability during storage as compared with non-coated nanocurcumin. Non-coated nanocurcumin and chitosan-coated curcumin were loaded into the Carbopol934 hydrogel matrix to investigate the effect on curcumin permeability across the skin. The results revealed that nanocurcumin hydrogel formulations showed a significantly higher amount of curcumin permeated across the skin.

Effect of hypobaric hypoxia on the fiber type transition of skeletal muscle: a synergistic therapy of exercise preconditioning with a nanocurcumin formulation.

Hypobaric hypoxia (HH) leads to various adverse effects on skeletal muscles, including atrophy and reduced oxidative work capacity. However, the effects of HH on muscle fatigue resistance and myofiber remodeling are largely unexplored. Therefore, the present study aimed to explore the impact of HH on slow-oxidative fibers and to evaluate the ameliorative potential of exercise preconditioning and nanocurcumin formulation on muscle anti-fatigue ability. C2C12 cells (murine myoblasts) were used to assess the effect of hypoxia (0.5%, 24h) with and without the nanocurcumin formulation (NCF) on myofiber phenotypic conversion. To further validate this hypothesis, male Sprague Dawley rats were exposed to a simulated HH (7620m) for 7days, along with NCF administration and/or exercise training. Both in vitro and in vivo studies revealed a significant reduction in slow-oxidative fibers (p < 0.01, 61% vs. normoxia control) under hypoxia. There was also a marked decrease in exhaustion time (p < 0.01, 65% vs. normoxia) in hypoxia control rats, indicating a reduced work capacity. Exercise preconditioning along with NCF supplementation significantly increased the slow-oxidative fiber proportion and exhaustion time while maintaining mitochondrial homeostasis. These findings suggest that HH leads to an increased transition of slow-oxidative fibers to fast glycolytic fibers and increased muscular fatigue. Administration of NCF in combination with exercise preconditioning restored this myofiber remodeling and improved muscle anti-fatigue ability.

A Comparative In Vitro Study: Cytotoxicity Effect of Curcumin Vs Biodegradable Nanoparticles/PVA Against HNO97 And Effect of Curcumin on Cell Cycle Phases

Purpose: This study evaluated that curcumin and biodegradable nanoparticles with PVA as capping material, in terms of cytotoxicity followed by cell cycle analysis against the HNO97 cell line (tongue carcinoma). Material and methods: 5mg of pure curcumin, 7.5 mg of Nano curcumin formulation, and SRB for IC50 done on five concentrations. A flow cytometry test was carried on to evaluate the effect of curcumin on the HNO97 cell line. Results: This study postulated that curcumin had the highest cytotoxicity effect against cell line (HNO97) with IC50=1.59 µg/ml in comparison to curcumin nanoparticles/PVA showed low cytotoxicity owing to the pH (7.4) and surface charges of PVA. Also, the cell cycle test for curcumin showed that the percentage of cells in G2/M phase was significantly increased (p=0.003). Conclusion: The study showed that curcumin has a powerful anticancer effect against in vitro tongue carcinoma. Generally, these results reported that curcumin can in vitro inhibit cancer cells.

A Comparative In Vitro Study: Cytotoxicity Effect of Curcumin Vs Biodegradable Nanoparticles/PVA Against HNO97 And Effect of Curcumin on Cell Cycle Phases

Purpose: This study evaluated that curcumin and biodegradable nanoparticles with PVA as capping material, in terms of cytotoxicity followed by cell cycle analysis against the HNO97 cell line (tongue carcinoma). Material and methods: 5mg of pure curcumin, 7.5 mg of Nano curcumin formulation, and SRB for IC50 done on five concentrations. A flow cytometry test was carried on to evaluate the effect of curcumin on the HNO97 cell line. Results: This study postulated that curcumin had the highest cytotoxicity effect against cell line (HNO97) with IC50=1.59 µg/ml in comparison to curcumin nanoparticles/PVA showed low cytotoxicity owing to the pH (7.4) and surface charges of PVA. Also, the cell cycle test for curcumin showed that the percentage of cells in G2/M phase was significantly increased (p=0.003). Conclusion: The study showed that curcumin has a powerful anticancer effect against in vitro tongue carcinoma. Generally, these results reported that curcumin can in vitro inhibit cancer cells.

