Thymoquinone Glucuronide Conjugated Magnetic Nanoparticle for Bimodal Imaging and Treatment of Cancer as a Novel Theranostic Platform.

BACKGROUND: Theranostic oncology combines therapy and diagnosis and is a new field of medicine that specifically targets the disease by using targeted molecules to destroy the cancerous cells without damaging the surrounding healthy tissues. OBJECTIVE: We aimed to develop a tool that exploits enzymatic TQ release from glucuronide (G) for the imaging and treatment of lung cancer. We added magnetic nanoparticles (MNP) to enable magnetic hyperthermia and MRI, as well as 131I to enable SPECT imaging and radionuclide therapy. METHODS: A glucuronide derivative of thymoquinone (TQG) was enzymatically synthesized and conjugated with the synthesized MNP and then radioiodinated with 131I. New Zealand white rabbits were used in SPECT and MRI studies, while tumor modeling studies were performed on 6-7- week-old nude mice utilized with bioluminescence imaging. RESULTS: Fourier-transform infrared spectroscopy (FTIR) and nuclear magnetic resonance (NMR) spectra confirmed the expected structures of TQG. The dimensions of nanoparticles were below 10 nm and they had rather polyhedral shapes. Nanoparticles were radioiodinated with 131I with over 95% yield. In imaging studies, in xenograft models, tumor volume was significantly reduced in TQGMNP-treated mice but not in non-treated mice. Among mice treated intravenously with TQGMNP, xenograft tumor models disappeared after 10 and 15 days, respectively. CONCLUSION: Our findings suggest that TQGMNP in solid, semi-solid and liquid formulations can be developed using different radiolabeling nuclides for applications in multimodality imaging (SPECT and MRI). By altering the characteristics of radionuclides, TQGMNP may ultimately be used not only for diagnosis but also for the treatment of various cancers as an in vitro diagnostic kit for the diagnosis of beta glucuronidase-rich cancers.

Synthesis, characterization and radiolabeling of folic acid modified nanostructured lipid carriers as a contrast agent and drug delivery system.

Nanostructured lipid carriers (NLCs) are the new generation of solid lipid drug delivery systems. Their suitability as contrast agents for gamma scintigraphy is an attracting major attention. The aim of current study was to prepare surface modified nanostructured lipid carrier system for paclitaxel (PTX) with active targeting and imaging functions. In accordance with the purpose of study, PTX loaded nanostructured lipid carriers (NLCs) prepared, modified with a folate derivative and radiolabeled with technetium-99m tricarbonyl complex (99mTc(CO)3+). Cellular incorporation ratios of radiolabeled nanoparticles (99mTc(CO)3-PTX-NLC) were investigated in vitro on three cancer cell lines. Additionally in vivo animal studies conducted to evaluate biological behavior of 99mTc(CO)3-PTX-NLC on female Wistar Albino rats. Biodistribution results showed that the folate derivative modified 99mTc(CO)3-PTX-NLC had considerably higher uptake in folate receptor positive organs. The data obtained from present study could be useful in the design of biodegradable drug carriers of PTX and folate receptor based tumor imaging agents.

Nonionic, water self-dispersible "hairy-rod" poly(p-phenylene)-g-poly(ethylene glycol) copolymer/carbon nanotube conjugates for targeted cell imaging.

The generation and fabrication of nanoscopic structures are of critical technological importance for future implementations in areas such as nanodevices and nanotechnology, biosensing, bioimaging, cancer targeting, and drug delivery. Applications of carbon nanotubes (CNTs) in biological fields have been impeded by the incapability of their visualization using conventional methods. Therefore, fluorescence labeling of CNTs with various probes under physiological conditions has become a significant issue for their utilization in biological processes. Herein, we demonstrate a facile and additional fluorophore-free approach for cancer cell-imaging and diagnosis by combining multiwalled CNTs with a well-known conjugated polymer, namely, poly(p-phenylene) (PP). In this approach, PP decorated with poly(ethylene glycol) (PEG) was noncovalently (π-π stacking) linked to acid-treated CNTs. The obtained water self-dispersible, stable, and biocompatible f-CNT/PP-g-PEG conjugates were then bioconjugated to estrogen-specific antibody (anti-ER) via -COOH functionalities present on the side-walls of CNTs. The resulting conjugates were used as an efficient fluorescent probe for targeted imaging of estrogen receptor overexpressed cancer cells, such as MCF-7. In vitro studies and fluorescence microscopy data show that these conjugates can specifically bind to MCF-7 cells with high efficiency. The represented results imply that CNT-based materials could easily be fabricated by the described approach and used as an efficient "fluorescent probe" for targeting and imaging, thereby providing many new possibilities for various applications in biomedical sensing and diagnosis.

