i BỘ CÔNG THƯƠNG ĐẠI HỌC CÔNG NGHIỆP THÀNH PHỐ HỒ CHÍ MINH BÁO CÁO TỔNG KẾT ĐỀ TÀI KHOA HỌC KẾT QUẢ THỰC HIỆN ĐỀ TÀI NGHIÊN CỨU KHOA HỌC CẤP TRƯỜNG Tên đề tài TỔNG HỢP XANH MỘT SỐ AMINOPHOSPHATE MỚI D[.]
BỘ CÔNG THƯƠNG ĐẠI HỌC CÔNG NGHIỆP THÀNH PHỐ HỒ CHÍ MINH BÁO CÁO TỔNG KẾT ĐỀ TÀI KHOA HỌC KẾT QUẢ THỰC HIỆN ĐỀ TÀI NGHIÊN CỨU KHOA HỌC CẤP TRƯỜNG Tên đề tài TỔNG HỢP XANH MỘT SỐ AMINOPHOSPHATE MỚI DỰA TRÊN CARBAZOLE VÀ HOẠT TÍNH SINH HỌC Mã số đề tài: 20/1.5CNH03 Chủ nhiệm đề tài: TS TRẦN NGUYỄN MINH ÂN Đơn vị thực hiện: KHOA CÔNG NGHỆ HĨA HỌC TP HỒ CHÍ MINH, 06.2020 Tp Hồ Chí Minh, … i DANH MỤC HÌNH ẢNH Hình Hợp chất 3c cho thấy khả kháng nấm C.albincans mạnh 200 µg.mL-1(377,78 µM) so sánh với Fluconazole nồng độ 1mg.mL-1 (3265µM) 93 Hình Hợp chất 3h cho thấy khả kháng nấm mạnh S cerevisiae nồng độ 200 µg.mL-1 (455.38 µM) so sánh với Fluconazole, nồng độ mg mL-1 (3265 µM), 3h cho thấy hoạt tính kháng nấm tốt nồng độ 50 100 µg.mL-1 93 Hình Hợp chất 3b cho thấy khả kháng khuẩn tốt, B.megaterium nồng độ 200 µg mL-1(388 µM), kháng trung bình nồng độ 100 µg.mL-1, khơng kháng nồng độ 50 µg mL-1 so sánh với Ampicillin nồng độ mg.mL-1 (5724 µM) 94 Hình Hợp chất 3j cho thấy khả kháng trung bình trở lại B cereus tất nồng độ so sánh với Ampicillin nồng độ mg mL-1 94 Hình Giá trị IC50 3a–j, kháng tế bào ung thư MCF–7 .96 Hình Giá trị IC50 3a–j, kháng tế bào ung thư, A–549 96 Hình Giá trị IC50 3a–j, kháng tế bào ung thư, HeLa 96 Hình Khả ức chế (%) tế bào ung thư, MCF–7 3a-j, nồng độ từ 5-100 μM 97 Hình Khả ức chế (%) tế bào ung thư, A-549 3a-j nồng độ từ 5-100 µM .97 Hình 10 Khả ức chế (%) tế bào ung thư, Hela 3a-j nồng độ từ 5-100 µM 97 Hình 11 Cấu dạng ổn định 3c sau hồn thành docking với C.albincans, tính tốn AutoDockTools-1.5.6rc3 trình bày Discovery Studio 2019 Client .100 Hình 12 Các tương tác cấu dạng ổn định ligand, 3c receptor, 6TZM 101 Hình 13 Các vị trí liên kết cấu trúc 3c với vị trí hoạt động C.albincans, 6TZM, Discovery Studio 2019 101 Hình 14 Bản đồ ligand minh họa Molegro Molecular Viewer 102 Hình 15 Cấu dạng ổn định 3h sau hoàn thành docking với S cerevisiae, 3ET5 102 Hình 16 Các liên kết vị trí hoạt động 3ET5 cấu dạng ổn định 3h 102 Hình 17 Các vị trí liên kết cấu hình ổn định 3h với tâm hoạt động 3ET5 103 Hình 18 Bản đồ ligand 3h 3ET5 103 ii DANH MỤC BẢNG Bảng Hoạt tính kháng nấm hợp chất, 3a-j 77 Bảng Hoạt tính kháng khuẩn hợp chất 3a-j .78 Bảng Các giá trị IC50 3a–j vitro kháng lại dòng tế bào ung thư phổi, MCF–7 79 Bảng Các giá trị IC50 3a–j vitro kháng lại dòng tế bào ung thư vú, A–549 80 Bảng Các giá trị IC50 3a–j vitro kháng lại dòng tế bào ung thư tử cung, Hela .80 Bảng Các kết docking quan trọng pose docking, 3c, 3h, đối chứng dương silico docking 81 Bảng Các kết docking quan trọng docking poses (3a-j) với receptor dòng tế bào ung thư: MCF–7 (6VNN: PDB), A–549 (4ASD), HeLa (5HES) .84 Bảng Giá trị IC50 đầu vào (3a-j) kháng dòng tế bào ung thư người , MCF–7, A– 549, HeLa, thuốc Camptothecin vitro 95 Bảng Các kết docking quan trọng cấu dạng ổn định 3c thuốc Fluconazole với Candida albicans, 6TZM 104 Bảng 10 Các kết đáng tính tốn docking 3h, thuốc, Fluconazole kháng nấm Saccharomyces cerevisiae, 3ET5 .104 Bảng 11 Các docking pose quan trọng 3b and ampicillin kháng Bacillus megaterium bacterium, code, 6NVW .105 Bảng 12 Kết docking docking pose 3a–j đến tâm hoạt động receptor dòng tế bào ung thư vú, MCF–7(6VNN) 106 Bảng 13 Kết docking docking pose 3a–j đến vị trí hoạt động receptor dòng tế bào ung thư phổi, A–549 107 Bảng 14 Kết docking docking pose 3a–j đến vị trí hoạt động receptor dòng tế bào ung thư, HeLa.[a] 108 Bảng 15 Các giá trị IC50 đầu vào 3a-j đối chứng dương kháng trở lại dòng tế bào ung thư MCF–7, A–549, HeLa, vitro .110 iii DANH MỤC SƠ ĐỒ Sơ đồ Tổng hợp dẫn xuất α- amino phosphate (3a-j) từ carbazole .58 Sơ đồ Tổng hợp dẫn xuất trung gian 1, sản phẩm, 3a-j a) Et-Br, KOH, DMSO khô, xúc tác TBAHS, đung hồi lưu 24 h, 91% 1; b) Phản ứng Vilsmeier Haack: DMF (khan) CH2Cl2; POCl3 ClCH2CH2Cl, N, đun hồi lưu, 48 h, NaOH, 95% 2; c) Kabachnik-Fields: 9– ethyl–9H–carbazole–3–carbaldehyde (1 mmol), diethyl phosphite (1.3 mmol), analogue primary aromatic amine (1 mmol), PEG–400 catalyst (0.38 % mol), 100 oC, 6–7 h, 3a-j: 84-91% 67 iv HÌNH PHỤ LỤC Hình Cấu dạng ổn định Fluconazole sau hoàn thành docking với nấm C.albincans, (6TZM: PDB), xây dựng mơ hình DSC Client 2019 116 Hình Sự tương tác amino acid lại 6TZM với cấu dạng bền Fluconazole 116 Hình Sự tương tác amino acid lại, Thr-426, Asn-444, Asp-445 Asn-476 6TZM với Fluconazole 117 Hình Các tương tác nguyên tử hoạt tính receptor (6TZM) cấu dạng bền (Fluconazole) sơ đồ 2D 117 Hình Bản đồ ligand tương tác bậc hai Fluconazole 6TZM 118 Hình Cấu dạng Fluconazole sau hồn thành tính tốn docking với Saccharomyces cerevisiae 118 Hình Sự tương tác amino acid 3ET5 với cấu trúc bền Fluconazole 118 Hình Các tâm hoạt động liên kết ligand-receptor, 3ET5 sơ đồ 2D .119 Hình Sự tương tác amino amin cịn lại 3ET5: Ser-164, Val-165 Arg-147 với tâm hoạt động Fluconazole 119 Hình 10 Bản đồ ligand tương tác thứ cấp Fluconazole 3ET5 sơ đồ 2D 120 Hình 11 Cấu dạng ổn định 3b sau docking với Bacillus megatherium, 6NVW 120 Hình 12 Sự tương tác amino acid lại 6NVW với cấu trúc ổn định 3b 121 Hình 13 Các tâm hoạt động liên kết từ ligand, 3b đến (6NVW) gồm tương tác liên kết .121 Hình 14 Sự tương tác amino acid lại 6NVW cấu dạng 3b 122 Hình 15 Bản đồ ligand cho thấy tương tác thứ cấp 3b 6NVW .122 Hình 16 Cấu dạng ổn định Ampicillin sau docking với Bacillus megatherium .123 Hình 17 Sự tương tác amino acid cịn lại 6NVW với cấu trúc ổn định Ampicillin sơ đồ 3D 123 Hình 18 Các tâm hoạt động (6NVW) cấu dạng ổn định Ampicillin .124 Hình 19 Sự tương tác amino acid lại 6NVW : Gln-23, Asn-462, Arg-266 Arg381 với tâm hoạt động Ampicillin .124 Hình 20 Bản đồ ligand tương tác thứ cấp cấu dạng ampicillin 