text
string | predicted_class
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float16 |
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The somatic health of patients in mental health care can no longer be ignored. Changing lifestyle behaviours is difficult, but combining several successful components (e.g. motivational interviewing, stage-of-change, objective monitoring, self-management, support of peers and family etc.) into one multidimensional intervention might enhance successful, sustainable lifestyle changes.
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other
| 99.9 |
Breast cancer is the most common cancer in women worldwide, with more than 1.5 million new cases recorded every year; it is also the fifth highest cause of cancer death (Nathan and Schmid, 2017). Estrogens are steroid hormones that play a critical role in the regulation of growth, differentiation, and metabolism of mammary cells, including malignant cells. Due to the ability of estrogens to significantly stimulate the growth of mammary cells, these hormones are involved in the progression of breast cancer. For more than 40 years, the antiestrogen tamoxifen (ICI 46474) is considered as the absolute leader in the endocrine therapy of hormone-dependent breast cancers (Jameera Begam et al., 2017). Tamoxifen belongs to selective estrogen receptor modulators (SERMs) (Cosman and Lindsay, 1999; Jameera Begam et al., 2017), which, in certain circumstances, perform the role of estrogen agonists or antagonists and modulate the effect of hormones in the target cells; otherwise, SERMs are also called estrogen agonists/antagonists. Due to convenient dosage forms for oral use, high efficiency, and low cost of the prolonged course of therapy, tamoxifen is considered as the “gold standard” for the treatment of patients with ERα-positive breast cancer. On the other hand, the effectiveness of tamoxifen may be limited by the development of resistance, an increased risk of endometrial cancer, and individual drug intolerance (Scherbakov et al., 2006; Ali et al., 2016; Traboulsi et al., 2017). This is why the development of novel classes of agents that effectively inhibit the growth of ERα-positive tumors and have no severe side effects is a challenge (Tryfonidis et al., 2016).
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review
| 99.9 |
Synthetic steroids encompass a wide range of compounds with various specific anticancer activities, e.g., aromatase inhibitors such as formestane and exemestane (Carlini et al., 2001), antiproliferative agents such as 2-methoxyestradiol (Lakhani et al., 2003), androgen signaling inhibitors such as galeterone and abiraterone (Bryce and Ryan, 2012), the SERM compound PSK3471, the steroid sulfatase inhibitor EMATE (Purohit and Foster, 2012), and the selective estrogen receptor degrader (SERD) fulvestrant (Nathan and Schmid, 2017; Figure 1).
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review
| 99.8 |
Aromatase and steroid sulfatase inhibitors, SERDs, and SERMs synthetically derived from natural hormones are of great interest for the development of new breast cancer treatment regimens, especially for metastatic forms of the disease (Singer et al., 2006; Scherbakov et al., 2013; Secky et al., 2013; Boer, 2017; Kaklamani and Gradishar, 2017). Fulvestrant is an estrogen receptor degrader that binds with high selectivity to target cells, causes their degradation, resulting in the complete inhibition of the estrogen-mediated growth of breast cancer cells (Nathan and Schmid, 2017). First approved in the US in 2002, fulvestrant is not associated with tamoxifen-like agonist side effects, is not cross-resistant to tamoxifen or exemestane, and produces very high response rates in breast cancer patients. A combination of fulvestrant with other drugs seems to be very promising. In this regards, the combination of anastrozole and fulvestrant is superior to anastrozole alone or sequential anastrozole and fulvestrant for the treatment of ERα-positive metastatic breast cancer (Mehta et al., 2012).
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review
| 99.9 |
In fulvestrant, the alkylsulfinyl moiety is attached to the endogenous estrogen receptor ligand, 17β-estradiol, at the 7-position, providing a structure similar to that of natural hormones but showing reverse biological activity. Recently, we have demonstrated that the modification of 17β-estradiol with imidazo[1,2-a]pyridine pendant at the 17α-position has the same effect (Rassokhina et al., 2016). 17α-Imidazopyridine-17β-methoxyestradiol showed remarkable effects as a selective ERα receptor modulator. In this study, we turned to an investigation of the structure—ERα-modulator activity relationship for two novel classes of heterosteroids possessing an N-heterocycle attached at the 16-position and fused to the A/D ring of the steroid core. We report the unique derivatives of the androstene and estrane series containing A/D-ring annulated pyrimidine or linked dihydrotriazine moieties. The antiproliferative potential of all synthesized compounds was evaluated in the MCF-7 and MDA-MB-231 breast cancer cell lines. The compounds were also tested toward two prostate cancer cell lines PC3 and 22Rv1. Taking into account activities of the compounds against hormone-dependent breast cancer ERα was analyzed as possible target for this series. Four compounds proved to be active as ERα antagonists. Steroidal dihydrotriazine 4a was selected as the lead compound and analyzed by the ERα-reporter assay, immunoblotting, and docking simulation. Finally, the binding of compound 4a to the estrogen receptor pocket was discussed using different docking models.
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study
| 100.0 |
NMR spectra were acquired on Bruker Avance 600 and 300 spectrometers at 293, 303, and 333 K; the chemical shifts δ were measured in ppm relative to the solvent (1H: DMSO-d6, δ 2.50 ppm; 13C: DMSO-d6, δ 39.50 ppm). Splitting patterns are designated as s, singlet; d, doublet; t, triplet; q, quartet; m, multiplet; dd, double doublet; ddd, double double doublet; dt, doublet triplet. The coupling constants (J) are in Hertz. The structures of compounds were established using 1D NMR (1H, 13C) and 2D NMR (1H-1H COSY, 13C-1H HMBC, 13C-1H HSQC) spectroscopy. Infrared spectra were measured on a FT-IR spectrometer in KBr pellets. High-resolution mass spectra (HRMS) were measured using electrospray ionization (ESI) in positive ion mode (interface capillary voltage 4,500 V); the mass range was from m/z 50 to 3,000 Da; external/internal calibration was performed using an electrospray calibrant solution. A syringe injection was used for solutions in CH3CN (flow rate 3 ml/min). Nitrogen was applied as a dry gas and the interface temperature was set at 180°C. Melting points were measured on a Boetius capillary melting point apparatus and are uncorrected. Analytical thin-layer chromatography (TLC) was carried out on silica gel plates (silica gel 60 F254 aluminum supported plates); the visualization was accomplished with an UV lamp (254/365 nm) and using chemical staining with [Ce(SO4)2/H2SO4]. Column chromatography was performed on silica gel 60 (230–400 mesh, Merck). Androst-5-en-3β-ol-17-one-3β-acetate, 17β-hydroxy-5α-androstan-3-one, estrone, phosphorus oxychloride, guanidine salts, and acetimidamide hydrochloride were commercially available and were used as purchased. The spectroscopic data for steroidal chlorovinyl aldehydes 2a–c are consistent with those reported previously (Komkov et al., 2015; Volkova et al., 2016). All reactions were carried out in freshly distilled and dry solvents.
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study
| 100.0 |
Chloro-16-formyl-Δ1,3,5(10)-estratetraen-3-ol 2a (113 mg, 0.36 mmol) was added to a suspension of acetamidine hydrochloride (51 mg, 0.54 mmol) and potassium carbonate (120 mg, 0.89 mmol) in DMF (4 mL). The mixture was stirred at 80°C for 9 h until the complete conversion of the intermediates (TLC monitoring). The resulting mixture was cooled to room temperature and diluted with water (30 mL). The precipitate that formed was filtered, dried, and washed with water (3 × 5 mL). The product was isolated by column chromatography using petroleum ether/ethyl acetate, 1:1, to obtain colorless solid (34 mg, 30% yield). Rf 0.27 (petroleum ether:EtOAc, 1:2; visualized by UV light at 254 nm); m.p. 264–266°C. 1H NMR (600 MHz, DMSO-d6), δ: 0.92 (s, 3H, 18-CH3), 1.35–1.41 (m, 1H, 7-CH2), 1.50–1.56 (m, 1H, 11-CH2), 1.61–1.69 (m, 2H, 8-CH2, 12-CH2), 1.70–1.76 (m, 1H, 14-CH), 1.90–1.95 (m, 1H, 7-CH), 2.14–2.18 (m, 1H, 12-CH2), 2.21–2.27 (m, 1H, 9-CH), 2.37–2.42 (m, 1H, 11-CH2), 2.48–2.54 (m, 1H, 15-CH2), 2.56 (s, 3H, 2′-CH3), 2.73–2.85 (m, 3H, 6-CH2, 15-CH2), 6.47 (s, 1H, 4-CH), 6.53 (dd, J = 2.4, 8.4 Hz, 1H, 2-CH), 7.07 (d, 1H, J = 8.4 Hz, 1-CH), 8.46 (s, 1H, 4-CH), 9.00 (br.s, 1H, OH). 13C NMR (125 MHz, DMSO-d6), δ: 17.1 (18-CH3), 25.5 (2′-CH3), 25.8 (11-CH2), 26.9 (7-CH2), 27.0 (15-CH2), 28.9 (6-CH2), 32.7 (12-CH2), 37.2 (8-CH), 43.7 (9-CH), 45.8 (13-C), 54.3 (14-CH), 112.8 (2-CH), 115.0 (4-CH), 125.8 (1-CH), 130.1 (10-C), 130.5 (16-C), 137.0 (5-C), 151.8 (4′-CH), 155.0 (3-C), 165.1 (2′-C), 181.0 (17-C). IR (KBr), cm−1: 3179 (OH), 2986, 2929, 2859 (CH), 1607, 1585, 1555, 1501 (C = C, C = N). HRMS (ESI) for C21H25N2O ([M+H]+): calcd 321.1961, found 321.1951.
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study
| 100.0 |
17-Chloro-16-formyl-Δ1,3,5(10)-estratetraen-3-ol 2a (142 mg, 0.45 mmol) was added to a suspension of guanidine acetate (80 mg, 0.67 mmol) and potassium carbonate (180 mg, 1.34 mmol) in methanol (10 mL). The mixture was refluxed for 6 h until the complete conversion of the intermediates (TLC monitoring). The resulting mixture was cooled to room temperature and the solvent was removed under reduced pressure. The solid reside was washed with water (10 mL) and dried. The workup afforded the analytically pure product as colorless solid (129 mg, 89% yield). Rf 0.31 (CHCl3:MeOH, 5:0.2; visualized by UV light at 254 nm). The spectral data are consistent with those reported by Forgo and Vincze (2002); m.p. 285–287°C [m.p.lit (Forgo and Vincze, 2002) = 284–286°C]. 1H NMR (600 MHz, DMSO-d6), δ: 0.89 (s, 3H, 18-CH3), 1.32–1.36 (m, 1H, 7-CH2), 1.45–1.50 (m, 1H, 11-CH2), 1.54–1.64 (m, 2H, 8-CH2, 12-CH2), 1.65–1.68 (m, 1H, 14-CH), 1.86–1.90 (m, 1H, 7-CH2), 2.03–2.08 (m, 1H, 12-CH2), 2.20–2.25 (m, 1H, 9-CH), 2.31–2.38 (m, 2H, 11-CH2, 15-CH2), 2.62 (dd, J = 6.6, 14.4 Hz, 1H, 15-CH2), 2.69 (dt, J = 5.4, 16.2 Hz, 1H, 6-CH2), 2.77 (dt, J = 11.4, 16.2 Hz, 1H, 6-CH2), 6.36 (br.s, 2H, NH2), 6.40 (s, 1H, 4-CH), 6.45 (dd, J = 1.8, 8.4 Hz, 1H, 2-CH), 6.98 (d, 1H, J = 8.4 Hz, 1-CH), 8.00 (d, J = 1.8 Hz, 1H, 4-CH) (the signal of OH group was not observed in the 1H NMR spectrum). 13C NMR (125 MHz, DMSO-d6), δ: 17.1 (CH3), 26.0 (11-CH2), 26.6 (15-CH2), 27.1 (7-CH2), 29.1 (6-CH2), 32.9 (12-CH2), 37.4 (8-CH), 43.9 (9-CH), 45.8 (13-C), 54.4 (14-CH), 113.5 (2-CH), 115.6 (4-CH), 121.0 (16-C), 125.6 (1-CH), 128.2 (10-C), 136.7 (5-C), 152.6 (4′-CH), 157.5 (3-C), 162.9 (2′-C), 182.0 (17-C). IR (KBr), cm−1: 3361, 3181 (OH), 2930, 2858 (CH), 1637 (NH2), 1608, 1560 (C = C, C = N). HRMS (ESI) for C20H24N3O ([M+H]+): calcd 322.1914, found 322.1903.
