Cisplatin Solution Research Articles

Characterization and cytotoxicity studies of DPPC:M2+ novel delivery system for cisplatin thermosensitivity liposome with improving loading efficiency

A novel, metal ion-assisted drug-loading model, in which the metal ion was used to modify the microstructure of DPPC bilayers, has been developed to improve the drug-loading efficiency of cisplatin thermosensitivity liposomes. The reactions of dipalmitoyl phosphatidylcholine (DPPC) with diverse metal ions (Zn2+, Cu2+, Mn2+ and Mg2+) yield four typical liposomes, which have been characterized by FT-IR, Raman and fluorescence techniques; the mechanism for higher drug encapsulation efficiency has also been investigated. In these prepared liposomes, the conformation of DPPC is changed due to the electrostatic interaction between the metal ions and phospholipid acyl group, leading to a closer arrangement of the lipid hydrocarbon chains and higher Tm of DPPC. As a result, the encapsulation efficiency of metal ion-assisted loading liposome is significantly higher than that of metal ion-free state. While for the release time, all four metal ion-assisted liposomes could be released within 10min at 42±0.5°C, which approach to the phase transition temperature, indicating that the introduction of metal ions into the DPPC bilayer membranes has no influence on the thermosensitivity of the liposome. Furthermore, the higher cytotoxicity of metal ion-bounded liposomes than that of free cisplatin solution suggests that high encapsulation efficiency can cause cytotoxicity increase. Hence, this work highlighted that metal ion-assisted loading model increased the encapsulation efficiency and cell cytotoxicity of cisplatin in thermosensitive liposomes with no obvious effects on sustained and temperature-dependent drug release.

A Cisplatin Derivative that Inhibits Collagen Fibril‐Formation in vitro

Using an in vitro random screening of small-molecule compounds, we discovered cis-diamminedichloroplatinum(II) (cisplatin), an anticancer agent, as a potential inhibitor of collagen fibril-formation. The inhibitory effect was found only when cisplatin was dissolved in dimethylsulphoxide (DMSO), indicating that the active species were cisplatin derivatives formed in the DMSO solution. The cisplatin derivatives inhibited the formation of collagen fibrils in vitro without affecting the triple-helical conformation of the collagen molecules. Incubation with the cisplatin solution in DMSO also inhibited in situ deposition of collagen fibrils in a human umbilical vein endothelial cell (HUVEC) culture. In addition, the derivatization of cisplatin in DMSO abolished the cytotoxicity of the original compound. The platinum complex was further revealed to interact with specific sites on the collagen triple helix, and the binding sites were suggested to contain His and/or Met residues. Mass spectrometry analysis of the cisplatin solution in DMSO and a structure-activity relationship study strongly suggested that the active compound is [Pt(NH3 )2 (Cl)(DMSO)](+) . This platinum complex will be useful for investigating molecular mechanisms of collagen self-assembly and for drug development for the treatment of fibrotic diseases.

Improved cisplatin delivery in cervical cancer cells by utilizing folate-grafted non-aggregated gelatin nanoparticles

PurposeCisplatin is highly effective in the treatment of cervical cancer. However, in therapeutic doses, cisplatin induces several adverse effects due to undesirable tissue distribution. Therefore, it is worth targeting cisplatin in cervical cancer cells by implicating non-aggregated ligand-modified nanotherapeutics. Methods and resultsHere, we report the preparation of non-aggregated folic acid-conjugated gelatin nanoparticles of cisplatin (Cis-GNs-FA) by two-step desolvation method with mean particle size of 210.6±9.6nm and 140.5±10.9nm for Cis-GNs to improve the drug delivery in cervical cancer, HeLa cells. FTIR and DSC spectra confirmed the presence and stability of cisplatin in gelatin matrix. Furthermore, amorphization of cisplatin in nanoparticles was ascertained by PXRD. Drug release followed a first-order release kinetic at both pH ∼ 5.6 (cervical cancer pH) and pH ∼ 7.4. In addition, a significant (P<0.05) decrease in IC50 value (8.3μM) and enhanced apoptosis were observed in HeLa cells treated with Cis-GNs-FA as compared to Cis-GNs (15.1μM) and cisplatin solution (40.2μM). In contrast, A549 lung cancer cells did not discriminate between Cis-GNs-FA and Cis-GNs due to the absence of folate receptors-α (FR-α). Consistently, higher cellular uptake, 80.54±7.60% was promoted by Cis-GNs-FA significantly (two-way ANOVA, P<0.05) greater than 51.68±9.78%, by Cis-GNs. This was also illustrated by CLSM images, which indicated that Cis-GNs-FA preferably accumulated in the cytoplasm of HeLa cells nearby nucleus by following receptor-mediated endocytosis pathway as compared to Cis-GNs. ConclusionTherefore, Cis-GNs-FA warrants further in-depth in vitro and in vivo investigations to scale up the technology for clinical translation.