Gold nanoparticle probes: design and in vitro applications in cancer cell culture.

A new architecture has been designed by the conjugation of [(18)F]2-fluoro-2-deoxy-D-glucose ((18)F-FDG), gold nanoparticles (AuNPs), and anti-metadherin (Anti-MTDH) antibody which is specific to the metadherin (MTDH) over-expressed on the surface of breast cancer cells. Mannose triflate molecule is used as a precursor for synthesis of (18)F-FDG by nucleophilic fluorination. For the conjugation of (18)F-FDG and AuNPs, cysteamine was first bound to mannose triflate (Man-CA) before synthesizing of (18)F-FDG which has cysteamine sides ((18)FDG-CA). Then, (18)FDG-CA was reacted with HAuCl(4) to obtain AuNPs and with NaBH(4) for reduction of AuNPs. At the end of this procedure, AuNPs were conjugated to (18)F-FDG via disulphide bonds ((18)FDG-AuNP). For the conjugation of Anti-MTDH, 1,1'-carbonyl diimidazol (CDI) was bound to the (18)FDG-AuNP, and Anti-MTDH was conjugated via CDI ((18)FDG-AuNP-Anti-MTDH). This procedure was also performed by using Na(19)F to obtain non-radioactive conjugates ((19)FDG-AuNP-Anti-MTDH). Scanning electron microscopy (SEM) images demonstrated that synthesized particles were in nano sizes. (18)FDG-AuNP-Anti-MTDH conjugate was characterized and used as a model probe containing both radioactive and optical labels together as well as the biological target. The (18)FDG-AuNP-Anti-MTDH conjugate was applied to MCF7 breast cancer cell line and apoptotic cell ratio was found to be increasing from 2% to 20% following the treatment. Hence, these results have promised an important application potential of this conjugate in cancer research.

Enzymatic synthesis of (125/131)I labeled 8-hydroxyquinoline glucuronide and in vitro/in vivo evaluation of biological influence.

8-Hydroxyquinoline (8-OHQ) is a long-known molecule which due to its metal-complexation ability is frequently used for analysis. It is also called oxine. Oxine and derivatives have been investigated to process antitumor and antimicrobial activities. 8-Hydroxyquinolyl-glucuronide (8-OHQ-Glu) was enzymatically synthesized using microsome preparates separated from Hutu-80 cells, labeled with (125)I to perform a radionuclide labeled prodrug and investigated of its biological affinities on Hutu-80 (human duodenum intestinal adenocarcinoma), Caco-2 (human colorectal adenocarcinoma), Detroit 562 (human pharynx adenocarcinoma) cells and ACBRI 519 (primary human small intestine epithelial cells) in this work. UDP-glucuronyl transferase (UDPGT) rich microsome preparates, which are used for glucuronidation in enzymatic synthesis, were extracted from Hutu-80 cells. 8-OHQ-Glu components were labeled using iodogen method with (125)I and (131)I. Structural analyses were performed with LC/MS/MS, (1)H NMR and (13)C-MMR for identify and measure chemical constituents. Results confirmed expected molecular structure. 8-OHQ-Glu could successfully radioiodinated with (125/131)I according to iodogen method. (125)I-8-OHQ-glucuronide incorporated with human gastrointestinal cancer cells such as Detroit-562 (human pharynx adenocarcinoma) (12.6%), Caco-2 (human colorectal adenocarcinoma) (7.8%), Hutu- 80 (human duodenum intestinal adenocarcinoma) (9.5%) and ACBRI 519 (primary human small intestine epithelial cells) (6.40%). (131)I-8-OHQ-Glu was tested in mice bearing subcutaneously implanted Caco-2 colorectal adenocarcinoma cells. The results demonstrated that radioiodinated 8-OHQ-Glu may be promising anticancer prodrug.

A new approach for in vitro imaging of breast cancer cells by anti-metadherin targeted PVA-pyrene.

