Alkynyl-substitutedanalogues were prepared using three key steps, nucleophilic aromaticsubstitution with a 3-aminoalanine derivative, benzotriazole formation via a one-pot diazotization and cyclization process, anda Sonogashira cross-coupling reaction. E-Alkenyl-substitutedbenzotriazoles were accessed by stereoselective partial hydrogenationof the alkynes using zinc iodide and palladium catalysis. The alkynylanalogues were found to possess higher quantum yields and strongerbrightness and, a solvatochromic study with the most fluorogenic α-aminoacids demonstrated sensitivity to polarity.Fluorescent unnaturalα-amino acids are widelyused as probesin chemical biology and medicinal chemistry. While a variety of structuralclasses have been developed, there is still a requirement for newenvironmentally sensitive analogues that can closely mimic proteinogenicα-amino acids. Here, we report the synthesis and fluorescentproperties of highly conjugated, benzotriazole-derived α-aminoacids designed to mimic  To imp11 including the preparation of benzotriazole-derived α-aminoacids, as potential structural mimics of l-tryptophan.12 In a previous study, a Suzuki–Miyauracross-coupling reaction was used to extend the conjugation of thebenzotriazole side chain allowing access to 5-aryl analogues suchas compound 4  with 3-aminoalanine derivative 5. Following reductionof the nitro group, a one-pot diazotization and cyclization reactionunder mild conditions would yield 5-iodobenzotriazole 8. Sonogashira coupling of 8 with a range of aryl-substitutedalkynes would result in the preparation of the alkynyl targets. Stereo-and chemoselective reduction of the alkynyl compounds would then allowthe rapid synthesis of the second set of targets, the styryl analogues.As shown in N-Boc-l-3-aminoalanine α-methyl ester 5 were preparedfrom commercially available N-Boc-l-asparagine 9 as previously described by Piantanida and co-workers  usingtriethylamine gave adduct 10 in 78% yield. Chemoselectivenitro group reduction of 10 was then performed usingzinc and acetic acid. Reduction under mild acidic conditions and witha short reaction time of 0.75 h allowed full conversion, while maintainingboth the C–I bond and Boc-group protection of the amine. Thisgave aniline 7 in 91% yield. The one-pot activation ofthe amine as the diazo intermediate and in situ cyclizationto benzotriazole 8 was performed as previously describedby us.14 Formation of the diazo intermediate wasachieved under mild conditions, using a polymer-supported nitritereagent and p-tosic acid.16 The one-pot reaction was complete after 1 h and gave benzotriazole 8 in 73% yield. Again, the integrity of the Boc-protectinggroup was not affected by the acidic conditions. The synthesis ofthe α-amino acids with extended alkynyl-fused benzotriazolechromophores was then achieved using a Sonogashira reaction of iodobenzotriazole 8 with various electron-rich and electron-deficient aryl-substitutedacetylenes. Under standard conditions, this gave the coupled products 11a–f in 70–95% yields.18 Crucial to the success of the coupling reactions was the requirementof an initiation phase at high temperature (100 °C for 0.1 h),before conducting the remainder of the reaction at room temperature.Initial heating allows efficient formation of the active palladium(0)species from the pre-catalyst, while returning to room temperaturefor the coupling step prevents extensive Glaser–Hay couplingof the terminal alkyne reagent.19 Deprotectionto the parent α-amino acids was then performed using a two-stepapproach. Ester hydrolysis with cesium carbonate was followed by theremoval of the Boc-group under acidic conditions. Despite the presenceof electron-rich alkynes, acidic deprotection proceeded cleanly, withoutany significant byproducts. Purification by recrystallization gavethe amino acid hydrochloride salts 12a–f in good overall yields.Initially, gram quantities of -workers 2.13 The 8 and although successful,competing protodepalladation during the reaction resulted in a lowyield of the desired product (32%). A second attempt involved theSuzuki–Miyaura cross-coupling of 2-arylvinylboronic acids witha bromo-analogue of 8. However, this again gave the alkenyl-coupledbenzotriazole in low yield (25%). For these reasons, the reductionof the alkynyl-fused benzotriazoles 11 was consideredas a possible approach to the corresponding alkenyl analogues. Fromthe range of literature methods available, a chemo- and stereoselectivehydrogenation procedure at atmospheric pressure, reported by Jackowskiand co-workers, was considered.20 Thisreaction involves the combination of a palladium catalyst and zinc(II)iodide, which promotes syn-hydrogenation, followedby Z- to E-isomerization. Attemptedreduction of alkyne 11b using the standard conditions(5 mol % of Pd cat. and 25 °C) required a reaction time of 112h and gave a 2:1 mixture of E- and Z-alkenes isomers (Table S1). Optimizationstudies showed that by increasing both the amount of palladium catalyst(20 mol %) and reaction temperature (40 °C), the reaction timefor reduction of 11b could be reduced to 20 h, to givesolely E-isomer 13b in 64% yield  in similar yields. Ester hydrolysis with cesium carbonateand mild acid removal of the Boc-group gave the deprotected aminoacids in high overall yields.To access the corresponding alkenylanalogues, various approacheswere attempted. These included the Heck cross-coupling reaction ofstyrene with iodobenzotriazole 4% yield 3. Having4 (21 The ultraviolet/visible (UV/visible) absorptionand photoluminescence spectra of the α-amino acids were recordedin methanol at a concentration of 5 μM. As proposed, the extendedchromophores all displayed red-shifted absorption. Direct comparisonof the p-methoxy analogues shows absorption bandsat 321 nm for alkyne 12b and 320 nm for alkene 14bvs 256 nm for 5-aryl analogue 4. Strong fluorescence with emission maxima in the visible regionwas observed for benzotriazoles containing electron-rich aryl sidechains.22 In addition, the alkynyl seriesof compounds displayed the most favorable properties. For example, p-methoxyphenyl 12b possessed a quantum yieldof 34% and was significantly brighter than both structural analogues 4 and 14b. These results suggest that the aryl-substitutedalkynyl-benzotriazoles can readily adopt a flat conformation whichallows effective conjugation across the chromophore, compared to thebiaryl (4) and alkenyl (14b) systems thatcan relax via nonradiative decay pathways due toextra rotational and vibrational modes.23 In addition to the strong fluorescence of electron-rich analogues,naphthyl-substituted alkyne 12d possessed the highestquantum yield (42%) and good brightness. Overall, the strategy ofinserting alkynyl and alkenyl spacer units to extend the chromophoreshas yielded compounds with improved photophysical properties, particularlyabsorption maxima that no longer overlap with proteinogenic aminoacids. It should be noted that while this has resulted in compoundswith smaller Stokes shifts than the original series, these are stillsignificantly large .Following the synthesis of the alkynyl-and alkenyl-fused benzotriazole-derivedα-amino acids, the optical properties were measured for eachcompound and compared with 5-aryl analogue 4 1.12,21 Tp-methoxyphenyl- and naphthyl-substitutedalkynyl-fused benzotriazoles 12b and 12d were found to be the brightest α-amino acids, the propertiesof these compounds were further explored via a solvatochromicstudy.24 In contrast to the absorptionbands of both compounds, which were found to be independent of solventpolarity (see the Supporting Information), the emission maxima displayed significant bathochromic shift withincreasing polarity .25 The linearity of these plots confirmsthe general effect of solvent in the shift of emission bands. Boththe bathochromic shift of emission in aqueous solvents and the environmentsensitivity of these amino acids suggests potential as probes forchemical biology applications.As the polarity 4. For exE-alkeneswas achieved by a chemo- and stereoselective, palladium-catalyzedhydrogenation reaction. Investigation of the photophysical propertiesof both classes of α-amino acids revealed that extended conjugationresulted in compounds with red-shifted absorption bands. This meansthese compounds can be excited in the presence of fluorescent proteinogenicα-amino acids. The majority of compounds demonstrated strongfluorescent properties and large Stokes shifts, with the alkyne seriespossessing the highest quantum yields and brightest chromophores.A solvatochromic study with the brightest α-amino acids showedsignificant environment sensitivity to solvent polarity. Work is currentlyunderway to investigate the use of compounds 12b and 12d as probes in chemical biology applications.In summary, a series of alkyne-fused benzotriazole-derivedα-aminoacids have been prepared using nucleophilic aromatic substitutionwith a 3-aminoalanine derivative, a one-pot diazotization, and cyclizationto access the benzotriazole unit and a Sonogashira cross-couplingreaction for the introduction of the unsaturated side chain as thekey steps. Access to the corresponding 5 has been previously describedin the literature.13 All reagents and startingmaterials were obtained from commercial sources and used as received.Reactions were performed open to air unless otherwise mentioned. Allreactions performed at elevated temperatures were heated using anoil bath. Brine refers to a saturated aqueous solution of sodium chloride.Flash column chromatography was performed using silica gel 60 (40–63μm). Aluminum-backed plates precoated with silica gel 60F254 were used for thin-layer chromatography and were visualizedwith a UV lamp or by staining with potassium permanganate, vanillin,or ninhydrin. 1H NMR spectra were recorded on an NMR spectrometerat either 400 or 500 MHz and data are reported as follows: chemicalshift in ppm relative to the solvent as internal standard , multiplicity . The abbreviations br s and br d refer to broad singletand broad doublet, respectively. 