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study
| 100.0 |
3β-Acetoxy-17-chloro-16-formylandrosta-5,16-diene 2b (130 mg, 0.34 mmol) was added to a suspension of acetamidine hydrochloride (65 mg, 0.69 mmol) and potassium carbonate (190 mg, 1.36 mmol) in DMF (5 mL). The mixture was stirred at 80°C for 6.5 h until the complete conversion of the intermediates (TLC monitoring). The resulting mixture was cooled to room temperature and the solvent was removed under reduced pressure. The crude product was isolated by column chromatography using petroleum ether/ethyl acetate, 1:2, to obtain colorless solid (52 mg, 40% yield). Rf 0.47 (petroleum ether:EtOAc, 1:2; visualized by UV light at 254 nm). The spectral data are consistent with those reported by Gogoi et al. (2013). m.p. 169–171°C [m.p.lit (Gogoi et al., 2013) = 165–167°C]. 1H NMR (600 MHz, DMSO-d6at303K), δ: 0.91 (s, 3H, 18-CH3), 1.05 (s, 3H, 19-CH3), 1.07–1.13 (m, 2H, 1-CH2, 9-CH), 1.49 (dt, J = 4.2, 12.6 Hz, 1H, 12-CH2), 1.53–1.65 (m, 4H, 2-CH2, 11-CH2, 12-CH2, 14-CH), 1.67–1.73 (m, 2H, 7-CH2, 11-CH2), 1.74–1.82 (m, 2H, 2-CH2, 8-CH), 1.86 (dt, J = 3.6, 13.2 Hz, 1H, 1-CH2), 1.99 (s, 3H, CH3CO), 2.05–2.12 (m, 1H, 7-CH2), 2.28–2.32 (m, 2H, 4-CH2), 2.47 (dd, J = 15.0 Hz, 1H, 15-CH2), 2.54 (s, 3H, 2′-CH3), 2.72 (dd, J = 6.6,15.0 Hz, 1H, 15-CH2), 4.43–4.48 (m, 1H, 3-CH), 5.39–5.40 (m, 1H, 6-CH), 8.43 (s, 1H, H-4′). 13C NMR (125 MHz, DMSO-d6at303K), δ: 16.7 (18-CH3), 18.9 (19-CH3), 20.0 (11-CH2), 21.0 (CH3), 25.4 (2′-CH3), 27.3 (2-CH2), 27.3 (15-CH2), 30.1 (8-CH), 30.6 (7-CH2), 32.4 (12-CH2), 36.3 (10-C), 36.3 (1-CH2), 37.7 (4-CH2), 45.4 (13-C), 49.8 (9-CH), 55.0 (14-CH), 73.1 (3-CH), 121.6 (6-CH), 130.5 (16-C), 139.8 (5-C), 151.8 (4′-CH), 165.1 (2′-C), 169.7 (CO), 180.8 (17-C). IR (KBr), cm−1: 2941, 2903, 2858 (CH), 1732 (CO), 1589, 1556 (C = C, C = N). HRMS (ESI) for C24H33N2O2 ([M+H]+): calcd 381.2537, found 381.2535.
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study
| 100.0 |
A mixture of steroidal pyrimidine 3c (50 mg, 0.13 mmol) and potassium carbonate (200 mg, 1.44 mmol) in MeOH (8 mL) was refluxed for 3 h until the complete conversion of the starting material (TLC monitoring). The resulting mixture was cooled to room temperature and the solvent was removed under reduced pressure. The crude product was washed with water (6 mL) and dried to get colorless solid (42 mg, 95% yield). Rf 0.28 (CHCl3:MeOH, 5:0.1; visualized by UV light at 254 nm); m.p. 182–183°C. 1H NMR (600 MHz, DMSO-d6 + CCl4), δ: 0.92 (s, 3H, 18-CH3), 1.03 (s, 3H, 19-CH3), 0.99–1.09 (m, 2H, 1-CH2, 9-CH), 1.32–1.38 (m, 1H, 2-CH2), 1.48 (dt, J = 4.2, 12.6 Hz, 1H, 12-CH2), 1.50–1.55 (m, 1H, 14-CH), 1.58–1.73 (m, 4H, 2-CH2, 7-CH2, 11-CH2), 1.74–1.82 (m, 2H, 1-CH2, 8-CH), 2.04–2.19 (m, 4H, 4-CH2, 7-CH2, 12-CH2), 2.46 (dd, J = 14.4 Hz, 1H, 15-CH2), 2.55 (s, 3H, 2′-CH3), 2.72 (dd, J = 14.4 Hz, 1H, 15-CH2), 3.22–3.32 (m, 1H, 3-CH), 5.28–5.30 (m, 1H, 6-CH), 8.39 (s, 1H, H-4′) (the signal of OH group was not observed in the 1H NMR spectrum). 13C NMR (125 MHz, DMSO-d6 + CCl4), δ: 16.7 (18-CH3), 19.1 (19-CH3), 20.0 (11-CH2), 25.3 (2′-CH3), 27.4 (15-CH2), 30.2 (8-CH), 30.7 (7-CH2), 31.3 (2-CH2), 32.5 (12-CH2), 36.3 (10-C), 36.8 (1-CH2), 42.2 (4-CH2), 45.4 (13-C), 50.1 (9-CH), 55.2 (14-CH), 69.9 (3-CH), 119.8 (6-CH), 130.4 (16-C), 141.6 (5-C), 151.5 (4′-CH), 164.9 (2′-C), 180.7 (17-C). IR (KBr), cm−1: 3378 (OH), 2965, 2938, 2858, 2818 (CH), 1599, 1554 (C = C, C = N). HRMS (ESI) for C22H31N2O ([M+H]+): calcd 339.2431, found 339.2431.
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study
| 100.0 |
3β-Acetoxy-17-chloro-16-formylandrosta-5,16-diene 2b (122 mg, 0.32 mmol) was added to a suspension of guanidine acetate (58 mg, 0.48 mmol) and potassium carbonate (134 mg, 0.97 mmol) in methanol (10 mL). The resulting mixture was refluxed for 6 h until the complete conversion of the intermediates (TLC monitoring). The resulting mixture was cooled to room temperature. The precipitate that formed was filtered, washed with H2O (5 mL), and dried. The workup afforded the analytically pure product as colorless solid (92 mg, 84% yield). Rf 0.56 (CHCl3:MeOH, 5:0.2; visualized by UV light at 254 nm). The spectral data are consistent with those reported by Matsumoto et al. (2003). m.p. 342–344°C [m.p.lit (Matsumoto et al., 2003) = 308–312°C]. 1H NMR (600 MHz, DMSO-d6at333K), δ: 0.90 (s, 3H, 18-CH3), 1.03 (s, 3H, 19-CH3), 0.98–1.10 (m, 2H, 1-CH2, 9-CH), 1.34–1.42 (m, 1H, 2-CH2), 1.42–1.50 (m, 1H, 12-CH2), 1.44–1.52 (m, 1H, 14-CH), 1.55–1.83 (m, 6H, 1-CH2, 2-CH2, 7-CH2, 8-CH, 11-CH2), 1.99–2.09 (m, 2H, 7-CH2, 12-CH2), 2.09–2.22 (m, 2H, 4-CH2), 2.29 (dd, J = 6.0, 13.8 Hz, 1H, 15-CH2), 2.55 (dd, J = 6.0, 13.8 Hz, 1H, 15-CH2), 3.24–3.32 (m, 1H, 3-CH), 4.41 (br.s, 1H, OH), 5.30–5.32 (m, 1H, 6-CH), 6.11 (br.s, 2H, NH2), 7.98 (s, 1H, H-4′). 13C NMR (125 MHz, DMSO-d6at333K), δ: 16.4 (18-CH3), 18.8 (19-CH3), 19.9 (11-CH2), 26.6 (15-CH2), 30.1 (7-CH2), 30.5 (8-CH), 31.2 (2-CH2), 32.5 (12-CH2), 36.1 (10-C), 36.6 (1-CH2), 42.0 (4-CH2), 45.0 (13-C), 50.0 (9-CH), 55.1 (14-CH), 69.7 (3-CH), 119.6 (6-CH), 121.0 (16-C), 141.5 (5-C), 152.2 (4′-CH), 162.6 (2′-C), 181.5 (17-C). IR (KBr), cm−1: 3535 (OH), 3368, 3314, 3159 (NH2), 2935, 2893, 2844 (CH), 1647 (NH2), 1608, 1559 (C = C, C = N). HRMS (ESI) for C21H30N3O ([M+H]+): calcd 340.2383, found 340.2382. Anal. calcd for C21H29N3O:C, 74.30; H, 8.6; N, 12.38. Found: C, 73.83; H, 8.65; N, 12.15.
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study
| 100.0 |
3-Chloro-2-formyl-17β-formyloxy-5α-androstane 2c (108 mg, 0.30 mmol) was added to a suspension of guanidine acetate (53 mg, 0.45 mmol) and potassium carbonate (124 mg, 0.90 mmol) in methanol (10 mL). The mixture was refluxed for 4 h until the complete conversion of the intermediates (TLC monitoring). The resulting mixture was cooled to room temperature. The precipitate that formed was filtered, washed with water (5 mL), and dried. The workup afforded analytically pure product as colorless solid (91 mg, 89% yield). Rf 0.67 (CHCl3:MeOH, 5:0.2; visualized by UV light at 254 nm). The spectral data are consistent with those reported by De Ruggieri et al. (1962). m.p. > 350°C (m.p.lit > 300°C). 1H NMR (600 MHz, DMSO-d6), δ: 0.68 (s, 3H, 18-CH3), 0.69 (s, 3H, 19-CH3), 0.78–0.84 (m, 1H, 9-CH), 0.86–0.95 (m, 2H, 7-CH2, 14-CH), 0.98–1.06 (m, 1H, 12-CH2), 1.14–1.29 (m, 2H, 6-CH2, 15-CH2), 1.32–1.42 (m, 3H, 8-CH, 11-CH2, 16-CH2), 1.49–1.56 (m, 3H, 5-CH, 6-CH2, 15-CH2), 1.58–1.64 (m, 1H, 11-CH2), 1.64–1.70 (m, 1H, 7-CH2), 1.77–1.82 (m, 1H, 12-CH2), 1.83–1.89 (m, 1H, 16-CH2), 2.16 (d, J = 15.6 Hz, 1H, 1-CH2), 2.24 (dd, J = 12.6, 18.0 Hz, 1H, 4-CH2), 2.43–2.56 (m, 2H, 4-CH2, 1-CH2), 3.47 (t, J = 8.4 Hz, 1H, 17-CH), 4.18 (br.s, 1H, OH), 5.90 (br.s, 2H, NH2), 7.91 (s, 1H, H-4′). 13C NMR (125 MHz, DMSO-d6), δ: 10.8 (18-CH3), 10.9 (19-CH3), 20.2 (11-CH2), 22.7 (15-CH2), 27.8 (6-CH2), 29.6 (16-CH2), 30.5 (7-CH2), 34.5 (10-C), 35.0 (8-CH), 35.2 (4-CH2), 36.3 (12-CH2), 38.4 (1-CH2), 40.8 (5-CH), 42.1 (13-C), 50.4 (14-CH), 53.0 (9-CH), 79.8 (17-CH), 116.7 (2-C), 157.9 (4′-CH), 161.6 (2′-C), 164.2 (3-C). IR (KBr), cm−1: 3322, 3169 (NH2), 2969, 2923, 2905, 2849 (CH), 1659 (NH2), 1596, 1561 (C = C, C = N). HRMS (ESI) for C21H32N3O ([M+H]+): calcd 342.2540, found 342.2538.
