the modification of the dipeptide terminus of peptidoglycan cell wall precursors from d - ala - d - ala to d - ala - d - lac in resistant bacteria reduces the binding affinity of vancomycin for the ligand by 1000 - fold leading to a 1000 - fold loss in biological activity . it had earlier been shown that a modified peptide ligand possessing a methylene in place of the lactate oxygen restores 100 - fold of this binding affinity by removal of a destabilizing lone pair interaction . it is disclosed herein that removal of the residue 4 carbonyl in the vancomycin aglycon produces an analogue with enhanced affinity for d - ala - d - lac and restores much of the biological activity of the molecule that is lost with resistant bacteria . moreover and among the range of potential modifications that could be envisioned , that entailing the simple removal of the residue 4 carbonyl providing 5 are disclosed to bind d - ala - d - ala and d - ala - d - lac with similar affinities providing an analogue having equivalent effectiveness against sensitive ( d - ala - d - ala ) and resistant ( d - ala - d - lac ) bacteria . efforts were extended on the preparation of glycopeptide antibiotics to a total synthesis of the [ ψ [ ch 2 nh ] tpg 4 ] vancomycin aglycon ( 5 ) in which the residue 4 carbonyl has been replaced with a methylene . consistent with expectations and relative to the vancomycin aglycon , 5 exhibited a 40 - fold increase in affinity for d - ala - d - lac ( k a = 5 . 2 × 10 3 m − 1 ) and a corresponding 35 - fold reduction in affinity for d - ala - d - ala ( k a = 4 . 8 × 10 3 m − 1 ) providing a molecule with balanced , dual binding characteristics . compound 5 exhibited antimicrobial activity against a vana resistant organism that remodels its d - ala - d - ala peptidoglycan cell wall precursor to d - ala - d - lac upon glycopeptide challenge displaying a potency that reflects these binding characteristics . challenges and synthetic plan for [ ψ [ ch 2 nh ] tpg 4 ] vancomycin aglycon . the desired analogue 5 was anticipated to be prepared by a route analogous to that developed for vancomycin ( boger , d . l . ; et al . j . am . chem . soc . 1999 , 121 , 3226 ; boger , d . l . ; et al . j . am . chem . soc . 1999 , 121 , 10004 ), with notable modifications . thus , two aromatic nucleophilic substitution reactions with formation of the biaryl ethers are enlisted for cd and de macrocyclization , a key macrolactamization reaction are employed for cyclization of the ab ring system , and the defined order of cd , ab , and de ring closures permit sequential control of the atropisomer stereochemistry of each of the newly formed ring systems or their immediate precursors , fig2 . thus , in addition to any kinetic diastereoselection that may be achieved in the ring closures , this order is disclosed to permit the recycling of any undesired atropisomer for each newly introduced ring system by thermal equilibration providing a predictable control of the stereochemistry and dependably funneling all synthetic material into one of eight possible atropdiastereomers . key to recognition of this preferential order of ring closures was the establishment of the thermodynamic parameters of atropisomerism for the individual vancomycin ring systems : de ring system ( boger , d . l . ; et al . j . org . chem . 1997 , 62 , 4721 ; boger , d . l . ; et al . j . org . chem . 1996 , 61 , 3561 ; boger , d . l . ; et al . j . org . chem . 1999 , 64 , 70 ; boger , d . l . ; et al . bioorg . med . chem . lett . 1997 , 7 , 3199 ; boger , d . l . ; et al . bioorg . med . chem . lett . 1998 , 8 , 721 ; boger , d . l . ; et al . j . am . chem . soc . 1998 , 120 , 8920 ) ( e a = 18 . 7 kcal / mol )& lt ; ab biaryl precursor ( boger , d . l . ; et al . j . am . chem . soc . 1999 , 121 , 3226 ; boger , d . l . ; et al . j . am . chem . soc . 1999 , 121 , 10004 ) ( e a = 25 . 1 kcal / mol )& lt ; cd ring system ( boger , d . l . ; et al . j . org . chem . 1997 , 62 , 4721 ; boger , d . l . ; et al . j . org . chem . 1996 , 61 , 3561 ; boger , d . l . ; et al . j . org . chem . 1999 , 64 , 70 ; boger , d . l . ; et al . bioorg . med . chem . lett . 1997 , 7 , 3199 ; boger , d . l . ; et al . bioorg . med . chem . lett . 1998 , 8 , 721 ; boger , d . l . ; et al . j . am . chem . soc . 1998 , 120 , 8920 ) ( e a = 30 . 4 kcal / mol ). thus , the molecule was assembled by coupling the modified and fully functionalized abcd ring system 27 with the e ring tripeptide 28 followed by a diastereoselective aromatic nucleophilic substitution reaction for closure of the 16 - membered de ring system with formation of the biaryl ether linkage . notably , the activating nitro substituent additionally serves as the precursor functionality for aryl chloride introduction and the analogous vancomycin ring closures ( boger , d . l . ; et al . j . am . chem . soc . 1999 , 121 , 3226 ; boger , d . l . ; et al . j . am . chem . soc . 1999 , 121 , 10004 ; boger , d . l . ; et al . bioorg . med . chem . lett . 1995 , 5 , 3091 ; evans , d . a . ; watson , p . s . tetrahedron lett . 1996 , 37 , 3251 ; evans , d . a . ; et al . angew . chem ., int . ed . 1998 , 37 , 2700 ; evans , d . a . ; et al . angew . chem ,. int . ed . 1998 , 37 , 2704 ) were effected with preferential formation of the natural ( p )- atropisomer . the e ring tripeptide 28 was derived in the manner described for vancomycin ( boger , d . l . ; et al . j . am . chem . soc . 1999 , 121 , 3226 ; boger , d . l . ; et al . j . am . chem . soc . 1999 , 121 , 10004 ) except that the e ring subunit was prepared enlisting an improved route developed during a more recent total synthesis of the ristocetin aglycon ( crowley , b . m . ; et al . j . am . chem . soc . 2004 , 126 , 4310 ) employing α - hydroxypinanone ( solladié - cavallo , a . ; nsenda , t . tetrahedron lett . 1998 , 39 , 2191 ) as the chiral auxiliary for a diastereoselective aldol addition . the most significant deviations rest with the required modifications in the preparation of the abcd subunit which house the modified amide and include the use of a reductive amination coupling of residues 4 and 5 ( d and b rings ) with protection of the newly generated amine as a methyl carbamate and an experimentally - derived altered order to the assembly of the bcd tripeptide . a relatively small and robust amine protecting group was chosen to avoid the introduction of unfavorable steric interactions that affects the cd macrocyclic ring closure and that is stable throughout the synthesis , yet still compatible with a final stage global deprotection . cd macrocyclization enlisting a key aromatic nucleophilic substitution reaction for formation of 16 - membered biaryl ether followed by suzuki coupling of the a ring subunit and ab macrolactamization was employed to complete the preparation of the modified abcd ring system 27 enlisting a ring closure order that permits the sequential and selective thermal adjustment of the cd and ab ring system atropisomer stereochemistry . key unforeknown features of the approach include the feasibility of conducting the critical cd ring closure enlisting the residue 4 protected amine versus amide , the resulting unknown atropisomer stereochemical issues ( kinetic and thermodynamic diastereoselectivity ), and the impact the deep - seated structural change on the conformational features of the cd or abcd ring systems and those of the final molecule . finally , the subtle choices of a nitrile as a precursor to the residue 3 side chain carboxamide permits a final stage amide deprotection yet conveys stability throughout the synthesis to any projected thermal atropisomer equilibrations in its presence ( boger , d . l . ; et al . j . am . chem . soc . 1998 , 120 , 8920 ), and the use of a mem protected hydroxymethyl precursor ( vs a methyl ester ) to the c - terminus carboxylic acid enhances the rate of the projected ab macrolactamization ( boger , d . l . ; et al . j . am . chem . soc . 1999 , 121 , 3226 ; boger , d . l . ; et al . j . am . chem . soc . 1999 , 121 , 10004 ) and precludes inadvertent epimerization throughout the synthesis . synthesis of the bcd tripeptide . the b and d subunits 6 and 7 were prepared following previously optimized procedures ( crowley , b . m . ; et al . j . am . chem . soc . 2004 , 126 , 4310 ; boger , d . l . ; et al . j . org . chem . 1997 , 62 , 4721 ; boger , d . l . ; et al . j . org . chem . 1996 , 61 , 3561 ; boger , d . l . ; et al . j . org . chem . 1999 , 64 , 70 ; boger , d . l . ; et al . bioorg . med . chem . lett . 1997 , 7 , 3199 ; boger , d . l . ; et al . bioorg . med . chem . lett . 1998 , 8 , 721 ; boger , d . l . ; et al . j . am . chem . soc . 1998 , 120 , 8920 ). oxidation of alcohol 7 ( compound 7 is available in 6 steps ( 37 % overall ) from methyl gallate using 3 recrystallizations and was scaled to 300 g , ( crowley , b . m . ; et al . j . am . chem . soc . 2004 , 126 , 4310 )) ( 2 . 0 equiv of dess - martin periodinane , ch 2 cl 2 , 0 - 25 ° c ., 1 h , 100 %) was followed by immediate reductive amination coupling of the sensitive aldehyde 8 with 6 ( compound 6 is available in 5 steps ( 55 % overall ) from ( r )- 4 - hydroxyphenyl - glycine using 2 recrystallizations and was scaled to 60 g , ( boger , d . l . ; et al . j . org . chem . 1997 , 62 , 4721 )) ( 1 . 1 equiv , ch 3 oh , 3 å ms , 0 ° c ., 45 min ; 3 . 0 equiv of acoh , 3 . 0 equiv of nabh 3 cn , − 20 ° c ., 2 d ) to afford amine 9 in good yield ( 75 %) and excellent diastereoselectivity ( 12 : 1 ), fig3 . shorter reaction times ( 14 - 20 h ) at higher temperatures (− 15 to − 5 ° c .) led to substandard selectivities ( 4 : 1 to 9 : 1 ) and the use of less nabh 3 cn ( 1 . 5 - 2 . 0 equiv ) at lower temperatures (− 20 ° c .) led to incomplete reactions . longer reaction times ( 3 - 8 d ) led to only marginal increases in yield ( 82 % after 8 d ) and roughly equal diastereoselectivities . initial efforts to prepare amine 9 directly by displacement of the mesylate derived from alcohol 7 were ineffective as were attempts to conduct the reductive amination with the bc dipeptide and 8 . amine protection of 9 as the methyl carbamate ( 10 equiv of meococl , 10 equiv of k 2 co 3 , thf , 0 - 25 ° c ., 18 h , 85 %) followed by benzyl ether deprotection ( benzyl ether deprotection at higher temperatures ( 25 ° c .) may lead to competitive aryl bromide reduction although this was only observed in appreciable amounts when excess raney ni was employed .) ( raney ni , ch 3 oh , 0 ° c ., 5 h , 98 %) and saponification ( 3 . 