Non-aggregated protamine-coated poly(lactide-co-glycolide) nanoparticles of cisplatin crossed blood–brain barrier, enhanced drug delivery and improved therapeutic index in glioblastoma cells: in vitro studies

Background and objectives: Non-aggregated protamine impregnated poly(lactide-co-glycolide) nanoparticles of cisplatin (Pt-PLGA NPs) were synthesized to augment brain delivery. Methods and results: The mean particle size of Pt-PLGA NPs and PLGA NPs were observed to be 173.2 ± 7.9 nm and 140 ± 10.2 nm, respectively. The Pt-PLGA NPs significantly (p < 0.05, one-way analysis of variance; ANOVA) delivered higher amount (172.41 ± 15.04 μg) of cisplatin in comparison to 110.48 ± 4.71 μg by PLGA NPs and 20.83 ± 1.65 μg by cisplatin solution across in vitro bovine brain microvessel endothelial cells. Cisplatin bearing Pt-PLGA NPs was found to be highly cytotoxic to U87 glioblastoma cells with an IC50 of 2.1 μM as compared (one-way ANOVA, p < 0.05) to PLGA NPs (3.9 μM) and cisplatin alone (13.33 μM). Impregnation with Pt enhanced the uptake of PLGA NPs in U87 glioblastoma cells as compared to PLGA NPs by following endocytosis mechanism. Conclusion: Cisplatin-loaded Pt-PLGA NPs compel preclinical tumour regression study to further improve its utility against glioblastoma.

A label-free G-quadruplex DNA-based fluorescence method for highly sensitive, direct detection of cisplatin

Cisplatin is a widely used anticancer chemotherapeutic, and fast, convenient detection methods for it are highly desirable. Based on the interaction of cisplatin with G-quadruplex DNA, we developed a simple label-free fluorescence method for rapid cisplatin detection. The titration experiment showed that the cisplatin concentration and the fluorescence signal change ratios (F0/F) exhibited a consistent linear correlation within the 1 to 10μM range with a limit of detection of 720nM, which was even lower than the common concentration of cisplatin in chemotherapy patients’ urine (54.3 to 321μM). The equilibrium dissociation constant KD value for cisplatin binding was determined to be 1.19×10−5M. This result demonstrated that our method had relatively high affinity toward cisplatin and could bind micromolar concentrations of cisplatin in solution. Our method also shows obvious selectivity among tested drugs, even between cisplatin and oxaliplatin/carboplatin. We demonstrated the high sensitivity of this methodology in the direct detection of cisplatin in urine samples and the fluorescence imaging of cisplatin in living cells.

Caprylate-Conjugated Cisplatin for the Development of Novel Liposomal Formulation

Cisplatin, first (platinum) compound to be evolved as an anticancer agent, has found its important place in cancer chemotherapy. However, the dose-dependent toxicities of cisplatin, namely nephrotoxicity, ototoxicity, peripheral neuropathy, and gastrointestinal toxicity hinder its widespread use. Liposomes can reduce the toxicity of cisplatin and provide a better therapeutic action, but the low lipid solubility of cisplatin hinders its high entrapment in such lipid carrier. In the present investigation, positively charged reactive aquated species of cisplatin were complexed with negatively charged caprylate ligands, resulting in enhanced interaction of cisplatin with lipid bilayer of liposomes and increase in its encapsulation in liposomal carrier. Prepared cisplatin liposomes were found to have a vesicular size of 107.9 ± 6.2nm and zeta potential of -3.99 ± 3.45mV. The optimized liposomal formulation had an encapsulation efficiency of 96.03 ± 1.24% with unprecedented drug loading (0.21mg cisplatin / mg of lipids). The in vitro release studies exhibited a pH-dependent release of cisplatin from liposomes with highest release (67.55 ± 3.65%) at pH 5.5 indicating that a maximum release would occur inside cancer cells at endolysosomal pH. The prepared liposomes were found to be stable in the serum and showed a low hemolytic potential. In vitro cytotoxicity of cisplatin liposomes on A549 lung cancer cell line was comparable to that of cisplatin solution. The developed formulation also had a significantly higher median lethal dose (LD50) of 23.79mg/kg than that of the cisplatin solution (12mg/kg). A promising liposomal formulation of cisplatin has been proposed that can overcome the disadvantages associated with conventional cisplatin therapy and provide a higher safety profile.