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study
| 100.0 |
17-Chloro-16-formyl-Δ1,3,5(10)-estratetraen-3-ol 2a (100 mg, 0.32 mmol) was added to a suspension of acetamidine hydrochloride (150 mg, 1.3 mmol) and potassium carbonate (260 mg, 1.92 mmol) in DMF (4 mL). The mixture was stirred at 80°C for 8 h until the complete conversion of the intermediates (TLC monitoring). The resulting mixture was cooled to room temperature and diluted with water (30 mL). The precipitate that formed was filtered and washed with water (5 mL) and hot benzene (5 mL). The workup afforded the analytically pure product as colorless solid (40 mg, 32% yield). Rf 0.37 (petroleum ether:EtOAc, 1:2; visualized by UV light at 254 nm); m.p. 209–210°C. 1H NMR (600 MHz, DMSO-d6), δ: 0.84 (s, 3H, 18-CH3), 1.30–1.36 (m, 1H, 7-CH2), 1.37–1.50 (m, 3H, 8-CH, 11-CH2, 12-CH2), 1.58–1.62 (m, 1H, 14-CH), 1.75–1.83 (m, 2H, 7-CH2, 12-CH2), 1.80 (s, 3H, 4′-CH3), 1.82 (s, 3H, 6′-CH3), 1.95 (dd, J = 12.0, 14.4 Hz, 1H, 15-CH2), 2.14 (dd, J = 6.6, 14.4 Hz, 1H, 15-CH2), 2.18–2.22 (m, 1H, 9-CH), 2.31–2.36 (m, 1H, 11-CH2), 2.68–2.78 (m, 2H, 6-CH2), 5.35 (s, 1H, 2′-CH), 6.44 (d, J = 2.4 Hz, 1H, 4-CH), 6.51 (dd, 1H, J = 2.4, 9.0 Hz, 2-CH), 7.02 (d, 1H, J = 9.0 Hz, 1-CH), 9.03 (br.s, 1H), 9.37 (br.s, 1H). 13C NMR (150 MHz, DMSO-d6), δ: 15.0 (18-CH3), 20.2 (4′-CH3, 6′-CH3), 25.8 (11-CH2), 26.7 (7-CH2), 28.9 (15-CH2, 6-CH2), 33.6 (12-CH2), 36.9 (8-CH), 43.7 (9-CH), 47.9 (13-C), 53.1 (14-CH), 67.8 (2′-CH), 112.7 (2-CH), 115.0 (4-CH), 125.6 (1-CH), 130.2 (10-C), 135.4 (17-C), 137.0 (5-C), 138.6 (16-C), 151.6 (4′-C, 6′-C), 155.0 (3-C). IR (KBr), cm−1: 3198 (NH), 2929, 2857 (CH), 1703, 1611 (C = C, C = N), 1499, 1456, 1435, 1378, 1287, 1248. HRMS (ESI) for C23H29ClN3O ([M+H]+): calcd 398.1994, found 398.1995.
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study
| 100.0 |
3β-Acetoxy-17-chloro-16-formylandrosta-5,16-diene 2b (100 mg, 0.26 mmol) was added to a suspension of acetamidine hydrochloride (125 mg, 1.3 mmol) and potassium carbonate (220 mg, 1.6 mmol) in DMF (4 mL). The mixture was stirred at 80°C for 8 h until the complete conversion of the intermediates (TLC monitoring). The resulting mixture was cooled to room temperature and diluted with water (30 mL). The precipitate that formed was filtered, washed with water (5 mL), and dried. The crude product was purified by column chromatography using chloroform/MeOH, 10:1, to obtain colorless solid (45 mg, 30% yield). Rf 0.45 (petroleum ether:EtOAc, 1:2; visualized by UV light at 254 nm); m.p. 158–160°C. 1H NMR (600 MHz, DMSO-d6), δ: 0.84 (s, 3H, 18-CH3), 1.01 (s, 3H, 19-CH3), 1.02–1.11 (m, 2H, 1-CH2, 9-CH), 1.32 (dt, J = 4.2, 12.6 Hz, 1H, 12-CH2), 1.36–1.41 (m, 1H, 14-CH), 1.43–1.49 (m, 1H, 11-CH2), 1.51–1.65 (m, 4H, 2-CH2, 7-CH2, 8-CH, 11-CH2), 1.69–1.73 (m, 1H, 12-CH2), 1.76–1.84 (m, 2H, 1-CH2, 2-CH2), 1.79 (s, 3H, 4′-CH3), 1.81 (s, 3H, 6′-CH3), 1.88 (dd, J = 14.4, 15.0 Hz, 1H, 15-CH2), 1.92–1.97 (m, 1H, 7-CH2), 1.98 (s, 3H, CH3CO), 2.07 (dd, 1H, J = 6.6, 15.0 Hz, 15-CH2), 2.26–2.31 (m, 2H, 4-CH2), 4.42–4.48 (m, 1H, 3-CH), 5.32 (s, 1H, 2′-CH), 5.34–5.36 (m,1H, 6-CH), 9.40 (br.s, 1H, NH). 13C NMR (125 MHz, DMSO-d6), δ: 14.8 (18-CH3), 18.8 (19-CH3), 19.9 (11-CH2), 20.3 (4′-CH3, 6′-CH3), 20.9 (CH3COO), 27.3 (2-CH2), 29.2 (15-CH2), 29.9 (8-CH), 30.3 (7-CH2), 33.4 (12-CH2), 36.3 (1-CH2), 36.3 (10-C), 37.6 (4-CH2), 47.5 (13-C), 49.8 (9-CH), 53.6 (14-CH), 67.4 (2′-CH), 73.1 (3-CH), 121.8 (6-CH), 135.4 (17-C), 138.5 (16-C), 139.8 (5-C), 152.0 (4′-C, 6′-C), 169.7 (CO). IR (KBr), cm−1: 3183 (NH), 2945, 2857 (CH), 1735 (COO),1704, 1629 (C = C, C = N). HRMS (ESI) for C26H37ClN3O3 ([M+H]+): calcd 458.2569, found 458.2558.
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study
| 100.0 |
3-Chloro-2-formyl-17β-formyloxy-5α-androstane 2c (120 mg, 0.34 mmol) was added to a suspension of acetamidine hydrochloride (160 mg, 1.69 mmol) and potassium carbonate (280 mg, 2.0 mmol) in DMF (5 mL). The mixture was stirred at 60–65°C for 6 h until the complete conversion of the intermediates (TLC monitoring). The resulting mixture was cooled to room temperature and the solvent was removed under reduced pressure. The product was purified by column chromatography using chloroform/MeOH, 6:1, to obtain colorless solid (41 mg, 29% yield). Rf 0.30 (CHCl3:MeOH, 5:0.3; visualized by UV light at 254 nm); m.p. 210–212°C. 1H NMR (600 MHz, DMSO-d6), δ: 0.62 (s, 3H, 18-CH3), 0.68 (s, 3H, 19-CH3), 0.65–0.71 (m, 1H, 9-CH), 0.78–0.88 (m, 2H, 7-CH2, 14-CH), 0.90–0.96 (m, 1H, 12-CH2), 1.11–1.18 (m, 2H, 6-CH2, 15-CH2), 1.22–1.39 (m, 5H, 8-CH, 11-CH2, 16-CH2), 1.40–1.50 (m, 3H, 5-CH, 6-CH2, 15-CH2), 1.60 (d, J = 12.0 Hz, 1H, 7-CH2), 1.64 (d, J = 16.8 Hz, 1H, 1-CH2), 1.71 (d, J = 12.6 Hz, 1H, 12-CH2), 1.80 (s, 3H, 4′-CH3), 1.82 (s, 3H, 6′-CH3), 2.00–2.05 (m, 1H, 4-CH2), 2.06 (d, J = 16.8 Hz, 1H, 1-CH2), 2.14–2.19 (m, 1H, 4-CH2), 3.39–3.43 (m, 1H, 17-CH), 4.40 (br.s, 1H), 5.48 (s, 1H, 2′-CH), 9.50 (br.s, 1H). 13C NMR (150 MHz, DMSO-d6), δ: 11.1 (18-CH3), 11.6 (19-CH3), 20.3 (4′-CH3, 6′-CH3), 20.4 (11-CH2), 23.0 (15-CH2), 27.2 (6-CH2), 29.8 (16-CH2), 30.6 (7-CH2), 34.1 (10-C), 35.0 (8-CH), 36.4 (12-CH2), 38.1 (4-CH2), 39.1 (1-CH2), 42.4 (13-C), 42.5 (5-CH), 50.4 (14-CH), 53.1 (9-CH), 70.7 (2′-CH), 80.0 (17-CH), 123.0 (3-C), 134.0 (2-C), 152.2 (4′-C, 6′-C). IR (KBr), cm−1: 3205 (NH), 2926, 2872 (CH), 1708, 1665 (C = C, C = N), 1498, 1469, 1444, 1380, 1380, 1338, 1250. HRMS (ESI) for C24H37ClN3O ([M+H]+): calcd 418.2620, found 418.2610.
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| 100.0 |
The MCF-7 and MDA-MB231 human breast cancer cell lines and the PC3 and 22Rv1 prostate cancer cell lines were purchased from the ATCC collection. Cells were cultured in standard high glucose DMEM medium (Hyclone) supplemented with 10% fetal calf serum (FCS) (HyClone) and 0.1 mg/ml sodium pyruvate (Santa Cruz) at 37°C, 5% CO2 and 80–85% humidity (NuAir CO2 incubator). The cell growth was evaluated by the modified MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) (Applichem) test (Iselt et al., 1989) as described in Volkova et al. (2016). Briefly, the cells were seeded at a density of 2.5 × 104 cells per well in 24-well plates (Corning) in 900 μL of the medium. The tested compounds were dissolved in DMSO (Applichem) to 10 mM before experiments and then were diluted in the medium to the required concentrations. Compounds with different concentrations in 100 μL of the medium were added 24 h after the seeding, and the cells were grown for 72 h. After incubation with the compounds, the medium was removed, and the MTT reagent dissolved in the medium was added to the final concentration of 0.2 mg/mL to each well and incubated for 3 h. Then the cell supernatants were removed and the MTT formazan purple crystals were dissolved in 100% DMSO (350 μL per well). Plates were gently shaken and the absorbance was measured at 571 nm with a MultiScan reader (ThermoFisher). The viability of the cells was assessed after subtraction of the blank value (the absorbance in the well w/o cells) from all wells. Dose-response curves were analyzed by regression analysis using sigmoidal curves (Log(concentration) vs. normalized absorbance). The half maximal inhibitory concentrations (IC50) were determined with GraphPad Prism.
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study
| 100.0 |
To determine the transcriptional activity of the estrogen receptor α (ERα), MCF-7 cells were transfected with the plasmids containing luciferase reporter gene under the control of the promoter containing estrogen responsive elements. Assay was performed in steroid-free conditions (phenol red-free DMEM medium supplemented with 10% DCC serum). The reporter plasmid ERE-TK-LUC used in this work was kindly provided by Reid et al. (2003). The transfection was carried out for 24 h at 37°C using Metafectene (Biontex Laboratories). To control the efficiency and potential toxicity of the transfection, the cells were co-transfected with the β-galactosidase plasmid. The tested compounds were added to phenol red-free DMEM medium supplemented with 10% DCC serum. The luciferase activity was measured according to a standard protocol (Promega) using a Infinite M200 Pro luminometer (Tecan). The β-galactosidase activity was analyzed using a substrate, p-nitrophenyl β-D-galactopyranoside (ONPG). Briefly, cell lysates were mixed with 0.1 M phosphate buffer (pH 7.5) containing 1.0 mM MgCl2, 3.3 mM ONPG and 53 mM β-mercaptoethanol. After incubation for 1 h at 37°C, the absorbances at 405 nm were measured on the MultiScan FC reader (ThermoFisher). The luciferase activity was calculated in arbitrary units evaluated as the ratio of the luciferase activity to the galactosidase activity.