13C NMR spectra were recordedon an NMR spectrometer at either 101 or 126 MHz and data are reportedas follows: chemical shift in ppm relative to tetramethylsilane orthe solvent as internal standard , multiplicity with respect to hydrogen . Infraredspectra were recorded on a Fourier transform infrared (FTIR) spectrometer;wavenumbers are indicated in cm–1. Mass spectrawere recorded using electrospray techniques. High-resolution massspectra (HRMS) were recorded using quadrupole time-of-flight (Q-TOF)mass spectrometers. Melting points are uncorrected. Optical rotationswere determined as solutions irradiating with the sodium D line (λ= 589 nm) using a polarimeter. [α]D values are givenin units 10–1 deg cm–1 g–1. UV–vis and fluorescence spectra were recorded on a fluorescenceand absorbance spectrometer. Absorbance spectra were recorded withan integration time of 0.05 s and a band pass of 5 nm. Fluorescencespectra were recorded with excitation and emission band pass of 5nm, an integration time of 2 s, and with detector accumulations setto 1. Quantum yield data were measured using anthracene and l-tryptophan as standard references.The synthesis of compound S)-2-[(tert-butoxycarbonyl)amino]-3-aminopropanoate(5)  in acetonitrile (50 mL) underargon were added 2-fluoro-5-iodonitrobenzene (6)  and triethylamine . The reactionmixture was stirred under reflux for 18 h. After cooling the reactionto ambient temperature, the solvent was removed in vacuo. The resulting residue was dissolved in ethyl acetate (50 mL) andwashed with water (3 × 50 mL) and brine (50 mL). The organiclayer was dried (MgSO4), filtered, and concentrated in vacuo. Purification by flash column chromatography elutingwith 0–10% ethyl acetate in dichloromethane gave methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-[(2′-nitro-4′-iodophenyl)amino]propanoate(10) as a yellow solid . Mp 103–105°C; IR (neat) 3364, 2978, 1744, 1709, 1611, 1500, 1233, 1159cm–1; [α]D21 +51.5 ; 1H NMR : δ 8.46 , 8.22 , 7.66 , 6.80 , 5.37 , 4.62–4.54 , 3.81 , 3.79–3.72, 3.68 , 1.45 ; 13C{1H} NMR :δ 170.7 (C), 155.2 (C), 144.4 (C), 144.3 (CH), 134.9 (CH), 133.5(C), 115.9 (CH), 80.7 (C), 75.3 (C), 53.0 (CH and CH3),44.8 (CH2), 28.3 (3 × CH3); MS (ESI) m/z 488 ; HRMS(ESI) m/z: [M + Na]+ calcdfor C15H20IN3O6Na 488.0289;found 488.0290.To a solution of methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-[(2′-nitro-4′-iodophenyl)amino]propanoate(10)  in methanol (40 mL) were addedzinc  and acetic acid . Thereaction mixture was stirred for 0.75 h at room temperature, thenfiltered through Celite, and concentrated in vacuo. Purification by flash column chromatography eluting with 10% ethylacetate in dichloromethane gave methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-[(4′-iodo-2′-aminophenyl)amino]propanoate(7) as a brown solid . Mp 176–180°C; IR (neat) 3348, 2987, 1795, 1697, 1503, 1395, 1250, 1066cm–1; [α]D18 +24.7 ; 1H NMR : δ 7.06 , 6.99 , 6.40 , 5.41 , 4.62–4.53 , 3.76 , 3.52, 3.44–3.31 , 1.44 ; 13C{1H} NMR : δ 171.6 (C), 155.6 (C), 136.6 (C), 136.5 (C), 129.1(CH), 124.7 (CH), 114.1 (CH), 80.9 (C), 80.5 (C), 53.6 (CH), 52.7(CH3), 46.5 (CH2), 28.3 (3 × CH3); MS (ESI) m/z 458 ; HRMS (ESI) m/z:[M + Na]+ calcd for C15H22IN3O4Na 458.0547; found 458.0545.To a solution of methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-[(4′-iodo-2′-aminophenyl)amino]propanoate(7)  in ethyl acetate (35 mL) at−10 °C was added p-toluenesulfonic acid and polymer-supported nitrite . The reaction mixturewas stirred for 1 h, filtered, and the resin washed with ethyl acetate(50 mL). The organic layer was washed with a saturated solution ofaqueous sodium hydrogen carbonate (50 mL) and brine (50 mL), dried(MgSO4), filtered, and concentrated in vacuo. Purification by flash column chromatography eluting with 0–5%ethyl acetate in dichloromethane gave methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-(5′-iodo-1H-benzo[d][1.2.3]triazol-1′-yl)propanoate(8) as a gray solid . Mp 113–116°C; IR (neat) 3676, 2972, 1701, 1395, 1250, 1163, 1066 cm–1; [α]D19 +21.9 ; 1H NMR : δ 8.43 , 7.73 , 7.32 , 5.31, 5.09 , 4.77 , 3.78, 1.42 ; 13C{1H} NMR : δ 169.5 (C), 155.0 (C), 147.5 (C), 136.2(CH), 133.3 (C), 129.1 (CH), 111.0 (CH), 87.6 (C), 80.8 (C), 53.9(CH), 53.2 (CH3), 48.8 (CH2), 28.2 (3 ×CH3); MS (ESI) m/z 469; HRMS (ESI) m/z: [M + Na]+ calcd for C15H19IN4O4Na 469.0343; found 469.0359.To a solution of methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-(5′-iodo-1H-benzo[d][1.2.3]triazol-1′-yl)propanoate(8)  in N,N′-dimethylformamide(3 mL) were added copper iodide  and bis(triphenylphosphine)palladium(II)dichloride . Phenylacetylene  was dissolved in degassed triethylamine (7 mL) and added tothe reaction mixture. The solution was heated to 100 °C for 0.1h and stirred at room temperature for 2 h. The solution was concentrated in vacuo, dissolved in ethyl acetate (20 mL), washed withwater (5 × 10 mL) and brine (2 × 10 mL), dried (MgSO4), and concentrated in vacuo. Purificationby flash column chromatography eluting with 0–5% ethyl acetatein dichloromethane gave methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-[5′-(phenylethynyl)-1H-benzo[d][1.2.3]triazol-1′-yl]propanoate(11a) as a yellow oil . IR (neat) 3357,2978, 2360, 1745, 1708, 1498, 1162, 756 cm–1; [α]D19 +21.1 ; 1H NMR : δ 8.43 , 7.63 , 7.57 , 7.50, 7.39–7.34 , 5.39, 5.12 , 4.79 , 3.78, 1.43 ; 13C{1H} NMR : δ 169.5 (C), 155.0 (C), 145.6 (C), 133.5(C), 131.7 (2 × CH), 131.2 (CH), 128.5 (CH), 128.4 (2 ×CH), 123.3 (CH), 122.9 (C), 119.4 (C), 109.5 (CH), 89.5 (C), 88.7(C), 80.7 (C), 53.9 (CH), 53.2 (CH3), 48.7 (CH2), 28.2 (3 × CH3); MS (ESI) m/z 443 ; HRMS (ESI) m/z: [M + Na]+ calcd for C23H24N4O4Na 443.1690; found 443.1687.To a solution of methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-{5′-[(4‴-methoxyphenyl)ethynyl]-1H-benzo[d][1.2.3]triazol-1′-yl}propanoate(11b) was synthesized as described for 11a using methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-(5′-iodo-1H-benzo[d][1.2.3]triazol-1′-yl)propanoate(8) , N,N′-dimethylformamide(7 mL), copper iodide , bis(triphenylphosphine)palladium(II)dichloride , 4-methoxyphenylacetylene , and triethylamine (14 mL). Purification by flash columnchromatography eluting with 5% ethyl acetate in dichloromethane gavemethyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-{5′-[(4‴-methoxyphenyl)ethynyl]-1H-benzo[d][1.2.3]triazol-1′-yl}propanoate(11b) as a yellow solid . Mp 155–158°C; IR (neat) 3369, 2976, 2357, 1748, 1714, 1605, 1514, 1250,1171, 1033, 833 cm–1; [α]D18 +17.3 ; 1H NMR :δ 8.19 , 7.61 ,7.53–7.47 , 6.90 ,5.32 , 5.11 , 4.79 , 3.84, 3.78 , 1.43 ; 13C{1H}NMR : δ 169.6 (C), 159.8 (C), 155.0(C), 145.7 (C), 133.4 (C), 133.1 (2 × CH), 131.2 (CH), 123.0(CH), 119.8 (C), 115.0 (C), 114.1 (2 × CH), 109.4 (CH), 89.6(C), 87.4 (C), 80.7 (C), 55.3 (CH3), 53.9 (CH), 53.2 (CH3), 48.7 (CH2), 28.3 (3 × CH3);MS (ESI) m/z 473 ; HRMS (ESI) m/z: [M + Na]+ calcd for C24H26N4O5Na 473.1795; found 473.1792.Methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-{5′-[(4‴-dimethylaminophenyl)ethynyl]-1H-benzo[d][1.2.3]triazol-1′-yl}propanoate(11c) was synthesized as described for 11a using methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-(5′-iodo-1H-benzo[d][1.2.3]triazol-1′-yl)propanoate(8) , N,N′-dimethylformamide(18 mL), copper iodide , bis(triphenylphosphine)palladium(II)dichloride , 4-dimethylaminophenylacetylene, and triethylamine (42 mL). Purification by flashcolumn chromatography eluting with 80% diethyl ether in hexane gavemethyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-{5′-[(4‴-dimethylaminophenyl)ethynyl]-1H-benzo[d][1.2.3]triazol-1′-yl}propanoate(11c) as a yellow oil . IR (neat) 3318,2927, 2207, 1746, 1709, 1606, 1510, 1365, 1164, 757 cm–1; [α]D17 +12.9 ; 1HNMR : δ 8.17 , 7.60 , 7.53–7.47 , 6.90, 5.33 , 5.10 , 4.79 , 3.77 , 3.00 ,1.43 ; 13C{1H} NMR : δ 169.6 (C), 155.0 (C), 150.3 (C), 145.8 (C), 133.1(C), 132.8 (2 × CH), 131.3 (CH), 122.5 (CH), 120.4 (C), 111.9(2 × CH), 109.6 (C), 109.3 (CH), 90.9 (C), 86.7 (C), 80.7 (C),53.9 (CH), 53.1 (CH3), 48.7 (CH2), 40.2 (2 ×CH3), 28.3 (3 × CH3); MS (ESI) m/z 486 ; HRMS(ESI) m/z: [M + Na]+ calcdfor C25H29N5O4Na 486.2112;found 486.2114.Methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-{5′-[ethynyl]-1H-benzo[d][1.2.3]triazol-1′-yl}propanoate(11d) was synthesized as described for 11a using methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-(5′-iodo-1H-benzo[d][1.2.3]triazol-1′-yl)propanoate(8) , N,N′-dimethylformamide(9 mL), copper iodide , bis(triphenylphosphine)palladium(II)dichloride , 2-ethynylnapthalene , and triethylamine (21 mL). The reaction mixture was stirredat room temperature for 4 h. Purification by flash column chromatographyeluting with 40% ethyl acetate in hexane gave methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-{5′-[ethynyl]-1H-benzo[d][1.2.3]triazol-1′-yl}propanoate(11d) as a yellow solid . Mp 180–182°C; IR (neat) 2980, 2360, 1746, 1708, 1503, 1366, 1163, 753 cm–1; [α]D21 +5.5 ; 1H NMR : δ 8.24 , 8.08 , 7.84–7.79 , 7.67 , 7.60 , 7.53–7.47 , 5.45 , 5.11 , 4.79 , 3.78 , 1.44 ; 13C{1H} NMR : δ169.6 (C), 155.1 (C), 145.6 (C), 133.6 (C), 133.0 (C), 132.9 (C),131.6 (CH), 128.3 (CH), 128.1 (CH), 127.8 (3 × CH), 126.8 (CH),126.7 (CH), 123.3 (CH), 120.2 (C), 119.5 (C), 109.6 (CH), 90.0 (C),89.1 (C), 80.7 (C), 54.0 (CH), 53.2 (CH3), 48.7 (CH2), 28.3 (3 × CH3); MS (ESI) m/z 493 ; HRMS (ESI) m/z: [M + Na]+ calcd for C27H26N4O4Na 493.1846; found493.1844.Methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-{5′-[(4‴-fluorophenyl)ethynyl]-1H-benzo[d][1.2.3]triazol-1′-yl}propanoate(11e) was synthesized as described for 11a using methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-(5′-iodo-1H-benzo[d][1.2.3]triazol-1′-yl)propanoate(8) , N,N′-dimethylformamide(6 mL), copper iodide , bis(triphenylphosphine)palladium(II)dichloride , 4-fluorophenylacetylene , and triethylamine (14 mL). Purification by flash columnchromatography eluting with 0–5% ethyl acetate in dichloromethanegave methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-{5′-[(4‴-fluorophenyl)ethynyl]-1H-benzo[d][1.2.3]triazol-1′-yl}propanoate(11e) as an off-white solid . Mp 145–147°C; IR (neat) 3358, 2976, 2359, 1748, 1709, 1510, 1221, 1157,835 cm–1; [α]D18 +21.4 ; 1H NMR : δ 8.21 , 7.61 , 7.51–7.38, 7.07 , 5.33 , 5.12 ,4.79 , 3.78 , 1.43; 13C{1H} NMR : δ 169.5 (C), 162.7 , 155.0 (C), 145.6 (C), 133.6 ,133.5 (C), 131.1 (CH), 123.3 (CH), 119.2 (C), 119.1 , 115.8 , 109.6 (CH), 88.5 (C), 88.4 , 80.8 (C), 53.9 (CH), 53.2 (CH3), 48.7 (CH2), 28.2 (3 × CH3);MS (ESI) m/z 461 ; HRMS (ESI) m/z: [M + Na]+ calcd for C23H23FN4O4Na 461.1596; found 461.1592.Methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-{5′-[(4‴-cyanophenyl)ethynyl]-1H-benzo[d][1.2.3]triazol-1′-yl}propanoate(11f) was synthesized as described for 11a using methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-(5′-iodo-1H-benzo[d][1.2.3]triazol-1′-yl)propanoate(8) , N,N′-dimethylformamide(9 mL), copper iodide , bis(triphenylphosphine)palladium(II)dichloride , 4-cyanophenylacetylene , and triethylamine (21 mL). Purification by flashcolumn chromatography eluting with 5% ethyl acetate in dichloromethanegave methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-{5′-[(4‴-cyanophenyl)ethynyl]-1H-benzo[d][1.2.3]triazol-1′-yl}propanoate(11f) as a yellow solid . Mp 110–115°C; IR (neat) 3361, 2984, 2230, 1747, 1690, 1519, 1252, 1155,1109, 837 cm–1; [α]D22 +6.1 ; 1H NMR : δ8.26 , 7.69–7.62 , 7.54 , 5.33 , 5.13 , 4.79 , 3.80 , 1.43 ; 13C{1H} NMR: δ 169.6 (C), 155.1 (C), 145.6 (C),134.1 (C), 132.3 (4 × CH), 131.2 (CH), 128.0 (C), 124.1 (CH),118.6 (C), 118.4 (C), 111.9 (C), 109.9 (CH), 93.2 (C), 88.0 (C), 80.9(C), 54.0 (CH), 53.4 (CH3), 48.9 (CH2), 28.4(3 × CH3); MS (ESI) m/z 446 ; HRMS (ESI) m/z: [M + H]+ calcd for C24H23N5O4H 446.1823; found 446.1832.Methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-[5′-(phenylethynyl)-1H-benzo[d][1.2.3]triazol-1′-yl]propanoate(11a)  in a mixture of methanol(6 mL) and 1,4-dioxane (6 mL) was added a solution of cesium carbonate in water (3 mL). The reaction mixture was stirredat room temperature for 20 h and then concentrated in vacuo. The resulting residue was dissolved in water (50 mL) and acidifiedto pH 1 with 2 M aqueous hydrochloric acid. The aqueous layer wasextracted with dichloromethane (3 × 30 mL), and the combinedorganic layers were washed with water (30 mL), dried (MgSO4), and concentrated in vacuo to give (2S)-2-[(tert-butoxycarbonyl)amino]-3-[5′-(phenylethynyl)-1H-benzo[d][1.2.3]triazol-1′-yl]propanoicacid as a yellow solid . This was used for the nextreaction without any further purification. To a solution of (2S)-2-[(tert-butoxycarbonyl)amino]-3-[5′-(phenylethynyl)-1H-benzo[d][1.2.3]triazol-1′-yl]propanoicacid  in acetonitrile (0.1 mL) was added 2 Maqueous hydrochloric acid (3 mL). The reaction mixture was stirredat room temperature for 6 h and then concentrated in vacuo. Trituration with chloroform gave (2S)-2-amino-3-[5′-(phenylethynyl)-1H-benzo[d][1.2.3]triazol-1′-yl]propanoicacid hydrochloride (12a) as an off-white solid . Mp 245–247 °C (decomposition); IR (neat) 2920,2359, 1732, 1472, 1338, 686 cm–1; [α]D18 +4.9 ; 1H NMR :δ 8.22 , 7.85 , 7.75 , 7.61–7.51, 7.45–7.35 , 5.37 , 5.27 ,4.77 ; 13C{1H} NMR : δ 167.6 (C), 145.4(C), 133.2 (C), 131.3 (CH), 131.2 (2 × CH), 128.4 (CH), 128.3(2 × CH), 122.7 (C), 122.0 (CH), 120.0 (C), 110.3 (CH), 89.3(C), 87.8 (C), 52.2 (CH), 46.8 (CH2); MS (ESI) m/z 307 ; HRMS(ESI) m/z: [M + H]+ calcdfor C17H14N4O2H 307.1190;found 307.1191.To a solution of methyl(2S)-2-Amino-3-{5′-[(4‴-methoxyphenyl)ethynyl]-1H-benzo[d][1.2.3]triazol-1′-yl}propanoicacid hydrochloride (12b) was prepared as described for 12a using methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-{5′-[(4‴-methoxyphenyl)ethynyl]-1H-benzo[d][1.2.3]triazol-1′-yl}propanoate(11b)  and cesium carbonate . This gave (2S)-2-[(tert-butoxycarbonyl)amino]-3-{5′-[(4‴-methoxyphenyl)ethynyl]-1H-benzo[d][1.2.3]triazol-1′-yl}propanoicacid  as a yellow solid. This was used for the nextreaction without any further purification. To a solution of (2S)-2-[(tert-butoxycarbonyl)amino]-3-{5′-[(4‴-methoxyphenyl)ethynyl]-1H-benzo[d][1.2.3]triazol-1′-yl}propanoicacid  in dioxane (2 mL) was added 2 M aqueoushydrochloric acid (0.8 mL). The reaction mixture was stirred at roomtemperature for 24 h and concentrated in vacuo. Purificationby recrystallization from methanol and chloroform gave (2S)-2-amino-3-{5′-[(4‴-methoxyphenyl)ethynyl]-1H-benzo[d][1.2.3]triazol-1′-yl}propanoicacid hydrochloride (12b) as an off-white solid . Mp 274–276 °C (decomposition); IR (neat) 3395,2932, 2214, 1728, 1605, 1512, 1250, 1173, 1026, 826 cm–1; [α]D17 +6.0 ; 1H NMR : δ 8.13 , 7.96 ,7.72 , 7.54 , 7.01 ,5.22 , 5.17 , 4.58 , 3.81 ; 13C{1H} NMR : δ 168.3 (C), 159.7 (C), 145.2(C), 133.3 (C), 133.1 (2 × CH), 130.7 (CH), 121.9 (CH), 118.7(C), 114.5 (C), 114.0 (2 × CH), 110.5 (CH), 89.4 (C), 87.8 (C),55.3 (CH3), 52.0 (CH), 47.2 (CH2); MS (ESI) m/z 359 ; HRMS(ESI) m/z: [M + Na]+ calcdfor C18H16N4O3Na 359.1115;found 359.1105.(2S)-2-Amino-3-{5′-[(4‴-dimethylaminophenyl)ethynyl]-1H-benzo[d][1.2.3]triazol-1′-yl}propanoicacid hydrochloride (12c) was prepared as for 12a using methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-{5′-[(4‴-dimethylaminophenyl)ethynyl]-1H-benzo[d][1.2.3]triazol-1′-yl}propanoate(11c)  and cesium carbonate . This gave (2S)-2-[(tert-butoxycarbonyl)amino]-3-{5′-[(4‴-dimethylaminophenyl)ethynyl]-1H-benzo[d][1.2.3]triazol-1′-yl}propanoicacid  as an off-white solid. This was used for thenext step without further purification. To a solution of (2S)-2-[(tert-butoxycarbonyl)amino]-3-{5′-[(4‴-dimethlaminophenyl)ethynyl]-1H-benzo[d][1.2.3]triazol-1′-yl}propanoicacid  in acetonitrile (0.05 mL) was added 1M aqueous hydrochloric acid (1 mL). The reaction mixture was stirredat room temperature for 0.5 h and concentrated in vacuo. Purification by trituration with chloroform gave (2S)-2-amino-3-{5′-[(4‴-dimethylaminophenyl)ethynyl]-1H-benzo[d][1.2.3]triazol-1′-yl}propanoicacid hydrochloride (12c) as a yellow oil ;IR (neat) 3406, 2920, 2361, 1740, 1694, 1508, 1211, 1130, 841 cm–1; [α]D19–4.5 ; 1H NMR : δ 8.24 ,7.89 , 7.77 , 7.74 , 7.59 , 5.40 , 5.29 , 4.78 , 3.27 ; 13C{1H} NMR :δ 167.8 (C), 145.2 (C), 143.6 (C), 133.2 (2 × CH), 131.4(CH), 130.9 (C), 122.3 (CH), 119.6 (2 × CH), 111.5 (C), 110.6(CH), 109.7 (C), 89.5 (C), 87.8 (C), 52.2 (CH), 46.8 (CH2), 44.9 (2 × CH3); MS (ESI) m/z 350 ; HRMS (ESI) m/z: [M + H]+ calcd for C19H19N5O2H 350.1612; found 350.1616.(2S)-2-Amino-3-{5′-[ethynyl]-1H-benzo[d][1.2.3]triazol-1′-yl}propanoicacid hydrochloride (12d) was prepared as described for 12a using methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-{5′-[ethynyl]-1H-benzo[d][1.2.3]triazol-1′-yl}propanoate(11d)  and cesium carbonate . This gave (2S)-2-[(tert-butoxycarbonyl)amino]-3-{5′-[ethynyl]-1H-benzo[d][1.2.3]triazol-1′-yl}propanoicacid  as a yellow solid. This was used for the nextreaction without any further purification. To a solution of (2S)-2-[(tert-butoxycarbonyl)amino]-3-{5′-[ethynyl]-1H-benzo[d][1.2.3]triazol-1′-yl}propanoicacid  in 1,4-dioxane (0.1 mL) was added 2 Maqueous hydrochloric acid (1 mL). The reaction mixture was stirredat room temperature for 3 h and concentrated in vacuo. Purification by recrystallization from methanol and diethyl ethergave (2S)-2-amino-3-{5′-[ethynyl]-1H-benzo[d][1.2.3]triazol-1′-yl}propanoicacid hydrochloride (12d) as a yellow solid . Mp 195–198 °C; IR (neat) 3321, 2849, 2353, 1742,1487, 1235, 1057, 810 cm–1; [α]D23 +14.7 ; 1H NMR :δ 8.25 , 8.10 , 7.92–7.84 , 7.79, 7.60 , 7.56–7.50 , 5.38 , 5.28 , 4.81 ; 13C{1H} NMR : δ 167.6 (C), 145.5 (C), 133.1 (2 ×C), 131.3 (CH), 131.2 (CH), 127.9 (CH), 127.8 (CH), 127.5 (CH andC), 127.4 (CH), 126.7 (CH), 126.5 (CH), 122.1 (CH), 120.0 (2 ×C), 110.4 (CH), 89.7 (C), 88.1 (C), 52.2 (CH), 46.8 (CH2); MS (ESI) m/z 357 ; HRMS (ESI) m/z:[M + H]+ Calcd for C21H16N4O2H 357.1346; Found 357.1350.(2S)-2-Amino-3-{5′-[(4‴-fluorophenyl)ethynyl]-1H-benzo[d][1.2.3]triazol-1′-yl}propanoicacid hydrochloride (12e) was prepared as described for 12a using methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-{5′-[(4‴-fluorophenyl)ethynyl]-1H-benzo[d][1.2.3]triazol-1′-yl}propanoate(11e)  and cesium carbonate . This gave (2S)-2-[(tert-butoxycarbonyl)amino]-3-{5′-[(4‴-fluorophenyl)ethynyl]-1H-benzo[d][1.2.3]triazol-1′-yl}propanoicacid  as an off-white solid. This was used for thenext step without further purification. To a solution of (2S)-2-[(tert-butoxycarbonyl)amino]-3-{5′-[(4‴-fluorophenyl)ethynyl]-1H-benzo[d][1.2.3]triazol-1′-yl}propanoicacid  in acetonitrile (3 mL) was added 6 M aqueoushydrochloric acid (2 mL). The reaction mixture was stirred at roomtemperature for 3 h and concentrated in vacuo. Purificationby trituration with chloroform gave (2S)-2-amino-3-{5′-[(4‴-fluorophenyl)ethynyl]-1H-benzo[d][1.2.3]triazol-1′-yl}propanoicacid hydrochloride (12e) as a white solid . Mp 280–285 °C (decomposition); IR (neat) 2921, 2359,1742, 1508, 1219, 833 cm–1; [α]D17 +13.6 ; 1H NMR :δ 8.19 , 7.85 , 7.73, 7.59 , 7.14 , 5.36, 5.27 , 4.79–4.72 ; 13C{1H} NMR : δ 167.5 (C),162.8 , 145.4 (C), 133.4 , 133.2 (C), 131.2 (CH), 122.0 (CH), 119.9 (C),119.0 , 115.4 , 110.3 (CH), 88.2 (C), 87.5 (C), 52.2 (CH),46.8 (CH2); MS (ESI) m/z 347 ; HRMS (ESI) m/z: [M + Na]+ calcd for C17H13FN4O2Na 347.0915; found 347.0913.(2S)-2-Amino-3-{5′-[(4‴-cyanophenyl)ethynyl]-1H-benzo[d][1.2.3]triazol-1′-yl}propanoicacid hydrochloride (12f) was prepared as described for 12a using methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-{5′-[(4‴-cyanophenyl)ethynyl]-1H-benzo[d][1.2.3]triazol-1′-yl}propanoate(11f)  and cesium carbonate . This gave (2S)-2-[(tert-butoxycarbonyl)amino]-3-{5′-[(4‴-cyanophenyl)ethynyl]-1H-benzo[d][1.2.3]triazol-1′-yl}propanoicacid  as a yellow solid. This was used for the nextreaction without any further purification. To a solution of (2S)-2-[(tert-butoxycarbonyl)amino]-3-{5′-[(4‴-cyanophenyl)ethynyl]-1H-benzo[d][1.2.3]triazol-1′-yl}propanoicacid  in acetonitrile (0.1 mL) was added 2M aqueous hydrochloric acid (1 mL). The reaction mixture was stirredat room temperature for 3 h and concentrated in vacuo. Purification by recrystallization from methanol and diethyl ethergave (2S)-2-amino-3-{5′-[(4‴-cyanophenyl)ethynyl]-1H-benzo[d][1.2.3]triazol-1′-yl}propanoicacid hydrochloride (12f) as a yellow solid . Mp 215–218 °C; IR (neat) 2862, 2230, 1748, 1600,1497, 1250, 814 cm–1; [α]D20 +12.4 ; 1H NMR : δ 8.39 , 8.09 , 7.93 , 7.81–7.76 , 5.24–5.21 , 4.65 ; 13C{1H} NMR : δ 47.4 (CH2), 52.2 (CH), 88.2(C), 93.5 (C), 111.5 (C), 112.3 (CH), 117.8 (C), 118.9 (C), 123.4(CH), 127.5 (C), 131.2 (CH), 132.7 (2 × CH), 133.2 (2 ×CH), 134.3 (C), 145.4 (C), 168.9 (C); MS (ESI) m/z 332 ; HRMS (ESI) m/z: [M + H]+ calcd for C18H13N5O2H 332.1142; found 332.1140.(2S)-2-[(tert-butoxycarbonyl)amino]-3-[5′-(phenylethynyl)-1H-benzo[d][1.2.3]triazol-1′-yl]propanoate(11a)  in tetrahydrofuran (1.2 mL).A 1 M solution of zinc iodide  in tetrahydrofuranwas added and then the reaction mixture was degassed, purged withhydrogen, and stirred at 40 °C for 20 h. The reaction mixturewas allowed to cool to room temperature and diluted with ethyl acetate(40 mL), washed with water (3 × 20 mL), dried (MgSO4), and concentrated in vacuo. Purification by flashcolumn chromatography, eluting with 70% diethyl ether in hexane gavemethyl -2-[(tert-butyloxycarbonyl)amino]-3-propanoate(13a) as an off-white solid . Mp 170–172°C; IR (neat) 3368, 2978, 2361, 1755, 1690, 1501, 1161, 748 cm–1; [α]D22 +7.0 ; 1H NMR : δ 8.08 , 7.73 , 7.57–7.53, 7.50 , 7.44–7.34, 7.31–7.27 , 7.25 , 7.16 , 5.39 , 5.15–5.04 , 4.81 , 3.77 , 1.43 ; 13C{1H} NMR : δ169.6 (C), 155.1 (C), 146.4 (C), 137.0 (C), 134.1 (C), 133.6 (C),129.5 (CH), 128.8 (2 × CH), 127.91 (CH), 127.94 (CH), 126.6 (2× CH), 126.4 (CH), 117.7 (CH), 109.5 (CH), 80.7 (C), 53.9 (CH),53.1 (CH3), 48.7 (CH2), 28.3 (3 × CH3); MS (ESI) m/z 423 ; HRMS (ESI) m/z: [M + H]+ calcd for C23H26N4O4H 423.2027; found 423.2035.To a dry microwave vial containing zinc and bis(triphenylphosphine)palladium(II) dichloride, under argon, was added a solution of methyl-2-[(tert-butyloxycarbonyl)amino]-3-(5′-[(4‴-methoxyphenyl)ethenyl]-1H-benzo[d]triazol-1′-yl)propanoate(13b) was synthesized as described for 13a using zinc , bis(triphenylphosphine)palladium(II)dichloride , methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-{5′-[(4‴-methoxyphenyl)ethynyl]-1H-benzo[d][1.2.3]triazol-1′-yl}propanoate(11b) , tetrahydrofuran (1.7 mL),and a 1 M solution of zinc iodide  in tetrahydrofuran.Purification by flash column chromatography, eluting with 75% diethylether in hexane gave methyl -2-[(tert-butyloxycarbonyl)amino]-3-(5′-[(4‴-methoxyphenyl)ethenyl]-1H-benzo[d]triazol-1′-yl)propanoate(13b) as a yellow solid . Mp 164–168°C; IR (neat) 3306, 2967, 2361, 1740, 1701, 1605, 1512, 1300,822 cm–1; [α]D21 +15.0 ; 1H NMR : δ 8.05 , 7.71 , 7.53–7.46 , 7.12 , 6.92 , 5.37, 5.16–5.04 , 4.81, 3.85 , 3.77 , 1.44 ; 13C{1H} NMR : δ 169.7 (C), 159.5 (C), 155.1 (C), 146.4 (C), 134.5(C), 133.3 (C), 129.8 (C), 129.0 (CH), 127.8 (2 × CH), 126.3(CH), 125.8 (CH), 117.1 (CH), 114.2 (2 × CH), 109.5 (CH), 80.6(C), 55.4 (CH3), 53.9 (CH), 53.1 (CH3), 48.7(CH2), 28.3 (3 × CH3); MS (ESI) m/z 475 ; HRMS(ESI) m/z: [M + Na]+ calcdfor C24H28N4O5Na 475.1952;found 475.1954.Methyl -2-[(tert-butyloxycarbonyl)amino]-3-(5′-[(4‴-fluorophenyl)ethenyl]-1H-benzo[d]triazol-1′-yl)propanoate(13c) was synthesized as described for 13a using zinc , bis(triphenylphosphine)palladium(II)dichloride , methyl (2S)-2-[(tert-butyloxycarbonyl)amino]-3-(5′-[(4‴-fluorophenyl)ethynyl]-1H-benzo[d][1.2.3]triazol-1′-yl)propanoate(11e) , tetrahydrofuran (1.4 mL),and a 1 M solution of zinc