0 equiv of lioh , thf — h 2 o , 0 ° c ., 6 h , 100 %) provided 12 . unexpectedly , the order of these latter two deprotections proved important . saponification of 10 ( saponification of 11 was considerably slower than that of 10 and occasionally required additional lioh for complete conversion to 12 with little effect on the amount of epimer generated in the reaction .) under a variety of conditions ( lioh , thf — h 2 o or t - buoh — h 2 o , − 10 to 0 ° c . ; liooh , thf — h 2 o ; me 3 snoh , 1 , 2 - dichloroethane , 70 ° c .) led to variable amounts of an epimer ( 5 - 20 %) that was difficult to separate from the product . in contrast , benzyl ether deprotection of 10 followed by saponification of 11 reduced the amount of epimer ( 0 - 3 %) presumably due to preferential deprotonation of the phenols such that subsequent c α deprotonation at residue 5 was less facile ( saponification of 11 was considerably slower than that of 10 and occasionally required additional lioh for complete conversion to 12 with little effect on the amount of epimer generated in the reaction .). coupling of 12 with 13 ( compound 13 is available in 3 steps ( 45 % overall ) from 4 - fluoro - 3 - nitrobenzaldehyde and was scaled to 30 g , ( crowley , b . m . ; et al . j . am . chem . soc . 2004 , 126 , 4310 ).) ( 3 . 0 equiv of depbt ( fan , c .- x . ; et al . org . lett . 1999 , 1 , 91 ), 3 . 0 equiv of nahco 3 , dmf , 0 - 25 ° c ., 8 h ) gave tripeptide 14 in good yield ( 70 %) and excellent diastereoselectivity ( 14 : 1 ). a range of other more conventional coupling reagents ( edcl — hoat , hatu , fdpp ) also provided good conversions ( 65 - 80 %), but suffered from considerable competitive racemization . synthesis of the abcd ring system . this set the stage for a detailed examination of one of the critical reactions in the approach to 5 entailing the cyclization of 14 . after considerable optimization ( fig5 a and 5b ), cyclization of 14 ( 20 equiv of k 2 co 3 , 20 equiv of caco 3 , 3 wt equiv of 3 å ms , 12 mm thf , 75 ° c . bath temp , 12 h ) afforded 15 in good yield ( 54 %) and good atropodiastereoselectivity ( 2 . 5 : 1 , 15 ( 54 %) and 16 ( 22 %)) even when conducted on a large scale ( 2 . 7 g ), fig4 . the inclusion of caco 3 in the reaction mixture is critical and serves to trap the liberated fluoride arising from the aromatic nucleophilic substitution as an insoluble byproduct ( caf 2 ) preventing tbs ether deprotection and a subsequent competitive base - catalyzed retro aldol reaction of the free alcohol . nearly comparable results were obtained by promoting the ring closure of 15 with the stronger base t - buok ( 1 . 0 equiv , thf , − 20 ° c ., 18 h ) providing 15 and its atropisomer 16 in 57 % and 19 % ( 3 : 1 atropodiastereoselectivity ), respectively , under remarkably mild reaction conditions (− 20 ° c ., thf ). however , the use of t - buok proved more sensitive to the reaction parameters , suffered competitive racemization if excess base was employed , and proved more difficult to implement on a large scale than the reaction enlisting k 2 co 3 / caco 3 . the cyclization of 14 represents a considerable improvement over the analogous ring closure reaction enlisted in an earlier synthesis of vancomycin ( 50 - 65 %, 1 : 1 atropisomers vs 76 - 87 %, 2 . 5 - 3 : 1 atropisomers ) where both the overall conversion and atropodiastereoselectivity were lower illustrating that the closure of 14 may benefit from both the increased conformational flexibility of the cyclization substrate and the residue 4 amine small protecting group . unlike the vancomycin cd ring system in which the atropisomers could be thermally equilibrated at 120 - 140 ° c . permitting the recycling and productive use of the unnatural atropisomer , the atropisomers 15 and 16 could not be thermally interconverted even at temperatures as high as 210 - 230 ° c ., fig6 . reduction of the nitro group ( raney ni , 0 ° c ., ch 3 oh , 1 h ) followed by diazotization ( 1 . 3 equiv of hbf 4 , 1 . 3 equiv of t - buono , ch 3 cn , 0 ° c ., 30 min ) and sandmeyer substitution ( 50 equiv of cucl , 60 equiv of cucl 2 , h 2 o , 0 - 25 ° c ., 1 h , 70 % from 15 ) cleanly provided 17 without loss of the atropisomer stereochemistry inherent in starting 15 . the unnatural atropisomer 16 was also subjected to these conditions to cleanly give 18 ( 75 %) ( fig6 ). the stereochemical assignments of these two compounds and their relationship as atropisomers ( vs epimers ) were established by 2d roesy 1 h — 1 h nmr experiments and confirmed chemically by their reductive dechlorination ( h 2 , 10 %, pd / c ) to afford the identical product 19 ( fig6 ). suzuki coupling of 17 with the hindered a ring boronic acid 20 ( boger , d . l . ; et al . j . am . chem . soc . 1999 , 121 , 3226 ; boger , d . l . ; et al . j . am . chem . soc . 1999 , 121 , 10004 ) ( 0 . 3 equiv of pd 2 ( dba ) 3 , 1 . 5 equiv of ( o - tol ) 3 p , toluene - ch 3 oh - 1 n aq na 2 co 3 10 : 3 : 1 , 80 ° c ., 30 min ) proceeded in excellent yield ( 90 %) under remarkably effective conditions ( boger , d . l . ; et al . j . am . chem . soc . 1999 , 121 , 3226 ; boger , d . l . ; et al . j . am . chem . soc . 1999 , 121 , 10004 ) given the steric constraints of the substrate 20 providing a separable 1 : 1 . 3 mixture of atropisomers ( 21 : 22 ) slightly favoring the unnatural configuration . thermal equilibration of isolated 22 was carried out initially employing reported conditions for vancomycin ( o - dichlorobenzene , 120 ° c ., 18 h , 81 % recovery of material ) ( boger , d . l . ; et al . j . org . chem . 1997 , 62 , 4721 ; boger , d . l . ; et al . j . org . chem . 1996 , 61 , 3561 ; boger , d . l . ; et al . j . org . chem . 1999 , 64 , 70 ; boger , d . l . ; et al . bioorg . med . chem . lett . 1997 , 7 , 3199 ; boger , d . l . ; et al . bioorg . med . chem . lett . 1998 , 8 , 721 ; boger , d . l . ; et al . j . am . chem . soc . 1998 , 120 , 8920 ) to afford a 1 : 1 . 1 separable mixture permitting the recycling of this unnatural atropisomer . an examination of the parameters for this isomerization ( k = 0 . 12 h − 1 , t 1 / 2 = 5 . 9 h at 120 ° c . and k = 0 . 36 h − 1 , t 1 / 2 = 1 . 8 h at 135 ° c .) revealed that it proceeds with an energy of activation ( e a ) of 25 . 6 kcal / mol ( δh ‡ = 24 . 8 kcal / mol , δs ‡ =− 0 . 26 e . u ., δg ‡ = 24 . 9 kcal / mol ) essentially indistinguishable from that observed with the authentic vancomycin ab biaryl system , but it does not result in the analogues 3 : 1 thermodynamic preference for the natural atropisomer . however , the unusual and unexpected atropisomer stability of the cd ring system allowed us to improve on the recycling conditions . heating the mixture in a microwave reactor at an elevated temperature ( 210 ° c ., o - dichlorobenzene ) shortened the reaction time significantly ( 5 min vs 18 h ) and slightly improved the recovery of material ( 88 % vs 81 %). this improvement impacted the efficiency of the recycling of 22 by allowing multiple equilibrations to be run in a single day rather than over the course of a week . silyl ether deprotection of 21 ( 1 . 2 equiv of bu 4 nf , thf , 0 ° c ., 10 min ) followed by n - cbz removal ( h 2 , 10 % pd / c , 1 % cl 3 cco 2 h — ch 3 oh , 15 min , 95 %) and methyl ester hydrolysis ( 1 . 0 equiv of lioh , thf — h 2 o , 0 ° c ., 1 h , 96 %) gave amino acid 25 . notably , n - cbz removal in the absence of cl 3 cco 2 h ( boger , d . l . ; et al . j . am . chem . soc . 2000 , 122 , 7416 ; boger , d . l . ; et al . j . am . chem . soc . 2001 , 123 , 1862 ) was much slower ( 11 h ) and these conditions led to competitive chloride reduction ( use of raney ni for n - cbz removal was also successful , though lower recoveries ( 84 %) of the product were observed .). macrolactamization with closure of the ab ring system was effected by treatment of 25 with pybop ( 3 . 0 equiv , 6 . 0 equiv of nahco 3 , 0 . 001 m ch 2 cl 2 - dmf 5 : 1 , 0 - 25 ° c ., 12 h ) to afford the fully functionalized bicyclic abcd ring system 26 in good yield ( 70 %) with only trace amounts of competitive epimerization (& lt ; 3 %). alternative coupling reagents ( edcl and hoat or hobt , hatu ) and reaction conditions ( 10 - 100 % dmf — ch 2 cl 2 , 3 - 5 equiv of na 2 co 3 , − 5 to 0 ° ) led to lower conversions ( 30 - 52 %) or required extended reaction times ( 3 d ). n - boc deprotection ( hco 2 h — chcl 3 1 : 1 , 10 h , 84 %) gave the free amine 27 for coupling with the e ring tripeptide . confirmation of the atropisomer stereochemistry and amide conformational assignments for 26 were established by 2d roesy 1 h — 1 h nmr . diagnostic noe crosspeaks for 26 were observed between c 5 4 — oh / c 4b 4 — h ( s ), c 5 4 — oh / c 6 4 — ome ( s ), n 1 7 — h / c 4a 5 — h ( s ), n 1 7 — h / c 2 5 — h ( s ), n 1 7 — h / c 3 6 — h ( m ), n 1 7 — h / c 2 6 — h ( m ), c 5a 6 — h / c 3 6 — h ( s ), c 5a 6 — h / c 2 6 — h ( s ), c 5b 6 — h / n 1 6 — h ( m ), c 3 6 — oh / n 1 6 — h ( s ), c 5b 6 — h / c 3 6 — oh ( m ), c 6b 6 — h / c 5b 6 — h ( s ), c 6b 6 — h / c 4a 4 — h ( w ), n 1 4 — h / c 4b 4 — h ( m ), n 1 4 — h / c 4a 4 — h ( w ), c 4b 5 — h / c 5 5 — h ( s ), c 2 6 — h / c 4a 5 — h , c 4b 5 — h / c 1b 4 — h ( m ), c 4a 5 — h / c 6 7 — h ( w ), c 4a 5 — h / c 2 5 — h ( s ), c 5 5 — h / c 6 5 — ome ( s ), c 4 7 — h / c 2 7 — h ( s ), c 4 7 — h / c 1b 7 — h ( s ), c 4 7 — h / c 1a 7 — h ( w ), c 4 7 — h / c 5b 7 — ome ( s ), c 4 7 — h / c 6 7 — h ( w ), c 6 7 — h / c 2 5 — h ( w ), c 6 7 — h / c 5b 7 — ome ( s ), c 6 7 — h / c 5a 7 — ome ( s ), c 2 5 — h / c 3 6 — h ( m ), c 2 5 — h / c 2 6 — h ( s ), c 3 6 — h / c 2 6 — h ( m ), c 1 7 —( mem - ch 2 ) 1 / c 1a 7 — h ( s ), c 1 7 -( mem - ch 2 ) 1 7 -( mem - ch 2 ) 2 ( s ), c 2 7 — h / c 1b 7 — h ( s ), c 2 7 — h / c 1a 7 — h ( s ) and no noe crosspeaks were observed between c 5b 6 — h / c 3 6 — h , c 5b 6 / c 2 6 — h , c 2 6 — h / c 3 6 — oh , n 1 6 — h / n 1 7 — h , n 1 6 — h / c 2 5 — h , and n 1 6 — h / c 4a 5 — h . most important in this spectroscopic assessment was not only the expected confirmation of the cd and ab atropisomer stereochemistry , but also the establishment of a vancomycin - like conformation for 26 bearing a cis amide linking the residues 5 and 6 ( strong diagnostic c 2 5 — h / c 2 6 — h noe ) maintaining the spatial relationships and orientations of the ab ring system ( strong diagnostic c 2 5 — h / c 4a 5 — h and c 2 6 — h / c 4a 5 — h noes ) and cd ring systems . ( diagnostic c 6b 6 — h / c 4a 4 — h noe ). although this might be considered unusual on the surface , even the natural atropisomer of the isolated ab ring system of vancomycin , without the surrounding cd ring system , adopts a conformation incorporating this cis amide structure illustrating that it is the confines of the ab ring system , not that of the cd ring system , that defines this key cis amide conformational preference ( boger , d . l . ; et al . j . am . chem . soc . 1999 , 121 , 3226 ; boger , d . l . ; et al . j . am . chem . soc . 