Hazardous drug residue on exterior vial surfaces: evaluation of a commercial manufacturing process.

Hazardous drug residue on the exterior surface of drug vials poses a potential risk for exposure of health care workers involved in handling these products. The purpose of this article is to heighten the awareness of this serious issue and to evaluate a commercial manufacturing process for removing and containing hazardous drug (HD) residue on exterior vial surfaces. Additionally, findings from this study are interpreted, incorporated into the current body of evidence, and discussed by experts in this field. This study includes separate evaluations for the presence or absence of surface drug contamination on the vials of 3 HD products: 5-fluorouracil, cisplatin, and methotrexate. The drug products were packaged in vials using a patented prewashing/decontamination method, application of a polyvinylchloride (PVC) base, and use of clear glass vials. An additional step of encasing the vial in a shrink-wrapped sheath was used for 5-fluorouracil and cisplatin. Of all 5-fluorouracil (110 vials), methotrexate (60 vials), and cisplatin (60 vials) tested, only 2 had detectable amounts of surface residue. One 5-fluorouracil vial was found to have approximately 4 mg of 5-fluorouracil on the surface of the vial. The second contaminated vial was cisplatin, which was discovered to have 131 ng of platinum, equal to 200 ng of cisplatin or 0.2 μL of cisplatin solution, on the vial sheath. Using validated extraction and analytic methods, all but 2 of the 230 tested vials were found to be free of surface drug contamination. Pharmacy leaders need to take an active role in promoting the need for clean HD vials. Manufacturers should be required to provide their clients with data derived from externally validated analytic studies, reporting the level of HD contamination on the exterior of their vial products.

Biodegradable polymeric system for cisplatin delivery: Development, in vitro characterization and investigation of toxicity profile

Cisplatin is one of the most potent anticancer agent used in the treatment of various solid tumors, however, its clinical use is limited due to severe adverse effects including nephrotoxicity. In this investigation cisplatin loaded poly (lactic-co-glycolic acid) (PLGA) nanoparticles were developed and characterized for various in vitro characteristics including size distribution, zeta potential, drug loading and release profile. PLGA nanoparticles were successfully developed as investigated using scanning electron microscopy and exhibited average particles size and zeta potential as 284.8nm and −15.8mV, respectively. Fourier transform infrared spectroscopy and differential scanning calorimetry indicated an absence of any polymer–drug interactions. Cisplatin nanoparticles exhibited in vitro anticancer activity against A549 cells comparable to that of cisplatin solution. The biodistribution study in mice indicated that the kidney cisplatin level was significantly (p<0.01) lower with cisplatin nanoparticles than cisplatin solution. Following two cycles of cisplatin treatment, a week apart, blood urea nitrogen level was found to be higher in case of cisplatin solution as compared to cisplatin nanoparticles. Further, there was a significant (p<0.01) increase in plasma creatinine level in case of cisplatin solution as compared to cisplatin nanoparticles. Histopathological examination of kidney from cisplatin nanoparticles treated group revealed no kidney damage, however, a sign of nephrotoxicity was observed in the case of cisplatin solution. The results suggest that PLGA nanoparticle based formulation could be a potential option for cisplatin delivery.

Dietary inclusion of sorghum (Sorghum bicolour) straw dye protects against cisplatin-induced nephrotoxicity and oxidative stress in rats

