A novel oral insulin-like growth factor-1 receptor pathway modulator and its implications for patients with non-small cell lung carcinoma: A phase I clinical trial.

BACKGROUND: A phase Ia/b dose-escalation study was performed to characterize the safety, efficacy and pharmacokinetic properties of the oral small molecule insulin-like growth factor-1-receptor pathway modulator AXL1717 in patients with advanced solid tumors. MATERIAL AND METHODS: This was a prospective, single-armed, open label, dose-finding phase Ia/b study with the aim of single day dosing (phase Ia) to define the starting dose for multi-day dosing (phase Ib), and phase Ib to define and confirm recommended phase II dose (RP2D) and if possible maximum tolerated dose (MTD) for repeated dosing. RESULTS AND CONCLUSION: Phase Ia enrolled 16 patients and dose escalations up to 2900 mg BID were successfully performed without any dose limiting toxicity (DLT). A total of 39 patients were treated in phase Ib. AXL1717 was well tolerated with neutropenia as the only dose-related, reversible, DLT. RP2D dose was found to be 390 mg BID for four weeks. Some patients, mainly with NSCLC, demonstrated signs of clinical benefit, including four partial tumor responses (one according to RECIST and three according to PET). The 15 patients with NSCLC with treatment duration longer than two weeks with single agent AXL1717 in third or fourth line of therapy showed a median progression-free survival of 31 weeks and overall survival of 60 weeks. Down-regulation of IGF-1R on granulocytes and increases of free serum levels of IGF-1 were seen in patients treated with AXL1717. AXL1717 had an acceptable safety profile and demonstrated promising efficacy in this heavily pretreated patient cohort, especially in patients with NSCLC. RP2D was concluded to be 390 mg BID for four weeks. Trial number is NCT01062620.

[HPLC determination of 4-[4"-(2",2",6",6"-tetramethyl-1"-piperidinyloxy) amino]-4'-demethylepipodophyllotoxin in rat plasma and studies of its pharmacokinetics].

4-[4"-(2",2",6",6"-tetramethyl-1"-piperidinyloxy) amino]-4'- demethylepipodophyllotoxin (GP-7) is a new podophyllotoxin spin-labeled derivative. Its primary effect is the antitumor activity on transplanted mouse tumors and cultured tumor cells. This paper describes a method for its determination using HPLC with UV detection and the determination of its pharmacokinetic parameters in rats. A Shimadzu LC-6A liquid chromatograph equipped with a Shimadzu SPD-6AV multiwavelength detector and a Chromatopac C-R3A data processor was used. The separation was performed on a Zorbax-ODS column (5 microns, 4.6 mm x 150 mm) with a mobile phase of methanol--water--glacial acetic acid (59:41:0.6). The flow-rate was 1.0 ml.min-1 and detection was made at 285 nm. A plasma specimen (0.2 ml) was spiked with 22.6 micrograms.ml-1 internal standard (podophyllic acid piperidinyl hydrazone nitroxide radical, GP-1) and extracted with ether--dichloromethane (3:1). The extract was evaporated at 45 degrees C. The residue was taken up with 0.1 ml of the mobile phase and 20 microliters aliquots were injected into the system. The calibration curve was linear in the range from 2 to 200 micrograms.ml-1 with r = 0.9997. The detection limit was 0.2 microgram.ml-1 and the recovery of GP-7 from rat plasma was 94.3%-100.9%. The relative standard deviations for within- day and between-day were 2.29%-4.64% and 5.55%-7.70%, respectively. After iv injection of GP-7 10, 20 and 30 mg.kg-1, the concentrations of the drug in rat plasma were determined. The pharmacokinetic parameters of GP-7 were obtained by using MCPKP program on a COMPAC-486 computer. The data obtained fitted a two-compartment open model, and the mean T1/2 beta value was 39.8 +/- 10.8 min.

[Changes in serum concentration of podophyllotoxin after its topical application with liposome on rat skin].

OBJECTIVE: To investigate the changes of serum concentration of podophyllotoxin after topical application of liposome podophyllotoxin suspension on rat skin. METHODS: SD rats were used in this study, which were divided into test group (n=48) to receive application of liposome podophyllotoxin (0.5 % ) suspension and control group (n=48) treated with 0.5 % podophyllotoxin alcohol solution. Blood samples were obtained from the heart at 1, 2, 4, 6, 8, 10, 12 and 24 h respectively after drug application, and the serum concentration of podophyllotoxin was determined by spectrofluorometry. RESULTS: The area under the curve of plasma drug concentration of the control group was 2.3-fold greater than that of the test group. Eight hours after drug application, the serum concentration of podophyllotoxin reached the peak in the test group, while in the control group, only two hours was needed to reach the peak. The peak serum level of podophyllotoxin in the test group were significantly lower than that of the control group (166.395 +/- 14.634 ng/ml vs 378.603 +/- 26.105 ng/ml, P<0.001). CONCLUSION: The systemic absorption of podophyllotoxin in rats after its topical application in liposome suspension is significantly lower than that after application of 0.5 % podophyllotoxin alcohol solution, therefore the systemic toxicity may be reduced.