1999 , 121 , 10004 ). the lack of discernable noes to the methyl carbamate protecting the amine of the modified amide established that it extends out and away from the abcd ring system binding pocket . synthesis of the full carbon skeleton . coupling of 27 and 28 ( 2 . 0 equiv of . depbt ( fan , c .- x . ; et al . org . lett . 1999 , 1 , 91 ), 2 . 2 equiv of nahco 3 , thf , 0 - 25 ° c ., 14 h , 73 %) afforded 29 with excellent diastereoselectivity ( 12 : 1 ) arising from little competitive racemization , fig7 . these conditions were utilized based on experience with the teicoplanin ( boger , d . l . ; et al . j . am . chem . soc . 2000 , 122 , 7416 ; boger , d . l . ; et al . j . am . chem . soc . 2001 , 123 , 1862 ) and ristocetin ( crowley , b . m . ; et al . j . am . chem . soc . 2004 , 126 , 4310 ) aglycons and are superior to those originally reported for vancomycin ( boger , d . l . ; et al . j . am . chem . soc . 1999 , 121 , 3226 ; boger , d . l . ; et al . j . am . chem . soc . 1999 , 121 , 10004 ) ( edcl ) in terms of diastereoselectivity ( 12 : 1 vs 3 : 1 ). closure of the de ring system with formation of the key biaryl ether was accomplished by treatment of 29 with csf ( 10 equiv , 20 equiv of caco 3 ( both the added 3 å ms and caco 3 result in cleaner conversions to product . it is not yet clear whether the soluble base under these conditions is csf or cs 2 co 3 with precipitation of insoluble caf 2 . ), 3 å ms , dmf , 25 ° c ., 17 h ) to afford 30 in good yield ( 74 %) and good atropodiastereoselectivity ( 6 - 7 : 1 ). notably , the closure of 30 was conducted under milder conditions than those originally disclosed for vancomycin ( boger , d . l . ; et al . j . am . chem . soc . 1999 , 121 , 3226 ; boger , d . l . ; et al . j . am . chem . soc . 1999 , 121 , 10004 ; boger , d . l . ; et al . j . org . chem . 1997 , 62 , 4721 ; boger , d . l . ; et al . j . org . chem . 1996 , 61 , 3561 ; boger , d . l . ; et al . j . org . chem . 1999 , 64 , 70 ; boger , d . l . ; et al . bioorg . med . chem . lett . 1997 , 7 , 3199 ; boger , d . l . ; et al . bioorg . med . chem . lett . 1998 , 8 , 721 ; boger , d . l . ; et al . j . am . chem . soc . 1998 , 120 , 8920 ; boger , d . l . ; et al . bioorg . med . chem . lett . 1995 , 5 , 3091 ; evans , d . a . ; watson , p . s . tetrahedron lett . 1996 , 37 , 3251 ; evans , d . a . ; et al . angew . chem ., int . ed . 1998 , 37 , 2700 ; evans , d . a . ; et al . angew . chem ,. int . ed . 1998 , 37 , 2704 ) ( dmf vs dmso at 25 ° c . with added 3 å ms and caco 3 ) and approaches the kinetic atropisomer diastereoselectivity observed in earlier efforts ( boger , d . l . ; et al . j . am . chem . soc . 1999 , 121 , 3226 ; boger , d . l . ; et al . j . am . chem . soc . 1999 , 121 , 10004 ) ( 8 : 1 ), while surpassing that detailed in the related evans ( evans , d . a . ; et al . angew . chem ., int . ed . 1998 , 37 , 2700 ; evans , d . a . ; et al . angew . chem ,. int . ed . 1998 , 37 , 2704 ) efforts ( 5 : 1 ), and contrasts the closure detailed by nicolaou ( nicolaou , k . c . ; et al . angew . chem ., int . ed . 1998 , 37 , 2717 ; nicolaou , k . c . ; et al . angew . chem ., int . ed . 1998 , 37 , 2708 ; nicolaou , k . c . ; et al . angew . chem ., int . ed . 1998 , 37 , 2714 ; nicolaou , k . c . ; et al . angew . chem ., int . ed . 1999 , 38 , 240 ; nicolaou , k . c . ; et al . chem . eur . j . 1999 , 5 , 2584 ; nicolaou , k . c . ; et al . chem . eur . j . 1999 , 5 , 2602 ; nicolaou , k . c . ; et al . chem . eur . j . 1999 , 5 , 2622 ; nicolaou , k . c . ; et al . chem . eur . j . 1999 , 5 , 2648 ) ( 1 : 3 ) where the unnatural atropisomer predominated with an alternative substrate and method of ring closure . thus , consistent with the adoption of a vancomycin - like conformation by 26 , the amide modification in the abcd ring system of 29 had little impact on the ease or diastereoselectivity of the de ring closure . reduction of the nitro group ( reduction of the nitro group was very sensitive to the choice of solvent in terms of recovery and observance of side products .) ( h 2 , 10 % pd / c , thf , 8 h , 94 %) followed by diazotization of the resulting amine 32 ( 1 . 2 equiv of hbf 4 , 1 . 2 equiv of t - buono , ch 3 cn , 0 ° c ., 20 min ) and sandmeyer substitution ( 50 equiv of cucl , 60 equiv of cucl 2 , h 2 o , 0 - 25 ° c ., 1 h , 55 %) gave 33 , which embodies the full carbon skeleton of 5 . completion of the synthesis . with the full carbon skeleton in hand , attention was directed towards completion of the synthesis , fig7 . tbs ether protection of the secondary alcohols ( 65 equiv of cf 3 conmetbs , ch 3 cn , 55 ° c ., 22 h ; aq citric acid , 25 ° c ., 13 h , 96 %) followed by mem ether deprotection of 34 ( 12 equiv of b - bromocatecholborane ( bcb ), ch 2 cl 2 , 0 ° c ., 2 h ; 5 . 1 equiv of boc 2 o , 6 . 0 equiv of nahco 3 , dioxane - h 2 o 2 : 1 , 0 - 25 ° c ., 2 . 5 h , 80 %) and two - step oxidation of the resulting primary alcohol 35 ( 4 . 0 equiv of dess - martin periodinane , ch 2 cl 2 , 0 ° c ., 15 min then 25 ° c ., 1 h ; 9 . 0 equiv of 80 % aq naclo 2 , 7 . 0 equiv of nah 2 po 4 . h 2 o , t - buoh / 2 - methyl - 2 - butene 4 : 1 , 25 ° c ., 20 min , 82 %) provided the carboxylic acid 36 . hydrolysis of the residue 3 nitrile with formation of the carboxamide 37 ( 40 equiv of 30 % aq h 2 o 2 , 8 . 0 equiv of 10 % aq k 2 co 3 , dmso , 25 ° c ., 3 . 5 h , 87 %) ( boger , d . l . ; et al . j . am . chem . soc . 1999 , 121 , 3226 ; boger , d . l . ; et al . j . am . chem . soc . 1999 , 121 , 10004 ) set the stage for a final global deprotection ( node , m . ; et al . j . org . chem . 1980 , 45 , 4275 ; evans , d . a . ; ellman , j . a . j . am . chem . soc . 1989 , 111 , 1063 ). in a final key reaction , 37 was treated with albr 3 ( 35 equiv , etsh , 25 ° c ., 5 h , 80 %) to afford 5 arising from the remarkable deprotection of four aryl methyl ethers , the two tbs ethers , the n - terminus boc group , and the critical residue 4 methyl carbamate . assessment of [ ψ [ ch 2 nh ] tpg 4 ] vancomycin aglycon . a subtle element in the design of 5 and choice of simply removing the residue 4 carbonyl rests with the projected properties of the molecule . in principle , one might consider reengineering the capabilities of a reverse h - bond into the vancomycin structure removing the destabilizing lone pair interaction with d - ala - d - lac and reinstating the lost h - bond . such opportunities include amidine derivatives ( e . g . [ ψ [ c (═ nh ) nh ] tpg 4 ] vancomycin aglycon , fig9 ). significantly , such derivatives enhance d - ala - d - lac binding so as to approach the level of affinity observed with vancomycin and d - ala - d - ala . however , such derivatives also reduce binding to d - ala - d - ala . consequently , they are disclosed to gain antimicrobial activity against constitutively resistant bacteria endowed with a d - ala - d - lac peptidoglycan cell wall precursor ( e . g . vand ), but be inactive against sensitive and inducibly resistant bacteria ( vana and vanb ) that maintain or at least start with a d - ala - d - ala peptidoglycan cell wall precursor . the closest modified vancomycins that would be expected to reproduce the binding results observed in fig1 are those that replace the amide bond linking residues 4 and 5 with a methylene ( ch 2 ch 2 ) or ethylene ( ch ═ ch ) linker . such derivatives , by analogy with the results in fig1 , would be expected to enhance d - ala - d - lac affinity 100 - fold missing only the contribution to binding derived from the h - bond . the targeted analogue 5 incorporating an amine in the linkage of residue 4 with residue 5 not only removes the offending carbonyl and the destabilizing lone pair interaction with d - ala - d - lac , but it maintains a local polar environment ( protonated amine ) that better accommodates the binding of an electronegative group or atom ( nh of d - ala - d - ala amide or o of d - ala - d - lac ester ). it is disclosed herein that , while this does not bind d - ala - d - lac quite as well as derivatives such as 40 , it is better than 40 at binding d - ala - d - ala . the results of the assessment of 5 alongside vancomycin ( 1 ) and its aglycon 38 are compiled in fig1 . an additional analogue 41 , derived from n - boc deprotection of the synthetic intermediate 33 ( fig1 ), was also examined that bears the methoxycarbonyl protecting group on the residue 4 / 5 linking amine . the binding affinity of 5 for ac 2 - l - lys - d - ala - d - ala ( 2 ) and ac 2 - l - lys - d - ala - d - lac ( 4 ) was essentially equivalent ( 4 . 8 vs 5 . 2 × 10 3 m − 1 , respectively ) with the d - ala - d - lac binding being slightly better . this represented the desired results relative to the vancomycin aglycon where the enhancement for binding d - ala - d - lac is 43 - fold ( 5 . 2 × 10 3 vs 1 . 2 × 10 2 m − 1 ) and the reduction in binding affinity for d - ala - d - ala is 37 - fold ( 4 . 8 × 10 3 vs 1 . 7 × 10 5 m − 1 ). in addition , the comparison of 5 with 41 reflect the impact of the polar amine ( protonated ) versus its carbamate derivative where the binding affinity for d - ala - d - ala with 5 versus 41 increases 3 - fold ( 4 . 8 vs 1 . 6 × 10 3 m − 1 ) while the impact on d - ala - d - lac is a more marginal 1 . 2 - fold increase in affinity ( 5 . 2 vs 4 . 1 × 10 3 m − 1 ). although there are additional structural features in the comparison of 5 and 41 that might impact the absolute affinities measured , in both instances the binding increases with the free amine 5 and it is with 5 that the dual binding is balanced . the four compounds were compared in an antimicrobial assay against vana enterococcus faecalis ( bm4166 ) that is inducibly resistant to treatment by glycopeptide antibiotics including vancomycin and teicoplanin , fig1 . it is the most difficult of the resistant organisms to treat ( vs vanb ) and characteristic of such organisms , they grow unchallenged enlisting a d - ala - d - ala peptidoglycan cell wall precursor , but switch to d - ala - d - lac upon glycopeptide treatment . as such , it represents a superb test of whether 5 and related dual d - ala - d - ala / d - lac binding antibiotics might prove useful in the treatment of resistant bacteria . compound 5 as well as 41 exhibited mics of 31 μg / ml being roughly 40 - fold more potent than vancomycin or its aglycon ( mics = 2000 and 640 μg / ml ) correlating well with the ca . 40 - fold increase in binding affinity for d - ala - d - lac . moreover , this potency is roughly 30 - fold weaker than that observed with vancomycin and its aglycon against sensitive e . faecalis ( mics = 1 - 2 . 