Supporting Information

for

Angew. Chem. Int. Ed. Z53466

© Wiley-VCH 2004

69451 Weinheim, Germany

Synthetic Strategy for Achieving Skeletal Diversity via Solid-Supported and Otherwise Unstable Reactive Intermediates.

Steven J. Taylor, Alexander M. Taylor, and Stuart L. Schreiber

Howard Hughes Medical Institute Harvard Institute of Chemistry and Cell Biology (ICCB)

Department of Chemistry and Chemical Biology Harvard University, Cambridge, Massachusetts 02138

I. Synthetic Methods Materials and Methods:

Except as otherwise noted, reactions were carried out under nitrogen with dry, freshly purified solvents. Solvents were passed through a solvent column (activated alumina, and Q5 reactant) prior to use. Triethylamine and 2,6-lutidine were distilled over calcium hydride under a nitrogen atmosphere. NMR spectra were recorded at 600 MHz, 500 MHz, or 400 MHz using a Varian I-600, I-500, or M-400 instrument respectively. 13C spectra were recorded on a Bruker DMX-500 MHz spectrometer. 1H NMR chemical shifts are reported relative to residual CHCl3 (7.26 ppm) or d-6 DMF (8.02, 2.96, 2.88 ppm). 13C NMR data were recorded relative to the solvent residue. Flash chromatography was performed with the indicated solvent on E. Merck silica gel 60 (40-63 mesh) or neutral alumina as indicated. Separations were monitored on E. Merck silica gel 60 F254 plates (0.25mm), and visualized with a UV lamp, or by staining with phosphomolybdic acid or cerium sulfate-ammonium molybdate solutions followed by heating. Reverse-phase LC/MS data was obtained with a Gilson/Finnigan LC/MS system. LC/MS chromatography was performed on a SymmetryShieldTM RP8, 3.5 µM, 4.6 x 100mm column (Waters Corporation, Milford, MA, Batch #111) using a flow rate of 1 mL/min and a 10 min gradient of 20-80% CH3CN in water, constant 0.1% formic acid, with UV detection at 214 and 254 nm. Percent purity of the crude compounds cleaved off the solid-support was determined by LC/MS (total peak area at 214 nm, 254 nm, or TIC whichever greatest) prior to purification. Low resolution mass spectra were obtained with JEOL AX-505H, SX-102A (CI/EI), Micromass Platform II and LCT (APCI/ES/LCMS). Only molecular ions, fractions from molecular ions and other major peaks are reported. Molecular modeling was performed with the Spartan software package (Wavefunction) using Hartree-Frock, Semi-empirical (AM1) and molecular mechanics modeling methods with the 3-21* basis set. Brominated polystyrene (Br-PS, 2 mmol/g) was obtained from Polymer Labs and functionalized with the silicon-based linker according to the reported protocol. 2-brombenzylbromide, 3-pyridinepropanol, diphenylphosphorylazide, dimethylacetylenedicarboxylate, 4-fluorophenylisocyanate, and benzaldoximechloride were all distilled under vacuum prior to use. 7-hydroxylisoquinoline was recrystalized from ethanol and dried under high vacuum prior to use. All other organic reagents were purchased from Aldrich, Lancaster, or Acros and used without further purification. General Procedure for Solid-Phase Reactions: All solid phase reactions (with the exception of loading the alcohols) were conducted in oven dried glass vials under and atmosphere of dry argon. Loading of the alcohols on solid support was performed in fritted polypropylene reaction tubes equipped with septum under an atmosphere of dry argon. Reactions were agitated by placement of the vessels on a Thermolyne RotoMix type 5080 shaker. After washings the

resin was dried under house vacuum for 1-4 h followed by high vacuum overnight. Reactions were examined prior to HF-py cleavage with MAS 1H NMR analysis to determine conversion. General Procedure for HF/Py cleavage of the reagents off of the solid support, and analysis of purity: Either an Eppendorf tube or 20 mL polypropylene vial was charged with the resin followed by the cleavage cocktail solution (85/10/5 THF/py/HF-py by volume, 100 µL solution per 10 mg of resin) and the samples were vortexed for 1-2 h at rt. The samples were then treated with methoxytrimethylsilane (volume equivalent to the amount of cleavage cocktail solution used), and vortexed for an additional 30 min to quench excess HF. The samples were then filtered trough a pipette plugged with glass wool and filled with celite (~ 1 cm). The resin and celite were washed 2x with methylene chloride, 2x with acetonitrile, and 2x with methylene chloride. The filtrates were collected and concentrated in vacuo prior to analysis and purification. II. Synthesis of Compounds on Solid Support

Loading of 7-hydroxyisoquinoline (2) on PS-macrobeads:

3-[Diisopropyl(p-methoxyphenyl)silyl]propyl functionalized polystyrene beads (1) (macrobeads, 1.00 g, 1.4 mmol/g, 1.4 mmol) in a 15 mL plastic column equipped with a septum and a plastic 3-way stopcock were flushed with argon for 10 min and then left under an argon atmosphere. Dichloromethane (10 mL) was added to the vessel via syringe, and the resin was allowed to swell for 15 min. The solvent was then removed from the vessel via argon push, and a 5% (v/v) solution of chlorotrimethylsilane in dichloromethane (10 mL of solution, 3.93 mmol TMS-Cl) was added to the vessel. The beads were agitated on a shaker for 30 min. The beads were filtered, washed thrice with CH2Cl2 (5 min each), and suspended in a 3% (v/v) solution of trifluoromethanesulfonic acid (12.0 mL, 4.03 mmol TfOH, 3.9 equiv.) in CH2Cl2 for 40 min, during which the vessel was shaken periodically. The beads were filtered away from the solution via argon push and suspended in a 3% (v/v) solution of trifluoromethanesulfonic acid (13.0 mL, 4.37 mmol TfOH, 3.9 equiv.) in CH2Cl2 for 40 min, during which the vessel was shaken periodically. The beads were then filtered, washed twice with CH2Cl2 (15 mL, 15 min), and then swollen in a minimal amount of CH2Cl2. Freshly distilled 2,6-lutidine (1.34 mL, 11.5 mmol, 6 equiv.) was added to the beads, which turned from red to clear over the course of a few min. The 7-hydroxyisoquinoline (610 mg, 4.20 mmol, 3 equiv.) was then added via syringe in 7 mL of DMF and the resin gently rocked for 20 h. The beads were then filtered, rinsed, and suspended in DMF (rinsed 3x 5 min, suspended for 2x 20 min) and then filtered away from the solvents. This washing procedure was repeated with THF and CH2Cl2, and then the reaction beads were dried under house vacuum (2 h) followed by high vacuum overnight providing (2). Resin (100.0 mg) was then subjected to the cleavage conditions that yielded 32 mg of crude product, 17.4 mg purified (1.2 mmol