Validated LC-MS/MS assay for quantitative determination of deoxypodophyllotoxin in rat plasma and its application in pharmacokinetic study.

A rapid and sensitive liquid chromatography/tandem mass spectrometry method was developed and validated for the quantification of deoxypodophyllotoxin (DPT) concentration in rat plasma with diazepam as internal standard (IS). DPT and IS were extracted with ethyl acetate, and the chromatographic separation was accomplished by using a Waters Symmetry C18 analytical column (2.1mm×150mm, 5µm) with a mobile phase consisting of acetonitrile and deionized water (70:30, v:v) containing 0.1% formic acid at a flow rate of 0.2mL/min. Multiple Reaction Monitoring (MRM), using electrospray ionization in positive ion mode, was employed to quantitatively detect DPT and IS. The monitored transitions were set at m/z 399.05-231.00 and m/z 285.00-154.00 for DPT and IS, respectively. The calibration curve was linear over the concentration range of 7.8-1000ng/mL (R(2)≥0.9999). The intra- and inter-day precision values were less than 7%. Similarly, the mean intra- and inter-day accuracy were found to be within -2.8% to 1.9% of the interval, with all samples locating within general assay acceptability criteria for QC samples according to FDA guidelines. This method was further and successfully applied in the pharmacokinetics study of DPT in rat.

Determination of picropodophyllin and its isomer podophyllotoxin in human serum samples with electrospray ionization of hexylamine adducts by liquid chromatography-tandem mass spectrometry.

A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for determination of the new anticancer agent picropodophyllin (AXL1717) and its isomer podophyllotoxin levels in human serum has been developed. Monitoring of hexylamine adducts rather than proton adducts was used to optimize sensitivity. The chromatography system was an Acquity BEH C18, 2.1 mm × 50 mm 1.7 µm column with gradient elution (mobile phase A: 2.5 mM hexylamine and 5 mM formic acid in Milli-Q water and mobile phase B: methanol). The retention times were 1.4 min for picropodophyllin, 1.5 min for podophyllotoxin and 1.9 min for internal standard deoxypodophyllotoxin. The isomers were base-line separated. The analytes were detected after electrospray ionization in positive mode with selected reaction monitoring (SRM) with ion transitions m/z 516→102 for picropodophyllin and podophyllotoxin and m/z 500→102 for internal standard. The sample preparation was protein precipitation with acetonitrile (1:3) containing internal standard followed by dilution of the supernatant with mobile phase A (1:1). The limit of quantification (LOQ) was 0.01 µmol/L for picropodophyllin and podophyllotoxin. The limit of detection (LOD) at 3 times the signal to noise (S/N) was estimated below 0.001 µmol/L for picropodophyllin and podophyllotoxin. The quantification range of the method was between 0.01 µmol/L and 5 µmol/L for both isomers. The accuracy was within ±15% of the theoretical value for both picropodophyllin and podophyllotoxin and inter-assay precision did not exceed ±15%, except for the 0.016 µmol/L level of podophyllotoxin, which was 18%. The selectivity of the method was verified by analysis of two different product ions for each analyte and by analysis for interference of seven different batches of blank human serum. The combined recovery and matrix effects were about 83% for picropodophyllin and podophyllotoxin. The new LC-MS/MS method showed sufficient sensitivity and selectivity for determination of picropodophyllin and its isomer podophyllotoxin levels in human serum from subjects receiving therapeutic doses of AXL1717.

[Quantitative determination of podophyllotoxin in dermal and blood microdialysis samples of rats by liquid chromatography-tandem mass spectrometry].

OBJECTIVE: To develop and validate a sensitive method for quantitative analysis of podophyllotoxin in blood and dermal microdialysis samples of rats based on liquid chromatography-tandem mass spectrometry (UFLC-MS-MS). METHODS: The microdialysis samples were prepared by liquid-liquid extraction using ethyl acetate with etoposide as the internal standard (IS). Podophyllotoxin was separated with an Agilent ZORBAX XDB-C18 column (2.1 mmx50 mm, 3.5 microm). The mobile phase consisted of acetonitrile: 10 mmol/L ammonium acetate (40:60, V/V) at a flow rate of 0.3 ml/min and the analysis was performed at the ambient temperature. The UFLC-MS/MS system was operated in the mode of multiple reaction monitoring using the electrospray ionization technique in positive mode. RESULTS: Podophyllotoxin and etoposide responses were optimized at the transitions m/z 432.7-->397.3 and 589.5-->229.5, respectively. Calibration curves were linear over the range 2.0-1000 ng/ml. The lowest limits of quantification and detection values were 2.0 ng/ml and 0.7 ng/ml, respectively. The inter- and intra-day precision and accuracy were both less than 15%. CONCLUSION: This selective and sensitive method can be used to quantity podophyllotoxin in the blood and dermal microdialysates of rats.

