mu - opioid receptor ( mor ) agonists , such 
 as morphine , are the gold standard in the treatment of moderate to 
 severe pain , however , their therapeutic use is often limited due to 
 serious side effects including tolerance , dependence , respiratory 
 depression , and constipation .
numerous studies have shown promise 
 for ameliorating mor - related side effects through simultaneous action 
 at -opioid receptors ( dor ) .
dor agonists have been shown to 
 reduce the development of mor agonist tolerance and dependence , presumably due to the ability of dor agonists to potentiate mor 
 mediated analgesia .
dor agonists have 
 also been shown to block -opioid receptor ( kor ) mediated antinociception 
 which has possible implications in reversing kor related addictive 
 behaviors .
strong evidence has also accumulated 
 for the potential of dor antagonists as agents to mitigate mor - mediated 
 side effects .
recently , there have been multiple reports on the utility 
 and application of mixed - efficacy compounds that simultaneously produce 
 mor activation while acting as dor antagonists .
while the mechanism remains unclear , this profile has been shown 
 to elicit the desired analgesia of a mor agonist but with reduced 
 risk of tolerance and dependence in a number of preclinical models 
 of analgesia .
we have previously described a series of peptidomimetics 
 in which key opioid pharmacophore elements were transferred from a 
 peptide scaffold to a tetrahydroquinoline ( thq ) core , a smaller , more 
 drug - like scaffold . expanding upon this initial report
, we modified 
 the thq core in an effort to increase metabolic stability in two ways : 
 through n - acetylation of the thq nitrogen and through 
 replacement the thq core with a tetrahydronaphthalene ( thn ) core . through these modifications
, we found that n - acetylation of the thq core of the mixed - efficacy mor 
 agonist / dor antagonist peptidomimetics improves dor affinity , which 
 results in an overall better balance of affinities at mor and dor 
 and provides selectivity over the -opioid receptor ( kor ) .
although the optimal balance of mor affinity 
 and dor affinity is unknown , we reasoned that similar high affinity 
 profiles at both mor and dor was a reasonable goal , as this would 
 ensure that both mor and dor character would be similarly represented . 
 in the same report
, we proposed that an additional polar contact between 
 the carbonyl of the n - acetyl moiety and tyr of dor in transmembrane 3 ( tm3 ) helix is responsible for this increase 
 in dor affinity .
analogues in the present 
 structure  activity relationship ( sar ) campaign were therefore 
 designed ( 1 ) to include a carbonyl moiety in order to maintain the 
 high dor affinity seen in the initial analogue series and ( 2 ) to incorporate 
 various aliphatic , cyclic , aromatic , and heteroatom - containing functionalities 
 ( figure 1 ) to probe 
 the effect of such modifications on the binding affinity and efficacy 
 profiles not only at dor but also at mor and kor .
peptidomimetic scaffold 
 and n - substitutions ; * previously published in ref ( 8) , * * previously published 
 in ref ( 9 ) .
analogues 4a and 4b have been 
 previously reported and are included here to serve 
 as standards of comparison for the additional benzyl pendant n - substituted series .
the majority of the remaining peptidomimetics 
 ( 4c4j ) reported herein were prepared 
 in four steps starting from the dihydroquinolinone 1 , 
 which was prepared as previously reported ( scheme 1 ) .
compound 1 was subjected to acylation using either an acid anhydride 
 or acyl chloride to form intermediates 2c32j .
it should be noted that the low yield for the synthesis 
 of 2d was due to the use of pyridine , which was originally 
 included to help catalyze the reaction .
however , we found that running 
 the reactions in neat anhydride at temperatures between 90 and 120 
 c led to better product conversion , and for the remainder of 
 syntheses , we excluded pyridine in this reaction step .
next , intermediates 2c2j were treated with ( r)-t - butanesulfinamide and ti(oet)4 to 
 yield imines in situ , which were reduced with nabh4 to 
 form the desired r - stereochemistry of intermediates 3c3j .
the ellman auxiliary 
 was cleaved using concentrated hydrochloric acid , forming hydrochloride 
 salts , which were then coupled to di - boc 
 protected 2,6-dimethyl - l - tyrosine ( di - boc - dmt ) 
 and subsequently deprotected using trifluoroacetic acid ( tfa ) to yield 
 analogues 4c4j .
as we have previously 
 shown , the ellman chemistry employed here produces the desired ( r)-stereocenter with high enantiomeric excess , as evidenced in the current syntheses by the 
 absence of nmr - detectable disateromer after coupling to diboc - dmt .