Context: Sorghum straw (dried leaves and stem fiber) extracts and infusion are employed in the management of several ailments in folklore, and it is also a natural dye source used in food preparation.Objective: This study sought to investigate the modulatory effect of dietary inclusion of Sorghum straw dye on cisplatin-induced nephrotoxicity and antioxidant status in rats.Materials and methods: Adult male rats were randomly divided into four groups of six animals each. Groups I (normal rats) and II (control rats) were fed with basal diet while Groups III and IV were fed with diets containing 0.5% and 1% sorghum straw dye, respectively. Nephrotoxicity was induced in Groups I–IV on the 20th day by the administration of a single dose of cisplatin solution (7 mg/kg body weight, i.p.) and the experiment was terminated 3 d after. Thereafter, the kidney and plasma of the rats were analyzed for kidney function (creatinine, urea, uric acid, and blood urea nitrogen) and antioxidant indices [superoxide dismutase (SOD), catalase, glutathione-S-transferase (GST), malondialdehyde (MDA), vitamin C, and reduced glutathione (GSH)].Results: The average feed intake of the rats in all the groups ranged from 9.0 to 9.5 (g/rat/day). Furthermore, the result indicated that administration of cisplatin caused significant (p < 0.05) elevation in plasma creatinine (2.2 mg/dL), uric acid (39.3 mg/dL), urea (81.4 mg/dL), and blood urea nitrogen (38.0 mg/dL) as well as a concomitant decrease in kidney antioxidant indices in control rats as against the normal rats. However, diets supplemented with 0.5 and 1.0% sorghum straw dye significantly reversed the plasma creatinine and the kidney antioxidant indices to near normal levels.Discussion and conclusion: The study suggests that dietary inclusion of sorghum straw dye as colorants could protect against oxidative stress and cisplatin-induced nephrotoxicity.

Study of claudin-4 in the diagnosis and treatment of endometrial carcinoma

To clarify the role of claudin-4 in endometrial tumorigenesis and explore claudin-4 be as potentially useful agent in the treatment of endometrial carcinoma. The expression of claudin-4 in 62 endometrioid endometrial carcinoma (EEC), 30 atypical hyperplasia endometrial tissue and 60 human normal endometrium was determined using immunohistochemistry and real-time PCR. Ninety female BALB/c mice were transplanted with Ishikawa endometrial cancer cells, which were divided into three groups with different intraperitoneal treatments with cisplatin, paclitaxel and saline solution. After the observation period, the tumors were extracted and stained with monoclonal antibody against claudin-4. The messenger RNA expression of claudin-4 was also detected using real-time PCR. Among the EEC samples, 34% (21/62) showed medium staining for claudin-4 and 66% (41/62) showed intense staining. In atypical hyperplasia group, 27% (8/30) showed weak staining, 53% (16/30) showed medium staining and 20% (6/30) showed intense staining for claudin-4. Of the normal endometrial tissue, 47% (28/60) showed weak staining and 53% (32/60) showed no staining for claudin-4. According to real-time PCR, the relative quantity of claudin-4 was 170 ± 12 in EEC group, 89 ± 15 in atypical hyperplasia group and 18 ± 3 in normal endometrium. Compared with those in atypical hyperplasia group and normal endometrium group, the protein and mRNA expression of claudin-4 were significantly increased in the group of EEC (all P < 0.05). In the study of Ishikawa xenografts, no significant changes in tumor volume and claudin-4 expression were shown in paclitaxel group compared with that in the control group. Nevertheless, a significant reduction of the tumor growth and a significant decrease in claudin-4 expression were observed in cisplatin group. After cisplatin treatment, the tumor volume was significantly decreased [(0.51 ± 0.21) versus (0.73 ± 0.12) cm(3)], and the mRNA expression of claudin-4 was also significantly decreased (153 ± 35 versus 273 ± 27). These results demonstrate that claudin-4 is strongly expressed in EEC, which may be a useful biomarker to monitor the effects of chemotherapy in patients with endometrial carcinoma.

