Validated assays for the determination of gemcitabine in human plasma and urine using high-performance liquid chromatography with ultraviolet detection

Freeman, Kelly B.; Anliker, Sally L.; Hamilton, Marta; Osborne, David R.; Dhahir, Patricia; Nelson, Robert L.; Allerheiligen, Sandra R. B.

doi:10.1016/0378-4347(94)00521-6

Cited by 66 publications

(54 citation statements)

References 6 publications

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“…Blood samples were collected from the mice under isoflurane anesthesia 2, 5, 15, 30, 60, and 120 min after drug administration in heparinized tubes containing tetrahydrouridine to prevent gemcitabine degradation by cytidine deaminase. Gemcitabine and its major plasma metabolite dFdU were determined using normal phase HPLC according to the published literature procedure (35). The amount of gold present in serum was quantified using ICP as described in the Supplementary Data.…”

Section: Methodsmentioning

confidence: 99%

Targeted Delivery of Gemcitabine to Pancreatic Adenocarcinoma Using Cetuximab as a Targeting Agent

et al. 2008

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One of the key challenges in anticancer therapy is the toxicity and poor bioavailability of the anticancer drugs. Nanotechnology can play a pivotal role by delivering drugs in a targeted fashion to the malignant cells that will reduce the systemic toxicity of the anticancer drug. In this report, we show a stepwise development of a nanoparticle-based targeted delivery system for in vitro and in vivo therapeutic application in pancreatic cancer. In the first part of the study, we have shown the fabrication and characterization of the delivery system containing gold nanoparticle as a delivery vehicle, cetuximab as a targeting agent, and gemcitabine as an anticancer drug for in vitro application. Nanoconjugate was first characterized physico-chemically. In vitro targeting efficacy, tested against three pancreatic cancer cell lines (PANC-1, AsPC-1, and MIA Paca2) with variable epidermal growth factor receptor (EGFR) expression, showed that gold uptake correlated with EGFR expression. In the second part, we showed the in vivo therapeutic efficacy of the targeted delivery system. Administration of this targeted delivery system resulted in significant inhibition of pancreatic tumor cell proliferation in vitro and orthotopic pancreatic tumor growth in vivo. Tumor progression was monitored noninvasively by measuring bioluminescence of the implanted tumor cells. Pharmacokinetic experiments along with the quantitation of gold both in vitro and in vivo further confirmed that the inhibition of tumor growth was due to targeted delivery. This strategy could be used as a generalized approach for the treatment of a variety of cancers characterized by overexpression of EGFR.

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Section: Methodsmentioning

confidence: 99%

Targeted Delivery of Gemcitabine to Pancreatic Adenocarcinoma Using Cetuximab as a Targeting Agent

et al. 2008

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“…1). CDA activity was measured with gemcitabine (Eli Lilly, Indianapolis, IN) as substrate using a modification of the assay of Miwa et al (11) with the high-performance liquid chromatography analysis of Freeman et al (12). Basal levels of enzyme activity for WT and variant allozymes were corrected for transfection efficiency on the basis of h-galactosidase activity measured with the h-Galactosidase Enzyme Assay System (Promega).…”

Section: Methodsmentioning

confidence: 99%

Gemcitabine Pharmacogenomics: Cytidine Deaminase and Deoxycytidylate Deaminase Gene Resequencing and Functional Genomics