( a ) 
 neat acid anhydride ( excess ) , 100 c , 24 h ( for compunds 2c , 2d ) ; ( b ) acid chloride ( eq ) , dcm , ( for 2e  j ) ; ( c ) ( r)-t - butanesulfinamide ( 23 equiv ) , thf , ti(oet)4 ( 46 equiv ) , 0 c , then reflux at 75 c ; 
 ( d ) nabh4 ( 6 equiv ) , thf , 50 c to rt , then 
 meoh , rt ; ( e ) hcl ( 6 equiv ) , dioxane , rt ; ( f ) diboc - dmt ( 1.05 equiv ) , 
 pybop ( 1 equiv ) , 6cl - hobt ( 1 equiv ) , dipea ( 10 equiv ) , dmf , rt ; ( g ) 
 1:1 tfa : dcm ( excess ) .
note : not all crude final product was purified ; 
 as such a yield was not calculated .
the synthesis of the remaining analogues 4k4 m began by protecting the nitrogen 
 in 1 with a tert - butyloxycarbonyl ( boc ) 
 group forming 5 ( scheme 2 ) , as previously described .
intermediate 5 was treated with ( r)-t - butanesulfinamide and ti(oet)4 to 
 yield an imine in situ , which was reduced with nabh4 to 
 give the desired r - stereochemistry of intermediate 6 . to ensure both the cleavage of the ellman auxiliary 
 and the removal of the boc group ,
6 was treated first 
 with tfa then with conc hcl in dioxane to ultimately yield the hydrochloride 
 salt 7 .
intermediate 7 was coupled to diboc - dmt , 
 forming 8 . from this intermediate , 8 , the 
 syntheses of 4k , 4l , and 4 m diverged .
the synthesis of 4k was completed by first 
 performing a finkelstein reaction on 2-chloroacetamide to form 2-iodoacetamide , 
 which was cannulated into a solution containing 8 and n , n - diisopropylethylamine ( dipea ) to give 9k .
intermediate 9k was treated with tfa to remove 
 the boc groups , yielding 4k . to complete the syntheses 
 of 4l and 4 m
, ethyl 2-bromoacetate was added 
 to intermediate 8 in the presence of potassium carbonate 
 ( k2co3 ) to give 9l .
intermediate 9l was then treated with tfa to remove boc groups and afford 
 compound 4l .
compound 4 m was completed by 
 first saponification of the ester of 9l with lithium 
 hydroxide , which was subsequently treated with tfa to remove the boc 
 groups and yield compound 4 m .
( a ) boc2o ( 1.22 
 equiv ) , dmap ( 0.1 equiv ) , dipea ( 1.22 equiv ) , dcm , 60 c ; 
 ( b ) ( r)-t - butanesulfinamide ( 23 
 equiv ) , thf , ti(oet)4 ( 46 equiv ) , 0 c , then 
 reflux at 75 c ; ( c)nabh4 ( 6 equiv ) , thf , 50 
 c to rt , then meoh , rt ; ( d ) 1:1 tfa : dcm ( excess ) ; ( e ) hcl ( 6 
 equiv ) , dioxane , rt ; ( f ) diboc - dmt ( 1.05 equiv ) , pybop ( 1 equiv ) , 
 6cl - hobt ( 1 equiv ) , dipea ( 10 equiv ) , dmf , rt ; ( g ) nal ( 2 equiv ) , 
 2-chloroacetamide ( 2 equiv ) , then dipea ( 1.5 equiv ) ; ( h ) ethyl 2-bromoacetate 
 ( 10 equiv ) , k2co3 ( 2 equiv ) , ch3cn , 
 24 h ; ( i ) lioh ( excess ) , etoh , 60 c , 2.5 h ; ( j ) 3 n hcl ( excess ) , 
 rt .
therefore , 
 only 1.6 mg of 95% pure product was isolated , which was sufficient 
 for opioid receptor binding and efficacy assays .
analogues 15a and 15b have been previously reported and are included here to serve as standards of comparison for the 
 additional 2-methylnaphthyl pendant n - substituted 
 series .
synthesis of the n - acylated peptidomimetics 
 containing the 2-methylnaphthyl pendant began by forming intermediate 10 , which was synthesized as previously described .
intermediate 10 was subjected to 
 suzuki cross - coupling to incorporate the 2-methylnaphthyl moiety and 
 yield 11 .