isoquinoline / g resin). > 95% pure by LC/MS (retention time 1.32 min), 1H NMR matches that of the commercially available 7-hydroxyisoquinoline.

2-(2-Bromo-benzyl)-7-hydroxy-isoquinolinium bromide (3): An oven dried amber 20 mL vial equipped with a Teflon-lined silicon septum was charged with 2 (1.2 mmol/g, 900.0 mg, 1.08 mmol), capped, and purged with argon for 5 min. A 10 mL flame-dried pear-shaped flask was charged with 2-bromobenzylbromide (2.90 g, 11.6 mmol, 10 equiv.), and diethyl ether (3 mL) was added to the flask. The resulting solution was transferred to the vial containing the resin, the flask was washed 2x with 1mL diethyl ether, and the washings were added to the resin. The reaction vial was sealed with Teflon tape and gently rocked overnight. After 16 h the solvents were removed via canula, and diethyl ether (10 mL) was added via syringe to the reaction. The solution stood for 5 min and then the ether was removed via canula. This washing procedure was repeated with diethyl ether (2x 5 min), toluene (2x 5 min), CH2Cl2 (2x 5 min), and again with diethyl ether (2x 5 min). The beads were then dried under house vacuum (2 h) followed by high vacuum overnight. The resin (56.7 mg) was exposed to the cleavage conditions, which yielded 26.0 mg of compound (0.06 mmol, 1.20 mmol/g 100%). 1H NMR (500 MHz, d-6 dmso): δ 9.95 (s, 1H), 8.50 (m, 2H), 8.27 (m, 1H), 7.83 (m, 1H), 7.85 (m, 1H), 7.68 (m, 1H), 7.49 (m, 1H), 7.42 (m, 1H), 7.36 (m, 1H), 6.03 (s, 1H), 6.01 (s, 1H) ppm; 13C NMR (125 MHz, d6-dmso): δ 160.0, 148.6, 148.5, 133.8, 132. 6, 132.2, 131.8, 131.7, 130.6, 129.1, 128.9, 126.3, 123.7, 110.7 63.7 ppm; LCMS: 4.17 min, 315.5, 94% of total peak area at 254 nm; HRMS (TOF ES) 314.0181 calcd for m/z (M+); 314.0184 (1.0 ppm) observed.

2-(2-Bromo-benzyl)-1-vinyl-1,2-dihydro-isoquinolin-7-ol (4): An oven dried amber 20 mL vial equipped with a Teflon-lined silicon septum was charged with 3 (1.2 mmol/g, 200.0 mg, 0.240 mmol), capped, and purged with argon for 5 min. The vial was equipped with a nitrogen inlet, and diethyl ether (3 mL) was added to the reaction vessel via a syringe. The solution was

cooled to -78 °C (dry ice, 2-propanol), and vinylmagnesium bromide (1.0M in THF 2.4 mL, 2.4 mmol, 10 equiv.) was added over 5 min. The mixture was allowed to react for 5 min at -78 °C and then placed into a refrigerator at 0 °C. The reaction was allowed to proceed with occasional stirring for 5 h. The vial was removed from the refrigerator and then placed into an ice/water bath. The solvents were removed from the reaction vessel via canula, and the beads were taken up in 5 mL of dry diethyl ether. The ether was allowed to swell the beads for 1 min and then removed via canula. This was repeated with diethyl ether (1x) and with dichloromethane (4x). The resin was dried under house vacuum for 5 h and then overnight under high vacuum. (600 MHz, CDCl3): δ 7.30 (brs, 2H), 6.0 (brs 2H), 5.75 (brs 1H), 5.30 (brs, 1H), 4.90 (brs, 1H, 4.80 (brs, 1H), 4.57 (brs, 1H), 4.30 (brs, 1H), 3.90 (brs, 1H), 2.50 (brs, 1H) ppm. Regions between 6.1-7.4 ppm and 0-1.5 ppm could not be accurately analyzed due to polystyrene peaks (matched compound synthesized independently in solution, see page 8 of the supporting information). The beads were subjected to the cleavage conditions, which resulted in the complete decomposition of the product.