Mitochondrial Ca2+ overload due to altered proteostasis amplifies apoptosis in C2C12 myoblasts under hypoxia: Protective role of nanocurcumin formulation.

Severe hypoxia triggers apoptosis leads to myofibers loss and is attributable to impaired intracellular calcium (iCa2+ ) homeostasis, resulting in reduced muscle activity. Hypoxia increases intracellular Ca2+ by activating the release of Ca2+ from iCa2+ stores, however, the effect of increased [iCa2+ ] on the mitochondria of muscle cells at high-altitude hypoxia is largely unexplored. This study examined mitochondrial Ca2+ overload due to altered expression of mitochondrial calcium uptake 1 (MICU1), that is, a gatekeeper of the mitochondrial Ca2+ uniporter, impaired mitochondrial membrane potential (ΔΨm). p53 stabilization and its translocation to the mitochondria were observed following disrupted mitochondrial membrane integrity in myoblasts under hypoxia. Furthermore, the downstream effects of p53 led to the upregulation of proapoptotic proteins (Bax, Caspase-3, and cytochrome C) in myoblasts under hypoxia. Nanocurcumin-pyrroloquinoline quinone formulation (NCF; Indian patent no. 302877), developed to address hypoxia-induced consequences, was found to be beneficial in maintaining mitochondrial Ca2+ homeostasis and limiting p53 translocation into mitochondria under hypoxia in muscle myoblasts. NCF treatment also modulates heat shock proteins and apoptosis-regulating protein expression in myoblasts. Conclusively, we proposed that mitochondrial Ca2+ overload due to altered MICU1 expression intensifies apoptosis and mitochondrial dysfunctionality. The study also reported that NCF could improve mitochondrial [Ca2+ ] homeostasis and antiapoptotic ability in C2C12 myoblasts under hypoxia.

A Nanocurcumin and Pyrroloquinoline Quinone Formulation Prevents Hypobaric Hypoxia-Induced Skeletal Muscle Atrophy by Modulating NF-κB Signaling Pathway.

Kushwaha, Asha D., and Deepika Saraswat. A nanocurcumin and pyrroloquinoline quinone formulation prevents hypobaric hypoxia-induced skeletal muscle atrophy by modulating NF-κB signaling pathway. High Alt Med Biol. 23:249-263, 2022. Background: Hypobaric hypoxia (HH)-induced deleterious skeletal muscle damage depends on exposure time and availability of oxygen at cellular level, which eventually can limit human work performance at high altitude (HA). Despite the advancements made in pharmacological (performance enhancer, antioxidants) and nonpharmacological therapeutics (acclimatization strategies), only partial success has been achieved in improving physical performance at HA. A distinctive combination of nanocurcumin (NC) and pyrroloquinoline quinone (PQQ) has been formulated (named NCF [nanocurcumin formulation], Indian patent No. 302877) in our laboratory, and has proven very promising in improving cardiomyocyte adaptation to chronic HH. We hypothesized that NCF might improve skeletal muscle adaptation and could be a performance enhancer at HA. Material and Methods: Adult Sprague-Dawley rats (220 ± 10 g) were divided into five groups (n = 6/group): normoxia vehicle control, hypoxia vehicle control, hypoxia NCF, hypoxia NC, and hypoxia PQQ. All the animals (except those in normoxia) were exposed to simulated HH in a chamber at temperature 22°C ± 2°C, humidity 50% ± 5%, altitude 25,000 ft for 1, 3, or 7 days. After completion of the stipulated exposure time, gastrocnemius and soleus muscles were excised from animals for further analysis. Results: Greater lengths of hypoxic exposure caused progressively increased muscle ring finger-1 (MuRF-1; p < 0.01) expression and calpain activation (0.56 ± 0.05 vs. 0.13 ± 0.02 and 0.44 ± 0.03 vs. 0.12 ± 0.021) by day 7, respectively in the gastrocnemius and soleus muscles. Myosin heavy chain type I (slow oxidative) fibers significantly (p > 0.01) decreased in gastrocnemius (>50%) and soleus (>46%) muscles by the seventh day of exposure. NCF supplementation showed (p ≤ 0.05) tremendous improvement in skeletal muscle acclimatization through effective alleviation of oxidative damage, and changes in calpain activity and atrophic markers at HA compared with hypoxia control or treatment alone with NC/PQQ. Conclusion: Thus, NCF-mediated anti-oxidative, anti-inflammatory effects lead to decreased proteolysis resulting in mitigated skeletal muscle atrophy under HH.