5 μg / ml ) correlating with the 35 to 40 - fold loss in binding affinity for d - ala - d - ala . these results suggest that regardless of the peptidoglycan cell wall precursor utilized by the organism , it remains equally sensitive to treatment by 5 and 41 . compound ( 9 ): a solution of 7 ( compound 7 is available in 6 steps ( 37 % overall ) from methyl gallate using 3 recrystallizations and was scaled to 300 g , ( crowley , b . m . ; et al . j . am . chem . soc . 2004 , 126 , 4310 ).) ( 16 . 85 g , 35 . 1 mmol ) in anhydrous ch 2 cl 2 ( 351 ml ) at 0 ° c . under ar was treated with dess - martin periodinane ( 29 . 73 g , 70 . 2 mmol , 2 . 0 equiv ) and the reaction mixture allowed to slowly warm to 25 ° c . and stirred for 1 h . after this time , the reaction mixture was diluted with et 2 o ( 500 ml ), quenched by addition to a cold solution of saturated aqueous nahco 3 ( 1 . 10 l ) and saturated aqueous na 2 so 3 ( 110 ml ) containing na 2 s 2 o 3 . 5h 2 o ( 24 . 2 g ), and stirred until two distinct layers were observed . the layers were separated and the aqueous phase extracted with et 2 o ( 3 × 700 ml ). the combined organic phases were washed with cold saturated aqueous nahco 3 ( 1 × 700 ml ) and cold saturated aqueous nacl ( 1 × 700 ml ), dried ( na 2 so 4 ), and the solvent was evaporated in vacuo to afford crude aldehyde 8 ( 16 . 78 g , 16 . 78 g theoretical , 100 %) as white foam that was carried directly to the next step . note : to prevent polymerization , the workup was carried out as quickly as possible ; the product was removed from the rotary evaporator immediately upon the formation of the foam and was not dried under high vacuum , and the crude aldehyde was immediately dissolved in anhydrous ch 3 oh ( 200 ml ) upon removal from the rotary evaporator . a solution of freshly prepared 6 ( compound 6 is available in 5 steps ( 55 % overall ) from ( r )- 4 - hydroxyphenyl - glycine using 2 recrystallizations and was scaled to 60 g , ( boger , d . l . ; et al . j . org . chem . 1997 , 62 , 4721 ).) ( 11 . 35 g , 41 . 4 mmol , 1 . 2 equiv ) and aldehyde 8 ( 16 . 78 g , 34 . 5 mmol ) in anhydrous ch 3 oh ( 351 ml ) at 0 ° c . under ar was treated with 3 å molecular sieves ( 52 g , 3 . 0 w / w , powder ) and the reaction mixture allowed to stir for 1 h . the solution was cooled to − 20 ° c . and treated dropwise with glacial acetic acid ( 5 . 76 ml , 103 . 5 mmol , 3 . 0 equiv ) to adjust the solution to ph 6 followed by portion - wise addition of nabh 3 cn ( 6 . 63 g , 103 . 5 mmol , 3 . 0 equiv , 4 equal portions , 15 min between additions ). the resulting reaction mixture was stirred at − 20 ° c . for 2 d . the reaction mixture was quenched by slow addition to cold saturated aqueous nahco 3 ( 1 . 0 l ) and the aqueous phase extracted with etoac ( 3 × 800 ml ). the combined organic phases were washed with saturated aqueous nacl ( 800 ml ), dried ( na 2 so 4 ), and concentrated in vacuo . flash chromatography ( sio 2 , 22 . 5 × 22 . 5 cm , 0 - 50 % etoac - hexanes ) gave 9 ( 19 . 40 g , 25 . 85 g theoretical , 75 %) as a light yellow solid : mp 61 ° c . ; [ α ] 25 d − 27 ( c 1 . 7 , chcl 3 ); maldi - ftms ( dhb ) m / z 757 . 2118 ( m + + na , c 38 h 43 brn 2 o 8 requires 757 . 2095 ). compound ( 10 ): a solution of 9 ( 15 . 40 g , 20 . 9 mmol ) in anhydrous thf ( 420 ml ) at 0 ° c . under ar was treated sequentially with k 2 co 3 ( 28 . 9 g , 209 mmol , 10 . 0 equiv ) and methyl chloroformate ( 16 . 2 ml , 209 mmol , 10 . 0 equiv ). the reaction mixture was allowed to warm to 25 ° c . and stirred for 18 h . after this time , the reaction mixture was diluted with cold h 2 o ( 300 ml ) and the aqueous phase was extracted with etoac ( 3 × 300 ml ). the combined organic phases were washed with saturated aqueous nacl ( 300 ml ), dried ( na 2 so 4 ), and concentrated in vacuo . flash chromatography ( sio 2 , 22 . 5 × 22 . 5 cm , 0 - 5 % etoac - ch 2 cl 2 ; then 15 % etoac - ch 2 cl 2 ) afforded 10 ( 14 . 12 g , 16 . 62 g theoretical , 85 %) as a white solid : mp 73 ° c . ; [ α ] 25 d − 47 ( c 0 . 5 , chcl 3 ); maldi - ftms ( dhb ) m / z 815 . 2150 ( m + + na , c 40 h 45 brn 2 o 10 requires 815 . 2150 ). compound ( 11 ): a solution of 10 ( 12 . 50 g , 15 . 7 mmol ) in ch 3 oh ( 525 ml ) at 0 ° c . was treated with raney nickel and the reaction mixture stirred under an atmosphere of h 2 at 0 ° c . for 5 h . the mixture was filtered through a pad of celite ( ch 3 oh , 50 ml ) and the solvent was evaporated in vacuo to give 11 ( 9 . 50 g , 9 . 69 g theoretical , 98 %) as a white solid that was & gt ; 98 % pure by 1 h nmr analysis : mp 91 ° c . ; [ α ] 25 - 25 ( c 0 . 2 , ch 3 oh ); esi - tof hrms m / z 613 . 1395 ( m + + h , c 26 h 33 brn 2 o 10 requires 613 . 1391 ). compound ( s1 ): a solution of 10 ( 0 . 22 g , 0 . 27 mmol ) in thf ( 14 ml ) at 0 ° c . was treated with 0 . 2 n aqueous lioh ( 1 . 65 ml , 0 . 33 mmol , 1 . 1 equiv ) and allowed to stir for 2 h . the reaction mixture was quenched by the addition of 0 . 2 n aqueous hcl until the ph of the solution reached 3 and the aqueous phase extracted with etoac ( 3 × 10 ml ). the combined organic phases were washed with saturated aqueous nacl ( 1 × 20 ml ), dried ( na 2 so 4 ), and concentrated in vacuo . flash chromatography ( sio 2 , 5 × 20 cm , 5 - 20 % ch 3 oh — ch 2 cl 2 ) afforded s1 ( 0 . 18 g , 0 . 18 g theoretical , 100 %) as a white solid : mp 97 ° c . ; [ α ] 25 d − 126 ( c 0 . 6 , chcl 3 ); maldi - ftms ( dhb ) m / z 778 . 2100 ( m + + na , c 39 h 43 brn 2 o 10 requires 778 . 6801 ). compound ( 12 ): a solution of 11 ( 9 . 50 g , 15 . 5 mmol ) in thf ( 310 ml ) at 0 ° c . was treated with 0 . 2 n aqueous lioh ( 93 . 0 ml , 46 . 5 mmol , 3 . 0 equiv ) and allowed to stir for 2 h . the mixture was quenched by the addition of 0 . 2 n aqueous hcl until the ph of the solution reached 3 and the aqueous phase extracted with etoac ( 3 × 200 ml ). the combined organic phases were washed with saturated aqueous nacl ( 1 × 200 ml ), dried ( na 2 so 4 ), and concentrated in vacuo to give 12 ( 9 . 26 g , 9 . 26 g theoretical , 100 %) as a white solid : mp 110 ° c . ; [ α ] 25 d − 57 ( c 0 . 3 , chcl 3 ); maldi - ftms ( dhb ) m / z 621 . 1068 ( m + + na , c 25 h 31 brn 2 o 10 requires 621 . 1054 ). compound ( 14 ): a solution of 12 ( 2 . 37 g , 3 . 95 mmol ) in anhydrous dmf ( 32 ml ) at 0 ° c . under ar was treated sequentially nahco 3 ( 1 . 00 g , 11 . 9 mmol , 3 . 0 equiv ), depbt ( 3 . 55 g , 11 . 9 mmol , 3 . 0 equiv ), and a solution of 13 [ compound 13 is available in 3 steps ( 45 % overall ) from 4 - fluoro - 3 - nitrobenzaldehyde and was scaled to 30 g , ( crowley , b . m . ; et al . j . am . chem . soc . 2004 , 126 , 4310 ).] ( 1 . 19 g , 4 . 35 mmol , 1 . 1 equiv ) in anhydrous dmf ( 8 . 0 ml ). the reaction mixture was allowed to slowly warm to 25 ° c . and stirred for 8 h . the reaction mixture was quenched by addition to saturated aqueous nahco 3 ( 60 ml ) and the aqueous phase extracted with etoac ( 3 × 60 ml ). the combined organic phases were washed with saturated aqueous nahco 3 ( 3 × 60 ml ), h 2 o ( 1 × 60 ml ), and saturated aqueous nacl ( 1 × 60 ml ), dried ( na 2 so 4 ), and concentrated in vacuo . flash chromatography ( sio 2 , 6 × 20 cm , 0 - 70 % etoac - ch 2 cl 2 ) afforded 14 ( 2 . 65 g , 3 . 79 g theoretical , 70 %) as a pale yellow solid : mp 121 ° c . ; [ α ] 25 d − 9 ( c 0 . 8 , chcl 3 ); maldi - ftms ( dhb ) m / z 975 . 2476 ( m + + na , c 41 h 54 brfn 4 o 14 si requires 975 . 2465 ). method a . a solution of 14 ( 2 . 65 g , 2 . 78 mmol ) in anhydrous thf ( 230 ml , additionally dried over 3 å ms for 18 h , then na for 12 h ) under ar was treated with k 2 co 3 ( 9 . 60 g , 69 . 5 mmol , 25 equiv , dried in vacuo at 130 ° c . for 18 h ), caco 3 ( 6 . 95 g , 69 . 5 mmol , 25 equiv , dried in vacuo at 130 ° c . for 18 h ), and 3 å molecular sieves ( 7 . 95 g , 3 . 0 w / w , powder , dried in vacuo at 130 ° c . for 18 h ). the reaction mixture was warmed at 75 ° c . ( bath temp .) and stirred for 12 h . after this time , the reaction mixture was cooled to 25 ° c . and filtered through celite ( eluted with thf ) and concentrated in vacuo . the remaining solid was dissolved in etoac ( 200 ml ), washed with saturated aqueous nh 4 cl ( 1 × 50 ml ) and saturated aqueous nacl , dried ( na 2 so 4 ), and concentrated in vacuo . flash chromatography ( sio 2 , 6 × 20 cm , 0 - 70 % etoac - ch 2 cl 2 ) afforded 15 ( 1 . 40 g , 2 . 59 g theoretical , 54 %) and 16 ( 0 . 56 g , 2 . 59 g theoretical , 22 %) as light yellow solids ( 76 % total conversion , 2 . 5 : 1 15 : 16 ): for 15 : mp 205 ° c . ; [ α ] 25 d − 32 ( c 1 . 1 , ch 3 oh ); maldi - ftms ( dhb ) m / z 955 . 2379 ( m + + na , c 41 h 53 brn 4 o 14 si requires 955 . 2403 ). for 16 : mp 207 ° c . ; [ α ] 25 d − 35 ( c 0 . 08 , ch 3 oh ); maldi - ftms ( dhb ) m / z 955 . 2401 ( m + + na , c 41 h 53 brn 4 o 14 si requires 955 . 2403 ). method b . a solution of 14 ( 9 . 0 mg , 9 . 4 μmol ) in anhydrous thf ( 1 . 57 ml , additionally dried over 3 å ms for 18 h , then na for 12 h ) under ar was cooled to − 78 ° c . and treated with a freshly prepared solution of potassium tert - butoxide ( 1 . 0 mg , 9 . 4 μmol ) in anhydrous thf ( 9 . 4 μl ), and the mixture was warmed to − 20 ° c . and stirred for 18 h . the reaction mixture was quenched by addition to cold saturated aqueous nh 4 cl ( 3 . 0 ml ) and the aqueous phase was extracted with etoac ( 3 × 3 ml ). the combined organic phases were washed with saturated aqueous nacl ( 1 × 3 ml ), dried ( na 2 so 4 ), and concentrated in vacuo . ptlc ( sio 2 , 2 . 5 % ch 3 oh — ch 2 cl 2 ) afforded 15 ( 5 . 0 mg , 8 . 8 mg theoretical , 57 %) and 16 ( 1 . 7 mg , 8 . 8 mg theoretical , 19 %) as light yellow solids ( 76 % total conversion , 2 . 9 : 1 15 : 16 ). a summary of conditions initially surveyed for the conversion of 14 to 15 may be found in fig5 a and selected conditions examined during the optimization of the reaction may be found in fig5 b . compound ( 17 ): a solution of 15 ( 1 . 54 g , 1 . 65 mmol ) in ch 3 oh ( 54 ml ) was treated with raney nickel at 0 ° c . and the reaction mixture was stirred under an atmosphere of h 2 at 0 ° c . for 2 h . the reaction mixture was filtered through a pad of celite ( eluted with ch 3 oh ) and the solvent was removed in vacuo to afford the crude aniline ( 1 . 49 g , 1 . 49 g theoretical , 100 %) as a light yellow foam solid that was carried on to the next step without purification . a solution of the crude aniline ( 1 . 49 g , 1 . 65 mmol ) in anhydrous ch 3 cn ( 26 ml , degassed ) at 0 ° c . under ar was treated with hbf 4 ( 0 . 1 m solution in ch 3 cn , 19 . 4 ml , 2 . 15 mmol , 1 . 3 equiv , 0 ° c .) for 10 min before the dropwise addition of t - buono ( 0 . 1 m solution in ch 3 cn , 19 . 4 ml , 2 . 15 mmol , 1 . 3 equiv , 0 ° c .). the reaction mixture was stirred at 0 ° c . for 10 min and then cooled to − 20 ° c . before addition to a vigorously stirring aqueous solution ( 52 ml , degassed ) containing cucl ( 7 . 63 g , 82 . 5 mmol , 50 equiv ) and cucl 2 ( 12 . 