Loading of 3-Pyridin-4-yl-propan-1-ol on PS macrobeads: 3-[Diisopropyl(p-methoxyphenyl)silyl]propyl functionalized polystyrene beads 1 (macrobeads, 1.00 g, 1.4 mmol/g, 1.4 mmol) in a 15 mL plastic column equipped with a septum and a plastic 3-way stopcock were flushed with argon for 10 min and then left under an argon atmosphere. Dichloromethane (10 mL) was added to the vessel via syringe and the resin was allowed to swell for 15 min. The solvent was then removed from the vessel via argon push, and then a 5% (v/v) solution of chlorotrimethylsilane in dichloromethane (10 mL of solution, 3.93 mmol TMS-Cl) was added to the vessel, followed by agitation of the beads on a shaker for 30 min. The beads were filtered, and washed thrice with CH2Cl2 (5 min each), and suspended in a 3% (v/v) solution of trifluoromethanesulfonic acid (12.0 mL, 4.03 mmol TfOH, 3.9 equiv.) in CH2Cl2 for 40 min during which the vessel was shaken periodically. The beads were filtered away from the solution via argon push and suspended in a 3% (v/v) solution of trifluoromethanesulfonic acid (13.0 mL, 4.37 mmol TfOH, 3.9 equiv.) in CH2Cl2 for 40 min during which the vessel was shaken periodically. The beads were then filtered, washed twice with CH2Cl2 (15 mL, 15 min), and then swollen in a minimal amount of CH2Cl2. Freshly distilled 2,6-lutidine (1.23 mL, 10.6 mmol, 6 equiv.) was added to the beads, which turned from red to clear over the course of a few min. The 3-pyridin-4-yl-propan-1-ol (531 mg, 3.87 mmol, 3 equiv.) was then added via syringe in 7 mL of DMF and the resin gently rocked for 20 h. The beads were then filtered, rinsed, suspended in DMF (rinsed 3 x 5 min, suspended for 2 x 20 min) and then filtered away from the solvents. This washing procedure was repeated with THF and CH2Cl2, and then the reaction beads were dried under house vacuum (2 h) followed by high vacuum overnight. The resin (100.0 mg) was then subjected to the cleavage conditions, which yielded 46 mg of crude product, 15.0 mg purified (1.05 mmol 3-pyridin-4-yl-propan-1-ol / bead). > 95% pure by LC/MS (retention time 0.82 min), 1H NMR matches that of the commercially available 3-pyridin-4-yl-propan-1-ol.

1-(2-Bromo-benzyl)-4-(3-hydroxy-propyl)-pyridinium bromide:

An oven-dried amber 20 mL vial equipped with a Teflon-lined silicon septum was charged with 2 (1.2 mmol/g, 870.0 mg, 1.08 mmol), capped, and purged with argon for 5 min. A 10 mL flame dried pear-shaped flask was charged with 2-bromobenzylbromide (2.21g, 8.87 mmol, 10 equiv.), and diethyl ether was added to the flask. The resulting solution was transferred to the vial containing the resin, and the flask washed 2 x with 1mL diethyl ether and the washings added to the resin. The reaction vial was sealed with Teflon tape and gently rocked overnight. After 16 h the solvents were removed via canula, and diethyl ether (10 mL) was added via syringe to the reaction. The solution stood for 5 min., and then the ether was removed via canula. This washing procedure was repeated with diethyl ether (2x 5 min), toluene (2x 5 min), CH2Cl2 (2x 5 min), and again with diethyl ether (2x 5 min). The beads were then dried under house vacuum (2 h) followed by high vacuum overnight. 56.7 mg of the resin was exposed to the cleavage conditions, which yielded 22 mg of compound (0.0504 mmol, 1.08 mmol/g 100%). 1H NMR (400 MHz, CD3CN); δ 8.73 (s, 1H), 8.71 (s, 1H), 7.94 (s, 1H), 7.92 (s, 1H), 7.74 (m, 1H), 7.57 (m, 1H), 7.51 (m, 1H), 7.42 (m, 1H), 5.87 (s, 2H), 3.54 (t, 2H), 3.01 ( t, 2H), 1.90 (m, 2H) ppm; 13C NMR (125 MHz, CD3CN): δ 149.2, 139.0, 137.7, 137.6, 137.2, 134.0, 133.5, 129.6, 68.6, 65.3, 37.5, 37.3 ppm; LCMS: 0.82 min. 308.0, 100% of the total peak area at 254nm; HRMS (TOF ES) 306.0494 calcd for m/z (M+H); 306.0485 (2.9 ppm) observed.

3-[1-(2-Bromo-benzyl)-2-vinyl-1,2-dihydro-pyridin-4-yl]-propan-1-ol (5): An oven-dried amber 20 mL vial equipped with a Teflon-lined silicon septum was charged with polystyrene bound 1-(2-bromo-benzyl)-4-(3-hydroxy-propyl)-pyridinium bromide (1.2 mmol/g, 200.0 mg, 0.210 mmol), capped, and purged with argon for 5 min. The vial was equipped with a nitrogen inlet, and diethyl ether (3 mL) was added to the reaction vessel via a syringe. The solution was cooled to -78 °C (dry ice, 2-propanol), and vinylmagnesium bromide (1.0M in THF 2.1 mL, 2.1 mmol, 10 equiv.) was added over 5 min. The reaction was allowed to react for 5 min at -78 °C and then placed into a refrigerator at 0 °C. The reaction was allowed to proceed with occasional stirring for 5 h. The vial was removed form the refrigerator and then placed into an ice/water bath. The solvents were removed from the reaction vessel via canula, and then the beads were taken up in 5 mL of dry diethyl ether. The ether was allowed to swell the beads for 1 min and then removed via canula. This was repeated with diethyl ether (1x), and with dichloromethane (4x). The resin was dried under house vacuum for 5 h then overnight under high vacuum. 1H MAS NMR (600 MHz, CDCl3): δ 7.50 (d, J= 24Hz, 1H), 6.0 (brs, 1H), 5.99 (m, 1H), 4.98 (d, J= 16.2 Hz, 1H), 4.92 (brs,

1H), 4.74 (brs, 1H), 4.59 (brs, H), 4.29 (brs, 2H), 4.08 (d, J= 15.0 Hz, 1H), 3.66 (brs, 2H), 2.50 (brs, 2H), 2.07 (brs, 2H), 1.68 (brs) ppm. Regions between 6.1-7.4 ppm and 0-1.5 ppm could not be accurately analyzed due to polystyrene peaks. (Matched compound synthesized independently in solution). The beads were subjected to the cleavage conditions, which resulted in the complete decomposition of the product.