intermediate 11 was treated with 
 tfa to remove the boc group , forming 12 .
intermediate 12 was treated with propionic anhydride to yield 13c , butyric anhydride to yield 13d , or methylchloroformate 
 to yield 13e .
intermediates 13c13e were treated with ( r)-t - butanesulfinamide and ti(oet)4 to form a chiral imine 
 in situ , which was reduced with nabh4 to form the desired r - stereochemistry for intermediate 14c14e .
the ellman auxiliary was cleaved using concentrated 
 hcl forming primary amines , which were then coupled to diboc - dmt and 
 deprotected to yield compound 15c15e ( scheme 3 ) .
( f ) 
 2-naphthalenylboronic acid , pd(dppf)cl2 , k2co3 , 3:1 acetone : h2o ; ( g ) 1:1 tfa : dcm ; ( h ) neat acid 
 anhydride , reflux , 24 h ( for 15c and 15d ) ; ( i ) mecoocl , dcm , rt , 16 h ( for 15e ) ; ( j ) ( r)-t - butanesulfinamide , thf , ti(oet)4 , 0 c , then reflux at 75 c ; ( k ) nabh4 , thf ,  50 c to rt , 3 h , then meoh , rt ; ( l ) hcl , dioxane , 
 rt , 3 h ; ( m ) diboc - dmt , pybop , 6cl - hobt , dipea , dmf , rt . * not all 
 of the crude compound was purified , so yield was not calculated or 
 appears low .
analogues 4c4 m and 15c15e were evaluated in in 
 vitro mor , dor , and kor binding and efficacy assays , as previously 
 described ( tables 1 and 2 ) .
binding affinities ( ki ( nm ) ) were obtained by competitive displacement 
 of radiolabeled [ h]diprenorphine in membrane preparations 
 from c6 cells stably expressing mor ( c6-mor ) or dor ( c6-dor ) or cho 
 cells stably expressing kor ( cho - kor ) , as previously described ( table 1 ) .
efficacy 
 data were obtained using agonist - induced stimulation of [ s]gtps binding to g protein in 
 the same cell preparations as previously mentioned .
efficacy data 
 are represented as percent maximal stimulation relative to standard 
 agonist ( damgo at mor , dpdpe at dor , and u69593 at kor ) , and potency 
 ( ec50 ( nm ) ) values are calculated as the concentration 
 of drug required to produce half - maximal stimulation ( table 1 ) .
briefly , all analogues in 
 both the benzyl and 2-methylnaphthyl series maintained high affinity 
 binding and efficacy at mor
. additionally , as predicted , all analogues 
 across both series maintained high binding affinity at dor , relative 
 to the unsubstituted analogues 4a and 15a , excluding 4 m , which displayed reduced affinity across 
 all three receptors .
interestingly , most of the n - substituted analogues in the benzyl pendant series produced increased 
 dor efficacy ( table 1 ) , creating varying degrees of mor agonist / dor agonist ligands in 
 the in vitro assays , while all analogues in the n - substituted 2-methylnaphthyl series displayed minimal or no efficacy 
 at dor , maintaining the mor agonist / dor antagonist profile ( table 2 ) .
lastly , while all 
 analogues maintained selectivity for mor and dor over kor , kor affinity 
 for many of the n - substituted analogues in the benzyl 
 series increased relative to 4a ( table 1 ) .
binding 
 affinities ( ki ( nm ) ) were obtained by 
 competitive displacement of radiolabeled [ h]diprenorphine 
 in membrane preparations .
efficacy is represented as percent 
 maximal stimulation relative to standard agonist damgo ( mor ) , dpdpe 
 ( dor ) , or u69,593 ( kor ) at 10 m .
potency ( ec50 ( nm ) ) 
 is calculated from the percent maximal stimulation as the concentration 
 of drug required to produce half the maximal stimulation all values are expressed as the mean 
 with sem in parentheses for n = 3 independent assays 
 in duplicate , unless otherwise noted .
dns : does not stimulate . published in ref ( 8) . published in ref ( 9 ) .
binding affinities 
 ( ki ( nm ) ) were obtained by competitive 
 displacement of radiolabeled [ h]diprenorphine in membrane 
 preparations .
efficacy is represented as percent maximal stimulation relative 
 to standard agonist damgo ( mor ) , dpdpe ( dor ) , or u69,593 ( kor ) at 
 10 m .
