Mono-6-O-(p-toluenesulfonyl)-beta-cyclodextrin

Synthesis was carried out as described in.1 In a 1L- double-necked, round-bottomed flask, β-CD (40.0 g, 35.2 mmol) was dissolved in deionized water (450 mL) by heating at 60 °C with vigorous stirring. To the resulting milky suspension, 1-(p-toluenesulfonyl)-imidazole (12 g, 54 mmol) was added. After 2 hrs stirring, a solution of sodium hydroxide (9 g, 225 mmol) in 25 mL of water was added slowly (in about 20 minutes). After further 20 minutes stirring, unreacted 1-(p-toluenesulfonyl)imidazole was separated by filtration through a sintered glass funnel. The reaction was quenched by the addition of ammonium chloride (24.1 g, 900 mmol) with swirling to dissolve all the solids. The resulting mixture was concentrated to about half of its originalvolume by blowing a stream of air over its surface. The resulting suspension was filtered througha large sintered-glass funnel and collected solid was washed with ice water (2x100 mL) andacetone (1x200 mL) and dried over CaCl2 in vacuum desiccators until constant weight. 20.63 g, 16 mmol, 45.39percent.Synthesis was carried out as described in [1]. In a 1L- double-necked, round-bottomed flask, β-CD (40.0 g, 35.2 mmol) was dissolved in deionized water (450 mL) by heating at 60 °C with vigorous stirring. To the resulting milky suspension, 1-(p-toluenesulfonyl)-imidazole (12 g, 54 mmol) was added. After 2 hrs stirring, a solution of sodium hydroxide (9 g, 225 mmol) in 25 mL of water was added slowly (in about 20 minutes). After further 20 minutes stirring, unreacted 1-(p-toluenesulfonyl)imidazole was separated by filtration through a sintered glass funnel. The reaction was quenched by the addition of ammonium chloride (24.1 g, 900 mmol) with swirling to dissolve all the solids. The resulting mixture was concentrated to about half of its original volume by blowing a stream of air over its surface. The resulting suspension was filtered through a large sintered-glass funnel and collected solid was washed with ice water (2x100 mL) and acetone (1x200 mL) and dried over CaCl2 in vacuum desiccators until constant weight. 20.63 g, 16 mmol, 45.39percent.TLC. Rf: 0.88 (i-Pr-OH:H2O:EtOAc:NH4OH/5:5:1:1), 1-H NMR (500 MHz, DMSO-d6) δ: 2.42 (s, 3 H), 3.20-3.65 (overlap with HDO, m, 40 H), 4.15-4.23 (m, 1 H), 4.30-4.38 (m, 2 H), 4.40-4.54 (m, 2 H), 4.50 (br s, 3 H), 4.76 (br s, 2 H), 4.84 (br s, 4 H), 5.62-5.84 (m, 14 H), 7.43 (d, 2 H, J = 7.75), 7.73 (d, 2 H, J = 7.75).The synthesis of mono-6-(p-toluenesulphonyl)-6-deoxy-β-cyclodextrin was carried out using a previously reportedprocedure.43, 44 In a 1 L double necked round-bottomed flask, β-cyclodextrin (40.0 g, 35.2 mmol) was dissolved in deionized water(450 mL) at 60 °C with vigorous stirring. 1-(p-Toluenesulphonyl)-imidazole (12 g, 54 mmol) was added to the resulting milkysuspension at room temperature and stirred for 2 h. After that, a solution of sodium hydroxide 9 g (225 mmol, in 25 mL water)was added dropwise over 20 min. After addition of thesodium hydroxide solution, the separation of unreacted 1-(ptoluenesulphonyl)imidazole was carried out by simple filtrationusing a sintered glass funnel. Finally the reaction was quenchedby the addition of ammonium chloride (24.1 g, 900 mmol), withagitation to dissolve the added ammonium chloride. The resultingmixture was concentrated to about half of its original volume byblowing a stream of air over its surface. The resulting suspensionwas filtered using a sintered-glass funnel and the collected solidwas washed twice using 200 mL ice-cold water and 100 mL ofacetone in one portion. The collected solid was well dried overnightin a vacuum at 60 °C and kept over CaCl2 in vacuum desiccatorsyielding 18.30 g, or 40.26percent.