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study
| 100.0 |
The cells were removed from the dishes with 1.2 ml of phosphate buffer, twice washed, and incubated for 10 min on ice in the modified lysis buffer containing 50 mM Tris-HCl, pH 7.5, 0.5% Igepal CA-630, 150 mM NaCl, 1 mM EDTA, 1 mM DTT, 1 mM PMSF, 0.1 mM sodium orthovanadate and aprotinin, leupeptin, pepstatin (1 μg/mL each) as described earlier (Scherbakov et al., 2006). The protein content was determined by the Bradford method.
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study
| 99.94 |
Cell lysates (40 μg protein) were separated in 10% SDS-PAGE under reducing conditions, transferred to a nitrocellulose membrane (SantaCruz), and processed according to the standard protocol. To prevent nonspecific absorption, the membranes were treated with 5% nonfat milk solution in TBS buffer (100 mM Tris, 150 mM NaCl, pH 7.5) with 0.1% Tween-20 and then incubated with primary antibodies overnight at 4°C.
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study
| 99.9 |
Primary antibodies to ERα were purchased from Sigma-Aldrich (Merck); the antibodies against α-tubulin (Cell Signaling Technology) were added to standardize loading. Goat anti-rabbit IgGs (Jackson ImmunoResearch) conjugated to horseradish peroxidase were used as secondary antibodies. Signals were detected using the ECL reagent as described in Mruk and Cheng (2011) and an ImageQuant LAS4000 system (GE HealthCare). ImageJ software (NIH) was used for densitometry.
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other
| 99.5 |
Each biology experiment was repeated three times. Statistical analysis was performed using Microsoft Excel and GraphPad Prism. Results were expressed as mean ± S.D. (standard deviation value). Student's t-test was used to evaluate the significance of differences in comparisons. P-value of < 0.05 was considered statistically significant.
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other
| 98.3 |
In silico docking was performed using Autodock Vina (Trott and Olson, 2010) run through PyRx interface to manage the workflow and PyMol to visualize the results. Ligands were prepared by generating the energy-minimized 3D structures using ChemBioDraw3D followed by processing with Autodock Tools 1.5.4 to assign Gasteiger charges, merge nonpolar hydrogens, and set torsional bonds. Initial docking runs were performed within a 25–30 Å cubic search space surrounding the binding pocket, with solutions found using an exhaustiveness of 8, and output modes ranked according to binding affinity (BA). For a detailed comparison, multiple runs with a reduced search space were run with an increased exhaustiveness of 16 and 32. The Autodock Vina produced ligand poses with the best fit and strongest BA (global minima) using a stochastic algorithm to explore surfaces/pockets of the rigid macromolecule, through an iterative series of local optimizations evaluating both intermolecular (hydrophobic interactions, repulsions, hydrogen bonding, etc.) and intramolecular (torsion, rotational torque) factors. SAR insights are greatly aided by molecular docking analysis but must be taken as putative due to the rigid modeling of the protein target and the potential for conformational bias (Bissantz et al., 2010).
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study
| 100.0 |
Our interest in the preparation of structurally diverse heterosteroids lead to a need for a facile flexible strategy, in which a common intermediate can be used in a conjunctive fashion to form an array of N-heterocycles attached or fused to the steroid core. Hence, we turned to β-chlorovinyl aldehydes, which are readily available by the Vilsmeier–Haack reaction (Tasneem, 2003) and proved to be highly reactive ambident electrophiles (Bera et al., 2008; Bezboruah et al., 2013; Brockmeyer et al., 2014; Kroger et al., 2015). Recently, we have reported the synthesis of steroidal pyridazines (Komkov et al., 2015; Volkova et al., 2016), thiadiazoles (Zavarzin et al., 2013), and 4,5-disubstituted pyrimidines (Komendantova et al., 2017) via condensation of β-chlorovinyl aldehydes with bis-nucleophiles such as oxamic acid thiohydrazides and amidines. Based on these results, we accomplished the efficient synthesis of heterosteroids possessing a six-membered N-heterocycle attached or fused to the A/D ring of the steroid core starting from readily available materials. Thus, the synthesis of derivatives of the androstene and estrane series containing A-/D-ring annulated pyrimidine (Schemes 1, 3a–f) or linked dihydrotriazine (Schemes 1, 4a–c) moieties was accomplished starting from natural hormones 1a–c (estrone, dihydrotestosterone, and dehydroepiandrosterone) by the general two-step sequence involving: (1) the Vilsmeier–Haack reaction giving steroidal β-chlorovinyl aldehydes 2a–c, (2) the condensation of the former with amidines (Scheme 1).
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study
| 99.94 |
Synthesis of A-/D-ring functionalized azasteroids of the androstene and estrane series. Steroidal β-chlorovinyl aldehydes 2 as ambident elecrophiles easily undergo cyclizations with bis-nucleophilic guanidine and acetimidamide under mild reaction conditions (2 equiv excess, under reflux in methanol with potassium carbonate) providing A-/D-fused steroidal pyrimidines 3a–f in 30–89% yields. The reaction of acetimidamide with 2 equiv excess of potassium carbonate in DMF produced heterosteroids 4a–c containing the dihydrotriazine substituent at the 16-C or 2-C position of the steroid core in 29–32% yields.
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study
| 100.0 |
The heterocyclization pattern was directed to dihydrotriazines by increasing amidine excess up to 4–5 equivs. The dihydrotriazine ring was constructed from two acetimidamide molecules and one steroid molecule via the nucleophilic attack of two amidine molecules on the formyl group. The reaction of acetimidamide with potassium carbonate in DMF produced heterosteroids 4a–c containing the dihydrotriazine substituent at the 16-C or 2-C position of the steroid core in 29–32% yields. The structural assignments for all compounds 3a–f and 4a–c were confirmed by 2D NMR (1H-1H COSY, 13C-1H HMBC, and 13C-1H HSQC, see Supplementary Material) techniques and HRMS.
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study
| 100.0 |
The antiproliferative activity of all the synthesized compounds was evaluated against the human estrogen-responsive MCF-7 breast cancer cell line and ERα-negative MDA-MB231 cells using the MTT assay (Figures S1, S2). Cisplatin, a standard chemotherapy drug, was used as the reference compound. All compounds were also tested for cytotoxicity toward prostate cancer cells (Figures S3, S4). AR-negative PC3 cells and AR-positive 22Rv1 cells were used in this assay. The corresponding inhibitory concentrations IC50 (IC50 is the half maximal inhibitory concentration) are given in Table 1.
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study
| 100.0 |
Most of the tested heterosteroids showed remarkable anticancer activity against ERα-positive MCF-7 cancer cells. Estranes 3a,b containing the D-ring-fused pyrimidine moiety proved to be inactive, while their androstene analogs 3c–e were active with the IC50 values in the range of 12.0–21.6 μM. It is remarkable that the IC50 value for compound 3d containing the 3-OH group is higher than that for compound 3c possessing the 3-OAc protected group. The solubility of steroidal A-ring annulated pyrimidine 3f in DMSO is too low to perform the MTT assay.
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study
| 100.0 |
Steroidal dihydrotriazines 4a–c proved to be more active against MCF-7 cancer cells compared to steroidal fused pyrimidines. Androstene derivative 4c bearing the dihydrotriazine moiety at C-2 had the IC50 value of 14.8 μM, while the IC50 value for compound 4b modified at 16-C reached 11.2 μM. The 16-C dihydrotriazine-modified estrane 4a was shown to be the most active derivative. Moreover, only steroidal dihydrotriazines 4a, 4b, and 4c were active against ERα-negative MDA-MB231 cells; their IC50 values vary in the range of 12.2–19.1 μM.
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study
| 100.0 |
All compounds were tested against 22Rv1 and PC3 prostate cancer cells. Among them, compounds 4a, 4b, and 4c displayed antiproliferative activity. Estrane derivative 4a inhibited the growth of PC3 and 22Rv1 prostate cancer cells with IC50 of 13.5 and 11.7 μM, respectively, while androstene derivative 4b displayed cytotoxicity comparable to that of cisplatin in hormone-dependent 22Rv1 prostate cancer cells. Androstene derivative 4c was less active against prostate cancer cells than compounds 4a and 4b, and revealed the IC50 value about 20 μM (Table 1).
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study
| 100.0 |
Considering indicated antiproliferative activity of compounds in ERα-positive MCF-7 breast cancer cells, ERα was analyzed as a possible target for these synthetic steroids. For this purpose the luciferase reporter assay was used to determine ERα activity in MCF-7 cells. The ERα-mediated reporter constructs were provided to express luciferase under the control of the promoter containing estrogen responsive elements (ERE-TK-LUC). Thus, ERα activity was correlated to luciferase activity measured in treated or control cells.
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study
| 100.0 |
As can be seen in Figure 2A, estranes 3a,b containing the D-ring-fused pyrimidine moiety did not inhibit ERα activity at 10 μM concentration. Moreover, these 3a and 3b stimulated ERα activity at low (10 nM) concentration acting as partial receptor agonists (Table S1). Androstene derivative 4b proved to be inactive as ERα inhibitor. Compound 4c showed weak inhibitory activity, while steroids 3c, 3d, 3e, and 4a highly inhibited E2-mediated ERα activity at 10 μM concentration. These compounds showed no ERα agonist activity in the luciferase reporter assay (Table S1).
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study
| 100.0 |
Evaluation of ERα activity and expression. (A) MCF-7 human breast cancer cells were transfected with the ERE-TK-LUC plasmid containing the luciferase reporter gene under the estrogen responsive element (ERE) and co-transfected with β-galactosidase plasmid. The media was removed 24 h after transfection and 10 nM of 17β-estradiol was used to induce ERα activity. Synthesized compounds at 10 μM concentration or vehicle control (cont) was added to phenol-free DMEM supplemented with 10% steroid-free serum (Hyclone). The luciferase and β-galactosidase activities were determined in 24 h. The luciferase activity was calculated in relative units evaluated as the ratio of the luciferase activity to the galactosidase activity. *P < 0.05 vs. E2-treated cells. (B) MCF-7 cells were treated with or without tamoxifen or compound 4a. The cell samples were subjected to Western blot analysis of ERα and α-tubulin as the loading control; Densitometry for ERα/α-tubulin ratio was carried out using ImageJ software with the densitometry protocol provided by The University of Queensland. *P < 0.05 vs. control cells, #P < 0.05 vs. tamoxifen-treated and control cells.
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study
| 100.0 |
Taking into consideration the two-fold gain in cytotoxicity of compound 4a against ERα-positive breast cancer cells vs. ERα-negative cells and its high activity as ERα inhibitor we performed immunoblotting of ERα in MCF-7 cells. The data obtained by immunoblotting confirmed that compound 4a exerted ERα inhibitory activity. The incubation of MCF-7 cells with compound 4a resulted in the partial suppression of ERα expression, as can be seen in Figure 2B. Tamoxifen was used as the standard reference drug and its application resulted in an increase in ERα expression, which may be attributed to tamoxifen-induced stabilization of inactive ERα in the cell cytoplasm as discussed in Wijayaratne and McDonnell (2001). As compared with tamoxifen, compound 4a was found to be active as the partial ERα downregulator (Figure 2B).