37 g , 99 mmol , 60 equiv ) at 0 ° c . under ar . the resulting reaction mixture was allowed to warm to 25 ° c . and stirred for 1 h . the reaction mixture was quenched by slow addition to a cold saturated aqueous solution of nahco 3 ( 100 ml ) and cold etoac ( 100 ml ) was added before filtration through a pad of celite . the layers were separated and the aqueous phase extracted with cold etoac ( 2 × 100 ml ). the combined organic phases were washed with cold saturated aqueous nahco 3 ( 2 × 50 ml ) and cold saturated aqueous nacl ( 1 × 50 ml ), dried ( na 2 so 4 ), and concentrated in vacuo . flash chromatography ( sio 2 , 6 × 20 cm , 0 - 70 % etoac - ch 2 cl 2 ) afforded 17 ( 1 . 07 g , 1 . 52 g theoretical , 70 %) as a light yellow solid : mp 195 ° c . ( dec ); [ α ] 25 d + 20 ( c 0 . 8 , chcl 3 ); maldi - ftms ( dhb ) m / z 944 . 2154 ( m + + na , c 41 h 53 brcln 3 o 12 si requires 944 . 2162 ). compound ( 18 ): a solution of 16 ( 5 . 1 mg , 5 . 5 mmol ) in ch 3 oh ( 1 . 8 ml ) was treated with raney nickel at 0 ° c . and the reaction mixture was stirred under an atmosphere of h 2 at 0 ° c . for 2 h . the mixture was filtered through a pad of celite ( eluted with ch 3 oh ) and the solvent was removed in vacuo to afford the crude aniline ( 5 . 0 mg , 5 . 0 mg theoretical , 100 %) as a light yellow solid that was carried on to the next step without purification . a solution of the crude aniline ( 5 . 0 mg , 5 . 5 μmol ) in anhydrous ch 3 cn ( 0 . 9 ml , degassed ) at 0 ° c . under ar was treated with hbf 4 ( 0 . 1 m solution in ch 3 cn , 0 . 65 ml , 71 . 7 μmol , 1 . 3 equiv , 0 ° c .) for 10 min before the dropwise addition of t - buono ( 0 . 1 m solution in ch 3 cn , 0 . 65 ml , 71 . 7 μmol , 1 . 3 equiv , 0 ° c .). the reaction mixture was stirred at 0 ° c . for 10 min and then cooled to − 20 ° c . before addition to a vigorously stirring aqueous solution ( 1 . 8 ml , degassed ) containing cucl ( 254 mg , 2 . 75 mmol , 50 equiv ) and cucl 2 ( 412 mg , 3 . 3 mmol , 60 equiv ) at 0 ° c . under ar and the reaction mixture was allowed to warm to 25 ° c . and stirred for 1 h . the reaction mixture was quenched by slow addition to a cold saturated aqueous solution of nahco 3 ( 4 ml ) and cold etoac ( 4 ml ) was added before filtration through a pad of celite . the layers were separated and the aqueous phase extracted with cold etoac ( 2 × 4 ml ). the combined organic phases were washed with cold saturated aqueous nahco 3 ( 2 × 2 ml ) and cold saturated aqueous nacl ( 1 × 2 ml ), dried ( na 2 so 4 ), and concentrated in vacuo . flash chromatography ( ptlc , sio 2 , 5 % ch 3 oh — ch 2 cl 2 ) afforded 18 ( 3 . 8 mg , 5 . 1 mg theoretical , 75 %) as a yellow solid : mp 190 ° c . ( dec ); [ α ] 25 d − 35 ( c 0 . 7 , chcl 3 ); maldi - ftms ( dhb ) m / z 944 . 2178 ( m + + na , c 41 h 53 brcln 3 o 12 si requires 944 . 2162 ). compound ( 19 ): from 18 : a solution of 18 ( 2 . 2 mg , 2 . 6 mmol ) in ch 3 oh ( 1 . 5 ml ) was treated with 10 % pd / c ( 0 . 4 mg , 0 . 2 w / w ) and placed under an atmosphere of h 2 in a parr shaker . the flask was pressurized with h 2 to 40 psi and shook for 16 h . the reaction mixture was filtered through celite , the pad was washed with ch 3 oh ( 5 ml ), and the solvent evaporated in vacuo to afford 19 ( 1 . 8 mg , 2 . 1 mg theoretical , 85 %) as a white solid : mp 180 ° c . ; [ α ] 25 d + 78 ( c 0 . 45 , chcl 3 ); maldi - ftms ( dhb ) m / z 832 . 3417 ( m + + na , c 41 h 55 n 3 o 12 si requires 832 . 3447 ). compound ( 19 ): from 17 : a solution of 17 ( 1 . 2 mg , 1 . 3 mmol ) in ch 3 oh ( 1 . 5 ml ) was treated with 10 % pd / c ( 0 . 3 mg , 0 . 2 w / w ) and placed under an atmosphere of h 2 in a parr shaker . the flask was pressurized with h 2 to 40 psi and shook for 16 h . the reaction mixture was filtered through celite , the pad was washed with ch 3 oh , and the solvent evaporated in vacuo to afford 19 ( 0 . 85 mg , 1 . 1 mg theoretical , 80 %) as a white solid identical in all respects to the material described above : mp 180 ° c . ; [ α ] 25 d + 75 ( c 0 . 4 , chcl 3 ); maldi - ftms ( dhb ) m / z 832 . 3452 ( m + + na , c 41 h 55 n 3 o 12 si requires 832 . 3447 ). compound ( 21 ): a suspension of 17 ( 820 mg , 0 . 89 mmol ), 20 ( boger , d . l . ; et al . j . am . chem . soc . 1999 , 121 , 3226 ; boger , d . l . ; et al . j . am . chem . soc . 1999 , 121 , 10004 )( 1 . 11 g , 2 . 40 mmol , 2 . 7 equiv ), pd 2 ( dba ) 3 ( 244 mg , 0 . 27 mmol , 0 . 3 equiv ), and tri - o - tolylphosphine ( 405 mg , 1 . 33 mmol , 1 . 5 equiv ) in toluene ( 6 . 34 ml , degassed ), ch 3 oh ( 1 . 90 ml , degassed ), and 1 m aqueous na 2 co 3 ( 0 . 19 ml , 0 . 19 mmol , degassed ) was warmed to 80 ° c . under ar and stirred vigorously for 30 min . the mixture was cooled to 0 ° c ., diluted with etoac ( 10 ml ) and h 2 o ( 10 ml ) and treated with 1 n aqueous hcl ( 2 . 0 ml ). the aqueous phase was extracted with etoac ( 3 × 10 ml ), and the combined organic phases were washed with saturated aqueous nacl ( 1 × 10 ml ), dried ( na 2 so 4 ), and concentrated in vacuo . flash chromatography ( sio 2 , 6 × 22 . 5 cm , 0 - 70 % etoac - ch 2 cl 2 ) provided 21 ( 443 mg , 1 . 13 g theoretical , 39 %) as a white solid and 22 ( 577 mg , 1 . 13 g theoretical , 51 %) as a white solid ( 1 . 13 g theoretical , 90 %; typically 75 - 90 %). for 21 : mp 145 ° c . ; [ α ] 25 d − 13 ( c 0 . 5 , chcl 3 ); maldi - ftms ( dhb ) m / z 1283 . 4874 ( m + + na , c 63 h 81 cln 4 o 19 si requires 1283 . 4845 ). for 22 : mp 139 ° c . ; [ α ] 25 d + 7 ( c 0 . 6 , chcl 3 ); maldi - ftms ( dhb ) m / z 1283 . 4824 ( m + + na , c 63 h 81 cln 4 o 19 si requires 1283 . 4845 ). thermal equilibration and recycling of 22 : a solution of 22 ( 260 mg , 0 . 21 mmol ) in o - dichlorobenzene ( 15 ml , dried over 3 å ms for 18 h ) under ar was placed into a 20 ml microwave reactor vial . the vial was loaded into a microwave reactor ( biotage initiator ) and heated to 210 ° c . for 5 min . flash chromatography ( sio 2 , 4 . 5 × 22 . 5 cm , 0 - 70 % etoac - ch 2 cl 2 ) afforded 21 ( 110 mg , 260 mg theoretical , 42 %) as a white solid and 22 ( 120 mg , 260 mg theoretical , 46 %) as a white solid ( 230 mg recovered , 260 mg theoretical , 88 % recovery ). compound ( 23 ): a solution of 21 ( 585 mg , 0 . 46 mmol ) in anhydrous thf ( 9 . 30 ml ) at 0 ° c . under ar was treated with bu 4 nf ( 1 . 0 m in thf , 567 μl , 0 . 56 mmol , 1 . 2 equiv ) and the reaction mixture was allowed to stir at 0 ° c . for 10 min . the reaction mixture was quenched with the addition of saturated aqueous nh 4 cl ( 15 ml ) and the aqueous phase was extracted with etoac ( 3 × 15 ml ). the combined organic layers were washed with saturated aqueous nacl ( 1 × 10 ml ), dried ( na 2 so 4 ), and concentrated in vacuo . flash chromatography ( sio 2 , 3 × 22 . 5 cm , 0 - 100 % etoac - ch 2 cl 2 ) afforded 23 ( 424 mg , 530 mg theoretical , 80 %) as a white solid : mp 152 ° c . ; [ α ] 25 d − 53 ( c 0 . 4 , chcl 3 ); maldi - ftms ( dhb ) m / z 1169 . 3964 ( m + + na , c 57 h 67 cln 4 o 19 requires 1169 . 3980 ). compound ( 24 ): a solution of 23 ( 392 mg , 0 . 34 mmol ) in anhydrous 1 % cl 3 cco 2 h — ch 3 oh ( 17 ml ) was treated with pd / c ( 10 %, 39 mg , 0 . 1 w / w ) and stirred under an atmosphere of h 2 at 25 ° c . for 15 min . the reaction mixture was filtered through a pad of celite , the pad washed with ch 3 oh — hoac ( 15 ml , 1 % hoac ), and the ph of the solution adjusted to 8 with the addition of solid nahco 3 . the solution was filtered through a pad of celite and the solvent evaporated in vacuo . flash chromatography ( sio 2 , 3 × 22 . 5 cm , 5 - 15 % ch 3 oh — ch 2 cl 2 ) afforded 24 ( 329 mg , 346 mg theoretical , 95 %) as a white foam : [ α ] 25 d − 17 ( c 0 . 4 , ch 3 oh ); esi - tof hrms m / z 1013 . 3798 ( m + + h , c 49 h 61 cln 4 o 17 requires 1013 . 3793 ). compound ( s2 ): a solution of 23 ( 23 . 7 mg , 20 . 7 mmol ) in thf ( 1 . 03 ml ) was treated with 0 . 2 n aqueous lioh ( 114 ml , 114 mmol , 1 . 1 equiv ) at 0 ° c . and the reaction mixture was allowed to stir for 1 h . the reaction mixture was diluted with h 2 o ( 1 . 0 ml ), quenched by addition of 0 . 2 n aqueous hcl until the ph of the solution reached 3 , and the aqueous phase extracted with etoac ( 3 × 1 . 0 ml ). the combined organic layers were washed with saturated aqueous nacl ( 1 × 1 . 0 ml ), dried ( na 2 so 4 ), and concentrated in vacuo . flash chromatography ( sio 2 , 0 . 5 × 18 cm , 5 - 15 % ch 3 oh — ch 2 cl 2 ) afforded s2 ( 23 . 4 mg , 23 . 4 mg theoretical , 100 %) as a white foam : [ α ] 25 d − 5 . 0 ( c 0 . 4 , ch 3 oh ); maldi - ftms ( dhb ) m / z 1155 . 3862 ( m + + na , c 56 h 65 cln 4 o 19 requires 1155 . 3824 ). 25 . a solution of 24 ( 312 mg , 0 . 31 mmol ) in thf — h 2 o ( 15 . 4 ml , 10 : 1 ) was treated with 0 . 5 n aqueous lioh ( 616 μl , 0 . 31 mmol , 1 . 0 equiv ) at 0 ° c . and the reaction mixture was allowed to stir for 1 h . the reaction mixture was quenched by addition of 0 . 2 n aqueous hcl until the ph of the solution reached 3 and then was concentrated in vacuo . flash chromatography ( sio 2 , 2 × 18 cm , 5 - 20 % ch 3 oh — ch 2 cl 2 ) afforded 25 ( 292 mg , 307 mg theoretical , 95 %) as a white foam : [ α ] 25 d + 70 ( c 0 . 4 , ch 3 oh ); maldi - ftms ( dhb ) m / z 999 . 3618 ( m + + h , c 48 h 59 cln 4 o 17 requires 999 . 3636 ). 26 . a cold solution of 25 ( 273 mg , 0 . 27 mmol ) in anhydrous ch 2 cl 2 - dmf ( 72 ml , 5 : 1 ) was added dropwise over the course of 1 . 5 h to a stirring solution of pybop ( 470 mg , 0 . 81 mmol , 3 . 0 equiv ) and nahco 3 ( 126 mg , 1 . 62 mmol , 6 . 0 equiv ) in ch 2 cl 2 - dmf ( 200 ml , 5 : 1 ) at 0 ° c . under ar . the reaction mixture was allowed to warm to 25 ° c . and stirred for 12 h . the reaction mixture was quenched by addition to saturated aqueous nh 4 cl ( 250 ml ) and the aqueous phase was extracted with etoac ( 3 × 250 ml ). the combined organic phases were washed with saturated aqueous nh 4 cl ( 5 × 100 ml ) and saturated aqueous nacl ( 1 × 80 ml ), dried ( na 2 so 4 ), and concentrated in vacuo . flash chromatography ( sio 2 , 3 . 5 × 18 cm , 10 % ch 3 cn — ch 2 cl 2 then 0 - 10 % ch 3 oh — ch 2 cl 2 ) afforded 26 ( 188 mg , 268 mg theoretical , 70 %) as a white solid : mp 188 ° c . ; [ α ] 25 d − 10 ( c 0 . 8 , chcl 3 ); esi - tof hrms m / z 981 . 3520 ( m + + h , c 48 h 57 cln 4 o 16 requires 981 . 3531 ). 29 . a solution of 26 ( 110 mg , 0 . 11 mmol ) in chcl 3 ( 5 . 6 ml ) was treated with hco 2 h ( 5 . 6 ml ) and stirred at 25 ° c . under ar for 10 h . the reaction was quenched by the addition of saturated aqueous nahco 3 until the ph of the solution reached 7 . 5 . the layers were separated and the aqueous phase was extracted with chcl 3 ( 3 × 10 ml ). the combined organic phases were washed with saturated aqueous nacl ( 1 × 10 ml ), dried ( na 2 so 4 ), and concentrated in vacuo to give the crude free amine 27 ( 82 . 