β-Cyclodextrin (35.0 g, 30.8 mmol) was dissolved in 350 mL HIn a 250-mL three-necked round-bottomed flask, bCD (5.0 g, 4.4 mmol) was dissolved in 112.5 mL of water by heating to60 C under vigorous stirring.28 After cooling to room temperature, finely powdered 1-(p-toluenesulfonyl)imidazole (3.9 g, 17.7 mmol)was added to the suspension. After 6 h, a solution of sodium hydroxide (2.3 g, 56.3 mmol) in 6.3 mL of water was added over20 min. After 10 min, unreacted 1-(p-toluenesulfonyl)imidazolewas removed by filtration. To the filtrate was added ammoniumchloride (6.1 g, 112.5 mmol) to quench the reaction. The resultingmixture was concentrated by blowing a stream of air across themixture and the product began to precipitate out of the solution.The suspension was filtered, and the solid was washed with icewater and acetone. The vacuum-dried tosyl bCD was obtained in28.2percent yield. The mono-tosylated bCD was treated with an equivalentamount of sodium azide in 16 mL of water at 80 C for 5 h.29After cooling, the solution was precipitated with acetone and thenlyophilized. Lyophilized azido bCD and triphenylphosphine (PPh3)(224 mg, 848 lmol) were dissolved in DMF (8 mL) and stirred for2 h at room temperature. After adding 1.6 mL of water, the solutionwas stirred for 3 h at 90 C, and the resulting product was precipitatedwith acetone. Amino bCD was purified by cation-exchangechromatography (CM-Sephadex C25) using 0.5-M ammoniumbicarbonate as a solvent and desalted with Bio-gel P2. The productwas confirmed using 1H NMR spectra.Tosyl -CD was obtained from the Microbial CarbohydrateResource Bank (MCRB) at Konkuk University, Korea. Tosyl -CDwas prepared according to the method in Byun (Byun, Zhong, &Bittman, 2000). In a 1 L three-necked round-bottom flask, -CD(10.0 g, 8.8 mmol) was dissolved in 250 mL of water by heating it to60C under vigorous stirring. After cooling to room temperature, finely powdered 1-(p-toluenesulfonyl)imidazole (6 g, 27 mmol)was added to the suspension. After 6 h, a solution of sodium hydrox-ide (4.5 g, 112.5 mmol) in 12.5 mL of water was added over 20 min.After 10 min, the unreacted 1-(p-toluenesulfonyl)imidazole wasseparated by filtration. Ammonium chloride (12.05 g, 225 mmol)was then added to the reaction mixture to quench the reaction.The resulting mixture was concentrated by blowing a stream of aironto the surface overnight, after which the product precipitatedfrom the solution. The suspension was filtered, and the solid waswashed with ice water and acetone twice. The vacuum-dried tosyl-CD was obtained at a 28.2percent yield.In a 1L three-necked round-bottom flask, β-CD (10.0g, 8.8mmol) was dissolved in 250mL of water by heating it to 60°C under vigorous stirring. After cooling to room temperature, finely powdered 1-(p-toluenesulfonyl)imidazole (6g, 27mmol) was added to the suspension. After 6h, a solution of sodiumhydroxide (4.5g, 112.5mmol) in 12.5mL of water was added over 20min. After 10min, the unreacted 1-(p-toluenesulfonyl)imidazole was separated by filtration. Ammonium chloride (12.05g, 225mmol) was then added to the reaction mixture to quench the reaction. The resulting mixture was concentrated by blowing a stream of air onto the surface overnight, after which the product precipitated from the solution. The suspension was filtered, and the solid was washed with ice water and acetone twice. The vacuum-dried tosyl β-CD was obtained at a 28.2percent yield.General procedure: Biotin tethering to cyclooligosaccharides starts from monotosylation on the primary hydroxylgroup, but the monotosylation method of both cyclooligosaccharides slightly differed from each other.For monotosylation of β-CD, β-CD (10.0 g, 8.8 mmol) was dissolved in water (250 mL) and