Determination of the new podophyllotoxin derivative NK 611 in plasma by high-performance liquid chromatography with ultraviolet detection.

A high-performance liquid chromatographic method has been developed for the determination of the new podophyllotoxin derivative NK 611 in plasma samples. A solid-liquid extraction procedure with C18 extraction columns was used for extraction of plasma samples containing NK 611. The adsorbed NK 611 was eluted from the extraction columns with methanol-acetonitrile (50:50, v/v). The elution liquid was injected into a reversed-phase system consisting of a Chrompack C18 column. The mobile phase was acetonitrile-20 mM phosphate buffer, pH 7 (30:70, v/v). The UV detection mode allows sensitive determination of NK 611 in plasma within phase I trials. The limit of detection was 10 ng/ml, the limit of quantitation 35 ng/ml (for 1 ml of extracted plasma and 20-microliters injection volume). The calibration curve is linear within the concentration range 100-1000 ng/ml. The recovery of NK 611 from spiked plasma samples was approximately 80%.

Podophyllotoxin in serum: absorption subsequent to three-day repeated applications of a 0.5% ethanolic preparation on condylomata acuminata.

High-performance liquid chromatography was used for quantitative determination of podophyllotoxin in serum subsequent to repeated applications of a 0.5% ethanolic preparation on condylomata acuminata. The drug was not detectable in the serum of ten men treated with < or = 50 microliters. In serum of seven patients receiving 100-1,500 microliters on extraordinarily large en-plaque lesions, peak levels of 1-17 ng/ml were measured within 1-2 hr. The drug did not accumulate in serum. Dispersion of 100 microliters gave rise to peak levels of < or = 5.0 ng/ml and to subsequent levels of < or = 3.0 ng/ml for 4 hr. Larger doses gave rise to traceable amounts in serum for < or = 12 hr. Investigations with 52 patients indicated that volumes of > 100 microliters are rarely required for topical treatment of condylomata. It is concluded that application of volumes of < or = 250 microliters twice daily for three days would satisfy the most stringent safety precautions and avoid any danger of systemic toxicity. Potential hazards associated with use of the nonstandardized 20% podophyllin preparations are emphasized.

High-performance liquid chromatographic assay for the determination of the novel podophyllotoxin derivative dimethylaminoetoposide (NK611) in human plasma.

A simple, rapid and reproducible plasma assay for the determination of the novel epipodophyllotoxin derivative, dimethylaminoetoposide (NK611, I) and its N-demethyl metabolite (II) is reported. The method involves solid-phase extraction using an isolute C18 cartridge and HPLC separation on a reduced-activity C18 column (8 cm long) with a mobile phase of acetonitrile-water-0.1 M phosphoric acid (23:76:1, v/v/v); peaks are detected at 205 nm. The intra- and inter-day precision and accuracy are within 5 and 4% for I and II, respectively. The sensitivity is 20 ng/ml for both I and II. The assay is applicable to clinical pharmacokinetic studies. In one cancer patient who received both an oral and an intravenous dose of 10 mg of I the bioavailability was 82% and the clearance 20.8 ml/min.

Simultaneous determination of NK611, a novel water-soluble derivative of etoposide, and its metabolite (DeNK611) in dog plasma by column-switching high performance liquid chromatography.

A simple, selective, reversed phase liquid chromatographic method using a column-switching technique has been developed for the simultaneous determination of a novel derivative of etoposide (NK611) and its O-demethyl metabolite in dog plasma. A good linear response was obtained for both drugs in the range 0.1-12.0 micrograms/mL. The mean recoveries were within 100 +/- 5%. The within- and between-day precisions were within 3.5% and 4.6%, respectively. This method was used in a pharmacokinetic study following intravenous and oral administration of NK611 to beagle dogs.

Drug level monitoring: cytostatics.

The present review on the quantification of cytostatic drugs has mainly been focussed on chromatographic techniques. Special attention has been paid to the precautions that have to be taken into account to ensure the selectivity and accuracy of the various methods. The various cytostatics that have been dealt with are: alkylating agents, antimetabolites, vinca alkaloids, antibiotics, cis-diamminedichloroplatinum, podophyllotoxine derivatives, and nitrosoureas.