8 mg , 98 . 8 mg theoretical , 84 %) that was carried on without further purification : maldi - ftms ( dhb ) m / z 903 . 2823 ( m + + na , c 43 h 49 cln 4 o 14 requires 903 . 2826 ). a solution of 27 ( 82 . 8 mg , 0 . 09 mmol ) and 28 ( boger , d . l . ; et al . j . am . chem . soc . 1999 , 121 , 3226 ; boger , d . l . ; et al . j . am . chem . soc . 1999 , 121 , 10004 ) ( 69 . 3 mg , 0 . 12 mmol , 1 . 3 equiv ) in anhydrous thf ( 1 . 88 ml ) at 0 ° c . was treated sequentially with nahco 3 ( 39 . 5 mg , 0 . 47 mmol , 5 . 0 equiv ) and depbt ( 84 . 3 mg , 0 . 28 mmol , 3 . 0 equiv ), and the reaction mixture was allowed to warm to 25 ° c . and stirred for 14 h . the reaction was quenched by addition of saturated aqueous nh 4 cl ( 10 ml ) and the aqueous phase was extracted with etoac ( 3 × 10 ml ). the combined organic phases were washed with saturated aqueous nh 4 cl ( 5 × 10 ml ) and saturated aqueous nacl ( 1 × 10 ml ), dried ( na 2 so 4 ), and concentrated in vacuo . flash chromatography ( sio 2 , 1 × 18 cm , 0 - 10 % ch 3 oh — ch 2 cl 2 ) afforded 29 ( 98 . 1 mg , 134 . 4 mg theoretical , 73 %) as a yellowish white solid : [ α ] 25 d + 19 ( c 0 . 1 , chcl 3 ); esi - tof hrms m / z 1430 . 5230 ( m + + h , c 68 h 81 clfn 9 o 22 requires 1430 . 5241 ). 30 . a solution of 29 ( 16 . 1 mg , 11 . 3 mmol ) in anhydrous dmf ( 1 . 88 ml , dried over 3 å ms for 18 h ) was treated sequentially with 3 å molecular sieves ( 48 mg , 3 w / w equiv ), caco 3 ( 22 . 5 mg , 0 . 23 mmol , 20 equiv ), and csf ( 17 . 1 mg , 0 . 11 mmol , 10 equiv ) at 25 ° c . under ar and allowed to stir for 17 h . the reaction mixture was quenched by the addition of saturated aqueous nh 4 cl ( 5 ml ) and the aqueous phase was extracted with etoac ( 3 × 5 ml ). the combined organic phases were washed with saturated aqueous nh 4 cl ( 5 × 5 ml ) and saturated aqueous nacl ( 1 × 5 ml ), dried ( na 2 so 4 ), and concentrated in vacuo . ptlc ( sio 2 , 7 . 5 % ch 3 o — ch 2 cl 2 ) afforded 30 ( 11 . 7 mg , 15 . 9 mg theoretical , 74 %) as a white solid and its atropisomer 31 ( 2 . 0 mg , 15 . 9 mg theoretical , 12 %) as a white film . for 30 : mp 244 ° c . ( dec ); [ α ] 25 d + 57 ( c 0 . 1 , chcl 3 ); esi - tof hrms m / z 1410 . 5205 ( m + + h , c 68 h 80 cln 9 o 22 requires 1410 . 5179 ). for 31 : [ α ] 25 d + 15 ( c 0 . 5 , chcl 3 ); esi - tof hrms m / z 1410 . 5181 ( m + + h , c 68 h 80 cln 9 o 22 requires 1410 . 5179 ). 33 . a solution of 30 ( 6 . 8 mg , 4 . 8 μmol ) in anhydrous thf ( 482 μl ) was treated with 10 % pd / c ( 1 . 4 mg , 0 . 2 w / w equiv ) and stirred under an atmosphere of h 2 in the dark for 8 h . the reaction mixture was filtered through a pad of celite , the pad was washed with thf ( 15 ml ), and the solvent was evaporated in vacuo at 4 ° c . to give crude aniline 32 ( 6 . 2 mg , 6 . 6 mg theoretical , 94 %) that was carried on without purification . a solution of 32 ( 6 . 2 mg , 4 . 5 μmol ) in ch 3 cn ( 448 μl , degassed ) at 0 ° c . was treated dropwise with hbf 4 ( 0 . 1 m solution in ch 3 cn , 57 μl , 5 . 7 μmol , 1 . 3 equiv , 0 ° c .) before the dropwise addition of t - buono ( 0 . 1 m solution in ch 3 cn , 57 μl , 5 . 7 μmol , 1 . 3 equiv , 0 ° c .). the reaction mixture was stirred at 0 ° c . for 10 min and then cooled to − 20 ° c . before addition to a vigorously stirring aqueous solution ( 900 μl , degassed ) containing cucl ( 22 . 2 mg , 225 μmol , 50 equiv ) and cucl 2 ( 21 . 7 mg , 270 μmol , 60 equiv ) at 0 ° c . under ar . the reaction mixture was warmed to 25 ° c . and stirred for 1 h . the reaction was quenched by addition to cold saturated aqueous nahco 3 ( 2 ml ) and the aqueous phase extracted with etoac ( 3 × 2 ml ). the combined organic phases were washed with cold saturated aqueous nahco 3 ( 1 × 2 ml ) and cold saturated aqueous nacl ( 1 × 2 ml ), dried ( na 2 so 4 ), and the solvent removed in vacuo . ptlc ( sio 2 , 7 . 5 % ch 3 oh — ch 2 cl 2 ) afforded 33 ( 1 . 9 mg , 3 . 6 mg theoretical , 54 %) as a white solid : [ α ] 25 d + 38 ( c 0 . 1 , chcl 3 ); esi - tof hrms m / z 1399 . 4924 ( m + + h , c 68 h 80 cl 2 n 8 o 20 requires 1399 . 4938 ). 34 . a solution of 33 ( 3 . 8 mg , 2 . 7 μmol ) in anhydrous ch 3 cn ( 270 μl ) under ar was treated with cf 3 conmetbs ( 42 μl , 0 . 18 mmol , 65 equiv ), warmed to 55 ° c ., and allowed to stir for 2 d . the reaction mixture was cooled to 25 ° c ., quenched by addition of etoac - 15 % aqueous citric acid ( 4 ml , 4 : 1 ), and stirred for 15 h . the solution was diluted with h 2 o ( 2 ml ) and the aqueous phase was extracted with etoac ( 3 × 2 ml ). the combined organic phases were washed with saturated aqueous nahco 3 ( 1 × 2 ml ) and saturated aqueous nacl ( 1 × 2 ml ), dried ( na 2 so 4 ), and concentrated in vacuo . ptlc ( sio 2 , 7 . 5 % ch 3 oh — ch 2 cl 2 ) afforded 34 ( 4 . 2 mg , 4 . 4 mg theoretical , 96 %) as a light yellow solid : [ α ] 25 d + 28 ( c 0 . 1 , chcl 3 ); esi - tof hrms m / z 1627 . 6685 ( m + + h , c 80 h 108 cl 2 n 8 o 20 si 2 requires 1627 . 6668 ). 35 . a solution of 34 ( 2 . 4 mg , 1 . 5 μmol ) in anhydrous ch 2 cl 2 ( 74 μl ) at 0 ° c . under ar in the dark was treated with b - bromocatecholborane ( 0 . 2 m solution in ch 2 cl 2 , 88 μl , 18 μmol , 12 equiv ) and the reaction mixture was allowed to stir for 2 h at 0 ° c . the reaction mixture was diluted with ch 2 cl 2 ( 1 ml ), quenched by addition of saturated aqueous nahco 3 ( 1 ml ), and the aqueous phase was extracted with etoac ( 3 × 1 ml ). the combined organic phases were washed with saturated aqueous nacl ( 1 × 1 ml ), dried ( na 2 so 4 ), and the solvent evaporated in vacuo . the crude amino alcohol was dissolved in dioxane - h 2 o ( 150 μl ), cooled to 0 ° c ., and treated sequentially with nahco 3 ( 0 . 7 mg , 9 . 0 μmol , 6 . 0 equiv ) and boc 2 o ( 1 . 6 mg , 7 . 5 μmol , 5 equiv ). the resulting reaction mixture was allowed to warm to 25 ° c . and stirred for 2 . 5 h . after this time , the reaction mixture was diluted with h 2 o ( 1 ml ) and the aqueous phase extracted with etoac ( 3 × 1 ml ). the combined organic phases were washed with saturated aqueous nacl ( 1 × 1 ml ), dried ( na 2 so 4 ), and the solvent evaporated in vacuo . ptlc ( sio 2 , 10 % ch 3 oh — ch 2 cl 2 ) afforded 35 ( 1 . 8 mg , 2 . 3 mg theoretical , 80 %) as a white solid : [ α ] 25 d + 18 ( c 0 . 3 , ch 3 oh ); esi - tof hrms m / z 1561 . 5984 ( m + + na , c 76 h 100 cl 2 n 8 o 18 si 2 requires 1561 . 5963 ). 36 . a solution of 35 ( 1 . 4 mg , 0 . 9 μmol ) in anhydrous ch 2 cl 2 ( 50 μl ) at 0 ° c . under ar was treated with dess - martin periodinane ( 1 . 5 mg , 3 . 6 μmol , 4 . 0 equiv ) and the reaction mixture was allowed to stir for 1 . 5 h . the reaction mixture was diluted with et 2 o ( 1 ml ), quenched by addition of saturated aqueous nahco 3 ( 1 ml ), and the aqueous phase extracted with et 2 o ( 3 × 1 ml ). the combined organic phases were washed with saturated aqueous nacl ( 1 × 1 ml ), dried ( na 2 so 4 ), and concentrated under a stream of n 2 . the crude aldehyde was dissolved in t - buoh - 2 - methyl - 2 - butene ( 50 μl , 4 : 1 ) and treated with a solution containing 80 % naclo 2 ( 1 mg , 8 . 1 μmol , 9 . 0 equiv ) and nah 2 po 4 . h 2 o ( 0 . 9 mg , 6 . 3 μmol , 7 . 0 equiv ) in h 2 o ( 8 . 0 μl ) and the reaction mixture was allowed to stir for 30 min at 25 ° c . the reaction mixture was diluted with h 2 o ( 1 ml ) and the aqueous phase extracted with etoac ( 3 × 1 ml ). the combined organic phases were washed with saturated aqueous nacl ( 1 × 1 ml ), dried ( na 2 so 4 ), and concentrated under a stream of n 2 . ptlc ( 10 % ch 3 oh — ch 2 cl 2 ) afforded 36 ( 1 . 1 mg , 1 . 4 mg theoretical , 80 %) as a white solid : [ α ] 25 d + 21 ( c 0 . 1 , ch 3 oh ); esi - tof hrms m / z 1553 . 5989 ( m + + h , c 76 h 98 cl 2 n 8 o 19 si 2 requires 1553 . 5936 ). 37 . a solution of 36 ( 1 . 0 mg , 0 . 64 μmol ) in dmso ( 160 μl ) was treated sequentially with h 2 o 2 ( 50 % aqueous solution , 2 . 0 μl , 27 . 2 μmol , 43 equiv ) and k 2 co 3 ( 10 % aqueous solution , 7 . 2 μl , 5 . 8 μmol , 9 . 0 equiv ) and allowed to stir for 3 . 5 h . the reaction mixture was diluted with etoac ( 1 ml ), quenched by the addition of 0 . 1 n aqueous hcl ( 580 μl ), and the aqueous phase extracted with etoac ( 3 × 1 ml ). the combined organic phases were washed with saturated aqueous nacl ( 1 × 1 ml ), dried ( na 2 so 4 ), and concentrated under a stream of n 2 . ptlc ( sio 2 , 10 % ch 3 oh — ch 2 cl 2 ) afforded 37 ( 0 . 9 mg , 1 . 0 mg theoretical , 87 %) as a white film : [ α ] 25 d + 26 ( c 0 . 3 , ch 3 oh ); esi - tof hrms m / z 1571 . 6046 ( m + + h , c 76 h 100 cl 2 n 8 o 20 si 2 requires 1571 . 6042 ). [ ψ [ ch 2 nh ] tpg 4 ] vancomycin aglycon ( 5 ). a vial containing 37 ( 0 . 7 mg , 0 . 45 μmol ) was treated with a solution of albr 3 ( 4 . 5 mg , 16 . 9 μmol , 38 equiv ) in etsh ( 23 μl ) and the reaction mixture was allowed to stir for 5 h . the reaction mixture was cooled to 0 ° c ., diluted with chcl 3 ( 100 μl ), quenched by the addition of ch 3 oh ( 10 μl ), and concentrated under a stream of n 2 . reverse phase chromatography ( c 18 , 100 % h 2 o then 50 % ch 3 cn — h 2 o ) afforded 5 ( 0 . 4 mg , 0 . 5 mg theoretical , 80 %) as a white film : 1 h nmr ( cd 3 od , 3 mm , 600 mhz ) δ7 . 70 ( br s , 1h ), 7 . 66 ( m , 1h ), 7 . 62 ( m 1h ), 7 . 42 ( m , 1h ), 7 . 31 ( m , 1h ), 7 . 25 ( br s , 2h ), 7 . 11 ( m , 1h ), 6 . 92 ( m , 1h ), 6 . 84 ( m , 1h ), 6 . 42 ( br s , 1h ), 5 . 50 ( s , 1h ), 5 . 46 ( s , 1h ), 5 . 37 ( m , 1h ), 5 . 33 ( br s , 2h ), 5 . 26 ( m , 1h ), 4 . 91 ( m , obscured by hod , 1h ), 4 . 52 ( m , 1h ), 4 . 21 ( m , 1h ), 4 . 11 ( m , 1h ), 4 . 07 ( br s , 1h ), 3 . 74 ( m , 1h ), 3 . 68 ( s , 4h ), 3 . 57 ( br s , 5h ), 2 . 81 ( m , 2h ), 2 . 77 ( s , 3h ), 2 . 02 ( m , 1h ), 2 . 18 ( m , 1h ), 1 . 66 ( m , 1h ), 1 . 59 ( s , 2h ), 0 . 94 ( m , 3h ), 0 . 88 ( m , 3h ); maldi - tof m / z 1129 . 2 ( m + + h , c 53 h 54 cl 2 n 8 o 16 requires 1129 . 3 ). binding constant determination . the binding constants for compounds 5 and 41 for association with the model ligands n , n ′- ac 2 - lys - d - ala - d - ala and n , n ′- ac 2 - lys - d - ala - d - lac were determined according to literature ( nieto , m . ; perkins , h . r . biochem . j . 1971 , 124 , 845 ; nieto , m . ; perkins , h . r . biochem . j . 1971 , 124 , 773 ; nieto , m . ; perkins , h . r . biochem . j . 1971 , 124 , 789 ) protocols . uv difference experiments were carried out on a cary 3e uv - vis spectrometer . uv scans were run with a baseline correction that consisted of 0 . 02 m sodium citrate buffer and covered a range from 200 to 345 nm . a solution of 5 or 41 ( 1 . 1 × 10 − 4 m in 0 . 02 m sodium citrate buffer ) was placed into a quartz uv cuvette ( 1 . 