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study
| 100.0 |
In order to gain insight into the structural basis of the observed ERα inhibitory effects of compound 4a, we performed in silico docking analysis using Autodock Vina. Low-energy binding poses were generated by evaluating the combined energetic contributions of torsion, steric repulsion, hydrogen bonding, and hydrophobic interactions between the ligand and the protein binding pocket. Using the crystal structures of ERα in complexes with estradiol [PDB: 1GWR (Warnmark et al., 2002), Figure 3A] and the weak agonists 17α-modified estradiol analogs TFMPV-E2 [PDB: 2P15 (Nettles et al., 2007), Figure 3B] and EEu [PDB: 2YAT (Li et al., 2011), Figure 3C], we found that the steroidal moiety of compound 4a in the docked poses differs from the estradiol moieties of the original ligands (Table 1, Figures 3A–C,E).
|
study
| 100.0 |
Crystal structures of agonist-bound ERα 1GWR (A), 2P15 (B), 2YAT (C) and antagonist-bound 3ERT (D) used in the docking analysis of ER modulator 4a. Their original ligands (E) are in purple, modulator 4a in orange, and helix 12 in red. Only the highest ranked poses with the strongest BA were selected: (A) +0.1 kcal/mol, (B) −8.1 kcal/mol, (C) −8.9 kcal/mol, (D) −8.9 kcal/mol.
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study
| 100.0 |
Compound 4a was found to be too big for the accommodation in the estradiol-binding pocket of ERa in the structure 1GWR. Meanwhile, the ligands TFMPV-E2 (PDB: 2P15) and EEu (PDB: 2YAT) are known to modulate the dynamics of the ERα helix 12 (shown in red, Figures 3A–D), resulting in an increase in the ligand-binding pocket surface of ER without changing the shape of the ligand-binding domain of ER due to the presence of bulky substituents at the 17α position of the estradiol core (Eignerova et al., 2010; Gryder et al., 2013). Compound 4a bearing the bulky dihydrotriazine ring at the 16-position is docked against the ERα crystal structures 2P15 and 2YAT with reasonable binding affinity (−8.1 and −8.9 kcal/mol, respectively). However, the major binding modes of compound 4a for 2P15 and 2YAT are as follows: the A-ring phenolic hydroxyl group points away the binding pocket of ER and the dihydrotriazine moiety points inward the binding pocket. The critical hydrogen-bonding interactions between the steroid estradiol, TFMPV-E2 and the A-ring phenolic hydroxyls of EEu with Arg-394/Glu-353 were not found for compound 4a (Figures 3A–C), which can be attributed to steric hindrance caused by the bulky substituent at the 16-position and spatial aliasing of the estrane core due to the addition of the C16 = C17 double bond.
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study
| 100.0 |
Alternatively, compound 4a docked against the crystal structure of ERα in complex with the ER antagonist 4-hydroxytamoxifen (PDB: 3ERT, Figures 3D,E) is able to fill the hydrophobic space and latch onto Arg-394/Glu-353. Tamoxifen prevents the helix 12 from closing properly upon the binding pocket of the ligand-binding domain, while compound 4a can extend the steroidal moiety through the opening left by the displaced helix 12. The D ring of steroid 4a modified at the 16-and 17-positions can go inside the pocket potentially stabilized by polar interactions with TRP-383, Phe-404, Leu-387, Met-388, and Leu-391. These docked structures may reflect the most probable mode of binding. Although the direct comparisons are speculative, our docking outputs are supported by our observation with compound 4a in ER luciferase reporter assays. The antagonist effect of compound 4a is interesting, because D-ring modifications of estradiol commonly result in weak ER agonists (Yang et al., 2000; Kreis et al., 2001).
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study
| 100.0 |
The initial synthesis of annulated steroidal pyrimidines by the groups of Clinton (Ackerman et al., 1964) and Ruggieri (De Ruggieri and Gandolfi, 1965; De Ruggieri et al., 1965, 1966a,b) dates back to the mid-1960s. Their synthetic approach was based on condensation of guanidines with activated α,β-unsaturated ketones, primarily β-enol ethers, and have become widely applied in chemistry of steroids due to a great diversity of obtainable products (Romo et al., 1968; Bajwa and Sykes, 1978; Hajos and Snatzke, 1989; Mallamo et al., 1992; Hasan et al., 1995; Forgo and Vincze, 2002). Unfortunately, this method suffers from drawbacks, such as harsh reaction conditions, moderate yields and high labor content/cost of preparing starting materials. Therefore, over the years a considerable effort has been directed toward the development of alternative methods for steroidal pyrimidines synthesis. Improved solid phase protocol of β-enol ethers heterocyclization was elaborated toward synthesis of steroidal A-ring fused pyrimidines (Barthakur et al., 2009). Although there is an example of condensation using β-enamino ketones (Xu et al., 2012). Boruah group have developed a range of methods, among which are three-component condensation of steroidal ketones with aromatic aldehydes and amidine derivatives in presence of potassium tert-butoxide (Saikia et al., 2014), Pd(OAc)2-catalyzed hetrocyclization of steroidal β-halo-α,β-unsaturated aldehydes with amidines (Gogoi et al., 2013) and SmCl3-catalyzed condensation of β-formyl enamide with urea under microwave irradiation (Barthakur et al., 2007). Baran group reported synthesis of 4,5-disubstituted pyrimidines from steroidal ketones and formamidine acetate (Baran et al., 2006). Here we have achieved high-yielding syntheses of novel A- and D-rings annulated steroidal pyrimidines via developed by us metal-free condensations of β-chlorovinyl aldehydes with amidines (Komendantova et al., 2017). These reactions are efficiently occur under mild conditions, with the added advantage that heterocyclization pattern can be easily switch to steroidal dihydrotriazines, previously unexplored class of heterosteroids.
|
study
| 99.7 |
Steroidal pyrimidines may be considered as promising compounds for the design of novel antitumor drugs. This line of research has been extensively developed in recent years. In 2017, Ke et al. designed novel steroidal[17,16-d]pyrimidines derived from dehydroepiandrosterone and evaluated their in vitro inhibitory activity against liver and gastric cancer cells (Ke et al., 2017). Briefly, 16 steroidal[17,16-d]pyrimidines derived from dehydroepiandrosterone were designed and synthesized via a sequence transformation, and their activities were assessed by MTT. Ke et al. found that some of these heterocyclic steroidal[17,16-d]pyrimidines showed antiproliferative activities against HepG2, Huh-7, and SGC-7901 cell lines compared to the reference 5-fluorouracil. Eight novel compounds synthesized by Ke et al. exhibited excellent inhibitory activities against all three cell lines with 70–82% growth inhibition at the concentration of 40 μg/mL. Thus, steroidal[17,16-d]pyrimidines might be used as promising compounds for discovery of novel anticancer drugs for treatment of liver and gastric cancers.
|
review
| 99.75 |
Other promising steroidal pyrimidines were discussed by Ali et al. (2017). The antitumor activity of the B-ring fused steroidal pyrimidines was tested in vitro against the MDA-MB231, HeLa, and HepG2 cancer cell lines and the non-cancer normal cell line PBMCs (peripheral blood mononuclear cells) by the standard MTT assay. The compounds showed moderate to good activity and proved to be nontoxic to normal PBM cells. One of the synthesized compounds was found to be active against all three cancer cell lines but more specific against the MDA-MB231 cells with IC50 of about 9 μM, which is similar to our data on the activity of the compounds against breast cancer cells. Finally, the authors discussed the ability of steroidal compounds to interact with the protein HSA involved in drug delivery.
|
review
| 67.75 |
Metastatic bone tumors occur at particularly high rates in cancers of the breast, prostate, and lung, accounting more than 70% of all patients. Treatment of skeletal metastasis and development of new specific “blockers” of bone resorption are relevant. Pyrimidine-fused betulinic acid may be considered as promising compounds for the design of novel inhibitors of osteoclast differentiation and bone resorption. Jun Xu et coworkers synthesized over 20 heterocyclic ring-fused betulinic acid derivatives and evaluated their inhibition on RANKL-induced osteoclast formation in preosteoclast RAW264.7 cells (Xu et al., 2012). Some compounds exhibited potent inhibitory activity on RANKL-induced osteoclast formation by TRAP assay.
|
study
| 92.5 |
The elucidation of the mechanism of action of compounds in target cells and understanding of their common metabolism in human body are of interest. The structural optimization will be performed and the molecular mechanism of novel steroidal pyrimidines will be investigated in due course. On the other hand, the activity of steroidal dihydrotriazines against cancer cells is less well known described, and our study is very relevant.
|
study
| 99.94 |
Here, we describe novel series of steroidal anticancer agents. In summary, this study demonstrates that the cyclization of steroidal β-chlorovinyl aldehydes with bis-nucleophilic amidines provides an easy approach to various novel heterosteroids. Natural hormones 1a–c (estrone, 3β-acetoxyandrostene, 3-keto-17β-hydroxyandrostane) were transformed into the corresponding A- and D-modified steroidal pyrimidines and dihydrotriazines in moderate to high yields (29–89%) using a two-step sequence involving the Vilsmeier–Haack reaction and condensation with amidines, such as guanidine and acetimidamide. The new compounds showed remarkable cytotoxic activity against breast and prostate cancer cells. Furthermore, lead compounds demonstrated selectivity toward ERα in MCF-7 breast cancer cells. Compound 4a inhibits 50% of ERα activity at its cytotoxic concentration. Using immunoblotting, partial ERα downregulation was observed in compound 4a-treated MCF-7 cells. Docking approaches confirmed the ability of compound 4a to bind to ERα. Thus, compound 4a may be considered as a candidate for future anticancer drug design, in particular, for ERα-positive breast cancers.
|
study
| 99.94 |
Despite a limited number of compounds in series, it provides significant novel insight into the structure–activity relationship of heterosteroids as anticancer agents. Biological studies show that annulation of androst-5-ene core with pyrimidine is efficient for development of novel selective compounds for treatment of hormone-dependent breast cancer. Moreover, installation of dihydrotriazine pendant at A- and D-rings of estrane and androst-5-ene cores results in strong antiproliferative activities against breast and prostate cancer cells comparable with cisplatine. The two-fold gain in cytotoxicity of 16-C dihydrotriazine-modified estrane against ERα-positive breast cancer cells vs. ERα-negative cells and its high activity as ERα inhibitor were shown while similar androstene derivative was less selective.
|
study
| 99.94 |
AS carried out the immunoblotting, performed the statistical analysis and the transient transfection, drafted and prepared the manuscript for submission, worked with cell cultures; AVK synthesized steroidal compounds; ASK synthesized steroidal compounds, prepared the supporting information; MY worked with cell cultures; OA performed the reporter analysis, worked with plasmids; VS managed the project; IZ managed the project; AH managed the project; YV wrote the manuscript, conceived of the study, in silico analysis, and managed the project; All authors read and approved the final manuscript.
|
other
| 99.94 |
Drug reaction with eosinophilia and systemic symptoms (DRESS) syndrome is a rare and potentially life-threatening idiosyncratic drug reaction. It presents with extensive rash, fever, lymphadenopathy, hematologic abnormalities (eosinophilia and/or atypical lymphocytosis) and internal organ involvement. The incidence of this syndrome ranges from 1/1,000 to 1/10,000 after drug exposure. Adults are more commonly affected than children . Approximately 50 drugs (sulfa derivates, antidepressants, nonsteroidal anti-inflammatory drugs) may induce DRESS syndrome, but it is most commonly induced by anticonvulsants (phenytoin, phenobarbital, carbamazepine) and antibiotics . It has been described in association with more than 50 drugs. To the best of our knowledge neither cefotaxime nor clindamycin have been previously reported to cause DRESS syndrome in children. Clindamycin was reported only in adults as an agent causing DRESS syndrome in the literature . We present a child with DRESS syndrome presenting 21 days after starting cefotaxime and clindamycin treatment.
|
clinical case
| 99.94 |
A 6-year-old boy was diagnosed with the left lower lobe pneumonia and pleural effusion and was put on parenteral cefotaxime and clindamycin treatment. He then improved clinically and was discharged 7 days later with oral amoxicillin clavulanate treatment. After four days, however, he was readmitted to the hospital with fever and cough. A chest X-ray revealed left lower lobe pneumonia and pleural effusion. As a case of complicated pneumonia was considered, we started parenteral cefotaxime and clindamycin treatment, 10 days after which his clinical and radiological findings were improved. At the 12th of day of hospitalization, however, he developed at 39°C, which we considered as a side effect of antibiotics. Therefore, antibiotic treatment was stopped. At the same day, he also developed a generalized maculopapular erythematous rash, which was considered an allergic reaction secondary to antibiotics (Figure 1).
