Synthesis and biological evaluation of novel propargyl amines as potential fluorine-18 labeled radioligands for detection of MAO-B activity.

The aim of this project was to synthesize and evaluate three novel fluorine-18 labeled derivatives of propargyl amine as potential PET radioligands to visualize monoamine oxidase B (MAO-B) activity. The three fluorinated derivatives of propargyl amine ((S)-1-fluoro-N,4-dimethyl-N-(prop-2-ynyl)-pent-4-en-2-amine (5), (S)-N-(1-fluoro-3-(furan-2-yl)propan-2-yl)-N-methylprop-2-yn-1-amine (10) and (S)-1-fluoro-N,4-dimethyl-N-(prop-2-ynyl)pentan-2-amine (15)) were synthesized in multi-step organic syntheses. IC(50) values for inhibition were determined for compounds 5, 10 and 15 in order to determine their specificity for binding to MAO-B. Compound 5 inhibited MAO-B with an IC(50) of 664 ± 48.08 nM. No further investigation was carried out with this compound. Compound 10 inhibited MAO-B with an IC(50) of 208.5 ± 13.44 nM and compound 15 featured an IC(50) of 131.5 ± 0.71 nM for its MAO-B inhibitory activity. None of the compounds inhibited MAO-A activity (IC(50) > 2 µM). The fluorine-18 labeled analogues of the two higher binding affinity compounds (10 and 15) (S)-N-(1-[(18)F]fluoro-3-(furan-2-yl)propan-2-yl)-N-methylprop-2-yn-1-amine (16) and (S)-1-[(18)F]fluoro-N,4-dimethyl-N-(prop-2-ynyl)pentan-2-amine (18) were both prepared from the corresponding precursors 9A, 9B and 14A, 14B by a one-step fluorine-18 nucleophilic substitution reaction. Autoradiography experiments on human postmortem brain tissue sections were performed with 16 and 18. Only compound 18 demonstrated a high selectivity for MAO-B over MAO-A and was, therefore, chosen for further examination by PET in a cynomolgus monkey. The initial uptake of 18 in the monkey brain was 250% SUV at 4 min post injection. The highest uptake of radioactivity was observed in the striatum and thalamus, regions with high MAO-B activity, whereas lower levels of radioactivity were detected in the cortex and cerebellum. The percentage of unchanged radioligand 18 was 30% in plasma at 90min post injection. In conclusion, compound 18 is a selective inhibitor of MAO-B in vitro and demonstrated a MAO-B specific binding pattern in vivo by PET in monkey. It can, therefore, be considered as a candidate for further investigation in human by PET.

Simultaneous determination of eight active components in Chinese medicine 'JiangYaBiFeng' tablet by HPLC coupled with diode array detection.

An effective, accurate and reliable method was developed for the simultaneous separation and determination of eight active components (baicalin, baicalein, sophoricoside, rutin, quercetin, genistein, pargyline and hydrochlorothiazide) in Chinese medicine 'JiangYaBiFeng' tablet (JYBF tablet) by high-performance liquid chromatography (HPLC) coupled with diode array detection (DAD). Due to the different UV characteristic of these components, different wavelengths were selected for analysis of different analytes, such as 210nm for pargyline, 256nm for sophoricoside, rutin, quercetin and genistein, and 280nm for baicalin, baicalein and hydrochlorothiazide. Excellent linear behaviors over the investigated concentration ranges were observed with the values of R(2) higher than 0.9990 for all analytes. The recovery rates and relative standard deviation (RSD) for all analytes at three different concentrations were 94.9-104.7% and 1.23-3.00%, respectively. The validated method was successfully applied to the simultaneously determination of these active components in 'JiangYaBiFeng' tablet from different production batches, indicating that the proposed method in this paper was particularly suitable for the routine analysis of JYBF tablet and its quality control.

Antioxidant properties of propargylamine derivatives: assessment of their ability to scavenge peroxynitrite.

A series of arylpropargylamines, variously substituted in the hydrogen in p-position and in the propargyl moiety, were studied as potential peroxynitrite scavengers. The scavenging activity of these compounds was evaluated through peroxynitrite (ONOO-)-mediated oxidation of dichlorofluorescin and linoleic acid by measuring the dichlorofluorescein formation and oxygen consumption, respectively. Among tested compounds, only 1-phenylpropargylamine (AP3) promoted concentration-dependent inhibition of ONOO(-)-induced dichlorofluorescin and linoleic acid oxidation with IC50 values of 637 and 63 microM, respectively. The AP3 spectral changes in UV-visible absorbance properties in the presence of peroxynitrite suggested the formation of a new compound. This was identified by gas-chromatograph-mass spectrometer analysis as phenylpropargyl alcohol. Structure-activity relationship analysis indicated that the scavenging activity of AP3 was due to the aminopropargyl moiety and availability of the nitrogen electron pair. This data suggested that AP3 could be considered a lead compound for the synthesis of new ONOO- scavenger derivatives.

Effect of application variables on emissions and distribution of fumigants applied via subsurface drip irrigation.