TH-A-141-03: High-Sensitivity L-Shell X-Ray Fluorescence CT Imaging of Cisplatin

Purpose: To increase the sensitivity of x-ray fluorescence computed tomography (XFCT) imaging by excitation of low-energy L-shell x-rays with higher interaction cross-section compared to K-shell x-rays. Methods: XFCT images of a 2-cm diameter acrylic phantom with 0.1–0.4% (or 1–4 mg/mL) Cisplatin solutions were simulated in a modified version of the EGSnrc/DOSXYZnrc Monte Carlo code. XFCT images were reconstructed based on platinum fluorescence x-rays excited by 15, 30, and 80 keV x-rays. Based on L-edge and K-edge energies of platinum, 15 and 30 keV XFCT images were reconstructed using L-shell x-rays (∼10 keV) and 80 keV images were reconstructed with K-shell x-rays (∼67 keV). XFCT images were reconstructed with simple filtered back-projection (FBP), maximum-likelihood expectation maximization (ML-EM) without and with attenuation correction to account for the low energy 15 and 30 keV excitation x-rays and L-shell fluorescence x-rays. Results: For a 2.5 mGy imaging dose, K-shell XFCT images reconstructed with simple FBP resulted in Cisplatin imaging sensitivity limit of 740 μg/mL, which was defined by contrast-to-noise ratio (CNR) value of 4. For the same dose, 15 keV L-shell XFCT imaging sensitivity increased by a factor of 3.8 for objects at the periphery of the phantom. However, contrast in L-shell images decreased drastically towards the center of the phantom due to attenuation of the excitation and fluorescence x-rays. Our ML-EM reconstruction algorithm with attenuation correction applied on the 15 keV L-shell XFCT image 1) significantly increased CNR and resulted in 2) a uniform signal across the phantom, and 3) accurate quantification of Cisplatin concentration. Conclusion: We have shown that L-shell XFCT imaging in connection with ML-EM and attenuation correction reconstruction algorithm resulted high imaging sensitivity and accurate quantification of imaging contrast. A clinically relevant concentration of 6 μg/mL of Cisplatin can be accurately detected with a low imaging dose of 18 mGy.

Overexpression of claudin-4 may be involved in endometrial tumorigenesis

To clarify the role of claudin-4 in endometrial tumorigenesis and to explore whether claudin-4 could be a potentially useful agent in the treatment of endometrial carcinoma, the expression of claudin-4 in endometrial carcinoma was investigated. The relationship between therapy with anti-neoplastic agents and the expression of claudin-4 was also analyzed using an endometrial carcinoma xenograft model. The expression of claudin-4 in endometrial endometrioid adenocarcinoma (EEC) and normal human endometrial tissue was determined using immunohistochemistry and real-time PCR. Ninety female BALB/c nu/nu mice were transplanted with Ishikawa endometrial cancer cells. The mice were divided into three groups with different intraperitoneal treatments: cisplatin, paclitaxel or saline solution. After the observation period tumors were extracted and stained with monoclonal antibody against claudin-4. The mRNA expression of claudin-4 was also detected using real-time PCR. Expression of claudin-4 was significantly increased at both protein and mRNA levels in the EEC group compared with the group of normal cyclic endometrium. In the study of Ishikawa xenografts, no significant changes in tumor volume and claudin-4 expression were shown in the paclitaxel group compared with the control group. A significant reduction of tumor growth and a significant decrease in claudin-4 expression were observed in the cisplatin group. These results demonstrate that claudin-4 is strongly expressed in EEC. Claudin-4 is a useful biomarker in the treatment of patients with endometrial carcinoma.

Abstract A38: Nanoreformulation of platinum-based drugs to address disparate cancer treatment outcomes.

Abstract Modest responses and systemic toxicities during treatment of advanced cancer limits cancer drug efficacy, especially in African Americans. We determined the utility of cisplatin-loaded biodegradable XPclad nanoparticles coated with folate-conjugated polycaprolactone-polyethylene glycol. XPclad nanoparticles under in vitro conditions exhibited sustained release of the encapsulated cisplatin. Prostate cancer (PCa) cell lines (PC3, LNCaP, PTEN-CaP2, etc.), but not normal prostatic epithelial cells (PrEC and RWPE-1), expressed high levels of folate receptor α (FRα/FR1). Similar to cancer cells, Treg cells but not T helper cells express high affinity folate receptor (FRδ/FR4). Treatment using 20 μM of cisplatin solution resulted in &amp;gt; 90% cell death of PCa cells, PrEC, and RWPE-1 cells as well as purified CD4+CD25HiFoxP3+ T cells (Tregs) and CD4+CD25LoFoxP3- T cells (T helper). However, XPclad nanoparticles, containing cisplatin (equivalent to a 200 nM dose), were taken up and induced the same level of apoptosis in PCa and Treg cells as cisplatin solution, but did not significantly affect PrEC or RWPE-1 cells or T helper cells. Finally, we show that PC3 tumor-bearing mice that received either cisplatin solution (8 mg/kg/week) or cisplatin-encapsulated folate-coated XPclad nanoparticles (equivalent cisplatin dose = 0.08 mg/kg/week) resulted in significant tumor regression. Mice receiving cisplatin solution had significantly higher kidney and liver toxicities than naïve or XPclad nanoparticle-treated mice. Our studies show XPclad nanoparticles enhance tumor therapy efficacy with significantly lower to no toxicity and suggest that FR-targeted nanotherapy modulates the destruction of both tumor cells and Treg cells for increased efficacy. GRANT SUPPORT: G12 RR03034, U54 CA118638 and P60MD00525 Citation Format: James W. Lillard, S. Singh, R. Singh. Nanoreformulation of platinum-based drugs to address disparate cancer treatment outcomes. [abstract]. In: Proceedings of the Fifth AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2012 Oct 27-30; San Diego, CA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2012;21(10 Suppl):Abstract nr A38.