6NVW 125 v Hình 21 Phổ 1H NMR 126 Hình 22 Mass spectrum 127 Hình 23 Phổ IR 128 Hình 24 Phổ khối lượng 129 Hình 25 Phổ khối 130 Hình 26 Phổ IR 3a 131 Hình 27 Phổ 1H NMR 3a .132 Hình 28 Phổ 1H NMR 3a, phổ giãn rộng: 1.00–2.70 ppm 133 Hình 29 Phổ 1H NMR 3a, giãn rộng: 3.40–5.30 ppm .134 Hình 30 Phổ 1H NMR 3a, giãn rộng: 6.60–8.50 ppm 135 Hình 31 Phổ 13C NMR 3a 136 Hình 32 Phổ 31P NMR 3a, at δ = 24.767 ppm 137 Hình 33 Phổ IR spectrum 3b 138 Hình 34 Phổ 1H NMR 3b 139 Hình 35 Phổ 1H NMR 3b, giãn rộng: 0.70–1.50 ppm 140 Hình 36 Phổ 1H NMR 3b, giãn rộng: 3.40–5.10 ppm 141 Hình 37 Phổ 1H NMR 3b, giãn rộng: 6.30–8.40 ppm .142 Hình 38 Phổ 13C NMR 3b 143 Hình 39 Phổ 31P NMR 3b, at δ = 24.189 ppm 144 Hình 40 Phổ IR 3c 145 Hình 41 Phổ 1H NMR 3c 146 Hình 42 Phổ 1H NMR 3c, giãn rộng: 0.90–2.50 ppm 147 Hình 43 Phổ 1H NMR 3c, giãn rộng: 3.40–5.20 ppm 148 Hình 44 Phổ 1H NMR 3c, giãn rộng: 6.20–8.70 ppm .149 Hình 45 Phổ 13C NMR 3c 150 Hình 46 Phổ 31P NMR 3c, δ = 24.265 ppm .151 Hình 47 Phổ IR 3d 152 vi Hình 48 Phổ 1H NMR 3d 153 Hình 49 Phổ 1H NMR 3d, giãn rộng: 0.85–2.05 ppm .154 Hình 50 Phổ 1H NMR 3d, giãn rộng: 3.00–6.40 ppm 155 Hình 51 Phổ 1H NMR 3d, giãn rộng: 6.50–9.00 ppm .156 Hình 52 Phổ 13 C NMR 3d 157 Hình 53 Hình 31P NMR 3d, δ =23.130 ppm .158 Hình 54 Phổ IR 3e 159 Hình 55 Phổ 1H NMR 3e 160 Hình 56 Phổ 1H NMR 3e, giãn rộng: 0.4–2.00 ppm 161 Hình 57 Phổ 1H NMR spectrum 3e, giãn rộng: 2.60–4.80 ppm .162 Hình 58 Phổ 1H NMR 3e, giãn rộng: 5.10–8.90 ppm 163 Hình 59 Phổ 13C NMR 3e 164 Hình 60 Phổ 31P NMR 3e, δ = 24.70 ppm 165 Hình 61 Phổ IR spectrum 3f .166 Hình 62 Phổ 1H NMR 3f 167 Hình 63 Phổ 1H NMR 3f, giãn rộng: 0.9–1.6 ppm 168 Hình 64 Phổ 1H NMR 3f, giãn rộng: 3.10–5.90 ppm 169 Hình 65 Phổ 1H NMR 3f giãn rộng: 6.20–8.30 ppm .170 Hình 66 Phổ 13C NMR 3f 171 Hình 67 Phổ 31P NMR 3f, δ = 23.435 ppm 172 Hình 68 Phổ IR 3g 173 Hình 69 Phổ 1H NMR c 3g 174 Hình 70 Phổ 1H NMR 3g, giãn rộng: 0.20–1.50 ppm .175 Hình 71 Phổ 1H NMR 3g, giãn rộng: 2.90–5.50 ppm .176 Hình 72 Phổ 1H NMR 3g, giãn rộng: 6.00– 8.30 ppm 177 Hình 73 Phổ 13 C NMR spectrum 3g 178 Hình 74 Phổ 31P NMR 3g, δ = 24.230 ppm 179 vii Hình 75 Phổ IR spectrum of compound 3h .181 Hình 76 Phổ 1H 3h 181 Hình 77 Phổ 1H NMR 3h, giãn rộng: 0.90– 1.50 ppm 182 Hình 78 Phổ 1H NMR 3h, giãn rộng: 3.20– 5.10 ppm 183 Hình 79 Phổ 1H NMR 3h, giãn rộng: 5.90– 8.30 ppm 184 Hình 80 Phổ 13C NMR 3h 185 Hình 81 Phổ 31P NMR 3h, δ = 23.560 ppm 186 Hình 82 Phổ IR spectrum 3i .187 Hình 83 Phổ 1H NMR 3i 188 Hình 84 Phổ 1H NMR spectrum 3i, giãn rộng: 0.60–2.10 ppm 189 Hình 85 Phổ 1H NMR 3i, giãn rộng: 3.40–5.40 ppm .190 Hình 86 Phổ 1H NMR 3i, giãn rộng: 6.40–8.40 ppm .191 Hình 87 Phổ 13C NMR spectrum giãn rộng 3i .192 Hình 88 Phổ 31P NMR 3i, δ = 23.680 ppm .193 Hình 89 Phổ IR spectrum 3j .194 Hình 90 Phổ 1H NMR 3j 195 Hình 91 Phổ 1H NMR 3j, giãn rộng: 0.30–1.50 ppm .196 Hình 92 Phổ 1H NMR 3j, giãn rộng: 2.40–5.60 ppm 197 Hình 93 Phổ 1H NMR 3j, giãn rộng: 6.10–8.30 ppm .198 Hình 94 Phổ 13C NMR 3j 199 Hình 95 Phổ 31P NMR 3j, δ = 24.642 ppm 200 viii MỤC LỤC DANH MỤC HÌNH ẢNH II DANH MỤC BẢNG III DANH MỤC SƠ ĐỒ IV HÌNH PHỤ LỤC V MỤC LỤC IX LỜI CÁM ƠN 55 PHẦN I THÔNG TIN CHUNG .56 PHẦN II BÁO CÁO CHI TIẾT ĐỀ TÀI NGHIÊN CỨU KHOA HỌC 64 CHƯƠNG 1: TỔNG QUAN 64 1.1 Tổng quan vấn đề nghiên cứu .64 1.2 Hiện trạng cơng trình nghiên cứu liên quan đến đề tài 65 CHƯƠNG 2: VẬT LIỆU VÀ PHƯƠNG PHÁP 67 2.1 Nguyên liệu hóa chất 67 2.2 Qui trình 67 CHƯƠNG 3: KẾT QUẢ VÀ THẢO LUẬN 72 3.1 Kết phân tích hóa lý 72 3.2 Hoạt tính sinh học: 77 CHƯƠNG 4: KẾT LUẬN VÀ KIẾN NGHỊ 111 REFERENCES .112 PHỤ LỤC 116 ix LỜI CÁM ƠN Chúng xin chân thành cảm ơn Quỹ nghiên cứu khoa học IUH, Ban lãnh đạo IUH, Lãnh đạo khoa Cơng nghệ Hóa học, Phịng thí nghiệm Khoa Cơng nghệ Hóa học, thành viên đề tài giúp tơi hồn thành đề tài nghiên cứu khoa học Cảm ơn thành viên nhóm nghiên cứu: PGS.TS Nguyễn Văn Cường, ThS.NCS Nguyễn Minh Quang, PGS.TS Trương Vũ Thanh, TS Mahboob Alam động viên giúp đỡ mặt tinh thần để hồn thành cơng trình nghiên cứu Trang 55 [IY[OU] ChemistrySelect Full Papers doi.org/10.1002/slct.202000855 z Organic & Supramolecular Chemistry Green Synthesis Using PEG-400 Catalyst, Antimicrobial Activities, Cytotoxicity and In Silico Molecular Docking of New Carbazole Based on -Aminophosphonate Tran Nguyen Minh An,*[a] Nguyen Van Cuong,[a] Nguyen Minh Quang,[a] Truong Vu Thanh,[b] and Mahboob Alam[c] New analog –amino phosphate series with high yield through an effective and environmentally friendly protocol using the PEG-400 catalyst via Kabachnik-Fields has been reported They were performed to screen antimicrobial activities by the dish diffusion method, anticancer against MCF-7, A-549, and HeLa human cancer cell lines by MTT assay, and carried out in silico molecular docking by Avogadro, AutoDockTools, Discovery Studio 2019 Client, and Molegro Molecular Viewer packages The 3c and 3h displayed excellent inhibition against Candida albicans and Saccharomyces cerevisiae fungi, respectively The molecular docking model indicated the reasonable explanations between the receptor and bioactive compounds in vitro of 3c, h, and 3b The 3c was shown as an excellent inhibitor Introduction The -amino phosphonate or diester of amino phosphonic acids were significant objects, valuable medical compounds because they had a structural analogy to amino acids, peptides, natural phosphates, and interesting biochemistry recently.[1] The strategy of synthesis and biological activities of some phosphorus organic derivatives have been reported in the literature.[2] The bioactivities that we performed as potential fungicides,[3] antibacteria, antitumor,[4] antioxidant,[5] inhibitors of enzymes involving in amino acid and peptide metabolism,[6] UDP-galactopyranose mutase,[7] human plasma renin.