heated to60 °C. After cooling to room temperature, 1-(p-toluenesulfonyl) imidazole (6 g, 27 mmol) was addedand mixed for 6 h. Sodium hydroxide (4.5 g, 112.5 mmol) solution in water (12.5 mL) was addeddropwise for 20 min. After filtration, the filtrate was quenched using ammonium chloride (12.05 g, 225 mmol), and the resulting mixture was subjected to air flow for drying. The precipitated productwas washed with water and acetone. The yield of monotosylated β-CD was 28percent. For monotosylatedCyS, CyS (1 g, 0.28 mmol) was dissolved in water (50 mL), and copper sulfate (1 g, 4 mmol) in water(100 mL), was added. NaOH (1 g, 25 mmol) solution in water (50 mL) was mixed and stirredfor 10 min. The suspension became dark blue, and p-toluenesulfonyl chloride (2.5 g, 13.2 mmol)dissolved in 0.1 mL of acetonitrile was added in a dropwise manner for 1 h. The suspension wasstirred at room temperature for 4 h then neutralized with ammonium chloride. After removing the precipitate, the filtrate was concentrated and desalted on a Bio-Gel P2 column. The sample wassubjected to semi-preparative HPLC (Agilent Technologies 1200 series, Santa Clara, CA, USA) on areverse-phase column (Eclipse XDB-C18, 9.4 x 250 mm, 5 μm) at room temperature. The yieldof monotosylated CyS was 25percent. Further azidation and amination were performed, as describedin our previous report [43, 44]. The mono-6-amino-β-CD or mono-6-amino-CyS were dissolved inDMF and reacted with biotinamidohexanoic acid N-hydroxysuccinimide ester for 24 h at 40 °C.After acetone precipitation, the samples were desalted and lyophilized. The overall synthetic yield fromoriginal cyclooligosaccharide to biotinylated one is about 8percent. The structures of the resulting biotinylcyclooligosaccharides were analyzed by NMR spectroscopy and MALDI-TOF mass spectrometry.25 g (0.022 mol; 1 eq.) of β-cyclodextrin (Roquette Freres SA) are suspended in 200 ml of distilled water, in a 500 ml Erlenmeyer flask, and then sodium hydroxide chips (8.8 g) are added in a single fraction. A 500 mL round-bottom flask equipped with a magnetic stirbar, a vacuum adapter and a septum was charged with a solution of dry β-cyclodextrin (8.530 g, 7.51 mmol) and 200 mL of dry pyridine. The solution was cooled to 0 °C before 1.29g (6.76 mmol) of tosyl chloride was added. The resulting solution was allowed to warm to room temperature overnight The pyridine was removed as much as possible in vacuo. The resulting residue was then recrystallized twice from 40 mL of hot water to yield 7.54 (88percent) of a white crystalline solid.Example 8 Synthesis of β-cyclodextrin-Tosylate, 16 (Melton, L. D., and Slessor, K. N., Carbohydrate Research, 18, p. 29 (1971))Synthesis of mono-6-deoxy-6-(p-tolylsulfonyl)^-cyclodextrin (CDOTs): A solution of p-toluenesulfonyl chloride (846.3 mg, 4.44 mmol) in 5 ml ACN was added to 80 ml aqueous solution of β-CD (5.0 g, 4.44 mmol) and NaOH (434.8 mg, 10.8 mmol) dropwise over 15 min. After stirring for 4 hrs at °C in NTo a solution of β-CD (2.5 g, 2.2 mmol) in water (110 mL), copper sulfate (1.65 g, 6.6 mmol) in water (165 mL) and sodium hydroxide (2.2 g, 55 mmol) in water (55 mL) were sequentially added. After 10 min, p-toluenesulfonyl chloride (3.3 g, 17.4 mmol) in acetonitrile (22 mL) was added drop by drop during 1 h. The reaction mixture was stirred for 4 h at ambient temperature, and then neutralized (pH = 12.5) with hydrochloric acid, the salts were eliminated by filtering and the volume of solution was decreased to 2/3 of its initial volume by lyophilization. _he β-CD-OTs was crystallized (three times) by dissolving in boiling water and washed with acetone (2* 7 mL), ether (2* 9 mL) and dried. After recrystallizations, pure β-CD-OTs (1.34 g, 46percent) was achieved (Baussanne et al., 2000).