Gilbert¹,

Salavaggione²,

Ji³

et al. 2006

Clinical Cancer Research

150

128

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Purpose: Gemcitabine is a nucleoside analogue with activity against solid tumors. Gemcitabine metabolic inactivation is catalyzed by cytidine deaminase (CDA) or, after phosphorylation, by deoxycytidylate deaminase (DCTD). We set out to study the pharmacogenomics of CDA and DCTD. Experimental Design:The genes encoding CDA and DCTD were resequenced using DNA from 60 African American and 60 Caucasian American subjects. Expression constructs were created for nonsynonymous coding single nucleotide polymorphisms (cSNP) and reporter gene constructs were created for 5V -flanking region polymorphisms. Functional genomic studies were then conducted after the transfection of mammalian cells. Results: CDA resequencing revealed 17 polymorphisms, including one common nonsynonymous cSNP, 79 A>C (Lys27Gln). Recombinant Gln27 CDA had 66 F 5.1% (mean F SE) of the wild-type (WT) activity for gemcitabine but without a significant decrease in level of immunoreactive protein. The apparent K m (397 F 40 Amol/L) for the Gln27 allozyme was significantly higher than that for the WT (289 F 20 Amol/L; P < 0.025). CDA 5V-flanking region reporter gene studies showed significant differences among 5V -flanking region haplotypes in their ability to drive transcription. There were 29 SNPs in DCTD, including one nonsynonymous cSNP, 172 A>G (Asn58Asp), in Caucasian American DNA. Recombinant Asp58 DCTD had 11 F 1.4% of WT activity for gemcitabine monophosphate with a significantly elevated level of immunoreactive protein. No DCTD polymorphisms were observed in the initial 500 bp of the 5V -flanking region. Conclusions: These results suggest that pharmacogenomic variation in the deamination of gemcitabine and its monophosphate might contribute to variation in therapeutic response to this antineoplastic agent.

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“…The concentration of gemcitabine in plasma and peritoneal fluid was measured according to a modification of a previously published high-performance liquid chromatography method (15). Briefly, following the addition of 2 ¶,3 ¶-dideoxycytidine as an internal standard, a solid-phase extraction step was used to eliminate interfering substances from plasma and peritoneal fluid.…”

Section: Methodsmentioning

confidence: 99%

Phase I Trial of Intraperitoneal Gemcitabine in the Treatment of Advanced Malignancies Primarily Confined to the Peritoneal Cavity

Morgan

Synold

et al. 2007

Clinical Cancer Research

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Purpose: To determine the maximally tolerated dose, toxicity, and pharmacokinetics of i.p.gemcitabine. Experimental Design: Patients had peritoneal carcinomatosis. Gemcitabine (40, 80, 120, or 160 mg/m 2 ) was administered into the peritoneal cavity in 2 L of warmed saline on days 1, 4, 8, and 12 of a 28-day cycle. Results:Thirty patients received 63 (median, 2; range, 0-6) courses.Tumors included ovary (14), uterus (2), colon (6), pancreas (3), and others (5). Dose-limiting toxicity included nausea, vomiting, diarrhea, dyspnea, fatal respiratory failure, and grade 3 elevation of alanine aminotransferase in three patients. Hematologic toxicity and pain were Vgrade 2. Three patients had decreased or resolved ascites. Of 19 patients evaluable for response, 10 had stable disease (median, 3.5 courses) and 9 had progressive disease. The median peak peritoneal concentration was1,116-fold (range, 456-1,886) higher than the peak plasma level. Plasma and peritoneal levels were undetectable within 8 to 12 h. At 120 mg/m 2 , the median peritoneal area under the concentration versus time curve (AUC) was 82,612 ng/mL Â h (range, 53,296-199,830) and the plasma AUC was 231 ng/mL Â h (range, 47.6-259.5). The mean peritoneal advantage (AUC peritoneal / AUC plasma ) was 847 (range, 356-1,385).

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Validated assays for the determination of gemcitabine in human plasma and urine using high-performance liquid chromatography with ultraviolet detection

Cited by 66 publications

References 6 publications

Targeted Delivery of Gemcitabine to Pancreatic Adenocarcinoma Using Cetuximab as a Targeting Agent

Targeted Delivery of Gemcitabine to Pancreatic Adenocarcinoma Using Cetuximab as a Targeting Agent

Gemcitabine Pharmacogenomics: Cytidine Deaminase and Deoxycytidylate Deaminase Gene Resequencing and Functional Genomics

Phase I Trial of Intraperitoneal Gemcitabine in the Treatment of Advanced Malignancies Primarily Confined to the Peritoneal Cavity

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