iodide  in tetrahydrofuran.Purification by flash column chromatography, eluting with 70% diethylether in hexane gave methyl -2-[(tert-butyloxycarbonyl)amino]-3-(5′-[(4‴-fluorophenyl)ethenyl]-1H-benzo[d]triazol-1′-yl)propanoate(13c) as a yellow solid . Mp 162–165°C; IR (neat) 3383, 2982, 2361, 1759, 1690, 1508, 1157, 829 cm–1; [α]D24 +6.2 ; 1H NMR :δ 8.16 , 7.90 , 7.85 , 7.68 , 7.43 , 7.40 , 7.24 , 5.08, 4.95 , 4.62 , 3.67 , 1.22 ; 13C{1H}NMR : δ 170.5(C), 162.2 , 155.4 (C), 146.3 (C), 134.1 , 133.9 (C), 133.6 (C),128.8 , 128.4 (CH), 128.0 (CH), 126.3 (CH), 117.2 (CH),116.1 , 111.4 (CH), 79.1 (C), 53.9 (CH), 52.8 (CH3), 48.3 (CH2), 28.4 (3 × CH3); MS (ESI) m/z 463 ; HRMS(ESI) m/z: [M + Na]+ calcdfor C23H25FN4O4Na 463.1752;found 463.1752.Methyl -2-[(tert-butyloxycarbonyl)amino]-3-propanoate (13a)  in a mixture of methanol (1.5 mL) and 1,4-dioxane(1.5 mL) was added a solution of cesium carbonate  in water (0.75 mL). The reaction mixture was stirred at roomtemperature for 20 h and then concentrated in vacuo. The resulting residue was dissolved in water (50 mL) and acidifiedto pH 1 with 1 M aqueous hydrochloric acid. The aqueous layer wasextracted with dichloromethane (3 × 30 mL), dried (MgSO4), and concentrated in vacuo to give -2-[(tert-butyloxycarbonyl)amino]-3-propanoicacid as a yellow solid . This was used for the nextreaction without further purification. To a solution of -2-[(tert-butyloxycarbonyl)amino]-3-propanoic acid  in 1,4-dioxane (0.1 mL) was added 2 M aqueous hydrochloricacid (2.5 mL). The reaction mixture was stirred at room temperaturefor 3 h and then concentrated in vacuo. This gave-2-amino-3-propanoicacid hydrochloride (14a) as an off-white solid . Mp 215–219 °C; IR (neat) 3368, 2955, 2361, 1748,1690, 1501, 1250, 1204, 1161, 691 cm–1; [α]D22 +5.6 ; 1H NMR :δ 8.09 , 7.93 , 7.82, 7.59 , 7.43–7.23 , 5.34 , 5.25 ,4.74 ; 13C{1H} NMR : δ 167.6 (C), 146.2 (C), 137.1 (C), 135.1 (C),133.1 (C), 129.7 (CH), 128.4 (2 × CH), 127.6 (CH), 127.2 (CH),126.7 (CH), 126.3 (2 × CH), 116.4 (CH), 110.0 (CH), 52.1 (CH),46.7 (CH2); MS (ESI) m/z 309 ; HRMS (ESI) m/z: [M + H]+ calcd for C17H16N4O2H 309.1346; found 309.1344.To a solution of methyl-2-Amino-3-(5′-[(4‴-methoxyphenyl)ethenyl]-1H-benzo[d]triazol-1′-yl)propanoicacid hydrochloride (14b) was synthesized as describedfor 14a using methyl -2-[(tert-butyloxycarbonyl)amino]-3-(5′-[(4‴-methoxyphenyl)ethenyl]-1H-benzo[d]triazol-1′-yl)propanoate(13b)  and cesium carbonate . This gave -2-amino-3-(5′-[(4‴-methoxyphenyl)ethenyl]-1H-benzo[d]triazol-1′-yl)propanoicacid as an off-white solid . This was used for thenext reaction without further purification using -2-amino-3-(5′-[(4‴-methoxyphenyl)ethenyl]-1H-benzo[d]triazol-1′-yl)propanoicacid , 1,4-dioxane (2 mL), and 6 M aqueoushydrochloric acid (1 mL). This gave -2-amino-3-(5′-[(4‴-methoxyphenyl)ethenyl]-1H-benzo[d]triazol-1′-yl)propanoicacid hydrochloride (14b) as a yellow solid . Mp 210–213 °C; IR (neat) 3372, 2932, 2361, 1728,1601, 1574, 1501, 1443, 1173, 1022 cm–1; [α]D21 +10.1 ; 1H NMR : δ 8.71 , 8.16 , 7.93–7.89, 7.58 , 7.36 , 7.29 ,6.98 , 5.24 , 5.17 ,4.69–4.63 , 3.79 ; 13C{1H} NMR : δ 168.9(C), 159.5 (C), 146.4 (C), 134.7 (C), 133.5 (C), 130.1 (C), 129.1(CH), 128.3 (2 × CH), 126.5 (CH), 126.1 (CH), 116.7 (CH), 114.7(2 × CH), 111.4 (CH), 55.7 (CH3), 52.2 (CH), 47.3(CH2); MS (ESI) m/z 361; HRMS (ESI) m/z: [M + Na]+ calcd for C18H18N4O3Na 361.1271; found 361.1270.-2-Amino-3-(5′-[(4‴-fluorophenyl)ethenyl]-1H-benzo[d]triazol-1′-yl)propanoicacid hydrochloride (14c) was synthesized as describedfor 14a using methyl -2-[(tert-butyloxycarbonyl)amino]-3-(5′-[(4‴-fluorophenyl)ethenyl]-1H-benzo[d]triazol-1′-yl)propanoate(13c)  and cesium carbonate . This gave -2-[(tert-butyloxycarbonyl)amino]-3-(5′-[(4‴-fluorophenyl)ethenyl]-1H-benzo[d]triazol-1′-yl)propanoicacid as an off-white solid . This was used for thenext reaction without further purification using -2-[(tert-butyloxycarbonyl)amino]-3-(5′-[(4‴-fluorostyryl)ethenyl]-1H-benzo[d]triazol-1′-yl)propanoicacid , 1,4-dioxane (0.2 mL), and 2 M aqueoushydrochloric acid (2.5 mL). This gave -2-amino-3-(5′-[(4‴-fluorophenyl)ethenyl]-1H-benzo[d]triazol-1′-yl)propanoicacid hydrochloride (14c) as an off-white solid . Mp 195–199 °C; IR (neat) 3372, 2951, 2361, 1728,1597, 1497, 1200, 826 cm–1; [α]D23 +7.3 ; 1H NMR : δ 8.21 , 7.97 , 7.94 ,7.69 , 7.42 , 7.25, 5.27 , 5.21 ,4.69–4.61 ; 13C{1H} NMR : δ 168.9 (C), 162.2 , 146.3(C), 134.2 (C), 134.1 , 133.7 (C), 128.9 , 128.4 (CH), 128.3 (CH), 126.6(CH), 117.2 (CH), 116.1 , 111.5 (CH), 52.2 (CH), 47.3 (CH2); MS (ESI) m/z 327 ; HRMS (ESI) m/z:[M + H]+ calcd for C17H15FN4O2H 327.1252; found 327.1260.(2


The same trend was found for the inhibitory action against acetylcholinesterase, with 1,2-NQ derivatives showing higher inhibition50µM than their 1,4-NQ homologues, with 4h being the most potent compound (Inhibition50µM = 85%). Docking studies were performed for the 1,2-NQ derivatives with the highest inhibitions, showing dual binding interactions with both CAS and PAS sites, while the less active 1,4-NQ derivatives showed interactions with PAS and the mid-gorge region.This work describes the design, sustainable synthesis, evaluation of electrochemical and biological properties against HepG2 cell lines, and AChE enzymes of different substituted derivatives of 1,4- and 1,2-naphthoquinones (NQ). A microwave-assisted protocol was optimized with success for the synthesis of the 2-substituted-1,4-NQ series and extended to the 4-substituted-1,2-NQ family, providing an alternative and more sustainable approach to the synthesis of naphthoquinones. The electrochemical properties were studied by cyclic voltammetry, and the redox potentials related to the molecular structural characteristics and the biological properties. Compounds were tested for their potential anti-cancer activity against a hepatocellular carcinoma cell line, HepG2, using MTT assay, and 1,2-NQ derivatives were found to be more active than their 1,4-NQ homologues ( Naphthoquinones (NQ) have played a crucial role in the drug discovery process been widely used as important pharmacophores or intermediates for the synthesis of bioactive compounds. These heterocyclic compounds are known to exhibit a diverse range of biological properties, such as antimicrobials, antitumoral, antioxidants, antimalarial, and neuroprotectants, among others ,2,3. TheDifferent types of cancers and several neurodegenerative disorders are often associated with intracellular redox imbalance and consequent oxidative damage ,11, makiThe NQ scaffold is undoubtedly important, and the quinone’s reactivity can be modulated by introducing atoms or groups into the central naphthoquinone ring, thus encouraging the design of derivatives and the exploration of their electrochemical and biological properties. Amine groups have been reported to play a key role in many bioactive compounds, and literature data have shown that their presence in the quinone backbone tunes its redox properties and induces oxidative stress in cells and DNA alkylation . AliphatThe NQ derivatives can be prepared by conventional methods with satisfactory yields, although in most cases, with time-consuming protocols and tedious workups. To overcome these restrictions, new, greener synthetic approaches have been explored ,29. MicrA set of compounds derived from 1,4-NQ and 1,2-NQ were studied  to evalu3a–f) was performed by microwave-assisted reactions in the presence of a basic catalyst with moderate to excellent yields. The same approach was used for the 4-substituted-1,2-NQ derivatives  in the optimized experimental conditions. The 4-substituted-1,2-NQ derivatives (4a–d and 4g) were obtained as previously reported by a biocatalytic strategy using CotA-laccase from Bacillus subtilis + at m/z 265, 266, 280, 275, 266 for 3a–d and 4h and m/z 230 for 3e,f and 4e,f, respectively. Generally, for all ions, the first fragmentation pattern [ESI(+)-MS2] is consistent with the correspondent typical loss of CO leading to signals [M+1−CO]+.The structures of synthesized compounds were assigned based on their pc or E1/2) and biological activities, demonstrating the relevance of electrochemical studies as tools for the comprehension of drug mechanism of action against various diseases, for the prediction of biological activities and for the design of potentially active compounds +; 265 [M+H]+, MS2 (m/z): 237 [M+H-CO]+, MS3 (m/z): 209 [M+H-CO-CO]+; ESI-MS negative mode: 263 [M−H]−. ESI-HRMS: m/z calcd. for [C16H13N2O2+H]+: 265.0972; found 265.0980.2-(4-aminophenol)-1,4-naphthoquinone (3b): orange solid (59%): 1H NMR : δ(ppm) = 8.11 , 8.02 , 7.80 , 7.70 , 7.17 , 6.87  and 6.0 . 13C NMR : δ(ppm) = 185.6 (C4), 182.9 (C1), 157.2 (C14), 135.9 (C7), 133.6 (C6), 132.1 (C10), 130.4 (C9), 127.4 (C5), 126.9 , 126.8 (C8), 117.1  and 101.7 (C3). ESI-MS positive mode: 266 [M+H]+, MS2 (m/z): 238 [M+H-CO]+. ESI-HRMS: m/z calcd. for [C16H11NO3+H]+: 266.0812; found 266.0810.2-(4-methoxyaniline)-1,4-naphthoquinone (3c): brown solid (57%): 1H NMR : δ(ppm) = 8.09 , 8.00 , 7.78 , 7.70 , 7.25 , 7.00 , 6.02  and 3.82 . 13C NMR : δ(ppm) = 185.6 (C4), 182.8 (C1), 159.4 (C14), 135.8 (C7), 133.6 (C6), 133.2 (C10), 132.1 (C9), 129.4 (C11), 127.4 (C5), 126.8 (C8), 126.7 , 115.8 , 101.9 (C3) and 56.0 (CH3). ESI-MS positive mode: 280 [M+H]+, MS2 (m/z): 252 [M+H-CO]+. ESI-HRMS: m/z calcd. for [C17H13NO3+H]+: 280.0968; found 280.0976.2-(4-aminobenzonitrile)-1,4-naphthoquinone (3d): orange solid (7%): 1H NMR : δ(ppm) = 8.31 , 8.05 , 7.82 , 7.65 , 7.48 , 7.35 , 6.22 . ESI-MS positive mode: 275 [M+H]+, MS2 (m/z): 247 [M+H-CO]+. ESI-HRMS: m/z calcd. for [C17H10N2O2+H]+: 275.0815; found 275.0819.1) or 1,2-naphthoquinone (2)  and alkyl amines (BA and DA) (50 mmol) were dissolved in acetonitrile (2 mL) and the microwave vial was sealed. Solution was stirred under microwave irradiation (200 W) at 150 °C for 5 min. After cooling, products were extracted with ethyl acetate. Solvent was evaporated under reduced pressure and residues purified by column chromatography . The 1,4-naphthoquinone (2-(butan-1-amine)-1,4-naphthoquinone (3e): orange solid (63%): 1H NMR : δ(ppm) = 8.01 , 7.75 , 7.66 , 5.68 , 3.23 , 1.66 , 1.44  and 0.98 . 13C NMR : δ(ppm) = 184.7 (C4), 182.6 (C1), 150.8 (C2), 135.8 (C7), 135.0 (C9), 133.3 (C6), 132.1 (C10), 127.3 (C8), 126.8 (C5), 99.9 (C3), 43.2 (C11), 31.1 (C12), 21.3 (C13) 14.1 (C14). ESI-MS positive mode: 230 [M+H]+. ESI-HRMS: m/z calcd. for [C14H15NO2+H]+: 230.1176; found 230.1180.2-(diethylamine)-1,4-naphthoquinone (3f): red solid (74%): 1H NMR : δ(ppm) = 7.98 , 7.76 , 7.68 , 5.86 , 3.62  and 1.32 . 13C NMR : δ(ppm) = 184.7 (C4), 184.4 (C1), 153.3 (C2), 135.0 (C7), 134.3 (C9), 134.1 (C10), 133.3 (C6), 125.9 , 105.0 (C3), 48.0  and 12.9 . ESI-MS positive mode: 230 [M+H]+. ESI-HRMS: m/z calcd. for [C14H15NO2+H]+: 230.1176; found 230.1177.4-(butan-1-amine)-1,2-naphthoquinone (4e): red solid (72%): 1H NMR : δ(ppm) = 8.12 , 8.05 , 7.80 , 7.70 , 5.90 , 3.52 , 1.76 , 1.48  and 1.00 . 13C NMR : δ(ppm) = 180.9 (C1), 175.3 (C2), 159.4 (C4), 135.8 (C7), 135.6 (C6), 132.9 (C7), 132.6 (C10), 131.9 (C9), 129.3 (C8), 124.6 (C5), 98.9 (C3), 44.9 (C11), 31.5 (C12), 21.3 (C13) 14.1 (C14). ESI-MS positive mode: 256 [M+Na]+ and 230 [M+H]+. ESI-HRMS: m/z calcd. for [C14H15NO2+H]+: 230.1176; found 230.1183.4-(diethylamine)-1,2-naphthoquinone (4f): red solid (75%): 1H NMR : δ(ppm) = 8.05 , 7.75 , 6.02 , 3.68  and 1.41 . 