[8] Many amino phosphonate synthesis methods were performed, but the synthesis approach via Kabachnik-Fields (K.F) reaction conducted the most effective and the reaction processes three components, amine, aldehyde and phosphites derivatives by magnesium perchlorate catalyst, which in high yielded in [a] Dr T N M An, Prof N V Cuong, Dr N M Quang Faculty of Chemical Engineering, Industrial University of Ho Chi Minh City, Ho Chi Minh city, Vietnam, 12 Nguyen Van Bao Road, Ward 4, Go Vap District, Ho Chi Minh City,Vietnam E-mail: trannguyenminhan@iuh.edu.vn [b] Prof T V Thanh Faculty of Chemical Engineering, Ho Chi Minh City University of Technology, VNU-HCM, Vietnam, 268 Ly Thuong Kiet, Ward 14, District 10, Ho Chi Minh City, Vietnam [c] Dr M Alam Division of Chemistry and Biotechnology, Dongguk University, Gyeongju 780–714, Republic of Korea Supporting information for this article is available on the WWW under https://doi.org/10.1002/slct.202000855 ChemistrySelect 2020, 5, 6339 – 6349 against Candida albicans, which was a new result in vitro and in silico molecular docking model The 3c, f, and 3i pointed out excellent inhibitions against HeLa cell lines and new anticancer results of +–amino phosphate compounds The docking studies of structures against receptors of three human cancer cell lines were conducted and recommended that the hydrogen bonds only formed from active sites of receptors to oxygen atom of the ethoxy group, nitrogen and hydrogen atoms of N H group, oxygen atom of the P=O double bond group, and the fluorine atoms of the CF3 group in 3i for calculated docking of the receptors of bacteria, fungi, and cancer cell lines to all ligands amino phosphonate products.[9] The heterogeneous catalyst, solid catalyst series applied reactions to obtain in high yields, for instance, antimony trichloride adsorbed on alumina, cobalt chloride,[10] iron(III) chloride,[11] zirconium(IV),[12] commercial titan dioxide under solvent-free conditions,[13] and sulfamic acid.[14] Some of lanthanide triflates-catalyzed in ion liquid formed products and conducted easily to reuse solid catalyst.[15] The chiral Brønsted acid made a catalyst for products with high enantioselectivities.[16] The super magnetic nano iron oxide in solvent-free conditions performed magnetically recyclable catalysts.[17] The sodium dodecyl sulfate in water, Lewis acid and surfactant supported to yield amino phosphonic acid diesters via K.F.[18] The homogeneous catalysts were used for K.F reaction as KHSO4 under the solvent-free condition at ambient temperature for moderate yields.[19] Recently, as extensive attention, the polyethylene glycol-400 (PEG-400) has been identified as a green catalyst, inexpensive, green media, non-toxics, reuse, short-time reaction, high yield, an efficient and eco-friendly reaction, greener approach, alternative reaction medium, and medium for the broad synthesis of transformation organic functional groups and biodegradable material.[20] The PEG-400 carried out the catalyst for Michael’s addition reaction as a non-ionic liquid solvent,[21] Suzuki crosscoupling reaction, Suzukie Miyaura cross-coupling reactions,[22] the nucleophile additions of phosphonic acid diesters to C=N, the double bond in K.F reaction.[23] As interested PEG-400, green approach and continuous report of new -amino phosphonate derivatives based on carbazole via K.F reaction, we carried out synthesis, anticancer, antimicrobial activities, 6339 © 2020 Wiley-VCH Verlag GmbH & Co KGaA, Weinheim ChemistrySelect Full Papers doi.org/10.1002/slct.202000855 and conducted in silico molecular docking, which discovered structure-activity relationship (SAR) and designed core structures having biological activities results Results and Discussion Reaction Mechanism Scheme The proposed reaction mechanism of Kabachnik–Fields: the reaction of components: 9–ethyl–9H–carbazole–3–carbaldehyde, analogue primary aromatic amine, and diethyl phosphite by a green catalyst, PEG-400 Scheme Synthesis of intermediate derivatives 1, 2, and products, a-j Reagents and conditions: a) Et–Br, KOH catalyst, dry DMSO, reflux 24 h, 91% 1; b) Vilsmeier–Haack reaction: DMF(dry) in CH2Cl2; POCl3 in ClCH2CH2Cl, N2, reflux, 48 h, NaOH, 95% 2; c) Kabachnik-Fields: 9–ethyl–9H–carbazole–3– carbaldehyde (1 mmol), diethyl phosphite (1.3 mmol), analog primary aromatic amine (1 mmol), PEG–400 catalyst (0.38% mol), 100 °C, 6–7 h, a-j: 84–91% Physical chemistry In 1H NMR as shown in the experimental section, the signals of the methine protons of HN–CH–P=O(OEt)2 groups showed the resonances as the doublet peaks in entries a–c, g–h, and j with chemical shifts () in region 4.88–4.95 ppm with coupling constants in the range of 23.5 –24.0 Hz, and doublet of doublet (dd) peaks in d–f and i with the chemical shifts from 4.89 – 5.25 ppm and coupling constants between 23.0 and 24.4 Hz, due to its near coupling with the hydrogen nucleus of the N–H group and phosphorus nucleus The proton signals of N–H bonds appeared as triplet peaks in d, f, and i with the chemical shifts in the range of 5.29 to 6.08 ppm and the coupling constants from 7.4–8.6 Hz They also showed quartet peaks in c and e with J and in the range of 5.4 –10.9 Hz and 4.88–6.19 ppm, respectively The proton signals of N–H ChemistrySelect 2020, 5, 6339 – 6349 bonds were resonance as triplets in d, f, and i, or quartet in c and e, because of coupling to occur with the nuclei of hydrogen and phosphorous nearby, HN–CH–P The proton signals of N H bonds in the others like a–b, g–h, and j disappeared because of the proton exchange in commercial CHCl3 solvent, which remained residual CHCl3 peak The signal for the exchangeable proton was merged into a single peak with an unpredictable chemical shift The methyl protons of N– ethyl groups were resonance as triplet signals from 1.04 to 1.23 ppm with coupling constants variety between 7.0 and 7.1 Hz in