β-CD (50.0 g, 44.0 mmol) was dissolved in 500 mL of a 0, 4 M aqueous sodium hydroxide solution and cooled to 4 °C. While stirring vigorously, p-toluenesulfonyl chloride (35.0 g, 184 mmol) was added within 5 minutes. The resulting suspension was stirred for additional 30 minutes and was then filtered. The filtrate was neutralized by addition of hydrochloric acid and stirred for an hour. The precipitate that was formed thereby was isolated via filtration, washed with water (3x) and dried at 60 °C overnight.Yield: 24.68 g (19.16 mmol, 43percent)71H-NMR (500 MHz, DMSO-d6): δ [ppm]= 7.77 (d, 2H), 7.45 (d, 2H), 5.74 (m, 14H), 4.79 (t, 7H), 4.50 (t, 7H), 3.18-3.78 (m, 42H), 2.45 (s, 3H).FT-IR [cm-1]: 3301 (b), 2924 (w), 1641 (w), 1554 (w), 1358 (w), 1021 (s), 836 (w).MALDI-TOF-MS (m/z)= 1311.4 [M+Na]+, 1327.5 [M+K]+, 1465.5 [disubst. CD + Na]+.To a NaOH solution (3.6 g, 0.09mol of NaOH in water120mL)was added β-CD (6.0 g, 5.3mmol)at 0-5. p-Tolylsulfonyl chloride (TsCl, 1.2 g, 6.3mmol)was added into the solution with vigorous stirring at 0-5. After5h at 0-5, the precipitate wasremoved by filtration. 10percent HCl wasadded into the filtrate, and thepH was adjusted to pH=7. The mixture was kept in the refrigerator overnight to afford a whitesolid product. The white solid was recrystallized in hot water to afford 2.3 gof product (yield 38.3percent).According to the published method [Journal of Chromatography A, 2009, 1216(2), 257-63], β-CD (6.0 g, 5.3 mmol) and NaOH (3.6 g, 0.09 mol) were stirred in a water solution (120 mL) at 0-5. Then p-tolylsulfonyl chloride (TsCl, 1.2 g, 6.3 mmol) was added into the solution at 0-5. After 5 h at 0-5, the precipitate was removed by filtration. 10percent HCl was added into the filtrate, and the pH was adjusted to pH=7. The mixture was kept in the refrigerator overnight to afford a white solid product. The white solid was recrystallized in hot water to afford 2.3 g of product (yield 38.3percent). The characteristic data were in accordance with literature [Journal of Chromatography A, 2009, 1216(2), 257-63]A solution of bCD was prepared by slowly adding 35.2 mL(88 mmol) of aqueous 2.5 N NaOH to 10 g (8.8 mmol) of bCD beingsuspended in 90 mL water. After 5 min, 2.52 g (13.22 mmol) of ptoluenesulfonylchloride was added in five portions and the formedsuspension was continuously stirred for 1 h at 25 C (400 rpm, magnetic stirrer). After filtration and washing of the residue(2 10 mL water), a total of 91 mL of pre-swollen cation exchangeresin 50W 4 20–50 mesh in the H+ form (100 meq, exchangecapacity 1.1 meq/mL) was added to the filtered solution. After20 min, the mixture was first filtered through a Büchner funnelprovided with glass wool to collect the exchange resin. Second, the precipitated white product was collected on filter paper. Afterwashing with 3 30 mL water, the product was lyophilized.Yield: 35percent (based on starting b-cyclodextrin).Melting point: 160–162 C.FTIR: 1600 cm1 (Ph-SO2–), 1230 cm1 (Ph-SO2-R), 815 cm1(Ph-SO2-O-R), 665 cm1 (Ph-SO2–).1H NMR (DMSO-d6): d = 7.77 (mc, 2H Ph), 7.43 (mc, 2H Ph), 5.83–5.63 (m, 14H, OH1 and OH3 Cyd), 4.85–4.77 (m, 7H, H1Cyd), 4.51–4.45 (m, 6H, OH6 CyD), 4.35 (mc, 2H, H60 CyD), 4.20(mc, 1H, H50 CyD), 3.75–3.40 (m, 25H, H3, H5 and H6 CyD), 3.40–3.15 (m, H2, H4 overlap with water), 2.44 (s, 3H, Ph-CH3) ppm.Derivatization Degree (DD): >98 mol percent.13C NMR (DMSO-d6): d = 144.77 (C Ts); 132.846 (C Ts); 129.98(C Ts); 127.55 (C Ts); 101.89 (C1); 98.14 (C10); 81.47(C4); 81.15(C40); 73.02, 72.40, 72.00 (C2, C3, C5); 69.67 (C50); 68.87 (C60);59.88 (C6); 21.17 (CH3 Ts) ppm.m/z (ESI): calculated 1289.1970, found 1289.3770 [M+H]+.Elemental analysis: calculated—C: 45.65; H: 