Ultrastructural effects of cisplatin on the inner ear and lateral line system of zebrafish (Danio rerio) larvae

Zebrafish, Danio rerio, has been a prominent model vertebrate for the study of chemical toxicity and human disease. Zebrafish hair cells (HCs) show significant structural, functional and molecular similarities to the mammalian inner ear HCs. We examined the effects of cisplatin, an anti-cancer drug, on HCs of the inner ear and on HCs and support cells (SCs) of neuromasts in zebrafish using transmission and scanning electron microscopy. Forty-five zebrafish larvae, 12 days post-fertilization, were assessed: 15 unexposed controls, 15 exposed to 10 µM cisplatin solution, and 15 exposed to 50 µM cisplatin solution. Hair cells in the cristae and maculae of the inner ear and of neuromasts were extremely sensitive to cisplatin. The drug was associated with vacuolization and the presence of myelinoid bodies in HC cytoplasm and with a condensation of the nuclear chromatin. The predominant pattern of injury was widespread degeneration of mitochondria, which appeared swollen and less electron-dense with disorganized or reduced cristae. Severity of damage seemed to be concentration-dependent, and the inner ear suffered more damage than the lateral line. Alterations similar to those in HCs were also observed in SCs of the neuromasts. Scanning electron microscopy showed loss of kinocilia in neuromasts of fish exposed to the higher concentration of cisplatin.

Abstract 4393: MRI-guided electroporation-mediated transarterial chemoembolization (E-TACE)

Abstract INTRODUCTION: Electropermeabilization involves the application of electrical pulses to increase cell membrane permeability; electrochemotherapy (ECT) takes advantage of this phenomenon to increase tumor uptake of drugs. The purpose of this study was to demonstrate the potential to use MRI for intra-procedural optimization of an electroporation-mediated TACE (E-TACE) approach intended to increase liver tumor drug uptake compared to transcatheter infusion alone. MATERIALS &amp; METHODS: Eight New Zealand White rabbits were used for our ACUC-approved experiments. Two VX2 tumor portions were implanted in the left medial hepatic lobe of each rabbit and a total of fourteen VX2 tumors were grown in these rabbits. Two tumor were grown in 6 rabbits (one as E-TACE treated tumor and other as control) and solitary larger tumors was grown in 2 rabbits (half of tumor treated with E-TACE, remaining half as control). Each rabbit was catheterized under DSA guidance with catheter placed in the left hepatic artery. Next, Baseline transcatheter intraarterial perfusion (TRIP) MRI (1.5T, Siemens Medical) was performed to generate tumor enhancement curves with an intra-arterial (IA) injection of Gd-DTPA. These contrast enhancement curves were generated to estimate time delay between IA bolus infusion and the period of maximum tumor uptake (for planning purposes to optimize delay interval, Td, between later IA Cisplatin infusion and application of electrical pulses). For E-TACE treated tumors (one tumor in each rabbit) bi-polar electrodes (1cm spacing) were inserted straddling the targeted tumor and 8 100μs 1300V pulses applied at the selected delay interval Td after the administration of Cisplatin solution through the catheter. Next, a post TRIP-MRI scan was performed to assess tumor perfusion alterations and treatment related tumor signal intensity changes. After allowing 3 hours for Cisplatin wash out, rabbits were euthanized and each tumor sectioned for ICP-MS measurement of intra-tumoral platinum concentrations. RESULTS: ICP-MS results demonstrated significantly increased Cisplatin uptake in E-TACE treated tumors, increases of 6.0-fold compared to transcatheter infusion alone in six of the rabbits (n=6/8, p=0.017). Follow-up T2W-TSE images showed significant intra-tumoral T2W signal decreases in five rabbits (n=5/8) whereas no such signal changes were observed in other rabbits (two of which showed negligible Cisplatin uptake increases). CONCLUSION: Our findings suggest that E-TACE may be effective for increasing tumor uptake of chemotherapeutic agents and that intra-procedural MRI should permit immediate confirmation of targeted drug delivery based upon signal changes only present in sufficiently treated tissues. Additional longitudinal studies are required to evaluate and compare treatment response between animals treated with E-TACE and those treated with conventional TACE approaches. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4393. doi:10.1158/1538-7445.AM2011-4393

Influence of time and chloride ions on the interaction of cisplatin with human albumin in-vitro.

