Es of BA labelled with greenemitting BODIPY (four,4difluoro4bora3a,4adiazasindacene) [28,29] and redemitting Rhodamine B [30] were synthesized to study its localization and trafficking in living cells. Within this operate, we synthesized and studied new derivatives of BA and BT labelled at C3 and C28 positions using a tiny blueemitting BODIPY dye. two. Materials and Solutions 2.1. Chemical Synthesis Aluminium silica gel sheets for detection in UV light (TLC Silica gel 60 F254, Merck, Darmstadt, Denmark) had been employed for thinlayer chromatography (TLC), subsequent visualization was proceeded by a diluted solution of sulfuric acid in methanol and plates were heated. Silica gel (300 , SiliTech, MP Biomedicals, Costa Mesa, CA, USA) was applied for column chromatography. NMR Spectra were recorded by AgilentMR DDR2 (Santa Clara, CA, USA). HRMS were measured by LTQ ORBITRAP VELOS with HESI/HESIionization (Thermo Scientific, Waltham, MA, USA). For microwave synthesis, an Initiator Classic 355,301 (Biotage, Uppsala, Sweden) was utilized. The following chemicals have been bought from TCI Europe (Zwijndrecht, Belgium): N,N,NtriethylamineEt3 N (99 ), 4dimethylaminopyridine4DMAP (99 ), 1(3dimethylaminopropyl)3ethylcarbodiimide hydrochlorideEDCI (98 ), N,N dicyclohexylcarbodiimideDCC (98 ), 1hydroxybenzotriazole monohydrateHOBt (97 ), triphenylphosphinePPh3 (95 ), ptoluenesulfonic acid monohydratepTsOH (98 ), and palladium on carbonPd/C (ten ). The following chemical compounds have been purchased from SigmaAldrich (Prague, Czech Republic): 3azidopropylamine (95 ), 1(2Nbocaminoethyl)piperazine (95 ), alanine (99 ). Betulinic acid (BA) was bought from Betulinines (St rnskalice, Czech Republic). r The solvents for column chromatography and reactions have been purchased from PENTA (Praha, Czech Republic) and had been utilised devoid of additional distillation. Compound Synthesis and Characterization 8N(3Azidopropyl)amino4,4difluoro4bora3a,4adiazasindacene (BODIPYN3 ) To a option of BODIPYSMe (205 mg, 0.86 mmol) in DCM (ten mL), 3azidopropylamine (95 mg, 0.95 mmol) was added and also the mixture was stirred for 30 min at RT. The solvents had been evaporated below reduced stress plus the residue was taken up with AcOEt and also the solution was precipitated by the addition of Teflubenzuron In stock hexanes. BODIPYN3 (243 mg, 0.83 mmol) was obtained as a yellowish strong in 97 yield. RF = 0.55 in hexanesAcOEt 1:1. 1 H NMR (400 MHz, CD3OD) ppm: 2.11 (quin, J = 6.7 Hz, two H), three.56 (t, J = 6.5 Hz, two H), 3.87 (t, J = 7.4 Hz, two H), 6.39 (br. s., 1 H), six.55 (br. s, 1 H), 7.32 (br. s, 1 H), 7.36 (br. s., 2 H), 7.57 (s, 1 H). 13 C NMR (101 MHz, CD3 OD) ppm: 27.00, 44.08, 48.76, 112.67, 113.99, 115.78, 123.16, 130.78, 133.76, 148.87. HRMSESI: calculated 290.12628 Da, identified m/z 291.13312 [MH] . 8N(3Aminopropyl)amino4,4difluoro4bora3a,4adiazasindacene (Pregnenolone 16α-carbonitrile web BODIPYNH2 ) To a answer of BODIPYN3 (150 mg, 0.52 mmol) in AcOEt (eight mL), was added Pd/C (80 mg) plus the mixture was stirred beneath hydrogen atmosphere for 2 h. The catalyst was filtered off plus the solvents were evaporated beneath reduced pressure. The residue was taken up with AcOEt and also the item was obtained immediately after precipitation with hexane. BODIPYNH2 (96 mg, 0.36 mmol) was obtained as yellow solid in 70 yield. RF = 0.15 in DCMMeOH 20:1 (v/v). 1 H NMR (400 MHz, CD3 OD) ppm: 1.98 (quin, J = 6.7 Hz, 2 H), 2.87 (t, J = six.7 Hz, two H), 3.83 (t, J = 7.0 Hz, 2 H), six.37 (br. s, 1 H), 6.52 (br. s, 1 H), 7.27 (br. s, 1 H), 7.30.36 (m, 2 H), 7.55 (br. s, 1 H). 13 C NMR (101 MHz, CD3 OD) ppm: 29.67, 39.12, 45.50, 1.