0 cm path length ) and the uv spectrum recorded versus a reference cell containing 0 . 02 m sodium citrate buffer . uv spectra were recorded after each addition of a solution of n , n ′- ac 2 - lys - d - ala - d - ala or n , n ′- ac 2 - lys - d - ala - d - lac in 0 . 02 m sodium citrate buffer to each cell from 0 . 1 to 140 . 0 equivalents . the absorbance value at the λ max was recorded and the running change in absorbance , δa x equiv ( a initial − a x equiv , ) measured . the number of ligand equivalents was plotted versus δa to afford the ligand binding titration curve . the break point of this curve is the saturation point of the system and its xy coordinates determined by establishing the intersection of the linear fits of the pre and postsaturation curves . δa saturation was calculated and employed to determine the concentration of free ligand in solution at each titration . δa was plotted versus δa / free ligand concentration to give a scatchard plot from which the binding constants were determined . fig1 illustrates the factors that determine the binding affinity of vancomycin and its analogs to the model tripeptide and the rationale for the omission of the carbonyl oxygen of amino acid 4 . thus , the binding affinity of vancomycin for 3 , which incorporates a methylene ( ch 2 ) in place of the linking amide nh of ac 2 - l - lys - d - ala - d - ala , was compared with that of ac 2 - l - lys - d - ala - d - ala ( 2 ) and ac 2 - l - lys - d - ala - d - lac ( 4 ). the vancomycin affinity for 3 was approximately 10 - fold less than that of 2 , but 100 - fold greater than that of 4 . this indicated that the reduced binding affinity of 4 ( 4 . 1 kcal / mol ) may be attributed to both the loss of a key h - bond and a destabilizing lone pair / lone pair interaction introduced with the ester oxygen of 4 ( 2 . 6 kcal / mol ) with the latter , not the h - bond , being responsible for the greater share ( 100 - fold ) of the 1000 - fold binding reduction . these observations can be employed for the reengineering of vancomycin to bind d - ala - d - lac . it is disclosed herein that redesign of vancomycin focuses principally on removing the destabilizing lone pair interaction rather than reintroduction of a h - bond and that this may be sufficient to compensate for two of the three orders of magnitude in binding affinity lost with d - ala - d - lac . thus , synthesis of a vancomycin analogue with removal of the residue 4 carbonyl and its destabilizing lone pair interaction restores much of the binding affinity of the antibiotic for the modified ligand . at present , such a deep - seated change in the molecule can only be achieved by total synthesis , since previous efforts to selectively modify the residue 4 carbonyl by selective reaction of the amide linking residues 4 and 5 within vancomycin aglycon derivatives have not yet been successful . fig2 illustrates the retrosynthetic steps used to map out the synthesis of this vancomycin analog . the desired analogue 5 was anticipated to be prepared by a route analogous to that developed for vancomycin ( boger , d . l . ; et al . j . am . chem . soc . 1999 , 121 , 3226 ; boger , d . l . ; et al . j . am . chem . soc . 1999 , 121 , 10004 ), with notable modifications . thus , two aromatic nucleophilic substitution reactions with formation of the biaryl ethers were then enlisted for cd and de macrocyclization , a key macrolactamization reaction were employed for cyclization of the ab ring system , and the defined order of cd , ab , and de ring closures permitted sequential control of the atropisomer stereochemistry of each of the newly formed ring systems or their immediate precursors . thus , in addition to any kinetic diastereoselection that may be achieved in the ring closures , this order permitted the recycling of any undesired atropisomer for each newly introduced ring system by thermal equilibration providing a predictable control of the stereochemistry and dependably funneling all synthetic material into one of eight possible atropdiastereomers . key to recognition of this preferential order of ring closures was the establishment of the thermodynamic parameters of atropisomerism for the individual vancomycin ring systems : de ring system ( boger , d . l . ; et al . j . org . chem . 1997 , 62 , 4721 ; boger , d . l . ; et al . j . org . chem . 1996 , 61 , 3561 ; boger , d . l . ; et al . j . org . chem . 1999 , 64 , 70 ; boger , d . l . ; et al . bioorg . med . chem . lett . 1997 , 7 , 3199 ; boger , d . l . ; et al . bioorg . med . chem . lett . 1998 , 8 , 721 ; boger , d . l . ; et al . j . am . chem . soc . 1998 , 120 , 8920 ) ( e a = 18 . 7 kcal / mol )& lt ; ab biaryl precursor ( boger , d . l . ; et al . j . am . chem . soc . 1999 , 121 , 3226 ; boger , d . l . ; et al . j . am . chem . soc . 1999 , 121 , 10004 ) ( e a = 25 . 1 kcal / mol )& lt ; cd ring system ( boger , d . l . ; et al . j . org . chem . 1997 , 62 , 4721 ; boger , d . l . ; et al . j . org . chem . 1996 , 61 , 3561 ; boger , d . l . ; et al . j . org . chem . 1999 , 64 , 70 ; boger , d . l . ; et al . bioorg . med . chem . lett . 1997 , 7 , 3199 ; boger , d . l . ; et al . bioorg . med . chem . lett . 1998 , 8 , 721 ; boger , d . l . ; et al . j . am . chem . soc . 1998 , 120 , 8920 ) ( e a = 30 . 4 kcal / mol ). fig3 is a scheme showing the synthesis of the bcd “ tripeptide .” the b and d subunits 6 and 7 were prepared following previously optimized procedures ( see main text for references ). oxidation of alcohol 7 ( compound 7 is available in 6 steps ( 37 % overall ) from methyl gallate using 3 recrystallizations and was scaled to 300 g , ( crowley , b . m . ; et al . j . am . chem . soc . 2004 , 126 , 4310 )) ( 2 . 0 equiv of dess - martin periodinane , ch 2 cl 2 , 0 - 25 ° c ., 1 h , 100 %) was followed by immediate reductive amination coupling of the sensitive aldehyde 8 with 6 ( compound 6 is available in 5 steps ( 55 % overall ) from ( r )- 4 - hydroxyphenyl - glycine using 2 recrystallizations and was scaled to 60 g , ( boger , d . l . ; et al . j . org . chem . 1997 , 62 , 4721 )) ( 1 . 1 equiv , ch 3 oh , 3 å ms , 0 ° c ., 45 min ; 3 . 0 equiv of acoh , 3 . 0 equiv of nabh 3 cn , − 20 ° c ., 2 d ) to afford amine 9 in good yield ( 75 %) and excellent diastereoselectivity ( 12 : 1 ). shorter reaction times ( 14 - 20 h ) at higher temperatures (− 15 to − 5 ° c .) led to substandard selectivities ( 4 : 1 to 9 : 1 ) and the use of less nabh 3 cn ( 1 . 5 - 2 . 0 equiv ) at lower temperatures (− 20 ° c .) led to incomplete reactions . longer reaction times ( 3 - 8 d ) led to only marginal increases in yield ( 82 % after 8 d ) and roughly equal diastereoselectivities . amine protection of 9 as the methyl carbamate ( 10 equiv of meococl , 10 equiv of k 2 co 3 , thf , 0 - 25 ° c ., 18 h , 85 %) followed by benzyl ether deprotection ( benzyl ether deprotection at higher temperatures ( 25 ° c .) may lead to competitive aryl bromide reduction although this was only observed in appreciable amounts when excess raney ni was employed .) ( raney ni , ch 3 oh , 0 ° c ., 5 h , 98 %) and saponification ( 3 . 0 equiv of lioh , thf — h 2 o , 0 ° c ., 6 h , 100 %) provided 12 . unexpectedly , the order of these latter two deprotections proved important . saponification of 10 ( saponification of 11 was considerably slower than that of 10 and occasionally required additional lioh for complete conversion to 12 with little effect on the amount of epimer generated in the reaction .) under a variety of conditions ( lioh , thf — h 2 o or t - buoh — h 2 o , − 10 to 0 ° c . ; liooh , thf — h 2 o ; me 3 snoh , 1 , 2 - dichloroethane , 70 ° c .) led to variable amounts of an epimer ( 5 - 20 %) that was difficult to separate from the product . in contrast , benzyl ether deprotection of 10 followed by saponification of 11 reduced the amount of epimer ( 0 - 3 %) presumably due to preferential deprotonation of the phenols such that subsequent c α deprotonation at residue 5 was less facile ( saponification of 11 was considerably slower than that of 10 and occasionally required additional lioh for complete conversion to 12 with little effect on the amount of epimer generated in the reaction .). coupling of 12 with 13 ( compound 13 is available in 3 steps ( 45 % overall ) from 4 - fluoro - 3 - nitrobenzaldehyde and was scaled to 30 g , ( crowley , b . m . ; et al . j . am . chem . soc . 2004 , 126 , 4310 ).) ( 3 . 0 equiv of depbt ( fan , c .- x . ; et al . org . lett . 1999 , 1 , 91 ), 3 . 0 equiv of nahco 3 , dmf , 0 - 25 ° c ., 8 h ) gave “ tripeptide ” 14 in good yield ( 70 %) and excellent diastereoselectivity ( 14 : 1 ). fig4 is a scheme for the synthesis of the abcd ring system starting from n - boc amino ester diamide 14 . after considerable optimization ( fig5 a and 5b ), cyclization of 14 ( 20 equiv of k 2 co 3 , 20 equiv of caco 3 , 3 wt equiv of 3 å ms , 12 mm thf , 75 ° c . bath temp , 12 h ) afforded 15 in good yield ( 54 %) and good atropodiastereoselectivity ( 2 . 5 : 1 , 15 ( 54 %) and 16 ( 22 %)) even when conducted on a large scale ( 2 . 7 g ). reduction of the nitro group ( raney ni , 0 ° c ., ch 3 oh , 1 h ) followed by diazotization ( 1 . 3 equiv of hbf 4 , 1 . 3 equiv of t - buono , ch 3 cn , 0 ° c ., 30 min ) and sandmeyer substitution ( 50 equiv of cucl , 60 equiv of cucl 2 , h 2 o , 0 - 25 ° c ., 1 h , 70 % from 15 ) cleanly provided 17 without loss of the atropisomer stereochemistry inherent in starting 15 . the unnatural atropisomer 16 was also subjected to these conditions to cleanly give 18 ( 75 %) ( fig6 ). suzuki coupling of 17 with the hindered a ring boronic acid 20 ( boger , d . l . ; et al . j . am . chem . soc . 1999 , 121 , 3226 ; boger , d . l . ; et al . j . am . chem . soc . 1999 , 121 , 10004 ) ( 0 . 3 equiv of pd 2 ( dba ) 3 , 1 . 5 equiv of ( o - tol ) 3 p , toluene - ch 3 oh - 1 n aq na 2 co 3 10 : 3 : 1 , 80 ° c ., 30 min ) proceeded in excellent yield ( 90 %) under remarkably effective conditions ( boger , d . l . ; et al . j . am . chem . soc . 1999 , 121 , 3226 ; boger , d . l . ; et al . j . am . chem . soc . 1999 , 121 , 10004 ) given the steric constraints of the substrate 20 providing a separable 1 : 1 . 3 mixture of atropisomers ( 21 : 22 ) slightly favoring the unnatural configuration . thermal equilibration of isolated 22 was carried out initially employing the reported conditions for vancomycin ( o - dichlorobenzene , 120 ° c ., 18 h , 81 % recovery of material ) ( boger , d . l . ; et al . j . org . chem . 1997 , 62 , 4721 ; boger , d . l . ; et al . j . org . chem . 1996 , 61 , 3561 ; boger , d . l . ; et al . j . org . chem . 1999 , 64 , 70 ; boger , d . l . ; et al . bioorg . med . chem . lett . 1997 , 7 , 3199 ; boger , d . l . ; et al . bioorg . med . chem . lett . 