2-(2-Bromo-benzyl)-1-vinyl-1,2,3,4-tetrahydro-isoquinolin-7-ol (6): An oven-dried vial was charged with (50 mg, 0.060 mmol) the resin bound enamine 4, and then the sodiumcyanoborahydride (124.0 mg, 0.300 mmol, 5 equiv.) was added to the vial. The vial was quickly capped with a septum and then purged with argon for 5 min. The vial was charged with CH2Cl2 (2.5 mL) and the beads allowed to swell for 5 min. After 5 min trifluoroethanol (2.5 mL) was added via syringe, the vial sealed with Teflon tape, and the reaction gently agitated overnight. After 15 h the solvents were removed via canula and the beads washed with CH2Cl2 (2x, 5 mL, 5 min), CF3CH2OH, (2x, 5 mL, 1 min.), 2:2:1 DMF, MeOH, TEA (2x, 5mL, 2 min) and then CH2Cl2 (3x, 5mL, 3 min.). The beads were then dried under house vacuum (2 h) then high vacuum overnight. The resin was then exposed to the cleavage conditions providing 16 mg (0.047 mmol, 0.93 mmol/g, 78%) of 6. 1H NMR (500 MHz, CDCl3): δ 7.61 (m, 1H), 7.54 (m, 1H), 7.29 (m, 1H), 7.11 (m, 1H), 6.98 (m, 1H), 6.67 (m, 1H), 6.61 (m, 1H), 5.90 (m, 1H), 5.33 (d, J= 10.0 Hz, 1H), 5.31 (d, J= 16.0 Hz, 1H), 4.11 (d, J= 8.5 Hz, 1H) 4.04 (d, J= 15.0 Hz, 1H), 3.65 (d, J= 15.5 Hz, 1H), 3.05 (m, 1H), 2.83 (m, 1H), 2.74 (m, 1H), 2.59 (m, 1H) ppm; 13C NMR (125 MHz, CDCl3): δ 155.8, 150.0, 141.2, 140.2, 134.9, 132.8, 132.3, 130.6, 129.8, 129.7, 129.5, 126.8, 120.9, 117.1, 116.3, 69.8, 60.4, 49.4, 30.7 ppm; LCMS: 4.41 min, 345 75% of the total peak area at 254nm; HRMS (TOF ES) 344.0605 calcd for m/z (M+H); 344.0644 observed.

4-(2-Bromo-benzyl)-1-phenyl-5-vinyl-3a,4,5,9b-tetrahydro-isoxazolo[5,4-c]isoquinolin-7-ol (7): An oven-dried vial was charged 4 (50 mg, 0.07 mmol), sealed, and purged with Argon. CH2Cl2 (1 mL) was added via syringe to the reaction vessel. A second oven-dried vial was charged with benzaldoxime chloride (54 mg, 0.35 mmol, 5 equiv.) and CH2Cl2 (1 mL), sealed, and purged with Argon. Triethylamine (10 µL, 0.7 mmol, 10 equiv.) was added to the second vial, which was agitated 10 min at rt. The oxime chloride solution was then transferred to the vial containing 4 via canula. The oxime chloride vial was rinsed with CH2Cl2 (1 mL), and this rinse was added to the vial containing 4 via canula. The mixture was agitated 1h at rt. The solvent was removed via canula, and the beads were washed CH2Cl2 (3x), THF (3x), acetonitrile (3x), and CH2Cl2 (3x). The beads were then dried under house vacuum followed by high vacuum overnight. The resin was then exposed to the cleavage conditions providing 16.7 mg (0.0362 mmol, 0.72 mmol/g, 58%) of 7. 1H NMR (600 MHz, CDCl3): δ 7.55 (m, 3H), 7.39 (m, 4H), 7.27 (t, J= 7.2 Hz, 7.2 Hz, 2H), 7.15 (t, 7.5, 7.5 Hz, 1H), 7.0 (d, J= 8.5 Hz, 1H), 6.54 (dd, J= 8.5, 2.5 Hz, 1H), 6.51 (s, 1H), 6.22 (m, 1H), 6.01 (d, J= 9.5 Hz, 1H), 5.21 (d, J= 10.2 Hz, 1H), 5.08 (d, J= 17.4 Hz, 1H), 4.75 (d, J= 9 Hz, 1H), 4.32 (s, 2H), 4.25 (d, J= 7.8 Hz, 1H) ppm; 13C NMR δ (500 MHz, CDCl3): 136.6, 135.2, 134.5, 130.6, 128.1, 127.8, 127.1, 126.4, 126.3, 126.0, 125.8, 125.4, 124.8, 124.7, 114.6, 112.4, 112.1, 111.4, 93.2, 59.8, 52.2, 44.3, 28.3 ppm; LCMS 9.07 min, 460.6 m/z LCMS: 58% of total peak area monocyclized product, 32% of total peak area dicyclized, 254nm; HRMS (TOF ESI) 461.0864 calcd for m/z (M+H); 461.0872 (1.7) observed.