The interaction of cis-dichlorodiammineplatinum (II) (cisplatin) with human serum albumin (HSA), dissolved in phosphate buffer with or without sodium chloride (0.1 M) has been examined at pH 7.4 and mu = 0.154. Equal volumes of cisplatin and HSA solutions were incubated at 37 degrees C for various times and filterable platinum concentrations versus time measured by flameless atomic absorption spectrophotometry. Binding kinetics differed depending on the buffer solutions used and on the time elapsing between cisplatin dissolution and outset of incubation with HSA. Experimental data were fitted to a theoretical equation used to calculate the number of nucleophilic sites per HSA molecule. Titrations of the HSA sulphydryl group content before and after incubation with a cisplatin solution were made, from which it was shown that the lone SH-group of the HSA macromolecule is involved in cisplatin binding. We also studied HSA's sensitivity towards denaturing agents when it was complexed with cisplatin. This sensitivity was decreased upon cisplatin binding. Also, the binding capacities of HSA and the HSA-Pt(II) complex to both tryptophan and warfarin were compared to determine the possible influence of cisplatin upon the binding to HSA of other drugs; this influence was negligible.

Transarterial Chemoembolization With Cisplatin as Second-Line Treatment for Hepatocellular Carcinoma Unresponsive to Chemoembolization With Epirubicin-Lipiodol Emulsion

The purpose of this retrospective study was to investigate the efficacy of transarterial chemoembolization (TACE) using cisplatin as a second-line treatment for advanced hepatocellular carcinoma (HCC) unresponsive to TACE using epirubicin-Lipiodol emulsion at our institution. Between January 2006 and March 2009, 51 patients with unresectable HCC underwent TACE using cisplatin. All patients had shown persistent viable tumor or tumor progression after at least 2 sessions of TACE using epirubicin-Lipiodol emulsion. TACE procedures consisted of arterial injection of a mixture of Lipiodol and cisplatin (30-100mg [mean 57±21]) (n=29) or arterial infusion of cisplatin (30-100mg [mean 87±19]) solution (n=22) followed by injection of 1-mm porous gelatin particles. Early tumor response was assessed by contrast-enhanced computed tomography (CT) according to Response Evaluation Criteria in Solid Tumors (RECIST) and European Association for the Study of the Liver (EASL) criteria. Overall survival and progression-free survival was calculated using the Kaplan-Meier method. Toxicity was assessed according to NCI-CTCAE version 3 criteria. Response rates were 11.8 and 27.5% by RECIST and EASL criteria, respectively. Overall survival rates were 61.9, 48.2, and 28.9% at 1, 2, and 3years, respectively, and the median survival time was 15.4months. Progression-free survival rate was 35.2% at 1year, and median progression-free survival time was 3.1months. No major complications were observed, and the occurrence of postembolization syndrome was minimal. Grade 3 to 4 toxicities included thrombocytopenia (5.8%), increased aspartate aminotransferase (AST) level (35.3%), and increased alanine aminotransferase (ALT) level (23.5%). Switching the TACE anticancer drug from epirubicin to cisplatin might be the feasible option for advanced HCC, even when considered resistant to the initial form of TACE.

Optimized intrapleural cisplatin chemotherapy with a fibrin carrier after extrapleural pneumonectomy: A preclinical study