potency ( ec50 ( nm ) ) is calculated from the 
 percent maximal stimulation as the concentration of drug required 
 to produce half the maximal stimulation all values are expressed as the mean with sem in parentheses 
 for n = 3 independent assays in duplicate , unless 
 otherwise noted .
dns : does not stimulate . published in ref ( 8) . published in ref ( 9 ) .
although the benzyl pendant n - substituted series displayed varying degrees of stimulation 
 at dor , all analogues still exhibited both high affinity and efficacy 
 at mor ; consequently , we elected to test select analogues ( 4e4k ) for in vivo efficacy in the wwtw assay using 
 a cumulative dosing procedure .
additionally , because all new compounds 
 in the 2-methylnaphthyl pendant series displayed the desired mor agonist / dor 
 antagonist profile , compounds 15c15e were also tested for in vivo efficacy in the same assay . briefly
, 
 withdrawal latencies were determined by placing a mouse into a cylindrical 
 plastic restrainer and immersing 23 cm of the tail tip into 
 a water bath maintained at 50 c .
the latency to withdrawal or 
 to rapidly flick the tail back and forth was recorded with a maximum 
 cutoff time of 20 s to prevent tissue damage .
acute antinociceptive 
 effects were determined using a cumulative dosing procedure , as previously 
 described .
each animal received an intraperitoneal 
 ( ip ) injection of saline , and then 30 min later , the baseline withdrawal 
 latencies were recorded . following baseline determinations , three 
 increasing doses ( 1 , 2.2 , and 6.8 mg / kg ) of a peptidomimetic ( dissolved 
 in either deionized water or 1:1:8 solution of ethanol : alkumuls : deionized 
 water , as was the case for 15c15e ) were given ip at 30 min intervals to provide final doses of 1 , 
 3.2 , and 10 mg / kg .
thirty minutes after each injection , the tail withdrawal 
 latency was measured as described above . to determine the duration 
 of antinociceptive action
, the tail - withdrawal test was performed 
 at varying times following administration of a peptidomimetic ( 10 
 mg / kg , ip ) . of all compounds tested in vivo , only two of the 
 novel n - substituted analogues tested displayed in 
 vivo efficacy after ip administration ( figure 2a ) .
analogue 4f produced partial 
 antinociception with an 11 s latency to tail flick at the 10 mg / kg 
 dosage following ip administration , while 4h produced 
 full antinociception , reaching the 20 s latency to tail flick cutoff 
 time point at the same dosage . because 4h displayed full 
 antinociception , we performed a time course assay to determine the 
 duration of antinociceptive action .
as can be seen in figure 2b , 4h has a similar 
 duration of action to that of 4a , producing maximal antinociception 
 for approximately 1 h following a 10 mg / kg ip dose .
( a ) cumulative antinociceptive 
 dose response curves of select benzyl pendant analogues ( n = 3 for all analogues ) in the mouse wwtw assay following ip administration . 
 plotted as average  sem . * * * * , p < 0.0001 
 for 4a and 4h for the 10 mg / kg dose when 
 compared to baseline , p
( b ) time course 
 of antinociception of 4a and 4h ( n = 3 ) in the mouse wwtw assay following ip administration 
 of 10 mg / kg . plotted as average  sem .
all new analogues across both series sets , excluding 4 m , maintained high binding affinity at mor and exhibited an increased 
 dor binding affinity when compared to the unsubstituted parent compounds 4a and 15a , presumably due to the carbonyl moiety 
 incorporated into each of the n - substitutions , as 
 had been previously observed for 15b .
the only exception to this trend , 4 m , contains 
 a carboxylic acid moiety and displayed significantly decreased affinity 
 across all three opioid receptors .
the poor binding affinity of 4 m could be potentially due to electrostatic repulsion between 
 its carboxylic acid moiety and a conserved negatively charged residue 
 in tm5 ( asp in dor , asp in kor , and glu in mor ) , which may also be complemented by acidic glu in kor . on the basis of these initial in vitro results 
 and previously reported computational modeling , it appears that a carbonyl - containing moiety is beneficial 
 in maintaining higher binding affinity at dor while a negatively charged 
 species is not well tolerated .