4-(2-Bromo-benzyl)-1-phenyl-5-(3-phenyl-4,5-dihydro-isoxazol-4-yl)-3a,4,5,9b-tetrahydro-isoxazolo[5,4-c]isoquinolin-7-ol: An oven-dried vial was charged 4 (50 mg, 0.07 mmol), sealed, and purged with Argon. CH2Cl2 (1 mL) was added via syringe to the reaction vessel. A second oven dried vial was charged with benzaldehydeoxime chloride (54 mg, 0.35 mmol 5equiv.) and CH2Cl2 (1 mL), sealed, and purged with Argon. Triethylamine (10 µL, 0.7 mmol, 10 equiv) was added to the second vial, which was agitated 10 min at rt. The oxime chloride solution was then transferred to the vial containing 4 via canula. The oxime chloride vial was rinsed with CH2Cl2 (1 mL), and this rinse was added to the vial containing 4 via canula. The mixture was agitated overnight at rt. The solvent was removed via canula, and the beads were washed CH2Cl2 (3x), THF (3x), acetonitrile (3x), and CH2Cl2 (3X). The beads were then dried under house vacuum followed by high vacuum overnight. The resin was then exposed to the cleavage conditions providing 15.8mg (0.0272 mmol, 0.54 mmol/g 45%) of the biscycloadduct. 1H NMR (600 MHz, CDCl3): δ 6.7-7.6 (m, 17H), 6.01 (d, J= 10.2 Hz, 1H), 5.42 (dt, J= 10.2, 3.0 Hz, 1H), 4.74 (d, J= 10.2 Hz, 1H), 4.57 (d, J= 15.0 Hz, 1H), 4.52 (d, J= 14.4 Hz, 1H), 4.35 (d, J= 3.0 Hz, 1H), 3.35 (dd, J= 17.4, 10.8 Hz, 1H), 2.67 (dd, J= 17.4, 9.6 Hz, 1H) ppm; 13C NMR (500 MHz, CDCl3) δ 175.7, 136.9, 133.6, 130.9, 130.4, 129.4, 129.1, 128.9, 128.8, 128.2, 127.9, 126.9, 126.7, 115.7, 115.3, 115.0, 114.9, 95.5, 95.2, 83.1, 61.1, 60.9, 56.4, 46.4, 37.9 ppm; LCMS 9.25 min, 581.6 m/z, 68% of total peak area at 254nm; HRMS (TOF ESI) 580.1236 calcd for m/z (M+H); 580.1245 (1.6) observed;

[2-(2-Bromo-benzyl)-7-hydroxy-1-vinyl-1,4-dihydro-2H-isoquinolin-3-ylidene]-phosphoramidic acid diphenyl ester (8):

An oven-dried vial was charged with 4 (50 mg, 0.060 mmol), and the vial was sealed with a Teflon-lined septum and purged with argon for 5 min. After 5 min THF (500 µL) was added to the reaction, followed by the phosphorazidic acid diphenyl ester (129 µL, 0.600 mmol, 10.0 equiv). The reaction was placed on a mixer for 4 h. After 4 h the beads were transferred into a 2 mL biorad biospin tube and then filtered away from the solvents and reagents. The beads were washed with THF (3x, 1 min), then rotated in THF (15 min). The solvents were removed, and the washing protocol was repeated with CH2Cl2, followed by drying under house vacuum (2 h) and high vacuum overnight. The resin was then exposed to the cleavage conditions providing 21 mg (0.357 mmol, 0.713 mmol/g, 60%) of 8. 1H NMR (500 MHz, CDCl3): δ 8.06 (brs, 1H), 7.54 (m, 1H), 7.25 (m, 4H), 7.08 (m, 4H), 7.00 (m, 4H), 6.85 (m, 3H), 6.56 (m, 1H), 5.65 (m, 1H), 5.33 (d, J= 16.0 Hz, 1H), 5.07 (d, J= 10.0 Hz, 1H), 4.95 (d, J= 17.0 Hz, 1H), 4.67 (d, J= 6 Hz, 1H), 4.45 (d, J= 19.5 Hz, 1H), 4.19 (d, J= 16.5 Hz, 1H), 3.51 (d, J= 19.5 Hz, 1H) ppm; 13C NMR (125 MHz, CDCl3): δ 170.1, 158.8, 153.9, 136.9, 136.2, 136.0, 135.3, 131.9, 131.7, 131.4, 131.2, 130.3, 130.2, 130.1, 127.0, 126.9, 125.5, 123.1, 123.0, 118.5, 118.3, 114.9, 67.0, 53.5, 37.2 ppm; LCMS 9.05 min, 588.1 m/z 81% of the total peak area at 254nm; HRMS (TOF ES) 589.0847 calcd for m/z (M+H); 589.0908 observed.

2-(2-Bromo-benzyl)-9-hydroxy-1-vinyl-1,2-dihydro-benzo[c]azocine-4,5-dicarboxylic acid dimethyl ester (9): An oven-dried vial was charged with 4 (50 mg, 0.060 mmol), and the vial was sealed with a Teflon-lined septum and purged with argon for 5 min. After 5 min acetonitrile (500 µL) was added to the reaction, followed by dimethylacetylenedicarboxylate (109 µL, 1.20 mmol, 20.0 equiv.). The reaction was placed on a mixer and agitated for 16 h. The beads were transferred into a 2 mL biorad biospin tube and then filtered away from the solvents and reagents. The beads were washed with THF (3x, 1 min), then rotated in THF (15 min). The solvents were removed, and the washing protocol was repeated with CH2Cl2, followed by drying under house vacuum (2 h) and high vacuum overnight. The resin was exposed to the cleavage conditions providing 20.1 mg (0.041 mmol, 0.83 mmol/g, 69%) of 9. 1H NMR (500 MHz, rt d7-DMF): δ 9.55 (brs, 1H), 7.48 (brs 1H), 7.32 (d, J= 7 Hz, 1H), 7.18 (brs, 1H), 7.04 (brm, 1H), 6.94 (brs, 1H), 6.87 brs, 1H), 6.88 (brs, 1H), 6.47 (brs, 1H), 6.20 (brs, 1H), 5.36 (brs, 1H), 4.56 (brs, 2H) 4.36 (brd, J= 15.5 Hz, 1H), 3.27 (brs, 3H), 3.08 (brs, 4H) ppm; 1H NMR (500 MHz, -30 °C, d7-DMF. 4.5:1 ratio of resolved isomers): δ 10.05 (s) 9.95 (s), 7.52 (brs), 7.35 (d, J= 8 Hz), 7.28 (d, J= 7.5 Hz), 7.21 (d, J= 7.5 Hz), 7.06 (brt, J= 7.5 Hz), 6.95 (brt, J= 7.5 Hz), 6.89 (m), 6.85 (brs), 6.77 (s), 6.71 (d, J= 8 Hz), 6.52 (d, J= 8.5 Hz), 6.45 (d, J= 7.5 Hz), 6.24 (s), 6.17 (m), 6.09 (d, J= 7.5 Hz), 5.27 (dd, J= 12.0, 15.7 Hz), 4.96 (d, J= 16.5 Hz), 4.83 (d, J= 10.0 Hz), 4.54 (brt, J= 13.5 Hz), 4.14 (m), 4.37 (d, J= 15 Hz), 4.17 (d, J= 16.5 Hz), 3.46 (brd), 3.30 (brs), 3.22 (brs), 3.09 (brs), 3.02 (brs) ppm; 13C NMR (125 MHz, -30 °C, d7-DMF): δ 168.4, 157.7, 157.3, 138.4, 136.1, 135.8, 135.7, 133.5, 133.1, 132.8, 131.7, 131.1, 130.7, 130.2, 127.9, 127.6, 124.0, 121.2, 118.1, 115.2, 113.2, 66.1, 61.9, 51.3, 50.5 ppm; LCMS 8.11 min., 485.1, 84% of the total peak area at 254nm; HRMS (TOF ES) 484.0759 calcd for m/z (M+H); 484.0761 (0.4) observed.