Our objective was to evaluate whether platinum concentrations in chest wall tissue and in serum are optimized by intracavitary application of cisplatin loaded to a fibrin carrier compared with cisplatin solution in a randomized setting of a pig model. After left-sided pneumonectomy including parietal pleurectomy, pigs were randomly assigned to receive either 90 mg/m(2) cisplatin intracavitary solution (n = 6) or to receive 5 mg cisplatin-fibrin (n = 5) applied on a predefined area of the chest wall. Platinum concentration in serum as well as in chest wall tissue was determined at several early time points until day 5 after treatment. Platinum levels were measured by inductively coupled plasma sector field mass spectrometric detection with a matrix-matched calibration procedure. The dose- and surface-corrected (geometric) mean concentration of cisplatin in chest wall tissue 2 hours but also at day 5 after the application was doubled in animals treated with cisplatin-fibrin compared with the animals treated with cisplatin-solution. In serum, the dose- and surface-corrected exposure toward cisplatin (area under the curve(0-5d)) was significantly lower with cisplatin-fibrin than with cisplatin-solution (P < .0005). This is also reflected by significantly reduced serum creatinine and urea values in the cisplatin-fibrin group (P < .0001). Animals treated with cisplatin-fibrin additionally had a significantly better postoperative course as assessed by a well-being score (P < .001). After cisplatin-fibrin treatment, cisplatin tissue concentration was increased whereas systemic cisplatin concentrations were significantly reduced in comparison with cisplatin-solution treatment. This finding offers a clear advantage inasmuch as rate and severity of systemic adverse events can be reduced while local cytotoxic concentrations are at least maintained.

Cisplatin-Conjugated Porous Gelatin Particles: Assessment of Optimal Conditions for Binding and Release

This study was designed to evaluate the optimal conditions for binding cisplatin and porous gelatin particles (PGPs) and to establish in vivo drug release pharmacokinetics. PGPs were immersed in cisplatin solutions under different conditions: concentration, immersion time, and temperature. Thereafter, PGPs were washed in distilled water to remove uncombined cisplatin and were then freeze-dried. The platinum concentration (PC) in the PGPs was then measured. For the in vivo release test, 50 mg/kg of the cisplatin-conjugated PGPs was implanted subcutaneously in the abdominal region of two rabbits. PCs in the blood were measured at different time intervals. PCs significantly increased in direct proportion to the concentration and immersion time (p < 0.01). Although PC increased at higher solution temperature, it was not a linear progression. For the in vivo release test, platinum was released from cisplatin-conjugated PGPs after 1 day, and the peak PC was confirmed 2 days after implantation. Platinum in the blood was detected until 7 days after implantation in one rabbit and 15 days after administration in the other rabbit. Platinum binding with PGPs increased with a higher concentration of cisplatin solution at a higher temperature over a longer duration of time. Release of cisplatin from cisplatin-conjugated PGPs was confirmed in vivo.

The preparation and characterization of folate-conjugated human serum albumin magnetic cisplatin nanoparticles

Objective Nanoparticles are becoming an important method of targeted drug delivery. To evaluate the importance of folate-conjugated human serum albumin (HSA) magnetic nanoparticles (Folate-CDDP/HSA MNP), we prepared drug-loaded Folate-CDDP/HSA MNPs and characterized their features. Methods First, folate was conjugated with HSA under the effect of a condensing agent, and the conjugating rate was evaluated by a colorimetric method using 2, 4, 6 – trinitrobenzene sulfonic acid. Second, under N 2 gas, Fe 3O 4 magnetic nanomaterials were prepared and characterized by using transmission electron microscopy (TEM), SEM-EDS and X-ray diffraction (XRD). Finally, Folate-CDDP/HSA MNP was prepared by using a solvent evaporation technique. TEM was used to observe particle morphology. The particle size and distribution of the prepared complexes were determined by a Laser particle size analyzer. Drug loading volume and drug release were investigated by a high performance liquid chromatography method (HPLC) in vitro. Results We successfully prepared folate-conjugated HSA and its conjugating rate was 27.26 μg/mg. Under TEM, Fe 3O 4 magnetic nanoparticles were highly electron density and had an even size distribution in the range of 10-20 nm. It was confirmed by SEM-EDS and XRD that Fe3O4 magnetic nanoparticles had been successfully prepared. Under TEM, drug-loaded magnetic nanoparticles were observed, which had a round shape, similar uniform size and smooth surface. Their average size was 79 nm which was determined by laser scattering, and they exhibited magnetic responsiveness. Encapsulation efficiency was 89.75% and effective drug loading was calculated to be 15.25%. The release results in vitro showed that the half release time (t1/2) of cisplatin in cisplatin Solution and Folate-CDDP/HSA MNP was 65 min and 24 h respectively, which indicated that microspheres had an obvious effect of sustained-release. Conclusion: Folate-CDDP/HSA MNPs were prepared successfully. The preparation process and related characteristics data provided a foundation for further study, including the mechanism of the nanoparticles distribution in vivo and their intake by tumor cells.