an additional trend seen in the 
 benzyl series is that increasing the n - acyl chain 
 length or overall bulk increases dor stimulation , with the exception 
 of 4j , which contains a benzoyl moiety and produces no 
 stimulation at dor . as seen in table 1 , the unsubstituted compound 4a shows 
 very weak stimulation at dor ( 16% maximal stimulation ) while the n - acetyl analogue 4b increased maximal stimulation 
 at dor to 26% and the n - propionyl ( 4c ) and n - butyryl ( 4d ) analogues further 
 increased dor stimulation to between 40 and 60% .
analogues 4e4 g contain branched or cyclic n - substitutions , and in the case of 4f ( containing a 
 cyclopropanecarbonyl moiety ) and 4 g ( containing a cyclobutanecarbonyl 
 moiety ) , dor stimulation increased to 69% and 58% , respectively . the computational model of 4 g in the active conformation 
 of dor ( figure 3a ) 
 demonstrates the presence of a hydrophobic cavity in the ligand binding 
 pocket formed by residues from extracellular loop 2 ( leu and phe ) and tm5 ( thr and val ) , which is preserved in mor and kor .
it can be seen that the cyclobutanecarbonyl 
 moiety extends into this cavity forming favorable hydrophobic interactions . 

these interactions may contribute to the improved binding affinity 
 of compounds 4c4f toward the active 
 receptor conformation and to their increased efficacy at dor .
as also 
 seen in figure 3a , 
 compound 4j has a benzoyl moiety which appears to be 
 too bulky to fit this cavity in the active receptor conformation although 
 it can be easily accommodated in the slightly enlarged ligand pocket 
 of the inactive receptor state .
( a ) structure 
 of 4 g ( teal ) and 4j ( magenta ) bound in the 
 active conformation of dor with the area of the binding pocket occupied 
 by the acyl chain circled ( black ) . ( b ) structure of 4k ( green ) bound in the active conformation of kor with a potential 
 polar contact between e and the terminal amide of the 
 ligand which could be responsible for the increased kor affinity and 
 efficacy .
similarly , hydrophobic interactions 
 with the nonpolar cavity may also contribute to the improved binding 
 affinity toward kor of compounds 4c , 4f , 4 g , and 4j with long or bulky nonpolar
n - substitutions ( ki values ranging 
 from 8 to 56 nm , as compared to ki = 68 
 nm of 4a ) , which act as partial agonists .
however , this 
 can not explain the observed activity of 4k , which has 
 a polar amide group as an n - substitution .
compound 4k not only displays increased affinity at kor ( 4.3 vs 68 
 nm for 4a ) but also exhibits significant kor stimulation 
 ( 49% ) .
the docking of 4k into the model of the active 
 conformation of kor ( figure 3b ) suggests that formation of a hydrogen bond between n - substituted amide of 4k and glu from tm6 of kor may be the reason for the improved binding and agonist 
 properties of this compound . while analogues 4c and 4e  g all represent a subset 
 of analogues with a mor agonist / dor agonist in vitro profile , 4 g displayed balanced binding affinities ( ki = 0.23 nm at mor and 0.15 nm at dor ) , low nanomolar 
 potency ( ec50 = 2.1 nm at mor and 5.1 nm at dor ) , and high 
 efficacy at both mor and dor ( 94% and 58% , respectively ) , along with 
 weak binding and no efficacy at kor .
while this profile was not our 
 initial focus , several reports have shown that coadministration of 
 a dor agonist with a mor agonist can increase both the potency and 
 efficacy of the mor agonist .
these findings suggest 
 ligands producing a mixed - efficacy mor agonist / dor agonist profile 
 could allow for the effective management of pain with a decrease in 
 side effects seen with the administration of only a mor agonist .
indeed , 
 this has been observed with mmp-2200 , a mor agonist / dor agonist peptide 
 ligand that produces in vivo activity in mice with reduced side effects .
additionally , we have previously shown 
 that 15b , with a 2-methylnaphthyl pendant and an acetyl n - substitution produces in vivo antinociception for greater 
 than 3 h in the wwtw assay .
thus , we hypothesized 
 that compounds containing a 2-methylnaphthyl pendant with carbonyl - containing n - substitutions could produce similar , or longer lasting , 
 in vivo activity when compared to 15b .
we synthesized 
 a modest set of 2-methylnaphthyl analogues and found that as the acyl 
 chain length increases , the balanced - affinity mor agonist / dor antagonist 
 profile seen with 15b remains essentially constant , while 
 kor affinity slightly decreases , indicating that increased chain length 
 helps provide mor and dor selectivity over kor ( table 2 ) .