5.94; S: 2.49; detected—C: 45.71; H: 6.29; S: 2.84.11.35 g (10 mmol) of β-CD were dissolved in 200 mL of 0.6 M NaOH(aq) at 0 °C. After 30 min, 2.3 g (12.1 mmol) of 4-Toluenesulfonyl chloride (TsCl), finely powdered with a mortar and pestle, wereadded to the aqueous solution. The reaction mixture was magnetically stirred for 8 h at 0 °C. Theunreacted TsCl was removed by filtration onto a sintered glass funnel. The aqueous phase was acidifiedat 0 °C with 10 mL of HCl (37percent) added dropwise. The product precipitated as white solid. The mixturewas kept at 4 °C overnight, filtered on paper and washed with deionized water until neutral pH. In orderto reduce the content of unreacted β-CD, the solid was dispersed in 100 mL of hot water (65 °C), stirred for 10 min, and finally filtered. This procedure was qualitatively followed by TLC (eluent:2-propanol:H2O:EtOAc:NH4OH = 5:3:1:0.5) and repeated for 3 times until the β-CD spot on TLC (Rf:0.25) was negligible compared to the CD1 spot (Rf: 0.47). The final product was washed with acetone(100 mL × 3 times), dried in air for 24 h and under vacuum (<5 mbar) at 25 °C for 3 h. Yield: 35percent(4.50 g). The tosylation procedure gave also the formation of very small amounts of ditosylates (Rf = 0.58), according to the literature [Brady2000]. Due to the small ΔRf between the mono- and the di-tosylate noattempt of further separation was made. ESI-MS analysis performed on the final product confirmed thepresence of negligible amounts of pristine CD and di-tosylate (see Figure SI2).2.3 A three-liter, three-necked, round-bottomed flask equipped with a mechanical stirring and thermometer was charged with β-cyclodextrin (1) hydrate (50 g, 44 mmol) and a solution of 25 g (625 mmol) of sodium hydroxide in 1.0 liter of water. The solution was stirred at 0-5° C. in an ice-water bath and p-toluenesulfonyl chloride (TsCl) (20 g, 105 mmol) was added in one portion. The reaction mixture was stirred vigorously for 2 h at 0-5° C., and then another portion of TsCl (30 g, 157 mmol) was added and the reaction mixture stirred at this temperature for further 3 h. The reaction mixture was filtered in a fritted glass funnel to separate unreacted TsCl. The filtrate was cooled at 0-5° C. while 10percent aqueous hydrochloric acid (HCl, 150 ml) was added. The resulting solution was stored overnight in a refrigerator at 0° C., and then filtered. The product was dried and recrystallized from boiling water. Storage provided 14.0 g (25percent) of 2 as a white solid. TLC analysis of 2 performed on silica plates (EtOAc:2-propanol:conc. NHβ-CD (24 g) was suspended in 180 mL of water, NaOH (2.623 g) in 20 mL of water was added dropwise over 30 min, and reacted under vigorous agitation at 0 °C for a period of 1 h then p-toluenesulfonyl chloride (4.032 g) in 20 mL of acetonitrile was added dropwise over 1 h, causing immediate formation of a white precipitate. The detailed preparation for the picolinamide-modified β-cyclodextrin/Pd(II) complex (Pd(II)PCA-β-CD) is illustrated in Scheme 1. First, mono-6-tosyl β-cyclodextrin (1, Tos-β-CD) was obtained by the method of Khan etc (Khan & Pitchumani, 2016). Then, Tos-β-CD was reacted with NaNA solution of sodium hydroxide (6.0000g, 150mmol) in water (20mL) was added dropwise to a solution of β-CD (56.7490g, 50mmol) in water (500mL) with magnetic stirring at 10–15°C over about 15min. The solution became homogeneous, and then a solution of p-toluenesulfonyl chloride (11.4384g, 60mmol) in acetonitrile (30mL) was added dropwise at 10–15°C over about 45min forming white precipitate immediately. The resultant solution was kept stirring for 3.0h, and rose to room temperature. The precipitate formed was collected by suction filtration and then