13C NMR : δ(ppm) = 183.7 (C1), 177.6 (C2), 165.6 (C4), 135.0 (C6), 133.3 (C9), 132.4 (C5), 130.2 (C10), 129.9 (C9), 129.9 (C8), 128.7 (C7), 105.4 (C3), 47.9  and 13.0 . ESI-MS positive mode: ESI-MS positive mode: 256 [M+Na]+ and 230 [M+H]+. ESI-HRMS: m/z calcd. for [C14H15NO2+H]+: 230.1176; found 230.1176.Drug score values indicate the overall potential of a compound to be a drug candidate. Mol inspiration is a web-based tool used to predict the bioactivity score of synthesized compounds against regular human receptors such as GPCRs, ion channels, kinases, nuclear receptors, proteases, and enzymes . The eva2. The 3--2,5-diphenyltetrazolium bromide (MTT) method, described in Gaspar et al., ). A control reaction was carried out using 1% DMSO water solution, instead of the compound’s solution, and this initial rate was considered 100% of the enzymatic activity, Vcontrol. The percentage of AChE inhibition (I) for various concentrations of the different compounds was determined as the ratio of V[compound] and Vcontrol. For each compound, both the AChEIC50 value (the concentration that inhibits 50% of the AChE activity) and the percentage of inhibition at the compound concentration of 50 µM (Inhibition50µM) were calculated by plotting I for each compound concentration. All the assays were carried out in triplicate.The inhibition of acetylcholinesterase (AChE) enzymatic activity was measured using the Ellman’s colorimetric method with some alterations . BrieflyMolecular docking was performed using AutoDock Vina v.1.2.0 software, CCSB. Both the compounds and the enzyme (PBD ID: 4EY7) were prepared for docking simulations via DockPrep. This function of Chimera 1.16 software removed water molecules and added hydrogens to the compounds, which structure was obtained through ChemDraw software. Five models were generated for each compound, and the highest-scoring model was viewed and analyzed using the Discovery Studio software .A microwave-assisted protocol was successfully established for the synthesis of 2-substituted-1,4-NQ and 4-substituted-1,2-NQ derivatives, with good yields and completing the reactions in shorter times. The electrochemical properties showed to be driven by the reductive processes within the NQ moiety, and the 1,2-NQ series showed to be easier reduced than the corresponding 1,4-NQ series. Moreover, the trend in the redox potential within each series is clearly related to the electronic properties of the amine substituents. pc) were the most active against the HepG2 cancer cell lines and as AChE inhibitors. The observed trend suggests that the bioreduction may play an important role in the biological activity of these compounds, but other factors must also be considered. The potential biological activity of the compounds was assessed for their cytotoxicity against HepG2 cell lines and as AChE inhibitors, showing that 1,2-NQ series exhibit a dual action, both as promising cytotoxic agents and moderate AChE inhibitors. The results showed a dependence on chemical modifications in the NQ moiety with an increase in cytotoxic and AChE inhibitory effects, in particular, for electron-donating substituted aromatic amines. Comparison between biological activities and electrochemical parameters showed that the first wave reduction potential is an important parameter as follows: the most easily reduced naphthoquinones (>E4b and 4h), confirming that both compounds interact with amino acid residues in the PAS and CAS of the AChE active site through hydrogen binding and hydrophobic interactions. These interactions, also found for dual-binding cholinesterase inhibitors, such as donepezil, may be related to the good inhibition profiles found for the 4-substituted-1,2-NQ family.To elucidate the possible mechanism of AChE targeting action, docking simulations were performed for the more active compounds (


Mammea siamensis (Miq.) T. Anders.  showed anti-proliferative activity against human prostate carcinoma LNCaP cells (IC50 = 2.0 µg/mL). Two new coumarin-related polysubstituted benzofurans, mammeasins P (1) and Q (2), and a known polysubstituted coumarin mammea B/AC cyclo F (39) were isolated from the extract along with 44 previously reported polysubstituted coumarin constituents (3–38 and 40–47). The structures of two new compounds (1 and 2) were determined based on their spectroscopic properties derived from the physicochemical evidence including NMR and MS analyses and taking the plausible generative pathway into account. Among the coumarin constituents, mammeasins A  and B , sugangin B , kayeassamins E  and G , and mammeas E/BA , E/BB , and E/BC  showed relatively potent anti-proliferative activity.A methanol extract of the flowers of  Prostate cancer, a hormonally driven cancer, is the second most frequent malignancy in men worldwide ,2,3,4. IMammea siamensis (Miq.) T. Anders., which is the name recorded by the World Flora Online , four methylene moieties , a pair of cis-substituted olefinic protons [δ 5.51 and 6.65 ], and a hydrogen-bonded hydroxy proton [δ 14.12 ]. 1H−1H COSY experiments on 1 indicated the presence of partial structures coumarin, a common structure of many mammeacoumarins, via intramolecular displacements to the furocoumarins, acid-catalyzed double-bond migration, and lactone opening  . The 1H- opening  38,39].,39.1) wa2) was determined to be C23H28O6, showing a molecular ion peak at m/z 400.1880 , using high-resolution EI-MS measurements. The 1H- and 13C-NMR spectra (3) of 2 were similar to those of 1, except for the signals owing to a 2-methyl-butyryl moiety in C-8 position  instead of an n-butyryl moiety, as seen in 1. As shown in 2 was elucidated via 1H–1H COSY and HMBC experiments. 1H−1H COSY correlations indicated the presence of the following partial structures of 2, shown in bold lines: linkage of C-2’–C-3’; C-3‘–C-4’’; C-2’’’–C-5’’’. HMBC correlations revealed long-range correlations between the following proton and carbon pairs: H2-3 [δ 3.75 ] and C-2, 4, 4a, 1’; H2-2’ [δ 2.67 ] and C-4, 1’; H-3’’ [δ 5.51 ] and C-6, 2’’, CH3-2’’; H-4’’ [δ 6.65 ] and C-5, 7, 2’’; CH3-2’’ [δ 1.52 ] and C-2’’, 3’’; 8a-OH [δ 14.09 ] and C-4a, 8, 8a; H2-2’’’, H2-3’’’, H3-5’’’ and C-1’’’. Therefore, the structure of 2 was established.The molecular formula of mammeasin Q  and Q (2), and 45 coumarin constituents (3–47) , B , C , D , E , F , G , H , I , J , K , L , M , N , and O ; surangins B , C , and D ; kayeassamin A , 8-hydroxy-5-methyl-7--9-(2-methyl-1-oxobutyl)-4,5-dihydropyrano chromen-2-one , 8-hydroxy-5-methyl-7--9-(3-methyl-1-oxobutyl)-4,5-dihydropyrano chromen-2-one ; kayeassamins E , F , G , and I ; mammeas A/AA , A/AB , A/AC , A/AD [=mesuol ], A/AA cyclo D , A/AB cyclo D , A/AC cyclo D , A/AA cyclo F , A/AC cyclo F , B/AB cyclo D , B/AC cyclo D , B/AC cyclo F , E/BA , E/BB , E/BC , E/BC cyclo D , and E/BD cyclo D ; and deacetylmammeas E/AA cyclo D , E/BB cyclo D , and E/BC cyclo D . To characterize the active constituents, the anti-proliferative effects of the isolates against LNCaP cells were examined. However, evaluation of the coumarin constituents , including the new compounds 1 and 2, for which we did not have a sufficient sample amount to assess biological activity, could not be performed. As shown in 3, IC50 = 1.2 µM) and B , sugangin B , kayeassamins E  and G , and mammeas E/BA , E/BB , and E/BC , showed relatively potent anti-proliferative activity. The structural requirements of the coumarins for the activity were suggested as the following: (1) regardless of the structure of the substitutions at C-4, C-5, C-6, and C-8, coumarins with 7-OH group  were essential for the strong activity; (2) compounds with the 6-prenylcoumarin moieties showed stronger or equivalent activity to those with the geranyl moiety  or those forming the 2,2-dimethyl-2H-pyran structure with 5-OH group ; (3) compounds with the 4-propylcoumarin moieties with the 1’-acetoxy group showed stronger activity than those of the corresponding deacetylcoumarins  or those forming the 2-methyl-3,4-dihydro-2H-pyran structure with 5-OH group .Chemical studies on the flowers of s (3–47) . As has l  =  5 cm); UV spectra, Shimadzu UV-1600 spectrometer; IR spectra, IRAffinity-1 spectrophotometer ; 1H NMR spectra, JNM-ECA800 (800 MHz), JNM-LA500 (500 MHz), JNM-ECS400 (400 MHz), and JNM-AL400 (400 MHz) spectrometers; 13C NMR spectra, JNM-ECA800 (200 MHz), JNM-LA500 (125 MHz), JNM-ECA400 (100 MHz), and JNM-AL400 (100 MHz) spectrometers ; EI-MS and high-resolution EI-MS, JMS-GCMATE mass spectrometer ; HPLC detector, SPD-10Avp UV-VIS detector; and HPLC columns, Cosmosil 5C18-MS-II . For NMR, the samples were dissolved in deuterated chloroform (CDCl3) at room temperature with tetramethylsilane as an internal standard. Columns of 4.6 mm × 250 mm and 20 mm × 250 mm were used for analytical and preparative purposes, respectively.The following instruments were used to obtain spectroscopic data: specific rotation, JASCO P-2200 polarimeter   and silica gel RP-18 WF254S  (reversed-phase); and reversed-phase HPTLC, pre-coated TLC plates with silica gel RP-18 WF254S . Detection was performed by spraying with 1% Ce(SO4)2–10% aqueous H2SO4, followed by heating.The following chromatographic materials were used for column chromatography (CC): highly porous synthetic resin, Diaion HP-20 ; normal-phase silica gel CC, silica gel 60 N ; reversed-phase ODS CC, Chromatorex ODS DM1020T ; TLC, pre-coated TLC plates with silica gel 60FM. siamensis flowers were collected from Nakhonsithammarat Province, Thailand, in September 2006, as previously described  to produce 11 fractions , as previously reported [2O →MeOH→acetone] to yield nine fractions , as described previously  to give mammeasins P  and Q . Fraction 5 (4.15 g) was subjected to reversed-phase silica gel CC  to obtain six fractions , as previously reported  to give mammea B/AC cyclo F  together with mammeasins M  and O  and mammeas A/AA , A/AC , A/AA cyclo D , A/AC cyclo F , E/BC cyclo D , and E/BD cyclo D  [The methanolic extract  of the dried flowers of eviously . FractioFr. 2-9 3.8 mg], a Fr. 5-2 89.8 mg, Fr. 2-9 3.8 mg], a Fr. 6 6. g, Fr. 7ε)]: 247 (4.23), 267 (4.03), 354 (3.29); IR (film): 1743, 1622, 1456, 1167, 1120; 1H-NMR  δ: see 13C-NMR data  δC: see m/z (%): 386 , 371 (100); high-resolution EIMS m/z 386.1724 .Pale-yellow oil; UV : 247 (4.23), 268 (4.04), 352 (3.36); IR (film): 1743, 1622, 1456, 1169, 1123 cm–1; 1H-NMR  δ: see 13C-NMR data  δC: see m/z (%): 400 ; high-resolution EIMS m/z 400.1880 .Pale-yellow oil; [α]RPMI 1640 medium was purchased from FUJIFILM Wako Pure Chemical Industries , fetal bovine serum (FBS) from Biosera , other chemicals from FUJIFILM Wako Pure Chemical Industries , and 96-well microplates from Sumitomo Bakelite .2 environment. LNCaP cells were seeded in 96-well plates at a density of 5 × 103 cells/well in 100 µL/well medium, and 100 μL/well of medium containing a test sample was added after an initial incubation of 24 h. Cell viability was detected after 96 h of incubation using the Cell Counting Kit-8 (CCK-8). CCK-8 was purchased from Dojindo Molecular Technologies . The O.D. of the yellow-colored formazan solution was measured using a microplate reader at 450 nm (reference: 650 nm)  was calculated using the following formula:Experiments were performed in accordance with previously reported methods ,41, with 650 nm) . The IC5Each test compound was dissolved in DMSO and added to the medium .p) values less than 0.05 were considered significant.Values are expressed as the mean ± standard error (S.E.M.). One-way analysis of variance (ANOVA) followed by Dunnett’s test was used for statistical analysis. Probability (M. siamensis (Miq.) showed anti-proliferative activity against human prostate carcinoma LNCaP cells (IC50 = 2.0 µg/mL). Two new coumarin-related polysubstituted benzofurans, mammeasins P (1) and Q (2), were isolated, and their structures were elucidated based on their spectroscopic properties derived from the physicochemical evidence including NMR and MS analyses as well as considering the plausible generative pathway. We have already achieved the total syntheses of mammeasins C (5) and D (6), which were isolated as new compounds from the same plant material  (Among the isolates, seven coumarin constituents, including mammeasins A (hibitor]  27]. Th. Th3, IC