Poly(vinyl alcohol)-pyrene-anti-metadherin (PVA-Py-(Anti-MTDH)), a novel antibody based water soluble probe containing both fluorescent and target sites in the structure for in vitro imaging of breast cancer cells is reported here. Since breast cancer cells have an excess of MDTH protein expressed on the surface, a PVA-Py prepared by "Click chemistry" approach is targeted by Anti-MTDH antibody and applied to the MCF-7 cell line. After characterization, the designed architecture was evaluated in terms of cell incorporation efficiency and compared with a non-targeted structure (PVA-Py). Atomic force microscopy (AFM) and fluorescence microscopy images of cells after incubation of the probe molecules were also obtained to monitor the interaction of the probes with the cancerous cells.

Synthesis and biodistribution studies of two novel radiolabeled estrone derivatives.

BACKGROUND: Two 99mTc-DTPA attached estrone derivatives were synthesized and their radiopharmaceutical potential was determined using female albino Wistar rats. MATERIALS AND METHODS: Two novel radiolabeled estrone derivatives, 99mTc-2,2',2"2"'-(2,2'-(2-(3-methoxy-13-methyl-17-oxo-7, 8, 9, 11, 12, 13, 14, 15, 16, 7-decahydro-6H- cyclopenta[a]phenanthren-2-ylamino)-2- oxoethylazanediyl) bis(ethane-2,1-diyl)) bis(azanetriyl) tetraacetic acid (99mTc-2-DTPA-3-methoxy estrone) and 9mTc-2,2',2",2'"-(2,2'- (2-(3-methoxy-13-methyl-17-oxo- 7,8,9,11,12,13,14,15,16,17-decahydro- 6H-cyclopenta[a]phenanthren-4-ylamino)-2-oxoethylazanediyl) bis(ethane-2,1-diyl))bis(azanetriyl)tetraacetic acid (99mTc-4-DTPA-3-methoxy estrone) were synthesized starting from estrone (3-hydroxy-13-methyl-7,8, 9,11,12,13,15,16-octahydro-6H-cyclopenta[a]phenanthren-17(14H)-one) and DTPA anhydride (2-(bis(2-(2,6-dioxomorpholino)ethyl)amino)acetic acid) as potential estrogen receptor imaging agents. The products were crystallized in ethyl alcohol (95%), purified by high performance liquid chromatography (HPLC) and characterized by nuclear magnetic resonance (NMR) and infrared spectroscopy (IR). The effect of the radiolabeled compounds on the biological behaviour of the molecules was evaluated through biodistribution studies in female albino Wistar rats. The rats were sacrificed at various time intervals, their organs were removed, and the activities of organs were counted using a gamma counter equipped with a Cd(Te) solid state detector. RESULTS AND CONCLUSION: Organ uptake was calculated as activity/gram tissue and time versus activity curves were generated. The tissue distribution studies exhibited a receptor-mediated uptake in the target organs of the rats for each compound. Both 99mTc-2-DTPA-3-methoxy estrone and 99mTc-4-DTPA-3-methoxy estrone were stable in vitro and were mainly excreted through the hepatobiliary pathway. The biological data showed that the 99mTc-2-DTPA-3-methoxy estrone had higher uptake in the target tissues than the 99mTc-4-DTPA-3-methoxy estrone. The favourable in vitro/in vivo stability and biodistribution profiles suggest that these radioligands are good candidates for further exploration of their potential clinical applications.

Somatostatin with 99mTc and biodistribution studies in rats.

Somatostatin (SST) is a short-lived peptide hormone that regulates the endocrine system. The main use of the derivatives of SST is to diagnose diseases related to growth hormone and to use against some forms of cancer that involve growth hormone. Also, SST suppresses gastric acid secretion, gallbladder contractions, and pancreatic enzyme secretion. In this study, two different bifunctional chelating agents were used to examine the changes in the biologic half-life of SST. For this purpose, first D-penicillamine (D-PA) and diethylene triaminepentaacetic acid (DTPA) were used to label SST with (99m)Tc and then radiopharmaceutical potential of three (99m)Tc-labeled complexes, (99m)Tc-D-PA, (99m)Tc-D-PA-SST, and (99m)Tc-DTPA-SST, were compared with each other. Quality control for each labeled complex was established by using radiochromatographic methods. The radiolabeled complexes maintained their stabilities for 5 hours. Then, biodistribution studies were performed on Albino Wistar rats independently for three complexes. The results demonstrated that (99m)Tc-D-PA-SST exhibited long-term uptake in organs, and its clearance took longer than the (99m)Tc-DTPA-SST complex.