The cardioprotective effects of nano-curcumin against doxorubicin-induced cardiotoxicity: A systematic review.

Although the chemotherapeutic drug, doxorubicin, is commonly used to treat various malignant tumors, its clinical use is restricted because of its toxicity especially cardiotoxicity. The use of curcumin may alleviate some of the doxorubicin-induced cardiotoxic effects. Especially, using the nano-formulation of curcumin can overcome the poor bioavailability of curcumin and enhance its physicochemical properties regarding its efficacy. In this study, we systematically reviewed the potential cardioprotective effects of nano-curcumin against the doxorubicin-induced cardiotoxicity. A systematic search was accomplished based on Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines for the identification of all relevant articles on "the role of nano-curcumin on doxorubicin-induced cardiotoxicity" in the electronic databases of Scopus, PubMed, and Web of Science up to July 2021. One hundred and sixty-nine articles were screened following a predefined set of inclusion and exclusion criteria. Ten eligible scientific papers were finally included in the present systematic review. The administration of doxorubicin reduced the body and heart weights of mice/rats compared to the control groups. In contrast, the combined treatment of doxorubicin and nano-curcumin increased the body and heart weights of animals compared with the doxorubicin-treated groups alone. Furthermore, doxorubicin could significantly induce the biochemical and histological changes in the cardiac tissue; however, coadministration of nano-curcumin formulation demonstrated a pattern opposite to the doxorubicin-induced changes. The coadministration of nano-curcumin alleviates the doxorubicin-induced cardiotoxicity through various mechanisms including antioxidant, anti-inflammatory, and antiapoptotic effects. Also, the cardioprotective effect of nano-curcumin formulation against doxorubicin-induced cardiotoxicity was higher than free curcumin.

Oral nano-curcumin formulation efficacy in the management of mild to moderate outpatient COVID-19: A randomized triple-blind placebo-controlled clinical trial.

BackgroundCurcumin, a natural polyphenolic compound, is proposed as a potential treatment option for patients with coronavirus disease by inhibiting the entry of virus to the cell, encapsulation of the virus and viral protease, as well as modulating various cellular signaling pathways. In this study, the efficacy and safety of nanocurcumin oral formulation has been evaluated in patients with mild‐moderate Coronavirus disease 2019 (COVID‐19) in outpatient setting.MethodsIn this triple‐blind randomized placebo‐controlled clinical trial, sixty mild to moderate COVID‐19 patients in outpatient setting who fulfilled the inclusion criteria were randomly allocated to treatment (n = 30) group to receive oral nanocurcumin formulation (Sinacurcumin soft gel which contains 40 mg curcuminoids as nanomicelles), two soft gels twice a day after food for 2 weeks or placebo (n = 30) group. Patients’ symptoms and laboratory data were assessed at baseline and during follow‐up period and compared between two groups.ResultsAll symptoms except sore throat resolved faster in the treatment group and the difference was significant for chills, cough and smell and taste disturbances. The CRP serum level was lower in the treatment group at the end of two weeks and the lymphocyte count was significantly higher in treatment group. No significant adverse reaction reported in the treatment group.ConclusionOral nanoformulation of curcumin can significantly improve recovery time in patients with mild to moderate COVID‐19 in outpatient setting. Further studies with larger sample size are recommended.