The lead compound 4e could significantly enhance cancer cell death, but not the death of normal cells if PAs were added to the cell culture media. Thus, the bispidine derivative 4e 3-(3-methoxypropyl)-7-[3-(1H-piperazin-1-yl)ethyl]-3,7-diazabicyclo[3.3.1]nonane could become a potential anticancer drug substance whose mechanism relies on the induction of PA catabolism in cancer cells.Polyamine (PA) catabolism is often reduced in cancer cells. The activation of this metabolic pathway produces cytotoxic substances that might cause apoptosis in cancer cells. Chemical compounds able to restore the level of PA catabolism in tumors could become potential antineoplastic agents. The search for activators of PA catabolism among bicyclononan-9-ones is a promising strategy for drug development. The aim of the study was to evaluate the biological activity of new 3,7-diazabicyclo[3.3.1]nonan-9-one derivatives that have antiproliferative properties by accelerating PA catabolism. Eight bispidine derivatives were synthetized and demonstrated the ability to activate PA catabolism in regenerating rat liver homogenates. However, only three of them demonstrated a potent ability to decrease the viability of cancer cells in the MTT assay. Compounds  Polyamines (PAs) are ubiquitous aliphatic polycations (putrescine (Put), spermidine (Spd), and spermine (Spm)) that can interact with biological polyanionic macromolecules ,2,3,4. TBispidines (diazabicyclononanes) are well known in medicinal chemistry, providing a wide variety of biological activities. They are intensively studied as anticancer  or antiv3a–3e and 6a–6c readily formed complexes 4a–4e and 7a–7c with β-cyclodextrin, respectively.The synthesized 3,7-diazabicyclo[3.3.1]nonan-9-ones (bispidine derivatives) were tested as potential modulators of PA metabolism and anticancer agents. They were prepared in the Laboratory of Chemistry of Synthetic and Natural Medicinal Compounds of A.B. Bekturov Institute of Chemical Sciences. The scheme in 2a–2e) were synthesized (1a–1c) with paraformaldehyde and the primary amines 1-(3-aminopropyl)imidazole, 1-(2-aminoethyl)pyridine and 1-(2-aminoethyl)piperazine in an acetic-methanol medium.3,7-Diazabicyclo[3.3.1]nonan-9-ones (thesized A by simu3a–3e) were obtained by the Kizhner-Wolf reaction, reducing compounds 2a–2e under the action of hydrazine hydrate in the presence of KOH in triethylene glycol.The corresponding 3,7-diazabicyclo[3.3.1]nonanes  was used for purification of novel bicyclic ketones, nonanes, bicyclic oximes and O-benzoyl oximes. The completion of the reactions was monitored by TLC. 1H and 13C NMR spectroscopies were used to determine the structures of the synthesized substances.2a–2e), characteristic bands of stretching vibrations of the carbonyl group appear.In the IR spectrum of 3,7-diazabicyclo[3.3.1]nonan-9-ones (3a–3e.The formation of bicyclic amines was evidenced by the absence of the absorption band of the carbonyl group in the IR spectra of compounds 3a–3e) with the spectra of the starting bicyclic ketones (2a–2e), it can be seen that they lack the signal of the carbon atom characteristic of the carbonyl group, while in the upfield part of the spectrum, a triplet signal of the carbon atom of the methylene group appears in the 9th position.When comparing the 13C NMR spectra of 3,7-diazabicyclo[3.3.1]nonanes (5a–5c). The most intense band in 6a–6c corresponds to the benzoyloxy-carbonyl group. The 13C NMR spectra of oximes (5a–5c) and their esters (6a–6c) showed resonances for C-9. In the 1H NMR spectra, H1,5 protons were observed as separate signals. The weakest signal was assigned to the proton of the NOH group (5a–5c). Signals for the protons of the phenyl ring were observed in the spectrum of the O-benzoyloxime derivatives (6a–6c).The absorption bands of the C=N bond and the OH group are identified in the IR spectra of oximes  were synthesized. The complexes melted above 240 °C with decomposition.To study the biological properties of the novel derivatives of 3,7-diazabicyclo[3.3.1]nonan-9-ones, their water soluble complexes with β-cyclodextrin  have been studied for myelo-stimulating and plant growth-regulating activity nonan-9-one derivatives  demonstrated selectivity toward cancer HepG2 cells  of the compounds, HepG2 human liver carcinoma cells and WI-38 normal fibroblasts were cultivated in the presence of different concentrations of synthesized compounds, and cell viability and apoptosis induction were measured after 72 h of incubation. G2 cells A with siG2 cells B. Compou4c or 4e, and the induction of apoptosis was assessed by labeling cell membrane phosphatidylserine with annexin V-FITC and cell DNA with PI combined with flow cytometry. The results of the apoptosis evaluation were in good agreement with the results from the MTT test. Both compounds induced apoptosis more efficiently in cancer HepG2 cells than in normal WI-38 cells: 56% and 20% of cells remained alive after incubation with 4c or 4e, respectively , such as hydrogen peroxide and acrolein) are capable of inducing cell death via apoptosis ,26,32,33G2 cells  is not e4c may be the reason for its low activity in living cells. The tested compounds were converted into complexes with β-cyclodextrins that are widely used in medicinal chemistry to improve water solubility and the transportation of hydrophobic drugs to the target cell membrane barriers. The penetration of such complexes inside the cell may occur via two main pathways -3,7-diazabicyclo[3.3.1]nonane can be subjected to PA-based drug development.As demonstrated in 1a) were purchased from Aldrich . IR spectra were recorded on a Nicolet 5700 instrument between KBr plates. 1H and 13C NMR spectra were recorded on a JNM-ECA Jeol 400 spectrometer  using DMSO-d6 solvent. The elemental analysis data were consistent with the calculated values. Column chromatography and thin-layer chromatography were carried out on alumina (Al2O3) of the third degree of activity, and Rf compounds were given for this type of plate. The spots were developed in iodine vapors. The IR and NMR spectra for the synthesized compounds are presented in the Primary amines, paraform and 1-Boc-piperidin-4-one (1-(3-Hydroxypropyl)piperidin-4-one(1b). A total of 385 mL of absolute toluene and 16.26 g (0.707 mol) of metallic Na were placed in a three-necked flask equipped with a mechanical stirrer, reflux condenser with a calcium chloride tube, and dropping funnel. The reaction mixture was heated to 110 °C in an oil bath until the sodium dissolved (to obtain suspensions of sodium in toluene) for 25–30 min. Then, the reaction mixture was cooled to 75–80 °C, then 128 mL of methanol was added dropwise and it was stirred at 75 °C for 2 h. Afterwards, a straight condenser was attached to the flask, and the reaction mixture was heated. A solution of 174.61 g (0.707 mol) of diester was added using a dropping funnel at 110 °C, while the azeotropic mixture of toluene and methanol was simultaneously distilled off by direct distillation. The rate of dropping of the mixture of diester and methyl alcohol should be equal to the rate of distillation of the solvents. When the temperature reached 110 °C, becoming equal to the boiling point of toluene, the heating was stopped. While cooling (ice was used for cooling) and stirring, a solution of 301 mL of concentrated hydrochloric acid and 301 mL of distilled water was gradually added. The formed organic and aqueous layers were separated. The lower aqueous acidic layer was boiled at 100 °C for 8 h. The progress of the reaction was monitored using a solution of 1% FeCl3. The color changed to maroon. The solution was alkalized with NaOH to pH 12, extracted with chloroform, and dried over anhydrous MgSO4. The solvent was evaporated, and the residue was purified by column chromatography on Al2O3, benzene:dioxane = 5:1. A total of 21.04 g (19%) of 1-(3-hydroxypropyl)piperidin-4-one (1b) was obtained in the form of light yellow oil with Rf = 0.65 .8H15NO2.Found, %: C 61.11, H 9.58, N 8.90. CCalculated, %: C 61.14, H 9.55, N 8.92.−1: 1716 (C=О).IR spectrum, cm1H NMR spectrum, δ, ppm: bicyclononan-9-one: 1.56 , 2.07-2.45 , 2.85-3.22 , 3.47–3.59 , 3.81–3.89 . hydroxypropyl: 4.60 .13C NMR spectrum ppm: bicyclononan-9-one: 59.2 , 60.2 , 214.5 (C-9). hydroxypropyl: 27.5 (NCH2CH2CH2OH), 49.7, 63.7 (NCH2CH2CH2OH).1-Methoxypropyl-piperidin-4-one (1c). The procedure for the synthesis of 1-methoxypropyl-piperidin-4-one (1c) was described earlier [ earlier .3-Boc-7-[3-(1H-imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1]nonan-9-one (2a). The procedure for the synthesis of 3-Boc-7-[3-(1H-imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1]nonan-9-one (2a) was described earlier [ earlier .3-(3-Hydroxypropyl)-7-[3-(1H-imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1]nonan-9-one (2b). In a three-necked flask equipped with a stirrer, reflux condenser and dropping funnel, 40 mL of methanol was deoxygenated under a stream of nitrogen. After 30 min, a mixture of 9 mL (0.076 mol) of 1-(3-aminopropyl)imidazole), 9.17 g (0.304 mol) of paraform, 4 mL of concentrated hydrochloric acid, and 6 mL of glacial acetic acid was added and stirred for 15 min in an atmosphere of nitrogen. A solution of 12 g (0.076 mol) of 1-(3-hydroxypropyl)piperidin-4-one (1b) and 6 mL of glacial acetic acid in 43 mL of methanol was added dropwise. After 10 h of heating the reaction mixture at 60–65 °C, a second equivalent of paraform was added and held for another 10 h at the same temperature. The solvent was evaporated, and the residue was dissolved in 121 mL of water. The extraction of neutral products was carried out with diethyl ether. The aqueous layer was alkalinized to pH 12, and the organic part was extracted with chloroform and dried over MgSO4. The solvent was evaporated, and the obtained product was purified by column chromatography on Al2O3:benzene:dioxane 5:1. A total of 13.84 g (59%) of 3-(3-hydroxypropyl)-7-[3-(1H-imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1]nonan-9-one (2b) was obtained in the form of a light yellow oil with Rf = 0.41 .16H26N4O2.Found, %: C 62.72, H 8.52, N 18.27. CCalculated, %: C 62.75, H 8.50, N 18.30.