|
clinical case
| 100.0 |
Despite the administration of antihistaminic drugs, his clinical status quickly deteriorated with generalized edema, lymphadenopathies and hepatosplenomegaly. Laboratory tests showed a white blood cell count of 4,300/mm3, a lymphocyte count of 1,300/μl, a hemoglobin level of 11.2 gr/dl, a platelet count of 120,000/μl, an eosinophil ratio of 10% on peripheral blood smear, a C-reactive protein level of 20 mg/dl, a procalcitonin level of 23.94 ng/ml and an erythrocyte sedimentation rate of 48 mm/h. Anti-nuclear antibody, anti-double strain DNA, the serologic tests for Epstein Bar virus, herpes simplex virus, parvovirus, mycoplasma, toxoplasmosis, rubella, cytomegalovirus were all found negative. Bone marrow aspiration was consistent with an autoimmune reaction. An echocardiographic examination was found normal. A Thoracic computerized tomography revealed multiple enlarged axillary, supraclavicular and anterior mediastinal lymph nodes. As the patient met 8 out of 9 RegiSCAR criteria for the diagnosis of DRESS, pulse methyl prednisolone (30 mg/kg/day) was administered for three days followed by 2mg/kg/day. On the 2nd day fever resolved and cutaneous rash and edema improved. He was discharged ten days after the appearance of eruptions. The patient’s father gave consent for publication of his son’s pictures.
|
clinical case
| 100.0 |
DRESS syndrome was first defined in 1996 by Bocquet et al . It is characterized by fever, cutaneous eruption, internal organ involvement and hematologic abnormalities within 1-8 weeks after the exposure of the suspected drug . Anticonvulsants such as carbamazepine, lamotrigine, phenobarbital, phenytoin and allopurinol are the most common causes of DRESS syndrome . In a series of 172 cases of DRESS syndrome carbamazepine accounted for the greatest percentage of cases (27%), followed by allopurinol (11%), lamotrigine (6%), phenobarbital (6%), sulfasalazine (6%) .
|
review
| 99.8 |
The pathogenesis of DRESS syndrome has not been elucidated. Different mechanisms have been proposed, including detoxification defects leading to reactive metabolite formation and subsequent immunological reactions, slow acetylation and reactivation of Human Herpes Virus (HHV-6-7) and EBV . Severe DRESS syndrome has also been reported following infection with or reactivation of HHV-6 . CMV and paramyxovirus have been reported as the other viral agents associated with DRESS syndrome. Although it is thought that there is a genetic predisposition to adverse reactions, no genetic factor responsible for the syndrome has been identified yet .
|
review
| 99.9 |
Three different sets of criteria are used to diagnose of DRESS syndrome: RegiSCAR criteria, Bocquet’s criteria and Japanese consensus group to diagnose DIHS . RegiSCAR criteria include at least 3 of the following 7 characteristics: 1) skin eruption; 2) fever > 38°C; 3) lymphadenopathy involving at least 2 sites; 4) involvement of at least 1 internal organ; 5) lymphocytosis (> 4000/mm3) or lymphocytopenia (< 1500/mm3); 6) blood eosinophilia (> %10 or 700/mm3) and 7) thrombocytopenia (< 120.000/mm3 ) . Three of the four main criteria (fever > 38°C, lymphadenopathy at least 2 sites, involvement of at least 1 internal organ and blood abnormalities) are required for the diagnosis of DRESS syndrome. Additional criteria are hospitalization and drug induced reaction [6, 8, 9]. Our patient met 6 of 7 RegiSCAR criteria and he also had two additional criterias ( hospitalization and drug induced reaction).
|
clinical case
| 99.9 |
The reported mortality of the syndrome ranges between 10% and 40%. Liver damage secondary to eosinophilic infiltration is the most important cause of mortality. There are no consensus guidelines for the management of patients with DRESS syndrome. The most important steps for a proper management include the recognition of the syndrome and immediate discontinuation of the offending drug. The French Society of Dermatology recommends the use of systemic corticosteroids (prednisone 1 mg/kg/day) and intravenous immunoglobulin (2 gr/kg) for five days especially in patients with life threatening internal organ involvement, such as in renal or respiratory failure . Pulse methyl prednisolone treatment was also reportedly administered in a pediatric case of DRESS syndrome secondary to anticonvulsant use . Gancyclovir has been suggested in patients with severe signs and the confirmation of a major viral reactivation of HHV-6 .
|
review
| 99.8 |
The diagnosis of the DRESS syndrome should be highly suspected in patients with fever, skin rash, liver involvement, hypereosinophilia and lymphadenopathy that develop during the use of a culprit drug. We report the first case of DRESS syndrome associated with cefotaxime and clindamycin exposure in a 6-year-old boy. Early recognition of the signs of the DRESS syndrome and immediate cessation of the suspected drug are the most important steps for an appropriate management of the syndrome. Pediatricians should be careful about this potential complication associated with these commonly prescribed antibiotics in pediatric practice.
|
clinical case
| 99.9 |
In February 2015, the website on preconception care, ‘gezondzwangerworden.be’, was launched by the Flemish Minister of Welfare, Public Health and Family. The website is edited in Dutch and holds evidence-based information for both women and men planning a pregnancy and health care providers.
|
other
| 99.94 |
Since the MRC Vitamin Study Research Group in 1991 found that a daily dose of 400 μg folic acid reduced the risk of neural tube defects drastically, periconceptional supplementation of folic acid has been recommended internationally to women planning a pregnancy (1). However, the time-frame in which women are advised to take folic acid differs between countries (2). The ‘Superior Health Council’ in Belgium recommends intake one month before pregnancy until pregnancy week 12; nonetheless, a study in 2010 on the practice of periconception folic acid intake in Flanders (the northern, Dutch-speaking part of Belgium) shows that only 36% of women actually pursue this recommendation (3).
|
review
| 99.6 |
Because of the proven effect of folic acid in the prevention of neural tube defects and recommendations of public and health authorities, on the one hand, and the lack of compliance in women planning a pregnancy, on the other hand, a resolution proposal on folic acid intake before conception and during pregnancy was granted by the Flemish Parliament on 16 June 2010 (4).
|
other
| 99.94 |
Subsequently, several stakeholders were involved in a number of meetings to discuss folic acid intake in the periconceptional phase. The public health agency ‘Zorg & Gezondheid’ and the Royal Academy for Medicine of Belgium organized these meetings on the initiative of the Flemish Minister of Welfare, Public Health and Family. After the final meeting on 28 April 2011, a public procurement was released on folic acid intake and preconceptional advice. One of the goals of this procurement was the development of an evidence-based website on preconception care, with a particular focus on folic acid intake. A multidisciplinary team from Ghent University worked on this procurement from April 2013 to July 2014. The website was developed in a systematic and scientific manner, and numerous experts were involved in the content validation.
|
other
| 99.75 |
The content of the website is based on available guidelines on preconception and prenatal care. We searched in PubMed, Cochrane, Web of Science, Google, and Google Scholar using the following keywords: guideline(s), preconception, preconception care, preconception health, prepregnancy, before pregnancy, and before conception. We also visited international and national websites of professional organizations providing guidelines. Few guidelines on preconception care in healthy women were available. Some of the prenatal guidelines included information on the periconceptional phase, and some issues of the prenatal guidelines were relevant for preconception care as well. The guidelines perused were national guidelines from Domus Medica (Association of General Practitioners) (5) and KCE (Federal Centre of Knowledge for Public Health) (6), as well as international guidelines from CDC (www.cdc.gov/ncbddd/folicacid/recommendations.html), NHG (7), AJOG (8), ICSI (9), and NICE (10). The quality of the guidelines was assessed by three authors by means of AGREE II (the Appraisal of Guidelines for Research and Evaluation instrument is recognized as the gold standard for practice guidelines evaluation) (11).
|
review
| 99.9 |
All relevant topics of the selected guidelines were inventoried. The list was reviewed by a multidisciplinary committee of five internal experts (a gynaecologist, a general practitioner, two midwives, and a researcher with expertise on folic acid), and a first selection was made by this internal expert committee (IEC) (n = 5). Within the IEC, there was a consensus to include information on the items summed up in Table 1.
|
study
| 61.03 |
In the selection of topics, it was our goal to focus on preconception care rather than prenatal or antenatal care. Topics considered less relevant for the website on preconception care were: information on breastfeeding, information on salmonella and parvovirus, soil and water pollution, personal hygiene, information on natal and postnatal care, pregnancy-related conditions, breast and abdominal examination, bacterial vaginosis, group B streptococcus, and asymptomatic bacteriuria.
|
other
| 99.8 |
Subsequently, the list of topics was sent to an external multidisciplinary committee of experts (EEC) (n = 16) together with a questionnaire where the experts could indicate if they agreed with the inclusion of information on the topic within the website and where they could mark the level of importance of every topic. Furthermore, the experts provided relevant information. Sixteen external experts were involved in the selection of topics: six general practitioners with expertise in preconception care, four gynaecologists (two academic and two non-academic), three midwives with expertise in preconception care, one pharmacologist, one nutritional expert, and one staff member of the most important Flemish public service for the well-being of children.
|
study
| 99.4 |
As for healthy nutrition, experts indicated that basic information is known by professionals. Information on nutrition advice after bariatric surgery, malabsorption, and vitamin D is needed more for this group. According to the experts, it was necessary to provide information on women at risk of having a child with neural tube defects.
|
other
| 99.8 |
The few experts who did not consider it important to inform the public about the risk of toxoplasmosis reported that—in their opinion—the women were already sufficiently aware of the risk; however, most experts considered it necessary to include information on toxoplasmosis prevention. Information on smoking was considered very important because of the potential health benefit in smoking cessation. Experts advised to refer to smoking cessation guidance on the website. Information on physical exercise was considered important in the prevention of obesity with its adverse effect on fertility and pregnancy outcome. Experts considered that information on the optimal age for childbearing was not always relevant for this website. They argued that when people seek information on preconception care, they cannot change their age. Nonetheless, most experts agreed to include information on maternal age.
|
review
| 99.9 |
In total, another 40 experts were involved in the revision of the information on the website (a detailed list of experts can be found on the bottom of the page ‘gezondzwangerworden.be/over-de-website’). Because of the importance of this process, experts were carefully chosen after consultation with the commissioning party (Zorg & Gezondheid) and the IEC. Thus, a multidisciplinary group was chosen to revise the part of the contents corresponding to their field of expertise. As such, gynaecologists, general practitioners, and midwives with an expertise in preconception care were involved; as were specialists in smoking cessation, obesity, and genetics; nutritional specialists; pharmacists; members of public services on occupational health, sexually transmitted infections, and hygiene; and delegates of health insurance companies, federations of birth defects, and members of the Dutch health council.
|
other
| 99.9 |
All experts revised their part of the draft text thoroughly and formulated suggestions and corrections. The content was modified according to the feedback from the experts. In case of uncertainties, the external experts were consulted. Again, we kept our focus on information that was relevant for couples who were not pregnant yet, as the majority of Flemish women are followed up thoroughly by a health care professional once they are pregnant.
|
other
| 99.9 |
In the draft version, we advised that women should stop alcohol intake when they were pregnant. Because of this remark, other experts were consulted and more literature was reviewed. This resulted in a stricter message, on alcohol use, within the website (avoid alcohol when you want to become pregnant).
|
other
| 99.94 |
Work progress was discussed with the commissioning party (Z&G) on a three-monthly basis. Choices were motivated and adjusted if necessary. In these meetings, it was decided that the website would have two parallel parts: a part for the lay public and a part for professionals; however, everyone should have access to both parts. The style of writing was adjusted to the target audience, and the content differed according to specific information needs.