2-(2-Bromo-benzyl)-7-hydroxy-1-vinyl-1,2-dihydro-isoquinoline-4-carboxylic acid (4-fluoro-phenyl)-amide (10): An oven dried vial was charged with 4 (50 mg, 0.060 mmol), and the vial was sealed with a Teflon-lined septum cap and purged with argon for 5 min. After 5 min acetonitrile (500 µL) was added to the reaction, followed by 4-fluorophenylisocyanate (120 uL, 1.20 mmol., 20.0 equiv.). The reaction was placed into an oil bath at 90 °C and allowed to react for 5 h. The beads were transferred into a 2 mL biorad biospin tube and then filtered away from the solvents and reagents. The beads were washed with DCM (3x 5 mL), taken up in DCM (2 mL) and rotated for 15 min. The solvents were removed and the washing procedure was repeated 1x with acetonitrile and 1x with DCM. The resin was then taken up in DCM and rotated overnight for a total of 16 h.. The solvents were removed, and the resin was dried under house vacuum for 3 h, followed by high vacuum overnight. The resin was then exposed to the cleavage conditions providing 18.9 mg (0.039 mmol, 0.79 mmol/g, 66%) of 10. 1H NMR (500 MHz, CDCl3/1% TEA): δ 7.57 (m, 1H), 7.48 (m, 1H), 6.12-7.3 (m, 5H), 6.48 (d, J= 2.44 Hz, 1H), 5.96 (m, 1H), 5.16 (d, J= 17.1Hz, 1H), 5.09 (d, J= 9.7 Hz, 1H), 4.71 (d, J= 7.82 Hz, 1H), 4.60 (d, J= 16.1 Hz, 1H), 2.40 (d, J= 16.1 Hz, 1H) ppm; 13C NMR (100 MHz, CDCl3/1% TEA): δ 165.9, 154.1, 141.8, 134.8, 133.2, 129.5, 129.2, 127.9, 123.9, 122.3, 122.3, 121.7, 121.6, 115.9, 115.7, 115.6, 115.5, 114.1, 63.7, 56.9 ppm; LCMS: 8.91 min, 60% of total peak area at 254 nm; HRMS (TOF ES) 479.0769 calcd for m/z (M+H); 479.0769 (0.2) observed.

3-[1-(2-Bromo-benzyl)-6-vinyl-1,2,3,6-tetrahydro-pyridin-4-yl]-propan-1-ol (11a/b): An oven-dried vial was charged with 25 mg of the resin bound enamine 4, and then the sodiumcyanoborahydride (6.0 mg, 0.300 mmol, 5 equiv.) was added to the vial. The vial was quickly capped with a septum and then purged with argon for 5 min. The vial was charged with CH2Cl2 (500 µL), and the beads were allowed to swell for 5 min. After 5 min trifluoroethanol (500 µL) was added via

syringe, the vial sealed with Teflon tape, and the reaction gently agitated overnight. After 15 h the solvents were removed via canula and the beads washed with CH2Cl2 (2x, 2.0 mL, 5 min), 2:2:1 DMF, MeOH, TEA (2x, 5 mL, 2 min) and then CH2Cl2 (3x, 5 mL, 3 min). The beads were then dried under house vacuum (2 h) then high vacuum overnight. The resin was exposed to the cleavage conditions providing 5.0 mg (0.015 mmol, 0.59 mmol/g, 55%) of 11a and 11b (obtained as a 1:1.2 ratio of inseparable olefin isomers, 11a and 11b). 1H NMR (400 MHz, CDCl3): δ 7.50-7.57 (m, 4H), 7.28-7.52 (m, 4H), 7.23 (m, 2H), 5.91 (m, 1H), 5.77 (m, 1H), 5.39 (m, 1H), 5.11-5.26 (m, 5H), 3.92 (d, J= 15.1 Hz, 1H), 3.84 (d, J= 14.6 Hz, 1H), 3.67 (brs, 4H), 3.53 (d, J= 14.2 Hz, 1H), 3.46 (d, J= 15.1 Hz), 3.24 (m, 1H), 3.10 (brd, J= 15.1Hz, 1H), 2.97 (brd, J= 16.1 Hz, 1H), 2.84 (m, 1H), 2.36 (m, 1H), 2.27 (dt, J= 20.0, 6.3 Hz, 2H), 2.06, (m, 4H), 1.71 (m, 4H) ppm; 13C NMR (100 MHz, CDCl3): δ 140.5, 140.1, 132.6, 132.5, 130.7, 130.3, 128.1, 127.9, 127.2, 127.1, 122.9, 118.9, 116.9, 1166, 64.6, 62.8,62.7, 60.9, 57.8, 57.7, 49.9, 47.0, 34.7, 33.3, 33.2, 30.3, 30.2, 28.8 ppm; LCMS: 3.82 min, 85.3% of the total peak area at 215nm; HRMS (TOF ES) calcd for m/z (M+H); observed.

