Afatinib is an oral tyrosine kinase inhibitor (TKI) approved for treating advanced non-small cell lung cancer. It is necessary to develop a simple quantification method for TKIs in order to facilitate therapeutic drug monitoring (TDM) in clinical settings. This study sought to develop a simple and sensitive com-petitive enzyme-linked immunosorbent assay (ELISA) to quantify afatinib in plasma for routine phar-macokinetic applications. An anti-afatinib antibody was obtained using (S)-N-4-(3-chloro-4-fluor-ophenyl)-7-(tetrahydrofuran-3-yloxy)-quinazoline-4,6-diamine (CTQD), which has the same sub-structure as afatinib, as a hapten. Enzyme labeling of afatinib with horseradish peroxidase was similarly performed using CTQD. A simple competitive ELISA for afatinib was developed based on the principle of direct competition between afatinib and the enzyme marker for the anti-afatinib antibody, which had been immobilized on the plastic surface of a microtiter plate. Plasma afatinib concentrations below the limit of quantification of 30 pg/mL were reproducibly measurable. Also, the values of plasma afatinib levels measured from 20 patients were comparable with those measured by high-performance liquid chromatography, and there was a strong correlation between the values determined by both methods (Y = 0.976X – 0.207, r = 0.975). As indicated by its specificity and sensitivity, this newly developed ELISA for afatinib is an important tool for TDM and studies of the pharmacokinetics of afatinib.

A statistical study on effects of laser-puncture was carried out. In addition, an attempt was made to compare between the effect of acupuncture and this method. The treatments were under-taken for various chronic and acute cases with pain. In the laser-puncture treatment, two types of laser (i, e. low energy 10mW laser and 70mW sharp laser) were used. Laser-beam was given to 20-30 points which were located in painful areas, innervating nerve areas and remote areas for 40 sec for each.
In the open study, 10mW laser-puncture was effective (including ‘rather effective’) in 64% of the cases, and 70mW laser-puncture 73%. In the blind test, ‘the day’ evaluation showed that the laser-punctures were more effective than placebo treatment: 10mW laser-puncture was effective in 80% of the cases, and placebo in 67%. 70mW laser-puncture was effective in 87% of them, and placebo in 80%. In situ acupuncture was effective in 97%, and no effect was seen in 7%.
Only ‘the day’ evaluation in the open study showed that 70mW laser-puncture was more effective than 10mW one. In the blind best, 10mW and 70mW laser-puncture were more effective than placebo, and in situ acupuncture was more effective than the laser-punctures, but there were no significant differences among the three treatmets.