A methanol and protic ionic liquid Ugi multicomponent reaction path to cytotoxic α-phenylacetamido amides

The Ugi four component reaction of an aldehyde, amine, isocyanide and an ethanoic acid was effected smoothly in protic ionic liquids ethylammonium nitrate (EAN) and propylammonium nitrate (PAN) to afford analogues of α-phenylacetamido amides in good to excellent isolated yields. The corresponding reactions in [BMIM][PF₆] and the protic ionic liquid ethanolammonium nitrate (ETAN) failed. Microwave irradiation in EAN facilitated rapid access to three focused libraries, based on the parent isocyanide: cyclohexyl isocyanide, benzyl isocyanide and ethyl isocyanoacetate. Analysis of the structure activity relationship data suggested the presence of a bulky moiety originating from the isocyanide (cyclohexyl and benzyl) enhanced cytotoxicity. Removal of the acetylenic H-atom from the ethanoic acid moiety was detrimental to cytotoxicity. The most active analogues produced, N-(2-cyclohexylamino)-1-(4-methoxyphenyl)-2-oxoethyl-N-(3,5-dimethoxyphenyl)propiolamide, returned average GI₅₀ values of ≤1 μM across the cancer cell lines evaluated. Combined, these data suggest that analogues of this nature are interesting potential anti-cancer development leads.

The Ugi reaction (aldehyde, amine, isocyanide and an ethanoic acid) in the protic ionic liquids ethylammonium nitrate (EAN) and propylammonium nitrate (PAN) gave excellent yields of α-phenylacetamido amides.     

Development of pharmacodynamic biomarkers for ATR inhibitors

Background

ATR, which signals DNA damage to S/G2 cell cycle checkpoints and for repair, is an attractive target in cancer therapy. ATR inhibitors are being developed and a pharmacodynamic assay is needed to support clinical studies.

Methods

Phosphorylation of ATR targets, Chk1 and H2AX, was evaluated in MCF7 and K562 cells, human volunteer PBMCs and whole blood by Western blot, immunofluorescence microscopy and flow cytometry after DNA damage. The effect of cell cycle phase, ATR knockdown and inhibition on these phosphorylation events was determined.

Results

Hydroxyurea, UV and 4NQO induced Chk1 and H2AX phosphorylation in MCF7 and K562 cells. UV/4NQO activation of ATR was detectable in non‐cycling cells. Chk1 phosphorylation was reduced by ATR knockdown and reflects ATR activity for 3 h, H2AX phosphorylation after UV/4NQO is ATR‐dependent for 1 h but increasingly ATM and DNA‐PK‐dependent at later time points. In isolated PBMCs both phospho‐targets were detectable after UV/4NQO but in PBMCs from whole blood treated with 4NQO only H2AX was detectable.

Conclusion

PhosphoChk1 and H2AX are useful biomarkers for ATR inhibition using a variety of immuno‐detection methods, but timing may be critical. Importantly, ATR activity is detectable in non‐cycling PBMCs allowing them to be used as a surrogate tissue for biomarker measurement. In PBMCs from whole blood treated with 4NQO phosphoH2AX was the most useful biomarker of ATR activity and a clinically viable pharmacodynamic assay for ATR inhibitors has been developed.     

A comparison of physical activity from Actigraph GT3X+ accelerometers worn on the dominant and non‐dominant wrist

The purpose of this study was to evaluate the agreement between several activity measures using raw acceleration data from accelerometers worn concurrently on the dominant and non‐dominant wrist. Fifty‐five adults (31·9 ± 9·7 years, 26 males) wore two ActiGraph GT3X+ monitors continuously for 1 day, one on their non‐dominant wrist and the other on their dominant wrist. Paired t‐tests were undertaken with sequential Holm‐Bonferroni corrections to compare wear time, moderate‐vigorous physical activity (MVPA), time spent in 10‐min bouts of MVPA (MVPA_10 min) and the average magnitude of dynamic wrist acceleration (ENMO). Level of agreement between outcome variables from the wrists was examined using intraclass correlation coefficients (ICC, single measures, absolute agreement) with 95% confidence intervals and limits of agreement (LoA). Time spent across acceleration levels in 40 mg resolution were also examined. There were no significant differences between the non‐dominant and dominant wrist for ENMO, wear time, MVPA or MVPA_10 min. Agreement between wrists was strong for most outcomes (ICC ≥0·92) including wear time, ENMO, MVPA, MVPA_10 min and the distribution of time across acceleration levels. Agreement was strong in the low acceleration bands (ICC = 0·970 and 0·922) with a mean bias of 3·08 min (LoA ?55·18 to 61·34) and ?5·43 (LoA ?43·47 to 32·62). In summary, ENMO, MVPA, MVPA_10 min, wear time and the distribution of time across acceleration levels compared well at the group level. The LOA from the two lowest acceleration levels suggest further work over a longer monitoring period is needed to determine whether outputs from each wrist are comparable.     

Early diffusion weighted magnetic resonance imaging can predict survival in women with locally advanced cancer of the cervix treated with combined chemo-radiation

Objectives

To assess the predictive value of diffusion weighted imaging (DWI) for survival in women treated for advanced cancer of the cervix with concurrent chemo-radiotherapy.

Methods

Twenty women treated for advanced cancer of the cervix were recruited and followed up for a median of 26 (range <1 to 43) months. They each had DWI performed before treatment, 2?weeks after beginning therapy (midtreatment) and at the end of treatment. Apparent diffusion coefficient (ADC) values were calculated from regions of interest (ROI). All participants were reviewed for follow-up data. ADC values were compared with mortality status (Mann–Whitney test). Time to progression and overall survival were assessed (Kaplan–Meier survival graphs).

Results

There were 14 survivors. The median midtreatment ADC was statistically significantly higher in those alive compared to the non-survivors, 1.55 and 1.36 (×10^?3/mm²/s), respectively, P?=?0.02. The median change in ADC 14?days after treatment commencement was significantly higher in the alive group compared to non-survivors, 0.28 and 0.14 (×10^?3/mm²/s), respectively, P?=?0.02. There was no evidence of a difference between survivors and non-survivors for pretreatment baseline or post-therapy ADC values.

Conclusion

Functional DWI early in the treatment of advanced cancer of the cervix may provide useful information in predicting survival.

Key Points

? Diffusion weighted magnetic resonance imaging (DWI) is increasingly used in cervical cancer. ? Functional DWI early in treatment of cervical cancer may help predict survival. ? DWI may help clinicians to tailor or individualise treatment appropriately. ? This may limit toxicity from ineffective treatment and allow early alternative therapy.