−1: 1729 (C=О), 1071 (C-О-C).IR spectrum, cm1H NMR spectrum, δ, ppm: bicyclononan-9-one: 1.54 , 2.07–2.45 , 2.85–3.22 , 3.47–3.59 , 3.81–3.89 . 1H-imidazol-1-yl)propyl: 6.75–6.78, 6.82, 7.33 . Side chains: 1.54 ; 1.69–1.81 . hydroxypropyl: 4.60 .13C NMR spectrum ppm: bicyclononan-9-one: 59.4 , 61.4 , 214.5 (C-9). (1H-imidazol-1-yl)propyl: 119.0 (CHimidazol), 128.7, 137.2. Side chains: 29.3 (CH2CH2CH2), 44.9, 47.4 (CH2CH2CH2). hydroxypropyl: 27.5 (NCH2CH2CH2OH), 49.7, 63.7 (NCH2CH2CH2OH).3-(3-Methoxypropyl)-7-[3-(1H-imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1]nonan-9-one (2c). The procedure for the synthesis of 3-(3-methoxypropyl)-7-[3-(1H-imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1]nonan-9-one (2c) was described earlier [ earlier .3-(3-methoxypropyl)-7-[2-(pyridin-2-yl)ethyl]-3,7-diazabicyclo[3.3.1]nonan-9-one (2d). The procedure for the synthesis of 3-(3-methoxypropyl)-7-[2-(pyridin-2-yl)ethyl]-3,7-diazabicyclo[3.3.1]nonan-9-one (2d) was described earlier [ earlier .3-(3-Methoxypropyl)-7-[2-(piperazin-1-yl)]-3,7-diazabicyclo[3.3.1]nonan-9-one (2e). In a three-necked flask equipped with a stirrer, reflux condenser and dropping funnel, 50 mL of methanol was deoxygenated under a stream of nitrogen. After 30 min, a mixture of 7.55 g (0.058 mol) of 1-(2-aminoethyl)piperazine, 7 g (0.304 mol) of paraform, 3 mL of concentrated hydrochloric acid, and 4 mL of glacial acetic acid was added and stirred for 15 min in an atmosphere of nitrogen. A solution of 10 g (0.058 mol) of 1-(3-methoxypropyl)piperidin-4-one (1c) and 4 mL of glacial acetic acid in 43 mL of methanol was added dropwise. After 10 h of heating the reaction mixture at 60–65 °C, a second equivalent of paraform was added and held for another 10 h at the same temperature. The solvent was evaporated, and the residue was dissolved in 119 mL of water. The extraction of neutral products was carried out with diethyl ether. The aqueous layer was alkalinized to pH 12, and the organic part was extracted with chloroform and dried over MgSO4. The solvent was evaporated, and the obtained product was purified by column chromatography on Al2O3:benzene:dioxane 5:1. A total of 17.15 g (67%) of 3-(3-methoxypropyl)-7-[2-(piperazin-1-yl)ethyl]-3,7-diazabicyclo[3.3.1]nonan-9-one (2e) was obtained in the form of a light yellow oil with Rf = 0.32 .17H32N4O2.Found, %: C 62.89, H 9.81, N 19.79. CCalculated, %: C 62.96, H 9.88, N 19.75.−1: 1732 (C=О), 1089 (C-О-C).IR spectrum, cm1H NMR spectrum, δ, ppm: bicyclononan-9-one: 1.55 , 2.80–2.96 , 2.98 , 3.47–3.59 , 3.87 . 2-(Piperazin-1-yl)ethyl: 4.55–4.58 . methoxypropyl: 3.60–3.98 (NCH2CH2CH2OCH3).13C NMR spectrum, ppm: bicyclononan-9-one: 46.9 , 58.5 , 214.8 (C-9). 2-(Piperazin-1-yl)ethyl: 46.7 (CH2piperazin). methoxypropyl: 56.0, 27.8, 66.0, 71.4 (NCH2CH2CH2OCH3).3-Boc-7-[3-(1H-imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1]nonane (3a). The procedure for the synthesis of 3-Boc-7-[3-(1H-imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1]nonane (3a) was described earlier [ earlier .3-(3-Hydroxypropyl)-7-[3-(1H-imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1]nonane (3b). A mixture of 2.0 g (0.006 mol) of 3-(3-hydroxypropyl)-7-[3-(1H-imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1]nonan-9-one (2b) and 1 mL (0.031 mol) of hydrazine hydrate (99% solution) in 17.5 mL of triethylene glycol at 60 °C was added to 4.17 g (0.074 mol) of KOH. The reaction mixture was heated to 150 °C and stirred at this temperature for 5 h. At a temperature of 190–200 °C, water and excess hydrazine were removed by evaporation. After cooling, 29 mL of distilled water was added, extracted with diethyl ether, and dried over anhydrous MgSO4. The solvent was evaporated, and the obtained product was purified by column chromatography on Al2O3, benzene:dioxane 5:1. A total of 1.28 g (67%) of 3-(3-hydroxypropyl)-7-[3-(1H-imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1]nonane (3b) was obtained as a pale yellow oil with Rf = 0.36 .16H28N4O.Found, %: C 65.77, Н 9.61, N 19.20. CCalculated, %: C 65.75, Н 9.59, N 19.18.−1: 1121 (C-О-C).IR spectrum, cm1H NMR spectrum, δ, ppm: bicyclononane: 2.03–2.11 , 1.42–1.64 , 2.37 , 2.87–3.08 . 1H-imidazol-1-yl)propyl: 1.56 , 3.05–3.08, 3.86 , 7.45 , hydroxypropyl: 4.12 .13C NMR spectrum, δ, ppm: bicyclononane: 30.2 , 36.3 (C-9), 58.4 . (1H-imidazol-1-yl)propyl: 32.8 (CH2CH2CH2), 51.6 (CH2CH2CH2), 45.6 (CH2CH2CH2), 118.9, 128.7, 137.2 (CHimidazol). hydroxypropyl: 27.8 (NCH2CH2CH2OH), 51.6, 59.8 (NCH2CH2CH2OH).3-(3-Methoxypropyl)-7-[3-(1H-imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1]nonane (3c). The procedure for the synthesis of 3-(3-methoxypropyl)-7-[3-(1H-imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1]nonane (3c) was described earlier [ earlier .3-(3-Methoxypropyl)-7-[2-(pyridin-2-yl)ethyl]-3,7-diazabicyclo[3.3.1]nonane (3d). The procedure for the synthesis of 3-(3-methoxypropyl)-7-[2-(pyridin-2-yl)ethyl]-3,7-diazabicyclo[3.3.1]nonan-9-one (3d) was described earlier [ earlier .3-(3-Methoxypropyl)-7-[2-(piperazin-1-yl)]-3,7-diazabicyclo[3.3.1]nonane (3e). A mixture of 2.0 g (0.006 mol) of 3-(3-methoxypropyl)-7-[2-(piperazin-1-yl)ethyl]-3,7-diazabicyclo[3.3.1]nonan-9-one (2e) and 1 mL (0.031 mol) of hydrazine hydrate (99% solution) in 17.5 mL of triethylene glycol at 60 °C was added to 4.17 g (0.074 mol) of KOH. The reaction mixture was heated to 150 °C and stirred at this temperature for 5 h. At a temperature of 190–200 °C, water and excess hydrazine were removed by evaporation. After cooling, 29 mL of distilled water was added, extracted with diethyl ether, and dried over anhydrous MgSO4. The solvent was evaporated, and the obtained product was purified by column chromatography on Al2O3 and benzene:dioxane 5:1. A total of 1.21 g (63%) of 3-(3-methoxypropyl)-7-[2-(piperazin-1-yl)ethyl]-3,7-diazabicyclo[3.3.1]nonane (3e) was obtained as a pale yellow oil with Rf = 0.34 .17H34N4O.Found, %: C 65.79, H 10.81, N 18.00. CCalculated, %: C 65.81, H 10.97, N 18.06.−1: 1075 (C-О-C), 3374 (N-H).IR spectrum, cm1H NMR spectrum, δ, ppm: bicyclononan-9-one: 1.59 , 2.42 , 2.98 , 2.94 , 3.04 . 2-(Piperazin-1-yl)ethyl: 4.55–4.58 . methoxypropyl: 1.13 (NCH2CH2CH2OCH3).13C NMR spectrum, ppm: bicyclononan-9-one: 30.0 , 58.6 , 58.4 , 32,0 (C9). 2-(Piperazin-1-yl)ethyl: 54,8, 53,4, 54,8, 46,3 (7N-CH2CH2piperazine). methoxypropyl: 56.3, 27.6, 66.0, 72.4 (NCH2CH2CH2OCH3).Complex3-Boc-7-[3-(1H-imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1]nonane (4a). The procedure for the synthesis of the complex of 3-Boc-7-[3-(1H-imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1]nonane (4a) was described earlier [ earlier .Complex of 3-(3-hydroxypropyl)-7-[3-(1H-imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1]nonane (4b). Hot solutions of 0.5 g (0.002 mol) of 3-(3-hydroxypropyl)-7-[3-(1H-imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1]nonane (3b) in 25 mL of ethyl alcohol and 1.94 g (0.002 mol) of β-cyclodextrin in 30 mL of distilled water were mixed. The mixture was placed in a drying cupboard, and ethanol and water were evaporated at 50–55 °C to produce 2.25 g of compound 4b.58H98N4O36.Found, %: C 48.84, Н 6.84, N 3.95. CCalculated, %: C 48.81, Н 6.87, N 3.93.Complex of 3-(3-methoxypropyl)-7-[3-(1H-imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1]nonane (4c). The procedure for the synthesis of the complex of 3-(3-methoxypropyl)-7-[3-(1H-imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1]nonane (4c) was described earlier [ earlier .Complex of 3-(3-methoxypropyl)-7-[2-(pyridin-2-yl)ethyl]-3,7-diazabicyclo[3.3.1]nonane (4d). The procedure for the synthesis of the complex of 3-(3-methoxypropyl)-7-[2-(pyridin-2-yl)ethyl]-3,7-diazabicyclo[3.3.1]nonan-9-one (4d) was described earlier [ earlier .Complex of 3-(3-methoxypropyl)-7-[2-(piperazin-1-yl)]-3,7-diazabicyclo[3.3.1]nonane (4e). Hot solutions of 1.4 g (0.003 mol) of 3-(3-methoxypropyl)-7-[2-(piperazin-1-yl)]-3,7-diazabicyclo[3.3.1]nonane (3e) in 35 mL of ethyl alcohol and 4.0 g (0.003 mol) of β-cyclodextrin in 60 mL of distilled water were mixed. The mixture was placed in a drying cupboard, and ethanol and water were evaporated at 50–55 °C to produce 4.75 g of compound 4e.59H104N4O36.Found, %: C 49.01, H 7.14, N 3.91. CCalculated, %: C 49.03, H 7.20, N 3.88.Oxime of 3-Boc-7-[3-(1H-imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1]nonane-9-one (5a). Five grams (0.014 mol) of 3-Boc-7-[3-(1H-imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1]nonan-9-one (2a) in 85 mL of ethyl alcohol and 1.7 mL (0.021 mol) of pyridine were placed in a three-necked flask equipped with a mechanical stirrer, reflux condenser with a calcium chloride tube, and dropping funnel. While stirring, 2.53 g (0.036 mol) of hydroxylamine hydrochloride was added. The reaction mixture was heated at 110–120 °C for 27 h, the solvent was evaporated, and the residue was dissolved in 40 mL of water, alkalized with NaOH to pH 12, extracted with chloroform, and dried over anhydrous MgSO4. The solvent was evaporated, and the residue was purified by column chromatography on Al2O3, benzene:dioxane = 5:1. A total of 4.5 g (86%) of oxime of 3-Boc-7-[3-(1H-imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1]nonan-9-one (5a) was obtained in the form of light yellow oils with Rf = 0.16 .18H29N5O3.Found, %: C 59.52; H, 8.00; N, 19.30. CCalculated, %: C 59.50; H, 7.98; N, 19.28.−1: 1651 (C=N), 3380 (О-H).IR spectrum, cm1Н NMR spectrum, δ, ppm: bicyclononan-9-ketoxime: 2.69 , 2.07–4.10 , 2.60–4.72 , 6.96 . 1H-imidazol-1-yl)propyl: 1.56 , 3.13–3.22, 3.85 , 6.96–7.49 . Boc: 1.48 .13C NMR ppm, δ, ppm: bicyclononan-9-ketoxime: 36.9 (C-5), 50.5 , 60.1 , 154.6 (C-9). (1H-imidazol-1-yl)propyl: 30.8 (CH2CH2CH2), 54.1 (CH2CH2CH2), 44.1 (CH2CH2CH2), 120.0 (CHimidazol), 128.7, 137.9. Boc: 28.6 (CH3), 79.1 (O-C), 158.3 (C=O).Oxime of 3-(3-hydroxypropyl)-7-[3-(1H-imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1]nonane (5b). A total of 6.0 g (0.020 mol) of 3-(3-hydroxypropyl)-7-[3-(1H-imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1]nonan-9-one (2a) in 122 mL of ethyl alcohol and 2.37 g (0.030 mol) of pyridine were placed in a three-necked flask equipped with a mechanical stirrer, reflux condenser with a calcium chloride tube, and dropping funnel. While stirring, 3.61 g (0.052 mol) of hydroxylamine hydrochloride was added. The reaction mixture was heated at 110–120 °C for 29 h, the solvent was evaporated, and the residue was dissolved in 30 mL of water, alkalized with NaOH to pH 12, extracted with chloroform, and dried over anhydrous MgSO4. The solvent was evaporated, and the residue was purified by column chromatography on Al2O3, benzene:dioxane = 5:1. A total of 4.85 g (77%) of oxime of 3-(3-hydroxypropyl)-7-[3-(1H-imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1]nonan-9-one (5b) was obtained in the form of light yellow oils with Rf = 0.07 .16H27N5O2.Found, %: C 59.79, H 8.40, N 21.84. CCalculated, %: C 59.81, H 8.41, N 21.81.