Soil fumigation is useful for controlling soil-borne pests and diseases in high-cash-value crops. Fumigants are highly volatile, and approaches to reduce atmospheric emissions are required to protect human and environmental health. Application of fumigants through drip irrigation has been proposed as a means to decrease fumigant emissions, improve fumigant distribution in soil, and minimize worker exposure. These experiments were conducted to investigate the effect of the configuration of the drip system on the volatilization and distribution of the fumigants 1,3-dichloropropene (1,3-D), propargyl bromide (PrBr), and methyl isothiocyanate (MITC) in bedded systems. Results indicated that changing the drip emitter spacing and using multiple drip lines in each bed had little effect on the emissions and distribution of any fumigant. Increasing the depth of application from 15 to 30 cm reduced volatilization of MITC by approximately 20 to >90%; emissions were reduced due to a decrease in the flux from the bed top, and deeper injection did not change the amount of fumigant volatilized from the bed side slope and furrow. Increasing the application depth resulted in a slight decrease in the rate of fumigant dissipation in soil, indicating the potential for some improvement in pest-control efficacy with deeper application.

Effect of surface tarp on emissions and distribution of drip-applied fumigants.

Soil fumigants are used to control a wide variety of soil-borne pests in high-cash-value crops. Application of soil fumigants through drip irrigation systems is receiving increasing attention as a method to improve the uniformity of fumigant application. Little information is available on the emissions and soil distribution of fumigants following subsurface drip application, or the effect of plastic tarp on fumigant emissions in these systems. In these experiments, the fumigant compounds 1,3-dichloropropene (1,3-D), Vapam (a methyl isothiocyanate (MITC) precursor), and propargyl bromide (PrBr) were applied to soil beds via drip irrigation at 15 cm depth. Beds were tarped with either standard 1-mil high-density polyethylene (HDPE) or a virtually impermeable film (VIF), leaving the furrows bare. Cumulative emissions of 1,3-D, MITC, and PrBr in these tarped bedded systems was very low, amounting to <10% of the applied mass. These experiments were conducted in the winter months, with average air temperatures of 12-15 degrees C. Cumulative emissions of MITC and 1,3-D from a sandy loam field soil were decreased by > or =80% by tarping the bed with VIF rather than HDPE. A large fraction of the 1,3-D and PrBr flux was from the untarped furrows in VIF-tarped plots, indicating that inhibiting volatilization from the furrow will be important in further reducing emissions in these systems. Monitoring the fumigant distribution in soil indicated that tarping the bed with VIF resulted in a more effective containment of fumigant vapors compared to use of a HDPE tarp.

Gas-phase sorption-desorption of propargyl bromide and 1,3-dichloropropene on plastic materials.

The goal of this research was to provide information for choosing appropriate materials for studying gas-phase concentrations of propargyl bromide (3BP) and 1,3-dichloropropene (1,3-D) in laboratory experiments. Several materials were tested and found to sorb both gas-phase chemicals in the following order: stainless steel (SS) < Teflon polytetrafluoroethylene (PTFE-FEP) approximately flexible polyvinyl chloride (PVC) approximately acrylic < low-density polyethylene (PE) < vinyl approximately silicone < polyurethane foam (PUF). Sorption of SS was insignificant and PUF sorbed all the fumigant that was applied. For the other materials, linear sorption coefficients (Kd) for 3BP ranged from 3.0 cm3 g(-1) for PVC to 215 cm3 g(-1) for silicone. Freundlich sorption coefficients for 1,3-D ranged from 11.5 to 371 cm3 g(-1). First-order desorption rate constants in an open system ranged from 0.05 to 1.38 h(-1) for 3BP and from 0.07 to 1.73 h(-1) for 1,3-D. In a closed system, less than 2% of sorbed fumigant desorbed from vinyl while up to 99% desorbed from PVC within 24 h when equilibrated at the highest headspace concentration. Sorption of both fumigants was linearly related to the square root of time except for vinyl and silicone. This may indicate non-fickian diffusion of fumigant into the polymer matrix. Vinyl, silicone, PE, and PUF should be avoided for quantitative study of organic gases, except possibly as a trapping medium. Use of PTFE, PVC, and acrylic may require correction for sorption-desorption and diffusion.

A dynamic two-dimensional system for measuring volatile organic compound volatilization and movement in soils.

There is an important need to develop instrumentation that allows better understanding of atmospheric emission of toxic volatile compounds associated with soil management. For this purpose, chemical movement and distribution in the soil profile should be simultaneously monitored with its volatilization. A two-dimensional rectangular soil column was constructed and a dynamic sequential volatilization flux chamber was attached to the top of the column. The flux chamber was connected through a manifold valve to a gas chromatograph (GC) for real-time concentration measurement. Gas distribution in the soil profile was sampled with gas-tight syringes at selected times and analyzed with a GC. A pressure transducer was connected to a scanivalve to automatically measure the pressure distribution in the gas phase of the soil profile. The system application was demonstrated by packing the column with a sandy loam in a symmetrical bed-furrow system. A 5-h furrow irrigation was started 24 h after the injection of a soil fumigant, propargyl bromide (3-bromo-1-propyne; 3BP). The experience showed the importance of measuring lateral volatilization variability, pressure distribution in the gas phase, chemical distribution between the different phases (liquid, gas, and sorbed), and the effect of irrigation on the volatilization. Gas movement, volatilization, water infiltration, and distribution of degradation product (Br-) were symmetric around the bed within 10%. The system saves labor cost and time. This versatile system can be modified and used to compare management practices, estimate concentration-time indexes for pest control, study chemical movement, degradation, and emissions, and test mathematical models.