the high affinity binding and lack of 
 stimulation at dor corroborates our previous findings that additional 
 hydrophobic bulk in the pendant region of our peptidomimetic series 
 increases affinity to dor in the inactive conformation while being 
 incompatible with the smaller dor active state binding site .
this trend suggests that the differences in dor 
 efficacy between most of the compounds in the benzyl series ( 1670% ) 
 and the 2-methylnaphthyl series ( 015% ) is due to differences 
 in bulk between these two pendant moieties . while the in vitro profile 
 of the 2-methylnaphthyl analogues is very promising in that all analogues 
 are high affinity , potent mor agonists and high affinity dor antagonists , 
 none of the novel analogues in this series ( 15c15e ) produced antinociception in the wwtw assay .
finally , 
 as we have seen with other peptidomimetic series , in 
 vivo activity is not predictable from in vitro profiles and physiochemical 
 parameters .
this fact remains true for this series of n - substituted analogues where 4h is the only analogue 
 to emerge that produces full antinociception in the wwtw assay at 
 a 10 mg / kg dose .
4h is a mixed - efficacy peptidomimetic 
 with nearly balanced affinities at mor and dor ( ki = 0.19 and 0.51 nm , respectively ) and produces potent 
 ( ec50 = 0.78 ) , full stimulation at mor ( 95% ) , and moderately 
 potent ( ec50 = 14 nm ) stimulation at dor ( 40% ) and selectivity 
 for mor and dor over kor .
further , 4h produces in vivo 
 activity for 1 h , a similar duration of action to 4a . 
 planned pharmacokinetic and metabolic stability testing of key members 
 of our series are expected to shed light on the as yet unexplained 
 differences in in vivo activity . in conclusion , we have expanded 
 our opioid peptidomimetic library by incorporating various n - substitutions on our previously published , unsubstituted peptidomimetics 4a and 15a after observing that n - acetylation of 
 the thq nitrogen in 4a and 15a to give 4b and 15b increases dor affinity .
all of the n - substitutions in 
 the present study contain a carbonyl moiety , which appears to be beneficial 
 in maintaining high dor affinity , an important factor in the development 
 of bifunctional mor / dor ligands . through this synthetic campaign
, 
 we have also discovered that an increased acyl chain length or additional 
 bulk increases dor stimulation within the benzyl pendant series while 
 increasing acyl chain length on the 2-methylnaphthyl series did not 
 increase dor stimulation , suggesting that pendant moieties play an 
 important role in anchoring the ligand to dor in the inactive conformation . 

analogue 4h , which displays high efficacy at mor and 
 moderate efficacy at dor and produces full in vivo antinociception 
 in the wwtw assay at a 10 mg / kg dose , may serve as a lead in the development 
 of bifunctional mor / dor agonists .
all reagents and solvents were obtained from 
 commercial sources and used without additional purification .
suzuki 
 couplings were performed on a discover s - class ( cem ) microwave in 
 a closed vessel with maximum power input of 300 w and temperature 
 set at 110 c for 1060 min under the standard method 
 from their synergy software .
flash column chromatography was carried 
 out using p60 silica gel ( 230400 mesh ) either manually or 
 with a biotage isolera instrument . before flash column chromatograph 

was performed , crude mixtures were analyzed using thin - layer chromatography 
 using 2:3 ea : hex , 1:1 ea : hex , and/or 3:2 ea : hex .
the rf values of 
 products and impurities were calculated then copied into either the 
 linear gradient or stepwise gradient wizard on the biotage isolera 
 instrument .
the tlc wizard then determined the optimal purification 
 technique which was used to purify crude mixtures .
purification of 
 final compounds was performed using a waters semipreparative hplc 
 with a vydac protein and peptide c18 reverse phase column , using a 
 linear gradient of 0% solvent b ( 0.1% tfa in acetonitrile ) in solvent 
 a ( 0.1% tfa in water ) to 100% solvent b in solvent a at a rate 1% 
 per minute and monitoring uv absorbance at 230 nm .
purity of synthesized 
 compounds was determined on a waters alliance 2690 analytical hplc 
 instrument and a vydac protein and peptide c18 reverse phase column , 
 using a linear gradient of 0% solvent b in solvent a to 45% , 70% , 
 or 90% solvent b in solvent a in 45 , 70 , or 90 min , respectively , 
 and uv absorbance at 230 nm ( gradient a ) .
purities of the final compounds 
 used for testing were 95% , unless otherwise stated , as determined 
 by hplc .