|
other
| 99.94 |
A pilot study was carried out by the ‘thinking aloud’ method (12) in a group of 30 first degree midwifery students (all female) on the one hand, and 6 people of reproductive age on the other hand (5 female, 1 male). One researcher led the group discussion, and another researcher noted the remarks. Without further information, the participants were asked to look at the front page of the website and talk about their first impression. They were asked what target audience they believed the website was intended for and what idea they had of the purpose of the website. Subsequently, they were asked to formulate a preconception-related question. After some explanation about the development of the website, the participants were asked further to explore the website and to search for the answer to their question on the website. They were asked if the website was complete in their opinion, or if it was too exhaustive, if all information was clear, if the website was user-friendly, and if they thought the website was appealing.
|
study
| 99.94 |
For the participants of the pilot study, the target group was obvious, the main messages and the purpose of the website were clear, information was found swiftly, and the website was easy accessible, appealing, and structured. There were some remarks about the colour use (some participants thought the pink was too female, other participants thought the colour would not discourage men visiting the website). Furthermore, suggestions for the lay-out were made. There were some remarks on grammar, participants found a few inactive links, and the need to explain difficult words in the website was expressed. Participants commented on the ambiguous message about alcohol use (to avoid alcohol when you are thinking of getting pregnant and stop drinking alcohol once you know you are pregnant) and pointed out that symptoms of STIs were not always systematically given. Based on the input from both groups, the website was further refined and improved.
|
other
| 64.9 |
The above-described process resulted in the website, ‘gezondzwangerworden.be’ (in English: ‘getting pregnant in good health’) (Figure 2). The front page holds information on folic acid, alcohol use avoidance, and smoking cessation. There is a direct link to information for the partner and a section where relevant news is updated regularly. Furthermore, there are seven main tab pages: ‘Healthy lifestyle’, ‘Becoming pregnant’, ‘Infections’, ‘Medical information’, ‘Other’, ‘For the partner’, and ‘For the professional’.
|
other
| 99.94 |
The tab page ‘Healthy lifestyle’ holds information on healthy nutrition, vitamins (folic acid and vitamin A), vegetarian and vegan nutrition, weight, hand and food hygiene, oral hygiene, alcohol, smoking, drugs, and leisure activities (physical activity, travelling, hyperthermia). The tab page ‘Becoming pregnant’ holds information on the menstrual cycle, lifestyle factors influencing fertility, and advice for people failing to conceive. On the tab page ‘Infections’, information is included on toxoplasmosis, listeria, rubella, varicella, pertussis, cytomegalovirus, parvovirus (this information was included later), STIs, hepatitis C, and tuberculosis. ‘Medical information’ holds information on personal anamnesis, medication, and vaccination. On the tab page ‘Other’, information is found on the domestic situation (socio-economic issues), occupational situation, cosmetics (hair dye and colourants, deodorants, and other), paint, and radiation. Furthermore, the tab page ‘For the partner’ holds information for both the male partner (lifestyle issues, such as alcohol, smoking, and drugs, hyperthermia, medication, harmful factors such as radiation, and information on STIs and hepatitis C) and information for the female partner (lifestyle issues and STIs). Finally, the tab page ‘For the professional’ has the same structure as the information for the public, but more information on medical issues is included: the information is more thorough and has other accents. For the professionals, the link to the source of information (guideline) is included within the text. The public can also look up the sources by means of a header ‘sources’ at the top of the website. Next to this header, there is a header ‘news’, where updates on the website are maintained and where relevant news on preconception care is found, a header where links to relevant websites are found, and a header ‘contact’ where visitors are encouraged to contact their professional for questions or to fill in a contact form if they want to provide feedback on the website.
|
other
| 99.9 |
The website was launched on 23 February 2015 by the Flemish Minister of Welfare, Public Health and Family, together with a press release. The news was covered in most written media and was mentioned in the television news at noon. The release of the website was announced to professional organizations of a large number of stakeholders (general practitioners, gynaecologists, midwives, health insurance companies).
|
other
| 99.94 |
After one week, the website had attracted 8,850 users (there are approximately 30,000 births in Flanders per year to first-time mothers) of which almost 6,000 users visited the website on the day it was launched (in total we had 11,431 visitors during the first month). Most visitors (46%) were referred by news websites, 36% by direct entry, 10% by social media, and 8% by an organic search on search engines. We attracted users from all regions in Flanders.
|
other
| 99.9 |
The first week, we received 11 contact forms, of which 9 were from professionals. There was some feedback on the content, which was pursued when relevant. Some professionals asked for flyers to promote the website amongst their clients. Two care clients responded that the website was not easy to use on a smartphone or tablet, a problem which has been solved promptly by the webhost. After the first week and until now, only few contact forms were received.
|
other
| 99.94 |
Since this first week, which had much media coverage, and until today (a year after the launch) the website has been reaching a constant average of 100–200 visitors a day (more than 4,000 per month). Three out of four visitors were new, and 85% of the visitors were women. Table 3 shows the age distribution of visitors. People aged 25–34 years represented 62% of the visitors.
|
other
| 99.9 |
At the moment, 70% of visitors reach the website via an organic search on the internet (Table 4). People got to the website by searching on a variety of terms, such as ‘becoming pregnant’, ‘folic acid’, ‘healthy pregnancy’, ‘cycle’, and a combination of food products and ‘pregnancy’; 15% of visitors came via links from other websites. More than 77% of those visitors followed the link on the website of Kind & Gezin (kindengezin.be), which is the most important preventive organ for well-being of children in Flanders (the Flemish counterpart of ONE—l’Office de la Naissance et de l’Enfance). Amongst other things, people look at this website to find information on official childcare initiatives.
|
other
| 99.9 |
During this first year of existence, a number of updates have been carried out. First, contact information of a patient organization for people with an unfulfilled child wish has been added. Second, the information on vitamins and minerals has been updated according to the new nutritional recommendations of the Superior Health Council, and the information on ‘cosmetics’ has been extended due to a new report on anti-transpirants of this council. The advice of the same council to follow strict hygiene measurements in the prevention of CMV was added as well. Subsequently, a health care provider noticed the lack of information on parvovirus. After considering the evidence on this topic, information on the website was completed after review of three experts. Finally, information on the Zika virus was added with referral to an evidence-based and regularly updated website on tropical medicine (www.itg.be).
|
other
| 99.8 |
Google analytics provide a service to evaluate the use of a website to some extent; however, the possibilities are limited. It is, for example, not clear how many and to what extent professionals refer to the website. Nonetheless, google analytics show that a number of visitors found our website by using a link provided on the website of professionals. On the contact form it is mentioned that specific questions can and should be discussed with the health care provider. Until now, the contact form was only used to provide feedback on the website (as is indicated on the website); however, it is not clear to what extent people address preconception care with their health care provider due to information they read on the website. Although the page with information on folic acid is one of the most visited pages of the website, it is currently not clear to what extent folic acid supplementation is (more) successful. It would be of interest to monitor folic acid intake for example in birth registers.
|
other
| 99.9 |
Breast cancer is a widely known disease that mostly affects women around the world. With one of the highest incidence rates of female cancer, the success of treatment really depends on diagnosing the cancer in its earlier stages . Treatments have progressed to have lower secondary effects, but breast cancer surgery is still a reality for most patients. For decades, Mastectomy was prescribed for almost every breast cancer case with a high rate of success for removing the tumor; however, this surgical option comprises the removal of the entire breast and it has a profound impact on the aesthetic appearance and self-confidence of women . With the widespread of screening mammography, the average size of the detected tumors has decreased and breast conservative treatment may be appropriate for most of patients (50–75%) with breast cancer at early stages . Breast conserving surgery (BCS) is an important part of conservative treatment, comprising the excision of the tumor plus a margin of healthy tissues to eliminate cancerous cells. With better cosmesis results, BCS is nowadays the preferred alternative to mastectomy. Yet, a treatment plan is always tailored based on both medical and personal choices. Treatment options are conditioned by the biology of the tumor, the stage of breast cancer, the patient’s health conditions and preferences .
|
review
| 99.9 |
Several studies have shown that the survival rate is almost the same for both mastectomy and BCS, with the benefit that the second imposes less deformation on breast and a more satisfactory aesthetic outcome can be achieved . Still, despite the smaller deformation after BCS, it has been reported that up to 30% of patients are dissatisfied with their post-operative appearance . Actually, the final aesthetic outcome can be affected by so many different variables, from different surgical practices and expertise, to some breast specific characteristics, such as volume and density, tumor size and location, hardening the prediction and patient/surgeon communication about surgical procedure results. Patients are usually involved in the decision process regarding their surgery, but most of the time surgeons lack the means to provide visual clues about the post-surgery results of different alternatives. Even though, this is an important step, regarding the acceptance of the final outcome, as also the contribution of breasts to the sense of femininity and beauty of most women. In fact, follow-up studies after breast cancer treatment show the harmful impact of poor aesthetic results on the psychosocial health of women, who describe loss of self-esteem , sexual impairment and dislike towards their bodies after treatment . On the other hand, physicians have been recognizing the value of support decision systems for planning BCS, to compare the outcome of different surgical options and facilitate surgeon/patient communication. The value of such systems is further supported by studies confirming that women are more willing to deal with the aesthetic results when they are included in the decision process . These visual sensor tools could inform better the patient about the aesthetic consequences of the treatment and improve the feeling about all the process.
|
review
| 99.9 |
The development of a planning/simulation tool for surgery demands the creation of three dimensional (3D) models of the breast that can be deformed in a realistic fashion, to reproduce known deformations imposed by surgery; however, the creation of such models is a challenging task due to the deformable characteristics of the breast, the lack of landmarks to define its shape and the complex nature of the deformations imposed by the surgery. To the best of our knowledge, there are currently no tools, other than surgical experience and clinical judgment , to predict the impact of BCS on the shape and deformation of the treated breast . In fact, the available solutions usually rely on generic models, are mainly targeted to plastic surgery (namely breast augmentation) and do not comprise complex deformations, such as the ones resulting from BCS. Moreover, they usually demand expensive and large equipment to scan patient’s torso and require expertise to handle those scans .
|
study
| 98.5 |
Still, in literature, strategies to model breast deformations are abundant and designed to different applications: estimate pose transformation , assist registration tasks among different radiological imaging modalities , model breast deformation , guide surgery , predict the healing process of the breast after tumor removal , among others. In particular, we highlight the work of Vavourakis et al. , that proposed a 3D surgical simulator to predict a patient-specific outcome after BCS. This framework predicts the breast shape after surgery taking the wound healing process into account. The simulator relies on a coupled multiscale Finite Element (FE) numerical procedure to solve two mathematical models: a biochemical model for wound healing and angiogenesis, and a biomechanical model for soft tissues and pose estimation. The first considers both wound healing biochemical process and the formation of new blood vessels, while the second predicts the breast shape as function of the breast tissues mass density and the body force vector. The final shape of the breast is then predicted as an integration of both models.
|
review
| 99.44 |
The aforementioned applications have in common the use of biomechanical models to predict deformations that, due to some inherent limitations, is not the most suitable approach to include in a tool designed to be used in the daily clinical practice. First, the computational process for most algorithms might take hours, days or even almost a week, depending on the complexity of the models. As a principle aim, to ease the patient/physician communications during their consultations, the planning tool must provide the solution in an expected short time. However, the biomechanical models demand high-end requirements which are not accessible in all clinics. Considering the common computation power of the available machines in clinics, the biomechanical models might take days to provide the required planning, which is too late to satisfy the aforesaid aim. Therefore, faster methodologies should be considered instead of faster machines. Second, most models are simple representations of the breast biomechanics, using unverified parameters, posing fidelity concerns on the predictions. Third, the characterization of individual-specific parameters to create personalized models of the breast, alongside with precise representation of loading and boundary constraints during different clinical procedures, is still an unsolved challenge . The aforesaid applications are intended to provide patient-specific treatment solutions. Thus, the model parameters should be easily personalized with respect to patient and tumor characterizations during the consultations. Alternative strategies to model breast deformations encompass the fitting of parametric models , physical equations to describe known breast deformations , user-intuitive parameters to change breast shape or dataset of known cases to simulate breast surgery outcomes . These strategies model the breast with limited number of parameters and produce results in a timely manner more adequate to clinical practice, but the modelling of breast deformations has still to be improved for clinical surgery planning applications.
