Rociletinib (CO-1686), a 2,4-diaminopyrimidine derivative, is a highly potent tyrosine kinase inhibitor (TKI) that acts on epidermal growth factor receptor (EGFR) with L858R/T790M mutations. We supposed radioiodinated CO-1686 would function as a useful tool for monitoring EGFR L858R/T790M mutations. To aid in patient selection before therapy with EGFR-TKIs, this study aimed to develop a 125I-labeled derivative of CO-1686, N-{3-[(2-{[4-(4-acetylpiperazin-1-yl)-2-methoxyphenyl]amino}-5-(trifluoromethyl)pyrimidine-4-yl] amino}-5-([125I]iodophenyl)acrylamide ([125I]ICO1686) and evaluate its selectivity toward EGFR L858R/T790M. Radiosynthesis was performed by iododestannylation of the corresponding tributylstannyl precursor with [125I]NaI and N-chlorosuccinimide. The selectivity of the tracer for detecting EGFR L858R/T790M was evaluated using three relevant non-small cell lung cancer (NSCLC) cell lines—H1975, H3255 and H441 overexpressing the dual mutation EGFR L858R/T790M, active mutant EGFR L858R and wild-type EGFR, respectively. The nonradioactive ICO1686 and the precursor compound were successfully synthesized. A novel radiolabeled probe, [125I]ICO1686, was prepared with high radiochemical yield (77%) and purity (>99%). ICO1686 exhibited high cytotoxicity toward H1975 (IC50 0.20 ± 0.05 μM) and H3255 (IC50 0.50 ± 0.21 μM), which is comparable to that of CO-1686. In contrast, the cytotoxicity of ICO1686 toward H441 was 10-fold lower than that toward H1975. In the cell uptake study, the radioactivity uptake of [125I]ICO1686 in H1975 was 101.52% dose/mg, whereas the uptakes in H3255 and H441 were 33.52 and 8.95% dose/mg, respectively. The uptake of [125I]ICO1686 in H1975 was greatly reduced to 45.61% dose/mg protein by treatment with excess CO-1686. In vivo biodistribution study of the radiotracer found that its accumulation in H1975 tumor (1.77 ± 0.43% ID/g) was comparable to that in H3255 tumor (1.63 ± 0.23% ID/g) and the accumulation in H1975 tumor was not reduced by pretreatment with an excess dose of CO-1686. Although this radiotracer exhibited highly specific in vitro uptake in target cancer cells, structural modification is required to improve in vivo biodistribution.

AbstrakSusu kedelai dan tempe adalah pangan olahan yang digemari masyarakat Indonesia dan menjadi salah satu sumber protein nabati yang terbuat dari kedelai. Kedelai mengandung senyawa isoflavon glikosida yang bermanfaat untuk kesehatan sebagai pencegah penyakit kardiovaskuler, kanker, diabetes, hipertensi, osteoporosis, obesitas dan menopause. Isoflavon glikosida memiliki struktur yang lebih kompleks, sehingga perlu mengalami proses biokimiawi untuk menyederhanakan strukturnya agar mudah diserap oleh tubuh. Isoflavon glikosida akan disederhanakan menjadi bentuk aglikon selama proses fermentasi dan hidrolisis kimiawi. Genistein merupakan salah satu bentuk isoflavon aglikon yang banyak ditemui pada produk kacang-kacangan. Susu kedelai dan tempe merupakan produk olahan dari kedelai, perbedaannya adalah tempe telah mengalami proses fermentasi, sehingga diyakini memiliki kadar isoflavon aglikon yang tinggi. Penelitian ini bertujuan untuk menentukan kadar genistein pada ekstrak susu kedelai dan tempe dengan menggunakan spektrofotometri UV-Vis. Sampel diekstraksi dengan menggunakan etil asetat, sehingga diperoleh ekstrak kental, selanjutnya diukur kadar genistein dengan metode spektrofotometri UVVis, menggunakan baku standar. Hasil penelitian diperoleh kadar genistein pada ekstrak susu kedelai dan tempe yaitu masing-masing 0.613 dan 5.519% (b/b). Hal ini menunjukkan bahwa kadar isoflavon aglikon lebih tinggi pada tempe dibanding susu kedelai. Kata kunci: genistein, isoflavon aglikon, susu kedelai, tempe Abstract Soymilk and tempeh are a processed food that is popular with Indonesian society and becomes one of vegetable