[1-(2-Bromo-benzyl)-4-(3-hydroxy-propyl)-6-vinyl-3,6-dihydro-1H-pyridin-2-ylidene]-phosphoramidic acid diphenyl ester (12): An oven-dried vial was charged with 4 (46.3 mg, 0.114 mmol), sealed with a Teflon-lined septum and purged with argon for 5 min. After 5 min THF (500 µL) was added to the reaction followed by the phosphorazidic acid diphenylester (247 µL, 1.15 mmol., 10.0 equiv.). The reaction was placed on a mixer for 4 h. After 4 h the beads were transferred into a 2 mL biorad biospin tube and filtered away from the solvents and reagents. The beads were washed with THF (3x, 1 min), then rotated in THF (15 min). The solvents were removed, and the washing protocol was repeated with CH2Cl2, followed by drying under house vacuum (2 h) and high vacuum overnight. The resin was then exposed to the cleavage conditions providing 13.0 mg (0.025 mmol, 0.50 mmol/g, 50%) of 12. 1H NMR (500 MHz, CDCl3): δ 7.54 (m, 1H), 7.26 (m, 4H), 7.19 (m, 3H), 7.15 (M, 3H), 7.10 (m, 1H), 7.04 (M, 2H), 6.90 (m, 1H), 5.54 (m, 1H), 5.43 (brs, 1H), 5.24 (d, J= 16 Hz, 1H), 5.17 (d, J= 10 Hz, 1H), 5.11 (d, J= 16.5 Hz, 1H), 4.21 (brs, 1H), 4.18 (brs, 1H), 3.81 (d, J= 21.5 Hz, 1H), 3.63 (brs, 2H), 3.24 (d, J= 21 Hz, 1H), 2.15 (t, J= 7.5 Hz, 2H) 1.70 (m, 2H) ppm; 13C NMR (125 MHz, CDCl3): δ 168.0, 154.2, 137.5, 137.1, 136.4, 135.3, 131.8, 131.7, 131.2, 130.3, 130.1, 126.7, 126.6, 125.6, 123.1, 123.0, 120.5, 120.1, 64.4, 63.9, 52.1, 36.8, 36.7, 34.1, 32.4 ppm; LCMS: 8.82 min., 580.9m/z 85% of the total peak area at 254nm; HRMS (TOF ES) 581.1205 calcd for m/z (M+H); 581.1208 (0.5) observed.

1-(2-Bromo-benzyl)-6-(3-hydroxy-propyl)-8-vinyl-1,8-dihydro-azocine-3,4-dicarboxylic acid dimethyl ester (13): An oven-dried vial was charged with 4 (49.9 mg, 0.052 mmol), and the vial was sealed with a Teflon-lined septum and purged with argon for 5 min. After 5 min acetonitrile (500 µL) was added to the reaction, followed by dimethylacetylenedicarboxylate (47.7 uL, 0.524 mmol., 10.0 equiv.). The reaction was placed on a mixer and agitated for 16 h. The beads were transferred into a 2 mL biorad biospin tube and then filtered away from the solvents and reagents. The beads were washed with THF (3x, 1 min.), then rotated in THF (15 min). The solvents were removed and the washing protocol repeated with CH2Cl2, followed by drying under house vacuum (2 h) and high vacuum overnight. The resin was then exposed to the cleavage conditions providing 13.1 mg (0.027 mmol, 0.55 mmol/g, 53%) of 13. 1H NMR (600 MHz, CDCl3): δ 7.61 (s, 1H), 7.54 (m, 1H), 7.31-7.36 (m, 2H), 7.17, (m, 1H), 6.74 (s, 1H), 5.78 (m, 2H), 5.69 (m, 1H), 4.43 (d, J= 17.1 Hz, 1H), 4.32 (d, J= 16.6 Hz, 1H), 3.79 (s, 3H), 3.65 (m, 2H), 3.62 (s, 3H), 2.30 (t, J= 8.3Hz, 2H), 1.73 (m, 2H) ppm; 13C NMR (100 MHz, CDCl3): δ 175.8. 170.9, 151.0, 144.9, 142.5, 140.0, 137.7, 133.3, 129.3, 127.8, 120.8, 62.3, 59.6, 56.1, 52.5, 51.6, 32.3, 31.4 ppm; LCMS: 7.76 min, 88% of total peak area at 254 nm. 477.8 m/z; HRMS (TOF ES) 476.1072 calcd for m/z (M+H); 476.1067 (1.1) observed.