Quaternary ammonium poly (amidoamine) dendrimeric encapsulated nanocurcumin efficiently prevents cataract of rat pups through regulation of pro-inflammatory gene expression

Curcumin with its plethora of potential health benefits bears limitations of poor aqueous solubility and bioavailability. In order to alleviate this crucial drawback, the present study focuses development of a nanocurcumin formulation using functionalized poly amidoamine (PAMAM) dendrimer (third generation) with encapsulated curcumin and characterized. The efficiency of this nanocurcumin to prevent and or cure the selenite induced cataract in wistar rat pups compare to native curcumin was assessed through the expression of pro-inflammatory genes using in vivo and human infant lens epithelial (HLE-B3) cells in vitro systems.The solubility of functionalized nanocurcumin increased 345-fold and also in vitro release of curcumin prolonged nearly 17 times compared to native curcumin. Furthermore, nano-curcumin alleviated cataracts in orally dosed rat pups at 2.5 mg/kg body weight (b.w.) in both pre- and post-selenite treatment. This is in contrast to earlier studies that utilized much higher doses (75 mg/kg b. w.) of native curcumin that also fared with limited impact post-selenite treatment. In addition, nano-curcumin proves as effectively as native curcumin in reducing cell death, RNA degradation and gene expression of iNOS (inducible nitric oxide synthase) level induced by selenite.Thus, the formulated non-toxic encapsulated curcumin in nano-form using QPAMAM that can offset major limitations to its native counterpart with increased solubility, sustained release and increased bioavailability, prevented oxidant mediated cataract development and found promising.

Nano-Curcumin Regulates p53 Phosphorylation and PAI-1 Expression during Bleomycin Induced Injury in Alveolar Basal Epithelial Cells

Background: Bleomycin (BLM) is known to cause DNA damage in the Alveolar Epithelial Cells (AECs). It is reported that BLM is involved in the up-regulation of inflammatory molecules such as neutrophils, macrophages, chemokines and cytokines. The complex underlying mechanism for inflammation mediated progression of lung injury is still unclear. This investigation was designed to understand the molecular mechanisms associated with p53 mediated modulation of Plasminogen Activator Inhibitor-I (PAI-I) expression and its regulation by nano-curcumin formulation. Methods: A549 cells were treated with BLM to cause the cellular damage in vitro and commercially available nano-curcumin formulation was used as an intervention. Cytotoxic effect of nano-curcumin was analyzed using Methyl Thiazolyl Tetrazolium (MTT) assay. Protein expressions were analyzed using western blot to evaluate the p53 mediated changes in PAI-I expression. Results: Nano-curcumin showed cytotoxicity up to 88.5 % at a concentration of 20 μg/ml after 48 h of treatment. BLM exposure to the cells activated the phosphorylation of p53, which in turn increased PAII expression. Nano-curcumin treatment showed a protective role against phosphorylation of p53 and PAI-I expression, which in turn regulated the fibro-proliferative phase of injury induced by bleomycin. Conclusion: Nano-curcumin could be used as an effective intervention to regulate the severity of lung injury, apoptosis of AECs and fibro-proliferation during pulmonary injury.

Curcumin-Loaded Mesoporous Silica Particles as Wound-Healing Agent: An In vivo Study.