entries a–i except for j The methyl protons of diethyl phosphite groups coupled with methylene protons nearby, which formed two distinction triplet resonance signals in the range of 1.29–1.52 and 1.41–1.61 ppm with coupling constants from 6.8–9.0 and 7.1–7.2 Hz at all entries except for e, respectively The methylene protons of N–ethyl groups coupled with methyl protons nearby exposed the quartet peaks in the region of 4.34–4.38 ppm with the coupling constants of 6.8–7.2 Hz The methylene protons of diethyl phosphite groups indicated three differential resonance arranges, which corresponded multiplet peaks in ranges were listed 3.38–3.9, 3.78–4.11, and 4.09–4.29 ppm, which were one, one, and two of methylene protons, respectively, except that two signals of methylene groups at j and i that showed the quartet and triplet signals at 3.57 and 4.15 with coupling constants of 8.2 and 7.1 Hz, respectively The signals of protons of carbazole ring, H–4 protons exposed as singlet peaks in the region of 8.15–8.45 The remained resonance signals of aromatic protons appeared in the expected ranges 6340 © 2020 Wiley-VCH Verlag GmbH & Co KGaA, Weinheim ChemistrySelect Full Papers doi.org/10.1002/slct.202000855 The 13C NMR showed the signals of methine carbons bound to nitrogen and phosphorus atoms, HN–CH–P coupled near range with them so that the methine carbons established those double peaks in the range of chemical shifts from 56.1 to 57.7 with coupling constants between 151.0 and 152.5 Hz The resonance signals of the phosphorus nucleuses indicated in the range of 23.13–24.70 ppm The infrared absorption bands were identified typically strong functional groups, stretching vibrations in the ranges of 3268–3396 (N–H), 2977–2981 (C–H, alkyl), 1596–1617, 1516–1599, 1480–1493, 1385–1387(C=C),1233– 1234 (P=O), 1300–1336 (C–O), 1024–1054 (C–N), and 748– 752 cm (P–CH) In Silico Molecular docking model Antimicrobial activities The results of docking pose or ligand of the activity structures in vitro like c and h against fungi, Candida albicans, and Saccharomyces cerevisiae, respectively, the b against the bacterium, Bacillus megaterium and standard drugs were shown in Figures 5–12, Table 1–3, and Figures S.1.1–S.1.20 As shown Antimicrobial activities Antifungal activity The entry c and h exposed excellent activities against both S cerevisiae and C.albincans fungi at a concentration of 200 #g.mL compared to the standard drug, Fluconazole at a concentration of 1000 #g.mL The entry h also pointed out good inhibition again S cerevisiae at the concentrations of 50 and 100 #g.mL Antibacterial activity The entry b designated good antibacterial activity against B.megaterium at a concentration of 200 #g mL 1, moderate activity at a concentration of 100 #g mL 1, and no inhibition at a concentration of 50 #g mL The j showed moderate activity against B cereus at all concentrations Figure The entry c in the image exhibited excellent activity against C.albincans at a concentration of 200 #g.mL (377.78 #M) and compared to the standard drug, Fluconazole at a concentration of mg.mL (3265 #M) Anticancer activity The compounds a, c, and h showed excellent cytotoxicity activity against MCF–7 with the IC50 values of 9.78 0.93, 9.39 0.64, and 6.38 0.22 #M against breast cancer cell lines, MCF–7, respectively as shown in Table and Figure 13 The compounds c, e, and h showed the highest inhibitions with the IC50 values, 7.32 0.07, 3.82 0.01, and 2.29 0.02 #M against small lung cancer cell lines, A–549, respectively as shown in Table and Figure 14 For cervical cancer cell lines, HeLa, the compounds, c, f, and i exposed high inhibitions against HeLa cancer cell lines with the IC50 values, which performed 5.98 0.02, 5.16 0.04, and 7.19 0.02 #M, respectively as shown in Table and Figure 15 These results of cytotoxicity of c, f, and i against cervical cancer cell lines, which were reported the first time screening cytotoxicity activity for +-amino phosphate compounds The IC50 values and inhibition abilities of entries a-j against MCF–7, A–549, and HeLa cancer cell lines depicted in Figures 13 and 15 and Figures 16–18, respectively Figure The h image exposed excellent activity against S cerevisiae at a concentration of 200 #g.mL (455.38 #M) compared to the standard drug, Fluconazole at a concentration of mg mL (3265 #M) and the h also showed good activity at the concentrations of 50 and 100 #g.mL ChemistrySelect 2020, 5, 6339 – 6349 6341 © 2020 Wiley-VCH Verlag GmbH & Co KGaA, Weinheim ChemistrySelect Full Papers doi.org/10.1002/slct.202000855 Figure The entry b demonstrated good inhibition against B.megaterium at a concentration of 200 #g mL 1(388 #M), moderate activity at a concentration of 100 #g mL 1, and no inhibition at a concentration of 50 #g mL It was compared to Ampicillin at a concentration of mg.mL (5724 #M) Figure The compound j showed moderate activity against B cereus at all concentrations compared to standard drug–Ampicillin at a concentration of mg mL in Table 1, the maximum negative value of the energy of binding at the active sites of a receptor and the docking pose c and an inhibitor constant of it with the receptor 6TZM (6TZM: PDB) were performed –7.03 kcal.mol and 6.98 #M, ChemistrySelect 2020, 5, 6339 – 6349 Figure The most stable conformation c after completion of docking calculation with C.albincans, 6TZM, the value of the lowest negative free energy of binding (ligand-receptor, c–6TZM) of –7.03 kcal.mol and an inhibitor constant, Ki of 6.98 #M were calculated by AutoDockTools-1.5.6rc3 and visualized by Discovery Studio 2019 Client package Figure The active site atoms in a 2D diagram between a receptor 6TZM and the most stable conformation c including the hydrogen bond, Van der Waals, halogen, unfavorable positive–positive, pi–carbon, alkyl, and