−1: 1510 (C=N), 3240 (О-H).IR spectrum, cm1Н NMR spectrum, δ, ppm: bicyclononan-9-ketoxime: 1.60 , 2.06–2.40 , 2.99–3.42 , 6.40 . 1H-imidazol-1-yl)propyl: 1.60 , 3.42, 3.85–3.87 , 6.92, 7.23, 7.45 . hydroxypropyl 4.83 , 1.88–1.90 , 2.40, 3.68 .13C NMR ppm, δ, ppm: bicyclononan-9-ketoxime: 42.2 (C-5), 57.7 , 58.1 , 160.9 (C-9). (1H-imidazol-1-yl)propyl: 27.7 (CH2CH2CH2), 50.0 (CH2CH2CH2), 45.4 (CH2CH2CH2), 119.0 (CHimidazol), 128.8, 137.1. hydroxypropyl: 27.7 (NCH2CH2CH2OH), 51.5, 59.5 (NCH2CH2CH2OH).Oxime of 3-(3-methoxypropyl)-7-[3-(imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1]nonan-9-one (5c). A total of 3.5 g (0.008 mol) of 3-(3-methoxypropyl)-7-[3-(imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1]nonan-9-one (2c) in 65 mL of ethyl alcohol and 1.3 mL (0.018 mol) of pyridine were placed in a three-necked flask equipped with a mechanical stirrer, reflux condenser with a calcium chloride tube, and dropping funnel. While stirring, 2.15 g (0.027 mol) of hydroxylamine hydrochloride was added. The reaction mixture was heated at 110–120 °C for 27 h, the solvent was evaporated, and the residue was dissolved in 27 mL of water, alkalized with NaOH to pH 12, extracted with chloroform, and dried over anhydrous MgSO4. The solvent was evaporated, and the residue was purified by column chromatography on Al2O3, benzene:dioxane = 5:1. 3.1 g (76%) of oxime of 3-(3-methoxypropyl)-7-[3-(imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1]nonan-9-one (5c) was obtained in the form of light yellow oils with Rf = 0.21 .17H33N5O2.Found, %: C 60.86, H 9.72, N 20.95. CCalculated, %: C 60.90, H 9.85, N 20.89.−1: 1659 (C=N), 3365 (О-H).IR spectrum, cm1Н NMR spectrum, δ, ppm: bicyclononan-9-ketoxime: 2.54 , 2.12–3.78 , 2.30–43.57 , 6.96 . 1H-imidazol-1-yl)propyl: 1.53 , 3.16–3.20, 3.75 , 6.96–7.49 . methoxypropyl: 3.83 , 1.75 , 2.53, 3.74 .13C NMR ppm, δ, ppm: bicyclononan-9-ketoxime: 37.9 (C-5), 53.5 , 65.1 , 161.5 (C-9). (1H-imidazol-1-yl)propyl: 28.8 (CH2CH2CH2), 45.9 (CH2CH2CH2), 44.8 (CH2CH2CH2), 115.4, 119.7, 127.5. (CHimidazol). methoxypropyl: 29.2 (CH3), 73.9 (O-C), 149.9 (C=O).O-Benzoyloxime of 3-Boc-7-[3-(1H-imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1] nonan-9-one (6a). Then, 4.42 g (0.012 mol) of oxime of 3-Boc-7-[3-(1H-imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1]nonan-9-one (5a) was mixed with 84 mL of absolute benzene, and a mixture of 14 mL of absolute benzene and 2.8 mL (0.024 mol) of benzoyl chloride was added dropwise. The reaction took place at room temperature. Finally, 10 mL of distilled water was added to the reaction mixture and neutralized with potash to pH 10–11. The product was extracted with chloroform, and the combined extracts were dried over anhydrous MgSO4. The solvent was evaporated, and the residue was distilled in vacuo. A total of 2.66 g (50% of theory) of the O-benzoyloxime of 3-Boc-7-[3-(1H-imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1]nonan-9-one (6a) was obtained in the form of a yellow oil (16), Rf = 0.56 .25H33N5O4.Found, %: C 64.27; H, 7.04; N, 15.01. CCalculated, %: C 64.24; H, 7.07; N, 14.99.−1: 1725 (C=О).IR spectrum, cm1Н NMR spectrum, δ, ppm: ester of bicyclononan-9-ketoxime: 2.77 , 2.12–4.15 , 2.65–4.77 , 7.67–8.05 . 1H-imidazol-1-yl)propyl: 1.56 , 3.13–3.22, 3.85 , 6.96, 7.03, 7.49 . Boc: 1.48 .13C NMR spectrum, δ, ppm: ester of bicyclononan-9-ketoxime: 36.9, 39.9 , 45.5 60.0 , 164.7 (C-9). (1H-imidazol-1-yl)propyl: 29.4 (CH2CH2CH2), 50.1 (CH2CH2CH2), 47.9 (CH2CH2CH2), 120.5 (CHimidazol), 133.9, 136.3. Boc: 28.6 (CH3), 80.2 (O-C), 159.5 (C=O).O-Benzoyloxime of 3-(3-hydroxypropyl)-7-[3-(1H-imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1]nonan-9-one (6b). Oxime (0.014 mol) of 3-(3-hydroxypropyl)-7-[3-(1H-imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1]nonan-9-one (5b) was mixed with 100 mL of absolute benzene, and a mixture of 14 mL of absolute benzene and 3.3 mL (0.014 mol) of benzoyl chloride was added dropwise. The reaction took place at room temperature. Finally, 10 mL of distilled water was added to the reaction mixture and neutralized with potash to pH 10–11. The product was extracted with chloroform, and the combined extracts were dried over anhydrous MgSO4. The solvent was evaporated, and the residue was distilled in vacuo. A total of 4.01 g (70% of theory) of O-benzoyloxime of 3-(3-hydroxypropyl)-7-[3-(1H-imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1]nonan-9-one (6b) was obtained in the form of a yellow oil, Rf = 0.64 .23H31N5O3.Found, %: C 64.90, H 7.32, N 16.45. CCalculated, %: C 64.94, H 7.29, N 16.47.−1: 1737 (C=О).IR spectrum, cm1Н NMR spectrum, δ, ppm: ester of bicyclononan-9-ketoxime: 2.74 , 2.11–2.24 , 2.65–2.77 , 7.67–8.05 . 1H-imidazol-1-yl)propyl: 1.56 , 3.13–3.22, 3.85 , 6.96, 7.03, 7.49 . hydroxypropyl: 4.83 , 1.75 , 2.56, 3.76 .13C NMR spectrum, δ, ppm: ester of bicyclononan-9-ketoxime: 37.7–40.7 , 59.1–59.4 , 167.6 (C-9). (1H-imidazol-1-yl)propyl: 29.4 (CH2CH2CH2), 50.1 (CH2CH2CH2), 48.0 (CH2CH2CH2), 120.5 (CHimidazol), 133.9, 136.3. hydroxypropyl: 28.1 (NCH2CH2CH2OH), 52.4, 60.6 (NCH2CH2CH2OH).O-benzoyloxime of 3-(3-methoxypropyl)-7-[3-(imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1]nonan-9-one (6c). Three grams (0.009 mol) of oxime of 3-(3-methoxypropyl)-7-[3-(imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1]nonan-9-one (5c) was mixed with 90 mL of absolute benzene, and a mixture of 12 mL of absolute benzene and 2 mL (0.014 mol) of benzoyl chloride was added dropwise. The reaction took place at room temperature. Finally, 8 mL of distilled water was added to the reaction mixture and neutralized with potash to pH 10–11. The product was extracted with chloroform, and the combined extracts were dried over anhydrous MgSO4. The solvent was evaporated, and the residue was distilled in vacuo. A total of 3.05 g (64% of theory) of O-benzoyloxime of 3-(3-methoxypropyl)-7-[3-(imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1]nonan-9-one (6c) was obtained in the form of a yellow oil, Rf = 0.61 .17H33N5O2.Found, %: C 60.86, H 9.72, N 20.95. CCalculated, %: C 60.90, H 9.85, N 20.89.−1: 1734 (C=О).IR spectrum, cm1Н NMR spectrum, δ, ppm: ester of bicyclononan-9-ketoxime: 2.71 , 2.01–2.14 , 2.35–2.45 , 7.67–8.05 . 1H-imidazol-1-yl)propyl: 1.56 , 3.13–3.22, 3.85 , 6.96, 7.03, 7.49 . hydroxypropyl: 4.83 , 1.75 , 2.56, 3.76 .13C NMR ppm, δ, ppm: bicyclononan-9-ketoxime: 36.0 (C-5), 51.9 , 64.5 , 159.5 (C-9). (1H-imidazol-1-yl)propyl: 27.5 (CH2CH2CH2), 43.9 (CH2CH2CH2), 45.2 (CH2CH2CH2), 116.2, 118.4, 129.2. (CHimidazol). methoxypropyl: 27.4 (CH3), 75.1 (O-C).Complex of O-benzoyloxime of 3-Boc-7-[3-(1H-imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1]nonan-9-one (7a). Hot solutions of 1.0 g (0.002 mol) of O-benzoyloxime of 3-Boc-7-[3-(1H-imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1]nonan-9-one (6a) in 25 mL of ethyl alcohol and 3.124 g (0.002 mol) of β-cyclodextrin in 40 mL of distilled water were mixed together. The mixture was placed in a drying cupboard, and ethanol and water were evaporated at 50–55 °C to produce 3.57 g of compound 7a.67H103N5O39.Found, %: C 50.25, H 6.46, N 4.35. CCalculated, %: C 50.22, H 6.43, N 4.37.Complex of O-benzoyloxime of3-(3-hydroxypropyl)-7-[3-(1H-imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1]nonan-9-one (7b). Hot solutions of 1.5 g (0.003 mol) of O-benzoyloxime of 3-(3-hydroxypropyl)-7-[3-(1H-imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1]nonan-9-one (6b) in 35 mL of ethyl alcohol and 4 g (0.003 mol) of β-cyclodextrin in 60 mL of distilled water were mixed together. The mixture was placed in a drying cupboard, and ethanol and water were evaporated at 50–55 °C to produce 4.98 g of compound 7b.65H101N5O38.Found, %: C 50.01, H 6.45, N 4.51. CCalculated, %: C 50.03, H 6.48, N 4.49.Complex of O-benzoyloxime of3-(3-methoxypropyl)-7-[3-(1H-imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1]nonan-9-one (7c). Hot solutions of 2.5 g (0.006 mol) of O-benzoyloxime of 3-(3-methoxypropyl)-7-[3-(1H-imidazol-1-yl)propyl]-3,7-diazabicyclo[3.3.1]nonan-9-one (6c) in 45 mL of ethyl alcohol and 6.8 g (0.006 mol) of β-cyclodextrin in 100 mL of distilled water were mixed together. The mixture was placed in a drying cupboard, and ethanol and water were evaporated at 50–55 °C to produce 8.8 g of compound 7c.66H103N5O39.Found, %: C 49.81, H 6.52, N 4.31. CCalculated, %: C 49.84, H 6.48, N 4.40.g for 20 min at +4 °C, and the supernatant was used as a source of polyamine oxidases in their natural environment for the following in vitro evaluation of the action of bispidines on their activities.The regenerating rat liver was used as the source of the enzymes of PA catabolism . The stuDetermination of amine oxidase activity in the presence of the tested compounds was performed in rat liver homogenates, as described previously . A 30% lHumаn hepаtocellulаr cаrcinomа HepG2 cell line  wаs grown in RPMI-1640 medium . Normаl humаn fibroblаsts WI-38  were grown in DMЕM medium  and were used as control noncancer cells. Both cell lines were passage number 4. Аll mediа were supplemented with 5% fetаl bovine serum  аnd 1% of sodium pyruvаte . Cells were grown аt 5% CО2/95% аir in а humidified аtmosphere аt 37 °C to confluency 80–90%. To harvest, the cells were washed up to three times with 5 mL prewarmed phosphate-buffered saline  and detached with prewarmed 0.25% trypsin/EDTA . Cell lines were tested for mycoplаsmа contаminаtion before the experiment using Mycoplаsmа Detection Kit PlаsmoTest™ .4 cells per well. In 24 h, synthesized compounds within the rаnge of concentrаtions 1–50 µM were added and incubated for 72 h. Tetrаzolium sаlt -2,5-diphenyltetrаzolium bromide, Servа, Heidelberg, Germаny) solution (10 µL) was added to each well to reach a final concentration of 0.45 mg/mL and incubated at 37 °C for 4 hr. Аfter incubаtion, the formаzаn crystаls were dissolved in 100 µL dimethyl sulfoxide (DMSO), аnd the аbsorbаnce wаs meаsured аt 570 nm with а CLARIOstar Plus multiplаte reаder . The percentage of cell viability was calculated relative to nontreated control cells.To determine the influence of the compounds on cell viability, an MTT assay was performed . BrieflyThe mаximum nontoxic concentrаtion (MNTC) wаs considered the highest concentrаtion of the compound thаt produced no stаtisticаlly significаnt increаse in cell viability. Compound concentrаtions thаt produce 50% of cell viаbility (IC50) were cаlculаted from curves constructed by plotting cell viаbility (%) versus drug concentrаtion (µM) . To test4 cells аt eаch point wаs performed by flow cytometry with а MАCS Quаnt Аnаlyzer 10  аs it wаs previously described -3,7-diazabicyclo[3.3.1]nonane demonstrated the ability to activate PA catabolism in regenerating rat liver homogenates. This compound could significantly decrease the viability of cancer cells in vitro. Adding Spd or Spm as a substrate of oxidation to boost toxicity, provided convincing evidence for the anticancer potential of bispidines. Minimally toxic bispidines strongly synergized the potency of the two PAs in the tumor cell line but not in normal fibroblasts. We can conclude that the lead compound 4e can become a potential anticancer drug substance which mechanism relies on the induction of PA catabolism in tumor cells.