Background:Curcumin likely has wound-healing properties, but its poor pharmacokinetic attributes inhibit its potential. To overcome these limitations, a novel nanoformulation was previously developed, wherein curcumin was loaded into mesoporous silica particles.Objectives:The objective of the study is to assess the efficiency of this nanocurcumin formulation as a wound-healing agent in an animal model.Materials and Methods:Curcumin was loaded onto mesoporous silica particles. Eighteen healthy, test-naive male Wistar rats were randomly separated into two groups of 9: Group 1 (control) rats were treated topically with a standard drug (sulfadiazine) and Group 2 with 1% curcumin formulation. A circular excision wound was made, and topical application was performed twice a day. The excision diameters were measured on days 3, 6, 9, 12, 15, 18 and 21 of treatment. Three rats from each group were sacrificed on days 7, 14 and 21, and a cross-section from skin specimen in the excision injury was obtained for histological assessment of inflammation, angiogenesis, fibroblast proliferation, presence of collagen and reepithelization.Results:Wound contraction percentage in rats treated with curcumin nanoformulation was nonsignificantly higher than that in the control group (P > 0.05). In both groups, inflammatory reactions considerably reduced by day 21 of treatment, the angiogenesis process was almost complete by day 7, fibroblast proliferation noticeably rose by day 14, and a high degree of wound reepithelization was achieved by day 21, with no significant differences between the groups. Interestingly, by day 21, the level of collagen significantly increased in curcumin nanoformulation-treated rats compared with those treated with sulfadiazine.Conclusions:Curcumin nanoformulation likely enhanced wound repair by inhibiting the inflammatory response, stimulating angiogenesis, inducing fibroblast proliferation as well as enhancing reepithelization and synthesis of collagen. Therefore, the curcumin nanoformulation used in this study may have potential as a wound-healing ethnomedicine.

Nanocurcumin\u2013pyrroloquinoline formulation prevents hypertrophy\u2013induced pathological damage by relieving mitochondrial stress in cardiomyocytes under hypoxic conditions

This study investigates the therapeutic effect of a nanocurcumin formulation (NCF) containing nanocurcumin (NC) and pyrroloquinoline quinone (PQQ) on ameliorating hypoxia-induced stress in hypertrophied primary human ventricular cardiomyocytes (HVCM) under hypoxic conditions, as validated in a Sprague-Dawley rat model of chronic hypobaric hypoxia (cHH)-induced right ventricular hypertrophy (RVH). Based on our previous findings, here, we analyzed the improvement in the protective efficacy of NCF against mitochondrial damage. The electron transport chain Complexes’ activities were analyzed as a chief operational center for mitochondrial homeostasis, along with key gene and protein markers for mitochondrial biogenesis, redox function, fatty acid oxidation, bio-energetic deficit and cell survival. NCF supplementation imparts cyto-protection from hypoxia-induced hypertrophy and damage in both in vitro and in vivo models while maintaining mitochondrial homeostasis better than NC and PQQ alone. This study proposes the use of NCF as a potential candidate molecule for imparting protection from high altitude-induced maladies in ascendants.

Combinatorial therapy of exercise-preconditioning and nanocurcumin formulation supplementation improves cardiac adaptation under hypobaric hypoxia.

Chronic hypobaric hypoxia (cHH) mediated cardiac insufficiencies are associated with pathological damage. Sustained redox stress and work load are major causative agents of cardiac insufficiencies under cHH. Despite the advancements made in pharmacological (anti-oxidants, vasodilators) and non-pharmacological therapeutics (acclimatization strategies and schedules), only partial success has been achieved in improving cardiac acclimatization to cHH. This necessitates the need for potent combinatorial therapies to improve cardiac acclimatization at high altitudes. We hypothesize that a combinatorial therapy comprising preconditioning to mild aerobic treadmill exercise and supplementation with nanocurcumin formulation (NCF) consisting of nanocurcumin (NC) and pyrroloquinoline quinone (PQQ) might improve cardiac adaptation at high altitudes. Adult Sprague-Dawley rats pre-conditioned to treadmill exercise and supplemented with NCF were exposed to cHH (7620 m altitude corresponding to pO2~8% at 28±2°C, relative humidity 55%±1%) for 3 weeks. The rat hearts were analyzed for changes in markers of oxidative stress (free radical leakage, lipid peroxidation, manganese-superoxide dismutase [MnSOD] activity), cardiac injury (circulating cardiac troponin I [TnI] and T [cTnT], myocardial creatine kinase [CK-MB]), metabolic damage (lactate dehydrogenase [LDH] and acetyl-coenzyme A levels, lactate and pyruvate levels) and bio-energetic insufficiency (ATP, p-AMPKα). Significant modulations (p≤0.05) in cardiac redox status, metabolic damage, cardiac injury and bio-energetics were observed in rats receiving both NCF supplementation and treadmill exercise-preconditioning compared with rats receiving only one of the treatments. The combinatorial therapeutic strategy showed a tremendous improvement in cardiac acclimatization to cHH compared to either exercise-preconditioning or NCF supplementation alone which was evident from the effective modulation in redox, metabolic, contractile and bio-energetic homeostasis.