4-Benzyl-8-(2-bromo-benzyl)-10-(3-hydroxy-propyl)-9-vinyl-4,8-diaza-tricyclo[5.2.2.02,6]undec-10-ene-3,5-dione (14): A 4 mL oven-dried vial equipped with a Teflon-lined septum was charged with 5 (51.3 mg, 0.072 mmol), followed by the addition of N-benzylmaleimide (134 mg, 0.718 mmol). The reaction vessel was sealed and purged with argon. Toluene was added to the reaction via syringe, and the resulting mixture was heating at 50 oC for 24 h. The reaction was cooled to rt, and the solvent was removed via canula. The beads were washed with CH2Cl2 (3x) and dried under house vacuum, followed by high vacuum overnight. The resin was then exposed to the cleavage conditions, providing 15.2 mg (0.029 mmol, 0.58 mmol/g, 54%) of 14. 1H NMR (400 MHz, CDCl3): δ 7.51 (m, 1H), 7.42 (m, 1H), 7.20-7.30 (m, 7H), 7.10 (m, 1H), 5.81 (m, 1H), 5.49 (m, 1H), 5.11 (d, J= 17.2 Hz, 1H), 4.89 (d, J= 10 Hz, 1H), 4.57 (dd, J= 14.0, 4.4 Hz, 1H), 4.45 (dd, J=

14.0, 4.0 Hz, 1H), 3.89 (dd, J= 14.8, 4.0 Hz, 1H), 3.77 (m, 1H), 3.68 (dd, J= 14.6, 4.0 Hz, 1H), 3.51 (m, 1H), 3.41 (brs, 2H), 3.06 (m, 1H), 2.99 (brs, 1H), 2.90 (brs, 1H), 1.80 (m, 1H), 1.71 (m, 1H), 1.30 (m, 1H), 1.12 (m, 1H) ppm; 13C NMR (125 MHz, CDCl3): δ 177.7, 177.4, 141.9, 138.9, 137.9, 135.9, 132.7, 130.7, 129.0, 128.7, 128.6, 128.5, 127.8, 127.4, 124.6, 124.3, 115.6, 64.3, 62.3, 55.6, 52.1, 42.7, 42.6, 42.3, 39.7, 31.3, 29.0 ppm; LCMS: LCMS: 6.15 min. 520.8 m/z , 93% of total peak area, TIC; HRMS (TOF ES) 521.1440 calcd for m/z (M+H); 521.1421 (3.6) observed. III. Stability Comparison of Dihydroisoquinolines on Solid-Support and in Solution

2-(2-Bromo-benzyl)-1-vinyl-1,2-dihydro-isoquinoline: A flame dried round bottom flask was charged with the solid intermediate (100 mg, 0.264 mmol) and a stir bar, sealed, and purged with Argon. Dry THF (2.6 mL) was added to form a suspension, and the mixture was cooled to -78 oC. Vinylmagnesiumbromide (530 µL, 0.53 mmol) was added dropwise, and the solution was kept at -78 oC 15 min. The mixture was then warmed to 0 oC and stirred for 30 min. The reaction was quenched with NH4Cl (aq.) and extracted with ether. The extracts were combined, concentrated in vacuo to near dryness, and then purified immediately by column chromatography with alumina (Pentane:Triethylamine 50:1) to yield a clear oil. Yield not determined. 1H NMR (400 MHz, C6D6): δ 7.58 (m, 1H), 7.43 (m, 1H), 7.29 (m, 1H), 7.15 (m, 2H) 7.02 (m, 1H), 6.92 (m, 2H), 6.20 (dd, J= 7.2, 1.2 Hz, 1H), 6.09 (ddd, 1H), 5.31 (d, J= 7.2 Hz, 1H), 5.13 (dt, J= 17, 1.4 Hz, 1H), 5.05 (dt, J= 11.4, 1.4 Hz, 1H), 4.81 (d, J= 7.2 Hz, 1H), 4.50 (d, J= 16 Hz, 1H), 4.27 (d, J= 16 Hz, 1H) ppm. An enamine analogous to intermediate 5, 2-(2-bromo-benzyl)-1-vinyl-1,2-dihydro-isoquinoline, was synthesized in solution. The compound was stored both neat (50 mg) and frozen in d6-benzene (50 mg/400 µL) at 0 oC for 14 d. The samples were photographed and analyzed by 1H NMR (50 mg/400 µL) at 0 d, 1 d, 7 d, and 14 d. After each analysis, the solvent was removed in vacuo for the compound that was stored neat.

IV. Computational Calculations Computational calculations were performed with SPARTAN ’02 software package for Windows (Wavefunction, Inc.). Minimized structures were obtained utilizing a monte-carlo search (Semi-empirical AM1 method), followed by overlaying each structure at the atoms indicated.

Entry Compound Number Molecule Energy

(Kcal/mol) MW

(g/mol) Area (Å^2)

Volume (Å^3)

dipole (debye)

1 9 Isoq Isocy 12.9962 479.349 439.53 431.16 3.45 2 10 Isoq Eight 73.59299 484.346 447.5 436.92 3.82

3 8 Isoq Azide -65.30336 589.426 563.55 539.83 N/A

4 4 Isoq enami 67.5486 342.236 324.58 310.92 2.55

5 6 Isoq Reduc 36.6859 344.252 327.54 314.4 1.68

6 7 Isoq Oxime 97.60807 461.359 421.36 419.65 0.56

7 3 Isoq Alkyl 187.420826 315.19 289.91 276.18 2.68 8 2 Isoq 5.987679 145.161 162.949 164.655 1.34

9 N/A Isoq Double 129.034 580.482 518.5 529.58 1.86

10 5 Pyr Enami 23.60841 334.257 335.56 314.3 2.24

11a 11a Other pyred 2.228110 336.273 339.25 317.45 1.83

11 11 Pyr Reduc 2.641428 336.273 339.39 317.53 1.93

12 15 Pyr Eight -114.59718 476.367 463.89 440.41 3.56

13 12 Pyr Oxime 69.694966 453.38 431.99 422.76 0.82

14 13 Pyr Azide -98.705649 581.447 570.05 542.81 6.73

15 14 Pyr Diels a -11.36004 521.455 487.78 488.09 3.95

16 N/A Pyr Alkyl 146.16577 307.211 297.25 279.54 2.42 17 N/A Pyr -35.039267 137.182 181.304 138.375 1.62