ABSTRACT
Two novel structural types of tyrosyl-DNA phosphodiesterase 1 (TDP1) inhibitors with hexahydroisobenzofuran 11 and 3-oxabicyclo [3.3.1]nonane 12 scaffolds were discovered. These monoterpene-derived compounds were synthesized through preliminary isomerization of (+)-3-carene to (+)-2-carene followed by reaction with heteroaromatic aldehydes. All the compounds inhibit the TDP1 enzyme at micro- and submicromolar levels, with the most potent compound having an IC50 value of 0.65 µM. TDP1 is an important DNA repair enzyme and a promising target for the development of new chemosensitizing agents. A panel of isogenic clones of the HEK293FT cell line knockout for the TDP1 gene was created using the CRISPR-Cas9 system. Cytotoxic effects of topotecan (Tpc) and non-cytotoxic compounds of the new structures were investigated separately and jointly in the TDP1 gene knockout cells. For two TDP1 inhibitors, 11h and 12k, a synergistic effect was observed with Tpc in the HEK293FT cells but was not found in TDP1 -/- cells. Thus, it is likely that the synergistic effect is caused by inhibition of TDP1. Synergy was also found for 11h in other cancer cell lines. Thus, sensitizing cancer cells using a non-cytotoxic drug can enhance the efficacy of currently used pharmaceuticals and, concomitantly, reduce toxic side effects.
Subject(s)
Bicyclic Monoterpenes/chemistry , Drug Design , Phosphodiesterase Inhibitors/chemical synthesis , Phosphodiesterase Inhibitors/pharmacology , Phosphoric Diester Hydrolases/metabolism , Signal Transduction/drug effects , CRISPR-Cas Systems , Cell Proliferation/drug effects , Cell Survival/drug effects , Drug Synergism , Gene Knockout Techniques , HCT116 Cells , HEK293 Cells , HeLa Cells , Humans , Inhibitory Concentration 50 , Phosphodiesterase Inhibitors/chemistry , Phosphoric Diester Hydrolases/genetics , Topotecan/pharmacologyABSTRACT
The methods for quantification of highly potent analgesic agent (2R,4aR,7R,8aR)-4,7-dimethyl-2-(thiophen-2-yl)octahydro-2H-chromen-4-ol in rat whole blood and plasma were developed and validated using dried matrix spots (DMS) or fabric phase sorptive extraction (FPSE) techniques in combination with LC-MS/MS. 2-Adamantylamine hydrochloride was used as an internal standard (IS). Chromatographic separation was carried out on a reversed-phase column (2.0×75â¯mm, 5⯵m) using water containing 0.1% formic acid and methanol containing 0.1% formic acid as mobile phases in gradient mode at a flow rate of 200⯵L/min. The mass spectrometric detection was performed using electrospray ionization (ESI) in positive ion mode. MRM transitions were m/z 284.5â¯ââ¯137.2/157.4 for the analgesic agent and m/z 152.3â¯ââ¯93.1/107.2 for IS. Calibration curves were linear within 20-5000â¯ng/mL in dried plasma spots (DPS) or dried blood spots (DBS) experiments. The linearity was obtained in the range of 20-5000â¯ng/mL and 50-5000â¯ng/mL for plasma-FPSE and blood-FPSE experiments, respectively. The intra- and inter-day accuracy and precision did not exceed acceptable limits. The mean extraction recovery (%) was 26 for DPS, 25 for DBS, 38 for plasma-FPSE, 31 for blood-FPSE.
Subject(s)
Analgesics/blood , Chromatography, Liquid/methods , Dried Blood Spot Testing/methods , Tandem Mass Spectrometry/methods , Thiophenes/blood , Animals , Benzopyrans/blood , Limit of Detection , Linear Models , Rats , Reproducibility of ResultsABSTRACT
A simple and efficient method for synthesizing chiral heterocyclic compounds with the hexahydrofluoreno[9,1-bc]furan framework via interaction between trans-4-hydroxymethyl-2-carene and aromatic aldehydes containing methoxy and hydroxyl moieties in the presence of montmorillonite clay was found. One of the synthesized compounds exhibited a high cytotoxic activity against lymphoblastoid cell line MT-4 (CTD50 0.9µM), which was higher than that of the comparative drug Doxorubicin. Death of cancer cells in this case substantially occurs via induction of apoptosis.
Subject(s)
Antineoplastic Agents/chemical synthesis , Antineoplastic Agents/toxicity , Furans/chemical synthesis , Furans/toxicity , Apoptosis/drug effects , Cell Line, Tumor , Cell Proliferation/drug effects , Cell Survival/drug effects , Drug Screening Assays, Antitumor/methods , Fluorenes/chemical synthesis , Fluorenes/toxicity , Humans , Structure-Activity RelationshipABSTRACT
(1R,2R,6S)-3-Methyl-6-(prop-1-en-2-yl)cyclohex-3-ene-1,2-diol 1 possesses potent antiparkinsonian activity in both MPTP and haloperidol animal models. The use of compound 1 resulted in nearly full recovery of the locomotor and exploratory activities and was as effective as the comparator agent (levodopa). All eight stereoisomers of compound 1 have been synthesized and the influence of the absolute configuration on the antiparkinsonian activity of compound 1 was shown.