suspended in water (300mL) with magnetic stirring at room temperature for 3.0h. The precipitate collected by suction filtration was washed successively with acetone (100mL) and water (160mL), and then dried in vacuum at 80°C for 8.0h to afford white solid powder 8.1746g in 12.68percent yield. (0016) [α]β-cyclodextrin (120.0 g, 105.8 mmol) was suspended in 800 ml of water. NaOH (13.14 g, 328 mmol) in 40 ml water was added dropwise. The suspension became homogeneous before the addition was complete. p- Toluenesulfonyl chloride (20.16 g, 105.8 mmol) in 60 ml of acetonitrile was added dropwise. After 4 hours of reaction at room temperature the precipitate was removed by filtration and 8 mmol diluted HCl was added into the filtrate. The filtrate was then refrigerated overnight at 4°C. The resulting white precipitate was collected by filtration and dried, yielding the crude product. The pure product was obtained by recrystallization in hot water. Yield: 10percent. Take 210 g of β-cyclodextrin recrystallized from water, then added in portions with 1300 mL of water in a three-necked flask, and stirred at room temperature. Weigh 17.2 g of sodium hydroxide dissolved in 50 mL of water, and then slowly the aqueous sodium hydroxide solution was added dropwise to the β-cyclodextrin suspension until the reaction solution was gradually clarifies, and stirring was continued for 1.5 h. After the cyclodextrin was completely dissolved and clarified in the reaction solution, a solution of 26.0 g of p-toluenesulfonyl chloride in acetonitrile (80 mL) was slowly added dropwise to the reaction solution (dropwise over about 30 minutes). After stirring was continued for 2 hours, the solution was filtered off Insoluble matter, the filtrate was adjusted to pH = 7.5 with 2 mol / L hydrochloric acid, a large number of white precipitate precipitated. The precipitate was collected by suction filtration and the precipitate was dissolved in 450 mL of distilled water under heating. The insoluble material was filtered while hot and the filtrate was allowed to stand at room temperature for 12 h. The precipitate was collected and recrystallized several times with water until thin layer chromatography showed the product is Pure. Obtained 17g of final product, yield 8percent;β-Cyclodextrin hydrate (12.5 g, 11 mmol) was dissolved in a solution which was prepared by dissolving 6.25 g of sodium hydroxide in 375 mL of water in 1 L round bottom flask. The mixture was cooled to 0–5 °C and stirred at same temperature in an ice-water bath while p-toluenesulfonyl chloride (5 g, 26.25 mmol) was being added in small portions. The reaction mixture was stirred vigorously for 2 h at 0–5 °C and then p-toluenesulfonyl chloride (7.5 g, 39.25 mmol) was added in small portions and the mixture was stirred at 0–5 °C temperature for 3 h further. The reaction mixture was filtered to separate unreacted tosyl chloride. Filtrate was cooled at 0–5 °C while 10percent aqueous hydrochloric acid (87.5 mL) was being added. Resulting solution was stored for 8 h in a refrigerator at 0 °C, and then filtered. The product was dried to constant weight in a vacuum oven to yield a white solid of TsO–β-CD. Further the white solid was recrystallized three times by dissolving it in 40 mL of boiling water and then cooling to room temperature (Brady et al., 2000, 2004; Lovrinovic & Niemeyer, 2007). Yield 3.97 g, 28percent.

Molecular Weight:1289.2
XLogP3:-13
Hydrogen Bond Donor Count:20
Hydrogen Bond Acceptor Count:37
Rotatable Bond Count:10
Exact Mass:1288.3786145
Monoisotopic Mass:1288.3786145
Topological Polar Surface Area:586
Heavy Atom Count:87
Complexity:2260
Defined Atom Stereocenter Count:35
Covalently-Bonded Unit Count:1
Compound Is Canonicalized:Yes