Objective: The current study aims to determine total flavonoid content (TFC), total phenolic content (TPC), and total carotenoid content (TCC) of ethanolic extract of Purslane (Portulaca oleraceae L.). Material and Method: Herb of Purslane (Portulaca oleraceae L.) was extracted using ethanol. Determination of total flavonoid content (TFC), total phenolic content (TPC), and total carotenoid content (TCC) The method used spectrophotometer UV-VIS by the used standard curve to compare the total content by linear regression calculation. Result: The results of this study demonstrated the TPC, TFC, and TCC value was 2.00 mg HE/g DW, 0.53 mg EQ/g DW, 128 mg GAE/g DW, respectively. For anti-obesity, a 200 mg/kg BW sample group had the greatest percentage decrease compared to the other groups (21.868%). Conclusion: This study showed that Purslane is a good source of flavonoids, phenolic, and carotenoids. Meanwhile, the ethanol extract of this plant has prospect potential as anti-obesity.

Activating double mutations L858R/T790M in the epidermal growth factor receptor (EGFR) region are often observed as the cause of resistance to tyrosine kinase inhibitors (TKIs). Third-generation EGFR-TKIs, such as osimertinib and rociletinib (CO-1686), was developed to target such resistance mutations. The detection of activating L858R/T790M mutations is necessary to select sensitive patients for therapy. Hence, we aimed to develop novel radiobromine-labeled CO-1686 as a positron emission tomography (PET) imaging probe for detecting EGFR L858R/T790M mutations. Nonradioactive brominated-CO1686 (BrCO1686) was synthesized by the condensation of N-(3-[{2-chloro-5-(trifluoromethyl)pyrimidin-4-yl}amino]-5-bromophenyl) acrylamide with the corresponding substituted 1-(4-[4-amino-3-methoxyphenyl]piperazine-1-yl)ethan-1-one. The radiobrominated [77Br]BrCO1686 was prepared through bromodestannylation of the corresponding tributylstannylated precursor with [77Br]bromide and N-chlorosuccinimide. Although we aimed to provide a novel PET imaging probe, 77Br was used as an alternative radionuclide for 76Br. We fundamentally evaluated the potency of [77Br]BrCO1686 as a molecular probe for detecting EGFR L858R/T790M using human non-small-cell lung cancer (NSCLC) cell lines: H1975 (EGFR L858R/T790M), H3255 (EGFR L858R), and H441 (wild-type EGFR). The BrCO1686 showed high cytotoxicity toward H1975 (IC50 0.18 ± 0.06 µM) comparable to that of CO-1686 (IC50 0.14 ± 0.05 µM). In cell uptake experiments, the level of accumulation of [77Br]BrCO1686 in H1975 was significantly higher than those in H3255 and H441 upon 4 h of incubation. The radioactivity of [77Br]BrCO1686 (136.3% dose/mg protein) was significantly reduced to 56.9% dose/mg protein by the pretreatment with an excess CO-1686. These results indicate that the binding site of the radiotracers should be identical to that of CO-1686. The in vivo accumulation of radioactivity of [77Br]BrCO1686 in H1975 tumor (4.51 ± 0.17) was higher than that in H441 tumor (3.71 ± 0.13) 1 h postinjection. Our results suggested that [77Br]BrCO1686 has specificity toward NSCLC cells with double mutations EGFR L858R/T790M compared to those in EGFR L858R and wild-type EGFR. However, the in vivo accumulation of radioactivity in the targeted tumor needs to be optimized by structural modification.