pi–alkyl interactions respectively They were used to select and analyze the best conformation for docking with a receptor, which embedded in 6342 © 2020 Wiley-VCH Verlag GmbH & Co KGaA, Weinheim ChemistrySelect Full Papers doi.org/10.1002/slct.202000855 Figure The binding sites of the most stable conformation c with active sites of C.albincans, 6TZM, four hydrogen bonds forming between residual amino acids of 6TZM and the best docking poses of conformation c by Discovery Studio 2019 Client package Figure The most stable conformation h after completion of the docking calculation with S cerevisiae, 3ET5, the value of the lowest negative free energy of binding (ligand-receptor, h–3ET5) of –9.59 kcal.mol and an inhibitor constant, Ki of 0.094 #M Figure The ligand map indicated the extra secondary interactions like the hydrogen bonds, steric and overlap interactions between a docking pose c and the active sites of a receptor, 6TZM by Molegro Molecular Viewer package a receptor 6TZM in Figure The values of inhibition constant and the free energy of binding of the best conformation c and receptor–6TZM in Table were calculated and compared to those of the standard drug indicated that the compound c at a concentration of 200 #g.mL (377.78 #M) has stronger inhibition than that of the standard drug The value of the lowest negative of free energy of binding of the best docking pose, c to receptor 6TZM was smaller than that of a drug to the same receptor indicated these bonds of docking pose c to the receptor were more stable bonds than those bonds of docking pose of standard drug, Fluconazole to the receptor The value of free energy of binding (~G) calculated based on the difference between amounts of the total of the Val der ChemistrySelect 2020, 5, 6339 – 6349 Figure 10 The active site atoms between a receptor 3ET5 and the most stable conformation h on a 2D diagram including the secondary interactions as the hydrogen bonds, Van der Waals, carbon-hydrogen bond, pi–donor hydrogen bond, pi–sigma, alkyl, and pi–alkyl Waals forces, hydrogen bond, desolvation, electrostatic, total internal, torsional free energy, and energy of an unbound system.[24] As visualized in Figure 6, the residual amino acids around the most stable conformation c examined as hydrophobic acids: Cys–423 and Ile–421 The Cys–423 formed one hydrogen bond with atom oxygen of the P=O double bond The residual hydrophilic amino acids were observed like Gly– 422, Gly–424, Arg–528, Lys–425, Asn–476, Asp–474, Asp–445, Arg–475, Asn–444, Tyr–628, Asn–446, Gln–443, Thr–427, and 6343 © 2020 Wiley-VCH Verlag GmbH & Co KGaA, Weinheim ChemistrySelect Full Papers doi.org/10.1002/slct.202000855 Figure 11 The binding sites of the most stable conformation h with the active site of 3ET5, one hydrogen bond from residual amino acids of 3ET5 to the best docking pose of conformation h Figure 13 The IC50 values of entries, a–j against breast cancer cell lines, MCF–7 in vitro Figure 14 The IC50 values of entries, a–j against small lung cancer cell lines, A–549 in vitro Figure 12 The ligand map indicated the secondary interactions, which included the hydrogen bonds, steric, and overlap interactions between the best docking pose h and 3ET5 Thr–426 The Gly–422, Gly–424, Lys–425, and Thr–426, the hydrophilic amino acids bound to atom oxygen of P=O double and atom oxygen of O–ethyl group via hydrogen bonds (green lines), respectively Other interactions showed the van der Waals, carbon-hydrogen, halogen, unfavorable positive–positive, pi–cation, alkyl, and pi–alkyl All interactions in Figure determined the total amount of minimum energy of the best conformation c for docking As shown in Figure and Table 1, the docking pose c formed four hydrogen bonds with the receptor 6TZM (6TZM: PDB, the crystal structure of Candida albicans) All those hydrogen bonds started from active sites of residual amino acids as Glyn–422, Gly–424, Lys–425, and Thr– ChemistrySelect 2020, 5, 6339 – 6349 426 of A chain of protein structure, 6TZM to atom oxygen of the double bond of P=O (3 hydrogen bonds) and atom oxygen of the O-ethyl group It demonstrated that the phosphonate group in c formed in the K.F reaction, which conducted excellent inhibition against the receptor of Candida albicans fungus The bond lengths of all most hydrogen bonds, which created from ligand c to receptor were shorter than them forming from standard drug to the same receptor It proved that a shorter length of hydrogen bonds, stronger strength of bonds they formed The ligand map exposed the extra secondary interactions such as the hydrogen bonds, steric, and overlaps Those interactions implied the strength of ligand c and receptor 6TZM interactions The green lines noted effect steric, which conducted the conformation of the molecular binding process The size of yellow and green circles on atoms 6344 © 2020 Wiley-VCH Verlag GmbH & Co KGaA, Weinheim ChemistrySelect Full Papers doi.org/10.1002/slct.202000855 Figure 18 The percentage of inhibition of entries, a–j against Hela in vitro in the range of concentrations from 5–100 #M Figure 15 The IC50 values of entries, a–j against cervical cancer cell lines (HeLa) in vitro Table The significant results of a docking pose c and drug–Fluconazole to Candida albicans fungus, code 6TZM[a] Figure 16 The percentage of inhibition of entries, a–j against cancer cell lines, MCF–7 in vitro in the range of concentrations from 5–100 #M Figure 17 The percentage of inhibition of entries, a–j against cancer cell lines, A–549 in vitro in the range of concentrations from 5–100 #M ChemistrySelect 2020, 5, 6339 – 6349 Entry DG[b] Ki[c] Hydrogen bond[d] Property and bond length[e] 3c -7.03 6.98 Drug[f] -5.09 185.6 A:Gly422:H–3 c:O (2.25) A:Gly424:H–3 c:O (2.03) A:Lys425:H–3 c:O (2.48) A:Thr426:H–3 c:O (2.10) A:Thr426:O–drug:N (2.69) A:Asn444:N–drug:N (3.12) A:Asp445:N–drug:N (2.75) A:Asn476:N–drug:N (3.10) Drug:H–Asp445:O (2.12) [a] The crystal structure of Candida albicans was received from the protein data bank [b] The free energy of binding, receptor-ligand presented in the unit of kcal.mol and calculated by AutoDockTools-1.5.6rc3 package [c] The inhibition constant calculated by AutoDockTools-1.5.6rc3 package and reported in the unit of #M [d] The number of hydrogen bonds was indicated by Discovery Studio 2019 Client package [e] They have visualized in Discovery Studio 2019 Client package and the unit of Angstrom, respectively [f] The structure of Fluconazole was conducted the optimal energy of molecule by Avogadro package via the MMFF94 method and docking calculations identified the strength of overlap interactions and contributed to steric hindrance as shown in Figure As shown in Figure and Table 2, the best docking pose of h against S cerevisiae, 3ET5 has calculated the values of inhibition constant and the free energy of binding of docking, which obtained 0.094 #M and –9.59 kcal.mol 1, respectively They have compared their values to the values of the standard drug The h indicated a stronger inhibitor against Saccharomyces cerevisiae fungus than Fluconazole, owing to the value of lower negative free energy of binding and the inhibition constant, Ki of the most stable conformation h with a receptor benchmarking with the values of Fluconazole in Table 2, respectively As exposed in Figure 2, the compound h showed a higher inhibitor against Saccharomyces cerevisiae at a concentration of 200 #g.mL (455.38 #M) than a drug at a concentration of mg.mL (3265 #M) because of both the IC50 of h and the value of free 6345 © 2020 Wiley-VCH Verlag GmbH & Co KGaA, Weinheim Full Papers doi.org/10.1002/slct.202000855 ChemistrySelect Table The remarkable results of docking pose h and drug, Fluconazole to Saccharomyces cerevisiae fungus, 3ET5 [a] Table The important docking poses b and ampicillin to Bacillus megaterium bacterium, 6NVW code [a] Entry DG Ki Hydrgen bond Property and bond length Entry DG Ki Hydrgen bond Property and bond length 3h Drug[b] -9.59 -5.90 0.094 47.0 C:Ser164:O–3 h:O (3.10) C:Arg147:H–drug:F (2.24) C:Ser164:H–drug:O (2.25) C:Val165:H–Drug:O (1.76) Drug–C:Val165:O (2.04) 3b -8.03 1.30 Drug[b] -8.19 0.99 B:Arg381:H–3 b:O (2.23) b:H–B:Gln23:O (2.42) B:Arg266:H–drug:O (2.45) B:Arg381:H–drug:O (2.20) Drug:H–B:Gln23 (2.15) Drug:H–B:Gln23:O (2.26) Drug:H–B:Gln23:O (2.32) Drug:H–B:Asn462 (2.14) [a] The crystal structure of Saccharomyces cerevisiae fungus took from the protein data bank [b] The structure of Fluconazole was performed the optimal energy of molecule by Avogadro package via force field methodMMFF94 and docking calculations energy of binding of docking pose h and the receptor–3ET5 in silico docking were lower those of Fluconazole as shown in Table Only one hydrogen bond formed from atom oxygen of Ser–164 to atom oxygen of P=O double bond with a bond length of 3.10 Å as shown in Figure 11 and Table It proved the phosphonate group in h showed excellent inhibition against a receptor of Saccharomyces cerevisiae The active site atoms at the active sites of a receptor, 3ET5, and ligand h were conducted on a 2D diagram in Figure 10, which visualized one hydrogen bond (from atom oxygen of Ser–164 of C chain of receptor to atom oxygen of O=P double bond of h and other bonds were non-bonding like the Van der Waals, carbonhydrogen bond, pi–donor–hydrogen bond, pi–sigma, alkyl, and pi–alkyl All those interactions established the minimum negative energy of interaction system between ligand h and receptor 3ET5 as shown in Table and led to the most stable conformation h for good docking at active sites of the protein All residual amino acids made contacts with the active sites of h, which were relative to C chain except for Asn–363 of B chain The ligand map identified extra interactions between h and receptor, 3ET5 in Figure 12 like the hydrogen bonds, steric, and overlap interactions, which controlled the strength of conformation ligand h and receptor 3ET5 The green lines (13 steric interactions) determined the formation of confirmation of h The Van der Waals radius of atoms varied from 1.70 to 2.08 Å that controlled the strength of overlap interactions and participants of steric hindrance in Figure 12 The entry h pointed out an excellent activity inhibitor against Saccharomyces cerevisiae fungus at a concentration of 200 #g.mL or 455.38 #M and compared with the entry g in our former article.[5] The g at a concentration of 25 #M showed higher activity than h at a concentration of 455.38 #M against Saccharomyces cerevisiae It was explained based on docking poses and differential structures Entries h and g bound to hydrogen and fluorine atom of the phenyl group, respectively, and the h had no atom halogen bond In the best conformation g with a receptor in a 2D diagram, the fluorine atom made unfavorable bonds The halogen bonds with residual amino acids nearby like Gln–353, Glu–277, and Asn–350, which led to g was a stronger inhibitor than h at a low concentration, 25 #M As shown in Table 3, the values of b compared to those of standard drug, Ampicillin against Bacillus megaterium, 6NVW indicated that the best conformaChemistrySelect 2020, 5, 6339 – 6349 [a] The crystal structure of Bacillus megaterium bacterium was downloaded from the protein data bank [b] The structure of Ampicillin was calculated the optimal energy of molecule by Avogadro package via force field method- MMFF94 and docking calculations tion b made a weaker inhibitor than a drug, owing to the values of calculated inhibition constant and free energy of binding of the most conformation b with receptor–6NVW presented lower than those of a drug as shown in Table As shown in Table