Abstract A57: Curcumin nanoformulation: A new therapeutic approach for cervical cancer treatment

Abstract Underserved populations such as African Americans (AA) and American Indians (AI) are experiencing increasing prevalence and mortality rates of cervical cancer (CxCa) as compared to Caucasians (CA). CxCa is one of the most common and deadly cancers among women worldwide and is associated with persistent Human Papillomavirus (HPV) infection. A persistent oncogenic HPV infection itself is not enough to immortalize and transform the epithelial host cell, as over a course of time only a small proportion of women with chronic HPV infection progress to develop disease. Additional factors are needed to acquire an immortal, malignant and invasive phenotype. In addition to HPV infection, cigarette smoking is a known risk factor. Polycyclic aromatic hydrocarbons (PAHs), such as Benzo[a]pyrene (BaP), are among the numerous carcinogens associated with cigarette smoking. The level of BaP has been found to be elevated in the cervical mucus of women who smoke. Moreover, a number of studies have implicated BaP to affect both the viral life cycle and the cellular environment. BaP has been shown to stimulate high levels of virion synthesis in cell lines prolifically infected with HPV. Furthermore, in other cancers the primary metabolite of BaP, benzo(a)pyrene diol epoxide increases the expression of oncogenic transcription factors like NFκB and oncogenic cytokines such as Interleukin-6 (IL-6). IL-6 in turn is known to activate transcription factors such as STAT3 and enhance the malignant transformation of cells. This is achieved through the transcriptional activation of oncogenic microRNAs (miRNA) like miRNA-21. MiRNAs are small non-coding RNAs that regulate gene expression and their aberrant regulation/function has been reported in a large majority of cancers. miRNA-21 has been shown to enhance chemoresistance to established regimens of chemotherapy and radiation therapy. This resistance to chemo-radio therapy with prolonged treatment, resulting in an invasive form of cancer, requires the development of novel therapeutic modalities to conquer chemo resistance and improve overall life expectancy of patients. Nanotechnology provides targeted delivery of anti-cancer drugs and indicates a new approach for cancer diagnosis and treatment. Our lab has developed PLGA (poly [lactic-co-glycolic acid]), an FDA approved polymer coated nanoparticle(s) using a well-known anti-cancer drug Curcumin (Cur) for the enhanced bioavailability and improved therapeutic efficacy of Cur, known as nano-curcumin (NC). In comparison to pure Cur, NC effectively inhibited Caski/ SiHa cell growth and also induced apoptosis as determined by staining for Annexin V/7AAD. PI staining revealed that NC treatment arrests growth cycle of Caski/SiHa cells in the G2-M phase. Additionally, NC treatment caused a marked decrease in the levels of miRNA 21, an oncomiRNA associated with chemoresistance, and enhanced by BaP, with a concomitant increase in the tumor suppressor miRNA-214 (decreased by BaP). It was found to down-regulate the expression of oncogenic proteins and cytokines such as IL-6 and enhance the expression of tumor suppressor entities as evident from Western blots. Moreover, NC formulation effectively reduced the tumor burden in Non SCID gamma mice. Furthermore, it decreased the expression of oncogenic miRNA-21 in in vivo experiments as observed through ISH. Our findings show that PLGA based NC significantly inhibits Caski/SiHa CxCa cell tumorigenicity and regulates the expression of miRNAs, associated with CxCa. In vivo experiments show that NC is efficacious in reducing the tumor burden. Therefore, NC may be a novel chemo-preventive and therapeutic modality for the overall management of CxCa. Citation Format: Mohd Saif Zaman, Neeraj Chauhan, Diane Maher, Rishi Gara, Mohammed Sikander, Murali Mohan Yallapu, Sheema Khan, Meena Jaggi, Subhash Chauhan. Curcumin nanoformulation: A new therapeutic approach for cervical cancer treatment. [abstract]. In: Proceedings of the Seventh AACR Conference on The Science of Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; Nov 9-12, 2014; San Antonio, TX. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2015;24(10 Suppl):Abstract nr A57.