1998 , 8 , 721 ; boger , d . l . ; et al . j . am . chem . soc . 1998 , 120 , 8920 ) to afford a 1 : 1 . 1 separable mixture permitting the recycling of this unnatural atropisomer . silyl ether deprotection of 21 ( 1 . 2 equiv of bu 4 nf , thf , 0 ° c ., 10 min ) followed by n - cbz removal ( h 2 , 10 % pd / c , 1 % cl 3 cco 2 h — ch 3 oh , 15 min , 95 %) and methyl ester hydrolysis ( 1 . 0 equiv of lioh , thf — h 2 o , 0 ° c ., 1 . h , 96 %) gave amino acid 25 . macrolactamization with closure of the ab ring system was effected by treatment of 25 with pybop ( 3 . 0 equiv , 6 . 0 equiv of nahco 3 , 0 . 001 m ch 2 cl 2 - dmf 5 : 1 , 0 - 25 ° c ., 12 h ) to afford the fully functionalized bicyclic abcd ring system 26 in good yield ( 70 %) with only trace amounts of competitive epimerization (& lt ; 3 %). fig5 a is a table summarizing the conditions tested for the cyclization of 14 to 15 . the inclusion of caco 3 in the reaction mixture is critical and serves to trap the liberated fluoride arising from the aromatic nucleophilic substitution as an insoluble byproduct ( caf 2 ) preventing tbs ether deprotection and a subsequent competitive base - catalyzed retro aldol reaction of the free alcohol . the cyclization of 14 represents a considerable improvement over the analogous ring closure reaction enlisted in this inventor &# 39 ; s original synthesis of vancomycin ( 50 - 65 %, 1 : 1 atropisomers vs 76 - 87 %, 2 . 5 - 3 : 1 atropisomers ) where both the overall conversion and atropodiastereoselectivity were lower illustrating that the closure of 14 may benefit from both the increased conformational flexibility of the cyclization substrate and the residue 4 amine small protecting group . fig5 b is a table summarizing the conditions used for the cyclization of 14 to 15 after conditions in fig5 a were tried . fig6 is a short scheme showing the steps taken to attempt to recycle the undesired atropdiastereomers 15 and 17 by heating in solvent and how they were identified as atropisomers of 16 and 18 , respectively . these two compounds were shown to be atropdiastereomers of 16 and 18 , respectively , by conversion of 17 to 19 . the identity of compound 19 was confirmed by conversion from 18 and 17 by dechlorination / debromination . unlike the vancomycin cd ring system in which the atropisomers could be thermally equilibrated at 120 - 140 ° c . permitting the recycling and productive use of the unnatural atropisomer , the atropisomers 15 and 16 could not be thermally interconverted even at temperatures as high as 210 - 230 ° c . the corresponding chloro compounds 17 and 18 were not able to be interconverted either . fig7 shows the synthesis of the complete carbon skeleton of the vancomycin aglycon analog . coupling of 27 and 28 ( 2 . 0 equiv of depbt ( fan , c .- x . ; et al . org . lett . 1999 , 1 , 91 ), 2 . 2 equiv of nahco 3 , thf , 0 - 25 ° c ., 14 h , 73 %) afforded 29 with excellent diastereoselectivity ( 12 : 1 ) arising from little competitive racemization . closure of the de ring system with formation of the key biaryl ether was accomplished by treatment of 29 with csf ( 10 equiv , 20 equiv of caco 3 ( both the added 3 å ms and caco 3 result in cleaner conversions to product . it is not yet clear whether the soluble base under these conditions is csf or cs 2 co 3 with precipitation of insoluble caf 2 . ), 3 å ms , dmf , 25 ° c ., 17 h ) to afford 30 in good yield ( 74 %) and good atropodiastereoselectivity ( 6 - 7 : 1 ). thus , consistent with the adoption of a vancomycin - like conformation by 26 , the amide modification in the abcd ring system of 29 had little impact on the ease or diastereoselectivity of the de ring closure . reduction of the nitro group ( reduction of the nitro group was very sensitive to the choice of solvent in terms of recovery and observance of side products .) ( h 2 , 10 % pd / c , thf , 8 h , 94 %) followed by diazotization of the resulting amine 32 ( 1 . 2 equiv of hbf 4 , 1 . 2 equiv of t - buono , ch 3 cn , 0 ° c ., 20 min ) and sandmeyer substitution ( 50 equiv of cucl , 60 equiv of cucl 2 , h 2 o , 0 - 25 ° c ., 1 h , 55 %) gave 33 , which embodies the full carbon skeleton of 5 . tbs ether protection of the secondary alcohols ( 65 equiv of cf 3 conmetbs , ch 3 cn , 55 ° c ., 22 h ; aq citric acid , 25 ° c ., 13 h , 96 %) followed by mem ether deprotection of 34 ( 12 equiv of b - bromocatecholborane ( bcb ), ch 2 cl 2 , 0 ° c ., 2 h ; 5 . 1 equiv of boc 2 o , 6 . 0 equiv of nahco 3 , dioxane - h 2 o 2 : 1 , 0 - 25 ° c ., 2 . 5 h , 80 %) and two - step oxidation of the resulting primary alcohol 35 ( 4 . 0 equiv of dess - martin periodinane , ch 2 cl 2 , 0 ° c ., 15 min then 25 ° c ., 1 h ; 9 . 0 equiv of 80 % aq naclo 2 , 7 . 0 equiv of nah 2 po 4 . h 2 o , t - buoh / 2 - methyl - 2 - butene 4 : 1 , 25 ° c ., 20 min , 82 %) provided the carboxylic acid 36 . hydrolysis of the residue 3 nitrile with formation of the carboxamide 37 ( 40 equiv of 30 % aq h 2 o 2 , 8 . 0 equiv of 10 % aq k 2 co 3 , dmso , 25 ° c ., 3 . 5 h , 87 %) ( boger , d . l . ; et al . j . am . chem . soc . 1999 , 121 , 3226 ; boger , d . l . ; et al . j . am . chem . soc . 1999 , 121 , 10004 ) set the stage for a final global deprotection ( node , m . ; et al . j . org . chem . 1980 , 45 , 4275 ; evans , d . a . ; ellman , j . a . j . am . chem . soc . 1989 , 111 , 1063 ). in a final key reaction , 37 was treated with albr 3 ( 35 equiv , etsh , 25 ° c ., 5 h , 80 %) to afford 5 arising from the remarkable deprotection of four aryl methyl ethers , the two tbs ethers , the n - terminus boc group , and the critical residue 4 methyl carbamate . fig8 is a table that shows the conditions used for the cyclization of 29 to form 30 by catalyzing with a fluoride ion in the presence of added base . closure of the de ring system with formation of the key biaryl ether was accomplished by treatment of 29 with csf ( 10 equiv , 20 equiv of caco 3 ( both the added 3 å ms and caco 3 result in cleaner conversions to product . it is not yet clear whether the soluble base under these conditions is csf or cs 2 co 3 with precipitation of insoluble caf 2 . ), 3 å ms , dmf , 25 ° c ., 17 h ) to afford 30 in good yield ( 74 %) and good atropodiastereo - selectivity ( 6 - 7 : 1 ). fig9 is a drawing showing the different modifications in the vancomycin structure of analogs that are possible and what relative affinity they might have for either the d - ala - d - ala ligand or the d - ala - d - lac ligand . the targeted analogue 5 incorporating an amine in the linkage of residue 4 with residue 5 not only removes the offending carbonyl and the destabilizing lone pair interaction with d - ala - d - lac , but it maintains a local polar environment ( protonated amine ) that may better accommodate the binding of an electronegative group or atom ( nh of d - ala - d - ala amide or o of d - ala - d - lac ester ). while this might not bind d - ala - d - lac quite as well as derivatives such as 40 , it was better than 40 at binding d - ala - d - ala . in the best case , 5 might bind d - ala - d - ala and d - ala - d - lac with equal affinities making it effective for the treatment of sensitive or resistant bacteria regardless of the structure of the peptidoglycan cell wall precursor . fig1 is an n - boc deprotection of 33 to give 41 without deprotecting the methyl carbamate of residue 4 and removing the mem group . compound 41 was synthesized to test its binding affinity in comparison with vancomycin , 5 and 38 . fig1 is a table showing the results of the assessment of 5 alongside vancomycin ( 1 ) and its aglycon 38 and structure 41 . an additional analogue 41 , derived from n - boc deprotection of the synthetic intermediate 33 ( fig1 ), was also examined that bears the methoxycarbonyl protecting group on the residue 4 / 5 linking amine . the binding affinity of 5 for ac 2 - l - lys - d - ala - d - ala ( 2 ) and ac 2 - l - lys - d - ala - d - lac ( 4 ) was essentially equivalent ( 4 . 8 vs 5 . 2 × 10 3 m − 1 , respectively ) with the d - ala - d - lac binding being slightly better . impressively , this represented the anticipated results relative to the vancomycin aglycon where the enhancement for binding d - ala - d - lac is 43 - fold ( 5 . 2 × 10 3 vs 1 . 2 × 10 2 m − 1 ) and the reduction in binding affinity for d - ala - d - ala is 37 - fold ( 4 . 8 × 10 3 vs 1 . 7 × 10 5 m − 1 ). fig1 shows the structure of the vancomycin analog and its binding constant with the two model ligands . fig1 is a skatchard analysis of compound 5 with the n , n ′- ac 2 - lys - d - ala - d - ala ligand . the binding constants for compounds 5 and 41 for association with the model ligands n , n ′- ac 2 - lys - d - ala - d - ala and n , n ′- ac 2 - lys - d - ala - d - lac were determined according to literature ( nieto , m . ; perkins , h . r . biochem . j . 1971 , 124 , 845 ; nieto , m . ; perkins , h . r . biochem . j . 1971 , 124 , 773 ; nieto , m . ; perkins , h . r . biochem . j . 1971 , 124 , 789 ) protocols . fig1 is a skatchard analysis of compound 5 with the n , n ′- ac 2 - lys - d - ala - d - lac ligand . fig1 is a titration curve of 5 and the n , n ′- ac 2 - lys - d - ala - d - ala ligand . fig1 is a titration curve of 5 and the n , n ′- ac 2 - lys - d - ala - d - lac ligand . fig1 illustrates important modifications to the basic vancomycin analog structure . most of the modifications are in the peripheral portion of the molecule as the backbone of the vancomycin structure has been preserved with the exception of the carbonyl oxygen of the fourth amino acid . this carbonyl has been replaced by a methylene group eliminating an energetically unfavorable interaction with the lone pairs of the ester oxygen of the d - lac .
pg 4 ] glycopeptide antibiotic analogs are reengineered forms of glycopeptides that exhibit antimicrobial activity against both wild type and glycopeptide antibiotic resistant strains of microorganisms . for example , tpg 4 ] vancomycin aglycon is a reengineered form of vancomycin that exhibits antimicrobial activity against both wild type and vana resistant organism . the vana resistant organism achieves its resistance , upon glycopeptide antibiotic challenge , by remodeling its d - ala - d - ala peptidoglycan cell wall precursor to d - ala - d - lac . pg 4 ] glycopeptide antibiotic analogs have an altered glycopeptide backbone wherein the carbonyl of the fourth amino acid residue of the glycopeptide backbone has been replaced with a methylene . this alteration of the glycopeptide backbone imparts dual binding affinities for both d - ala - d - ala and d - ala - d - lac and dual antimicrobial activities for both wild type and resistant strains . for example , tpg 4 ] vancomycin aglycon displays a antimicrobial potency that reflects its altered binding characteristics .