|
review
| 99.9 |
Although ML methodologies have not been visited as a solution for real data, not only are they capable of performing the prediction in a meaningful time, but also they need less expertise for configuration. Besides the mentioned advantages, the ability of discovering the hidden correlation between different features distinguish ML from the other possible methodologies to solve this problem. In , Bessa et al. were able to use regression models to predict the deformation parameters of some of the physical equations proposed by Chen et al. and model the breast shape according to the degree of deformation defined by the user. Despite the promising results, it was limited by the usage of a completely synthetic dataset and by the dependence on previous knowledge of the physical equations of deformations. Here, the aim is to expand the described work by using real data, avoiding the usage of specific physical equations to model the breast deformations. Besides, to accomplish the requirement of providing a dataset with the demanded properties, an in-house dataset is generated using data from Magnetic Resonance Imaging (MRI) of real patients and an available breast cancer surgery simulator .
|
study
| 99.9 |
The rest of this paper is organized in five sections. Section 2 is focused on the design and construction of a dataset, with a brief description of the breast anatomy and a detailed explanation of the surgery simulator used to generate the dataset instances. In Section 3, the methodology followed in this work is described, from the feature engineering step to the machine learning approaches used. Implementation of the aforementioned regressions, together with numerical evaluations are explained in Section 4, indicating a promising prediction of the breast shape. Then, the complementary discussions will be expressed in Section 5. Finally, the explanations of the proposed methodology and generated dataset are wrapped up in Section 6, as conclusion.
|
study
| 89.6 |
Historically, several factors have been identified to have a significant impact on the shape deformations caused by BCS, which can be grouped into patient, tumor and surgery related factors . Hence, any model designed to predict breast deformations should take into account the influence of those characteristics. Since the main goal of this study is to learn the influence of different combinations of those factors, a large number of 3D data of patient’s breast before and after surgery (≈1 year) is mandatory. However, there is no available dataset that satisfies such requirements. As consequence, an in-house dataset was constructed, taking advantage of available MRI data, acquired before surgery and having some anatomical structures of interest annotated. Due to the lack of post-surgery data, the wound healing process was simulated using the BCS simulator proposed by Vavourakis et al. , which made the source code available for the scientific community.
|
study
| 100.0 |
In this Section, some insights on the breast anatomy and characteristics that condition shape deformations are provided, the framework designed by Vavourakis et al. used to simulate BCS results is explained and the construction of the dataset used to develop the proposed methodology is detailed.
|
other
| 73.4 |
Breasts are important organs in a women’s body, whose primary role is related to sexual attraction and production of milk to nourish children. These highly deformable organs are located on the anterior and lateral parts of the chest, overlying the pectoralis major and minor muscles. With an heterogeneous structure (Figure 1), consisting of mammary glands (fibroglandular tissue) and adipose tissues (fat), breasts are firmly attached to the skin and underlying structures by fibrous bands referred to as Cooper’s ligaments. These suspensory ligaments provide the function of support, hold the breasts in place and contribute to determine the shape and contour of the breast . Adipose tissue is fat-storing loose connective tissue, which determines the size of the breast, and mammary glands are modified sweat glands that are responsible for milk production.
|
review
| 99.56 |
The relative distribution of fibroglandular and fat tissues varies significantly depending on patient’s age, menstrual cycle, pregnancy/lactancy, hormone therapy and menopause, which affects the structure and morphology of the breast . The ratio of these two types of tissues defines the breast density, which among other factors, can be related to the risk of developing breast cancer. In fact, women with a higher breast density are more likely to develop breast cancer . Because fibroglandular and fat tissues have different mechanical behaviours, breast density also influences the aesthetical outcome of BCS, alongside with other breast characteristics, such as size and volume. Other factors that influence the deformations caused by BCS are the tumor size and location. Additionally, surgery-related factors such as the used technique, the placement of incisions, the volume of excised tissue or varying surgeon expertise lead to different types and extents of shape deformations . Although the extent of the scar, color alterations or roughness of the treated breast also weight in its visual appearance, in this work only shape and geometry related deformations are modelled.
|
study
| 99.94 |
Vavourakis et al. proposed a BCS simulator that models the 3D post-surgical shape of the breast by coupling a physiological model of tissue recovery with a biomechanical model of pose estimation. The former predicts breast contraction caused by wound healing while the latter uses a Mooney-Rivlin biomechanical model of the breast to simulate deformations for different patient’s positions. The main stages of the surgery simulation (Figure 2) comprise the construction of a patient-specific Finite Element Model (FEM) of the breast, the definition of the tumor and the wound healing simulation itself, which uses the multiscale biomechanical model of wound healing described above.
|
study
| 99.94 |
Each of the wound healing pipeline stages are carried out with the patient in different positions and thus, some pose transformations are also accounted for (Figure 3). In fact, MRI is acquired with the torso facing down (prone position), surgery is performed with the patient facing up (supine position), the simulation occurs in an unloaded stage and surgery results are evaluated in the upright position.
|
other
| 99.8 |
According to Vavourakis et al. , a 3D patient-specific biomechanical FEM of the breast can be created from MRI data (Figure 2A), that is segmented to delineate breast and background (Figure 2B). Fat and fibroglandular tissues can also be differentiated to create a more detailed FEM, in which distinct mechanical properties are assigned to elements according to the breast tissues they represent. Upon the segmentation of the structures of interest, a 2D surface mesh of FE that represents skin and a 3D mesh representing the interior of the breast shall be generated (Figure 2C).
|
study
| 100.0 |
The FEM generated from MRI data represents the breast in the prone position because it assumes the configuration of the patient during MRI acquisition. Yet, during the surgery simulation work-flow, some stages require the model to represent the patient in different positions. As consequence, two explicit pose transformations occur (Figure 2D,F), and a third one takes place during the wound healing step (Figure 2G). In detail, MRI data is acquired with the patient lying in prone position (Figure 3A), but the upright position (Figure 3G) is the most suitable pose to evaluate the natural shape of breast and, consequently, its deformations. Thus, wound healing results are outputted in the upright position (Figure 3F). On the other hand, BCS is performed with the patient lying in supine position (Figure 3D), implying that any surgical planning activity, such as the definition of tumor characteristics and the volume to excise, should be done in this position. The multiscale model used for surgery simulation can estimate pose transformations by computing the unloaded state (a gravity free reference state) and re-applying gravity stress in the desired direction. As result, the FEM, constructed in prone position, is converted to an unloaded state (Figure 3C), from which both supine (Figure 3D) and pre-surgery upright position (Figure 3G) can be estimated. The tumor is defined in supine position (Figure 3D) and the FEM is again converted from supine to the unloaded state (Figure 3E), where the wound healing simulation is carried out. The simulation result (post-surgery data) is returned in upright position (Figure 3F). With this pose transformation pipeline, both pre- and post-surgery data are represented in the upright position, and can be directly compared for predicting shape deformations caused by BCS.
|
study
| 100.0 |
The multiscale biomechanical model proposed by Vavourakis et al. simulates the wound healing process taking as input the volume excised during surgery. This volume depends on the tumor position and size, and therefore, a virtual surgery has to occur in which all FE inside the volume to excise are re-labelled as damaged and assigned with different physiological and mechanical parameters. In , this virtual surgery is simulated in the supine position: the surgeon identifies the tumor position, defines the incision lines and outlines the incision path inside the breast. The excision volume is then approximated by a cylinder that contains the lesion and whose axis is perpendicular to the chest-wall, extending from the skin to the pectoral muscle. All FE contained inside this cylinder are assigned with damaged tissue properties.
|
study
| 99.94 |
To create the dataset, a subset of MRI data from the PICTURE project (http://www.vph-picture.eu/) was used. In detail, T1-weighted MRI image sets were used, containing approximately 60 axial slices each, with an average voxel resolution of 0.59×0.59×3 mm, (x, y and z axes, respectively). Taking advantage of the manually annotated structures, 3D point clouds (PCLs) of patient’s torsos were created using the breast contour, Latissimus Dorsi muscles and the pectoral muscle, as frontal, lateral and posterior boundaries, respectively. However, considering that the computational cost of the wound healing simulation is decreased if performed for each breast individually, instead of using the entire torso as done by Vavourakis et al. , the torso point cloud (PCL) was vertically divided with a plane defined along the sternum. Performing this division, individual breasts PCLs were obtained, which provide breast shape variability in the dataset. Any breast PCL holding visible MRI coil compression are discarded from the simulation because the deformation is not reversible with the multiscale biomechanical model.
|
study
| 100.0 |
After segmentation, each resulting breast PCL (≈1900 surface points) was converted to a 3D triangulated surface mesh to model the skin, using the Ball-Pivoting algorithm , in MeshLab . The breast volume was next meshed in Gmsh , by inserting uniformly distributed points inside the object (≈2500 volume points), subsequently connecting them with tetrahedrons elements. To complete the creation of the FEM, distinct boundary conditions and material properties were assigned to the surface mesh - frontal surface, pectoral muscle (back surface), lateral limits, and top and bottom boundaries were defined - following the strategy proposed by Vavourakis et al. .
|
study
| 99.94 |
For the purpose of data augmentation, the number of dataset instances was increased by varying some input parameters related to breast and tumor characteristics, namely the breast density, and the tumor size and position, known to influence the aesthetical result after BCS.
|
study
| 99.7 |
To represent all categories of breast densities in the dataset, it was necessary to model other ratios of tissues than the ones represented in original MRI data, by varying the number of FE that are assigned with mechanical properties of fibroglandular and fat tissues. To avoid a segmentation step, the approach proposed by Del Palomar et al. was used to simplify the structural complexity of the breast, by assigning a weighted average value of the mechanical properties of each tissue type to all elements of the model. Hence, the ratios defined by the American College of Radiologists (ACR) classification system - the Breast Imaging Reporting and Data System (BI-RADS®) - can be considered for weighting the material property values described in , and represent several breast densities. This reporting system identifies 4 categories of breast density (A, B, C and D), which are described in Table 1. Following this strategy, the fibroglandular/fat ratios: A—10/90; B—35/65; C—60/40; D—85/15, were used to average material properties of each category, as detailed in Table 2.
|
study
| 100.0 |
Besides the breast shape and composition, it is also important to characterize the tumor location and size. Since the relationship between tumor position and the aesthetical outcome is generally defined using breast quadrants to discretize tumor locations, tumors are randomly positioned inside each quadrant instead of choosing any position inside the breast, which assures the representativeness of the dataset by guarantying that there are instances of tumors in every quadrant. There are 4 quadrants defined by a vertical and horizontal division of the breast through the nipple and in the upright position (Figure 4): Upper-Outer or superolateral Quadrant (UOQ), Upper-Inner or superomedial Quadrant (UIQ), Lower-Outer or inferolateral Quadrant (LOQ), Lower-Inner or inferomedial Quadrant (LIQ) .
|
study
| 99.94 |
The breast quadrants division is established in upright position. However, according to the surgery simulator pipeline (Figure 2), the tumor is defined in the supine position. Therefore, in an attempt to define quadrants in the horizontal position which correctly correspond the upright ones, the nipple position (manually annotated) is used as a reference point to compute the vertical and horizontal planes that define the quadrants boundaries. Three main planes are then sequentially defined, as seen in Figure 5. The first plane corresponds to the one along the pectoral muscle (Figure 5a) and is defined by its normal which is computed by the cross product of two vectors, defined by three corner points of the pectoral muscle (two on the top and one on the bottom). The second plane sets the superior-inferior boundary (red plane in Figure 5b) and is defined as parallel to the xy-plane (taking into account that the direction of the MRI acquisition is perpendicular to this plane), crossing the nipple. Finally, the third plane sets the lateral-medial boundary (green plane in Figure 5c), being perpendicular to the pectoral plane and crossing the nipple too.
|
study
| 99.94 |
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