Mitigating prolonged QT interval in cancer nanodrug development for accelerated clinical translation

BackgroundCardiac toxicity is the foremost reason for drug discontinuation from development to clinical evaluation and post market surveillance [Fung 35:293-317, 2001; Piccini 158:317-326 2009]. The Food and Drug Administration (FDA) has rejected many potential pharmaceutical agents due to QT prolongation effects. Since drug development and FDA approval takes an enormous amount of time, money and effort with high failure rates, there is an increased focus on rescuing drugs that cause QT prolongation. If these otherwise safe and potent drugs were formulated in a unique way so as to mitigate the QT prolongation associated with them, these potent drugs may get FDA approval for clinical use. Rescuing these compounds not only benefit the patients who need them but also require much less time and money thus leading to faster clinical translation. In this study, we chose curcumin as our drug of choice since it has been shown to posses anti-tumor properties against various cancers with limited toxicity. The major limitations with this pharmacologically active drug are (a) its ability to prolong QT by inhibiting the hERG channel and (b) its low bioavailability. In our previous studies, we found that lipids have protective actions against hERG channel inhibition and therefore QT prolongation.ResultsResults of the manual patch clamp assay of HEK 293 cells clearly illustrated that our hybrid nanocurcumin formulation prevented the curcumin induced inhibition of hERG K+ channel at concentrations higher than the therapeutic concentrations of curcumin. Comparing the percent inhibition, the hybrid nanocurcumin limited inhibition to 24.8% at a high curcumin equivalent concentration of 18 μM. Liposomal curcumin could only decrease this inhibition upto 30% only at lower curcumin concentration of 6 μM but not at 18 μM concentration.ConclusionsHere we show a curcumin encapsulated lipopolymeric hybrid nanoparticle formulation which could protect against QT prolongation and also render increased bioavailability and stability thereby overcoming the limitations associated with curcumin.

Polymeric curcumin nanoparticle pharmacokinetics and metabolism in bile duct cannulated rats.

The objective of this study was to compare the pharmacokinetics and metabolism of polymeric nanoparticle-encapsulated (nanocurcumin) and solvent-solubilized curcumin formulations in Sprague-Dawley (SD) rats. Nanocurcumin is currently under development for cancer therapy. Since free, unencapsulated curcumin is rapidly metabolized and excreted in rats, upon intravenous (i.v.) administration of nanocurcumin only nanoparticle-encapsulated curcumin can be detected in plasma samples. Hence, the second objective of this study was to utilize the metabolic instability of curcumin to assess in vivo drug release from nanocurcumin. Nanocurcumin and solvent-solubilized curcumin were administered at 10 mg curcumin/kg by jugular vein to bile duct-cannulated male SD rats (n = 5). Nanocurcumin increased the plasma Cmax of curcumin 1749 fold relative to the solvent-solubilized curcumin. Nanocurcumin also increased the relative abundance of curcumin and glucuronides in bile but did not dramatically alter urine and tissue metabolite profiles. The observed increase in biliary and urinary excretion of both curcumin and metabolites for the nanocurcumin formulation suggested a rapid "burst" release of curcumin. Although the burst release observed in this study is a limitation for targeted tumor delivery, nanocurcumin still exhibits major advantages over solvent-solubilized curcumin, as the nanoformulation does not result in the lung accumulation observed for the solvent-solubilized curcumin and increases overall systemic curcumin exposure. Additionally, the remaining encapsulated curcumin fraction following burst release is available for tumor delivery via the enhanced permeation and retention effect commonly observed for nanoparticle formulations.