and Figure S.1.13, the docking pose b formed hydrogen bonds, one from Arg–381 of B chain to atom oxygen of P=O double bond and another from atom hydrogen of secondary amine b to atom oxygen of Gln–23 of B chain From the results of docking poses of c, h, and b were briefly presented in Table 1–3 that proved all those hydrogen bonds formed from the active sites on receptors to O=P(OEt)2 groups or hydrogen atom of HN of secondary amine groups via K.F reaction It was very significant to design structures having bioactivities In vitro, a concentration of b against Bacillus megaterium was 388 #M, which was compared to a concentration of Ampicillin of 5724.0 #M was lower than that of Ampicillin, but the diameter of the inhibition zone of b was identical to that of ampicillin drug, which led to inhibition ability of b at a concentration of 200 mg.mL (388 #M) was a potential inhibitor The entry j as shown in Figure 4, indicated moderate activity against B cereus at all concentrations without explanations a docking model for this conformation Anticancer activities The notable results of docking of entries, a–j against three cancer cell lines like MCF–7 (6VNN), A–549 (4ADS), and HeLa (5HES) have been briefly presented in Tables and 6, and other information were reported in supporting information With MCF–7 cancer cell lines 6VNN, based on the calculated results of docking pose of the most stable conformations a–j, the docking abilities of the best conformations a–j were determined and presented in order i > c > h > Drug > f > g > j > b > d > e > a, owing to values of the maximum negative free energy of binding between the docking poses and a receptor of cancer cell lines 6VNN, the inhibitor constants and abilities of formed hydrogen bonds In vitro, the results of test compounds also showed that c, h, and a exposed excellent cytotoxicity activity against MCF–7 6346 © 2020 Wiley-VCH Verlag GmbH & Co KGaA, Weinheim Full Papers doi.org/10.1002/slct.202000855 ChemistrySelect Table The docking poses of entries a–j to active sites of a receptor of breast cancer cell line, MCF–7(6VNN)[a] Table The docking poses of entries a–j to activities sites of a receptor of cervical cancer cell lines, HeLa.[a] Entry DG Ki Hydro-gen bond Property and bond length Entry DG Ki Hydro-gen bond Property and bond length 3a 3b 3c -7.79 -8.53 -9.39 1.94 0.562 0.132 3a 3b 3c -9.60 -9.29 -8.96 0.092 0.154 0.207 0 3d 3e 3f 3g 3h -9.03 -8.56 -9.93 -9.73 8.92 0.239 0.530 0.052 0.074 0.289 0 1 No hydrogen bond b:H–A:Asp473:O (2.04) C:Ala430:N–3 c:O (2.83) C:Ser433:O–3 c:O (2.62) No hydrogen bond No hydrogen bond C:Ser433:O–3 f:O (2.89) C:Ser433:O–3 g:O (2.84) h:H–C:Ser433:O (2.21) D:Lys472:N–3 h:O (2.76) C:Ser433:O–3 h:O (2.67) C:Arg412:N–3 i:F (3.21) C:Ser433:O–3 i: N(3.00) i:H– C:Ser433:O(2.09) j:H - C:Asp426:O(2.43) C:Ser433:O–drug:N (2.80) D:Phe461:N–drug:O(3.12) D:Leu462:N–drug:O (2.74) 3d 3e -8.95 -7.68 0.275 2.36 3f -10.11 0.039 3g 3h 3i -9.60 -9.53 -9.87 0.092 0.104 0.059 3j Drug[b] -8.88 -9.11 0.308 0.208 No hydrogen bond No hydrogen bond A:Gly23:N–3 c:O (2.92) c:H–A:Asp92:O (2.18) A:Lys135:N–3 d:O (2.65) A:Tyr84:O–3 e:O (2.57) A:Tyr84:O–3 e:O (2.69) A:Tyr84:O–3 e:O (2.83) B:Arg284:N–3 e:O (3.03) B:Cys285:N–3 f:N (1.81) f:H–A:Tyr84:O (2.43) No hydrogen bond h:H–A:Arg137:O (2.25) A:Lys135:N–3 i:F (3.18) A:Arg137:N–3 i:F (2.75) A:Arg137:N–3 i:F (3.16) No hydrogen bond A:Thr82:O–drug:O (2.99) Drug:H–A:Thr82:O (2.03) 3i -9.51 0.107 3j Drug[b] -9.58 -8.90 0.096 0.298 [a] The crystal structure of 6VNN (6VNN: PDB,1.63 Å, and DOI: 10.2210/ pdb6VNN/pdb) was downloaded from the protein data bank The most important values were calculated by AutoDockTools-1.5.6rc3 and visualized by DSC The others were indicated in supporting information, Table S.4 [b] The standard drug was Camptothecin Table The docking poses of entries a–j to active sites of a receptor of lung cancer cell, A–549 [a] Entry DG Ki Hydro-gen bond Property and bond length 3a 3b 3c 3d -8.83 -9.86 -9.47 -10.24 0.335 0.059 0.145 0.031 0 3e 3f -8.68 -8.54 0.434 0.550 3g -8.69 0.425 3h -8.47 0.612 3i -8.49 0.597 3j Drug[b] -9.21 -9.38 0.178 0.133 No hydrogen bond No hydrogen bond c:H– A:Leu840:O (2.32) A:Val899:N–3 d:O (2.99) A:Asp1046:N –3 d:N (2.91) No hydrogen bond A:Asn923:N–3 f:O (2.56) A:Asn923:N–3 f: O (2.70) A:Asp1052:N–3 f:N (2.71) f:H–A:Leu840:O (2.16) A:Asn923:N–3 g:O (2.98) g:H–A:Leu840:O (2.05) A:Asn923:N–3 h:O (3.18) h:H–A:Leu840:O (2.15) A:Tyr1082:O–3 i:F (2.80) i:H–A:His1026:O (2.09) No hydrogen bond A:Cys919:N–Drug:O (2.58) [a] The crystal structure of 4ASD (4ASD: PDB, 2.03 Å, and doi.org/10.2210/ pdb4ASD/pdb) was downloaded from the protein data bank The most significant values were performed by AutoDockTools-1.5.6rc3 and visualized by DSC The others were indicated in supporting information in Table S.4 [b] The standard drug was Camptothecin The calculated results of docking pose conducted that the remarkable hydrogen bonds formed from the residual amino acids of a receptor, 6VNN to an oxygen atom of P = O double bond, an oxygen atom of ethoxy groups, hydrogen, and nitrogen atoms of N–H group via K.F’s reaction as shown in Table For docking poses of the most stable conformations of ligands a–j to a receptor 4ASD as shown in Table 5, the ChemistrySelect 2020, 5, 6339 – 6349 [a] The crystal structure of 5HES (5HES: PDB, 2.14 Å, and doi.org/10.2210/ pdb5HES/pdb) was received from the protein data bank The most fundamental values were conducted by AutoDockTools-1.5.6rc3 and visualized by DSC The others were indicated in supporting information in Table S.4 [b] The standard drug was Camptothecin sequence of the most stable conformations, which docked to a receptor described as d > c > drug > g > f > i > h > b > j > a > e To compare the results of the docking model and test in vitro, the h and c are