Evaluation of HPLC retention data by non-linear mapping, cluster analysis and varimax rotation. A comparative study.

The retention of 17 propargylamine derivatives was determined on a beta-cyclodextrin polymer (beta-CDP)-coated silica column using tetrahydrofuran-0.05 M K2HPO4 (6:4, v/v) as eluent. The inclusion complex formation between the propargylamine derivatives and a water-soluble beta-CDP was studied by charge-transfer chromatography carried out on reversed-phase TLC layers. The capacity factors were correlated with the various measured and calculated physicochemical parameters of the solutes using principal component analysis followed by non-linear mapping, varimax rotation and cluster analysis. Calculations proved that the hydrophobicity and steric parameters have the highest influence on the retention of propargylamine derivatives. It has been established that each statistical method can be used for the evaluation of similar retention data matrices, however, the results can be slightly different according to the method applied.

Determination of D,L-propargylglycine and N-acetylpropargylglycine in urine and several tissues of D,L-propargylglycine-treated rats using liquid chromatography-mass spectrometry.

An experimental animal model with cystathioninuria was obtained by the injection of D,L-propargylglycine into rats. The concentrations of D,L-propargylglycine in urine, several tissues and serum at different times after the injection were measured by liquid chromatography-mass spectrometry. The propargylglycine accumulated rapidly in several tissues and serum of the rats, and reached its maximum level at about 2 h after the injection. Approximately 21.2% of the administered propargylglycine was excreted in urine. N-Acetylpropargylglycine was identified as a new metabolite of propargylglycine in urine. The concentration of propargylglycine was 100 times that of N-acetylpropargylglycine in urine.

Enantiospecific analysis: applications in bioanalysis and metabolism.

Enantiospecific analysis has a significant role in modern drug development from discovery-chemistry to the clinical evaluation of novel compounds. Chromatographic techniques, involving the use of either chiral derivatizing agents or chiral stationary phases, represent the most commonly used approaches to enantiospecific analysis. The advantages and limitations of these two techniques are examined using the analysis of the enantiomers of the 2-arylpropionic acids (tiaprofenic acid and ibuprofen) and the chiral N-oxides of N-ethyl-N-methylaniline and pargyline, as representative examples for each approach. The potential of biosensors in enantiospecific analysis is addressed and some preliminary results on the development of an enantioselective biosensor for the analysis of (S)-warfarin are presented.

Biological characterization of 10-(2-propynyl) estr-4-ene-3, 17-dione (MDL 18,962), an enzyme-activated inhibitor of aromatase.

MDL 18,962 was shown to be a highly specific, potent (Ki = 3-4 nM), enzyme-activated inhibitor of aromatase with minimal intrinsic endocrine properties. The affinity of MDL 18,962 was higher for human and baboon placental aromatase than for rhesus placental or rodent ovarian aromatase. These species differences necessitated the development of a novel model of peripheral aromatase utilizing human enzyme. Human choriocarcinoma trophoblast xenografts in athymic nude mice were used for pharmacologic and pharmacokinetic evaluation of MDL 18,962. The ED50 for inhibition of aromatase activity in these trophoblast tumors at 6 h post-treatment was 1.4 mg/kg, s.c. and 3.0 mg/kg, oral. Preliminary results indicated that the ED50 for inhibition of peripheral aromatization of androgen by MDL 18,962 in female baboons was 0.01 mg/kg, i.v. and 4 mg/kg, oral.

Microsomal formation and chemical decomposition of pargyline N-oxide.

Pargyline undergoes metabolic N-oxidation in rat and rabbit liver microsomal preparations. The reaction requires oxygen and is NADPH dependent. N-oxidation and N-demethylation are equal in both control and induced rat liver microsomes, while N-oxidation is more dominant in rabbit tissue. Experiments investigating the CO-sensitivity and the effects of metyrapone suggest that cytochrome P-450 systems are involved in both reactions in the rat while an additional enzyme is responsible for the N-oxidation in the rabbit. Pargyline N-oxide is characterized by chemical instability and undergoes two consecutive rearrangements to yield propenal and Schiff bases, the latter undergoing hydrolysis to aldehydes and primary amines. Accordingly, due to the inherent instability of the N-oxide, metabolic N-oxidation of pargyline is, in addition to alpha-carbon oxidation, indicated as a metabolic route to benzaldehyde. Similarly the ease with which pargyline N-oxide generates propenal implicates N-oxidation as a metabolic route to be considered when evaluating the toxicity of pargyline.