Highly Stabilized Curcumin Nanoparticles Tested in an In Vitro Blood–Brain Barrier Model and in Alzheimer’s Disease Tg2576 Mice

The therapeutic effects of curcumin in treating Alzheimer's disease (AD) depend on the ability to penetrate the blood-brain barrier. The latest nanoparticle technology can help to improve the bioavailability of curcumin, which is affected by the final particle size and stability. We developed a stable curcumin nanoparticle formulation to test in vitro and in AD model Tg2576 mice. Flash nanoprecipitation of curcumin, polyethylene glycol-polylactic acid co-block polymer, and polyvinylpyrrolidone in a multi-inlet vortex mixer, followed by freeze drying with β-cyclodextrin, produced dry nanocurcumin with mean particle size <80 nm. Nanocurcumin powder, unformulated curcumin, or placebo was orally administered to Tg2576 mice for 3 months. Before and after treatment, memory was measured by radial arm maze and contextual fear conditioning tests. Nanocurcumin produced significantly (p=0.04) better cue memory in the contextual fear conditioning test than placebo and tendencies toward better working memory in the radial arm maze test than ordinary curcumin (p=0.14) or placebo (p=0.12). Amyloid plaque density, pharmacokinetics, and Madin-Darby canine kidney cell monolayer penetration were measured to further understand in vivo and in vitro mechanisms. Nanocurcumin produced significantly higher curcumin concentration in plasma and six times higher area under the curve and mean residence time in brain than ordinary curcumin. The P(app) of curcumin and tetrahydrocurcumin were 1.8×10(-6) and 1.6×10(-5)cm/s, respectively, for nanocurcumin. Our novel nanocurcumin formulation produced highly stabilized nanoparticles with positive treatment effects in Tg2576 mice.

FORMULATION OF NANOCURCUMIN USING LOW VISCOSITY CHITOSAN POLYMER AND ITS CELLULAR UPTAKE STUDY INTO T47D CELLS

Using of curcumin as anti cancer agent is restricted by its low solubility, therefore it has low bioavability. This obstacle can be solved by the development of curcumin nanoparticle. Nanoparticle technology has been started to be developed as an alternative solution to improve drug delivery pofile, especially for the less bio-available chemical. This study was aimed to develope nanocurcumin formulation with low viscosity chitosan as the matrix and to study its ability to be taken into the cells in vitro. Method used in the formulation of nanocurcumin in this study is by ionic gelation followed by freeze drying. Entrapment Efficiency then assayed, and its stability was tested by incubating the formula into artificial intestinal fluid (AIF). Furthermore, its toxicity was evaluated, also its cellular uptake ability into T47D cell line. It was found that the Entrapment Efficiency in acetate buffer at pH 4 is higher than at pH 5. This formula also has a good stability in AIF. For the cellular uptake study through fluorescence microscope, it was found that the complex has an ability to penetrate cellular membrane into the cytosol. The cytotoxicity study tell us that the nanocurcumin is non-toxic to normal cell line. For the characterization of the nanoparticles, the average size of this particle is 269.8 nm, its zeta-potential is +18.63 mV, with spherical particle morphology. From the result of this study, it is concluded that formulation of nanocurcumin using low viscosity chitosan polymer as the matrix has a great potential as an alternative for anticancer therapy. Key words: nanoparticle, curcumin, low viscosity chitosan, T47D cell line.