the best solutions and the h has been given to explain the good quantitative structure-activity relationship in the concepts of an inhibitor constant, Ki of 0.61 #M (in silico docking model) and the IC50 value of 2.29 #M (in vitro) as shown in Table and Table For anticancer against cervical cancer cell lines, HeLa, 5HES, the trends of docking poses were indicated as expression, f > i > h > drug > c > d > e > a > b > g > j in Table Based on docking poes, the docking abilities presented as f > i > c and compared to compounds, which showed potential inhibitions in vitro like f > c > i The f has Table The IC50 values of entries against MCF–7, A–549, HeLa, and drug– Camptothecin in vitro Entries Cancer cell lines IC50[a] MCF-7 A549 HeLa 3a 3b 3c 3d 3e 3f 3g 3h 3i 3j Drug[b] 9.78 0.93 45.94 0.66 9.39 0.64 124.78 0.63 613.99 0.39 44.45 1.45 120.96 1.27 6.38 022 41.61 0.23 22.14 0.04 2.84 1.33 88.40 0.36 629.56 2.45 7.32 0.07 767.03 2.35 3.82 0.01 10486.9 2.31 52922.39 1.61 2.29 0.02 123.07 0.05 27.58 0.81 0.58 0.17 22.41 0.35 23.89 0.40 5.98 0.02 16.59 0.20 18.47 0.31 5.16 0.04 102.90 048 16.33 0.26 7.19 0.02 115.89 0.31 2.15 0.32 [a] The values were in #M [b] The standard drug was Camptothecin 6347 © 2020 Wiley-VCH Verlag GmbH & Co KGaA, Weinheim ChemistrySelect Full Papers doi.org/10.1002/slct.202000855 explained the equivalent result in vitro and in silio docking model Except for a style of hydrogen bonds as explaining above, with the most stable conformation i, three hydrogen bonds formed from Lys 135, Arg 137 of A chain of a receptor, 5HES to F atoms of CF3 group, which bound to a phenyl group at 3-position as shown in Table Conclusions The new analog +–amino phosphate series were performed the green synthesis by PEG–400 as a green catalyst via Kabachnik– Fields reaction based on carbazole screened antimicrobial and cytotoxicity activities and conducted in silico molecular docking model The yield of reactions obtained from 85–91% (after completion of chromatography column) in a few hours by the effectively green catalyst and isolated entries The entry c and h exposed excellent inhibition against Candida albicans and Saccharomyces cerevisiae fungi at the same concentration of 200 #g.mL and obtained higher inhibitors than Fluconazole at mg.mL The entry b showed moderate activity against Bacillus megaterium bacterium at a concentration of 200 #g.mL The results of docking study of some active compounds indicated the reasonable explanations between the bioactivities in vitro of c, h, and b and their docking poses to receptors as activity expresses for antifungal activity: c and h > Fluconazole, for antibacterial activity: b < Ampicillin The entry c gave an excellent inhibitor against Candida albicans in vitro and in silico molecular docking that was a new result The h made benchmarking with g for antifungus, Saccharomyces cerevisiae in our former article.[5] The h made a weaker inhibitor than g This molecular docking model proved that the hydrogen bonds also formed from active sites of receptors to a diethyl phosphate group or an atom hydrogen of the N–H bond of secondary a amine group, which was established by K.F’s reaction It was fundamental to design activity structures The compounds c, f, and i pointed out excellent cytotoxicity against HeLa cancer cell lines and reported the new results in vitro The IC50 value of f against HeLa cancer cell lines in vitro and an inhibitor constant, Ki was equivalent among the highest activity compounds In docking study against cancer cell lines, the novel docking results exposed the hydrogen bonds only formed from almost the residual amino acids of receptors to an oxygen atom of O=P double bond, an oxygen atom of an ethoxy group, and a nitrogen or hydrogen atom of N–H group of (N–HP=O (OEt)2 group), which yielded via K.F reaction, or they formed from the residual amino acids of receptors to fluorine atoms of CF3 groups in i The a, c, and h showed excellent cytotoxicity against MCF–7 cancer cell lines The c, e, and h indicated highest inhibitions against A-549 cancer cell lines in vitro The next article will design, synthesize, screen in vitro and in silico bioactivities of new +-amino phosphate derivatives via Kabachnik-Fields using PEG–400 as a green media ChemistrySelect 2020, 5, 6339 – 6349 Supporting Information Summary The supporting information contains procedures of synthetic compounds of target compounds 3a-j, intermediate compounds, 1–2, and brief procedure to reuse PEG-400 catalyst The IR, 1H, 13C, and 31P NMR of compounds were presented in figures and explained spectral data The bioactivities assays were performed and the results of antimicrobial activities were shown in tables The procedure of molecular docking was briefly presented and the calculated results of a docking pose of structure 3b, the standard drug-Fluconazole, and Ampicillin with receptors were found, visualized, and indicated the key results in tables and figures Acknowledgments This research was an institute project that was supported by the research fund from Industrial University of Ho Chi Minh City, Ho Chi Minh City Vietnam (20/1.5 CNH03) Conflict of Interest The authors declare no conflict of interest Keywords: Amino acids · Antiproliferation · Green chemistry · Multicomponent reactions · 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tổng kết phương pháp tổng hợp hoạt tính sinh học với tần suất cao, dễ dàng tổng hợp, ... cứu Trang 55 PHẦN I THƠNG TIN CHUNG I Thơng tin tổng qt 1.1 Tên đề tài: Tổng hợp xanh số aminophosphate dựa carbazole hoạt tính sinh học 1.2 Mã số: 20/1.5CNH03 1.3 Danh sách chủ trì, thành viên... hợp, cô lập hiệu suất cao, hoạt tính sinh học đa dạng tiêu biểu[5,9-10] Các hoạt tính sinh học mà chúng tơi thực nghiên cứu có tính tiềm cao việc nghiên cứu tổng hợp số thuốc hữu phosphote có