h nmr and c nmr data were obtained 
 on either a 400 or 500 mhz varian spectrometer using cdcl3 or cd3od solvents .
the identity of each compound was 
 verified by mass spectrometry using an agilent 6130 lc 
ms mass 
 spectrometer in positive mode . to a flame - dried round - bottom flask under ar
the reaction vessel was placed back under vacuum 
 for 5 min then excess acid anhydride ( propionic or butyric ) was added 
 via syringe and the solution stirred for 5 min .
the round - bottom flask 
 was flooded with ar , equipped with a condenser , and placed in oil 
 bath at 90120 c .
the reaction stirred at reflux for 
 1220 h under ar and was monitored by tlc .
once the reaction 
 was complete , the solvent was removed under reduced pressure , yielding 
 a crude yellow oil which was purified using silica gel chromatograph 
 to obtain the pure product . to a flame - dried round - bottom flask with stir 
 bar under
to a round - bottom flask already containing dried , desiccated n - substituted dihydroquinolinone intermediate ( 1.0 equiv ) 
 was added ( r)-2-methylpropane-2-sulfinamide ( 3.0 
 equiv ) , then the round - bottom flask was placed under vacuum for 10 
 min . meanwhile , a reflux condenser was flame - dried under vacuum and 
 then flooded with ar .
next , anhyd thf ( 20 ml ) was added to 
 the reaction vessel containing starting reagents via syringe .
the 
 reaction solution allowed to stir under vacuum for 5 min and 
 then was flooded with ar .
the round - bottom flask was placed in ice 
 bath and allowed to equilibrate .
once addition was complete , the reaction 
 vessel was taken out of the ice bath and placed in an oil bath at 
 7075 c , equipped with condenser , and stirred for 1648 
 h under ar .
once sufficient conversion to the tert - butanesulfinyl 
 imine was observed , the reaction vessel was taken out of the oil bath 
 and cooled to ambient temperature .
meanwhile , an additional round - bottom 
 flask containing a stir bar was flame - dried under vacuum , then flooded 
 with ar , then nabh4 was added quickly , and then reaction 
 vessel was placed back under vacuum for 5 min .
minimal anhyd thf was 
 added ( 5 ml ) , and the vessel allowed to stir under vacuum 
 for 5 min , then was flooded with ar . the round - bottom flask 
 was placed in dry ice / xylenes bath and allowed to equilibrate .
contents 
 from the round - bottom flask containing the imine intermediate were 
 transferred to round - bottom flask containing nabh4 via 
 cannula .
once contents completely added , the reaction was taken out 
 of dry ice / xylenes bath and allowed to warm to room temperature . the 
 reaction stirred at ambient temperature for 23 h. once the 
 reaction was complete , meoh was added to quench .
the residue was 
 resuspended in dcm , solid remained , and the remaining solid was removed 
 by filtration through a cotton plug and the mother liquor was concentrated 
 and purified using silica gel chromatography to yield pure sulfinamide . to a round - bottom flask already containing sulfinamide intermediate 
 was added 1520 ml of dioxane followed by conc hcl ( 6.0 equiv ) . 

the reaction stirred at rt for up to 3 h. solvent was removed under 
 reduced pressure to yield slightly yellow , clear residue .
if a white solid precipitated 
 ( the hcl salt of the amine ) : solid was removed via filtration 
 as product without any further purification necessary .
if 
 a white solid did not precipitate , but residue remains as film on 
 flask : residue washed with fresh et2o ( 3  
 5 ml ) and dried without any further purification necessary .
the ( r)-amine intermediate ( 1.0 equiv ) and diboc - dmt ( 1.01.05 
 equiv ) and the coupling reagents pybop ( 1.0 equiv ) or hatu ( 1.0 equiv ) , 
 hobt - cl ( 1.0 equiv ) , were dissolved in dmf ( 1015 ml ) followed 
 by the addition of the and dipea ( 10.0 equiv ) .
after concentration under reduced 
 pressure , the crude residue was dissolved in a 1:1 mixture of dcm 
 and tfa ( 10 ml ) and stirred for 1 h. the mixture was concentrated 
 and purified by semipreparative hplc , then lyophilized to yield the 
 final compound .
2c was synthesized 
 according to a modified general procedure a using 1 ( 110 
 mg , 0.47 mmol , 1.0 equiv ) and excess propionic anhydride .
after reaction 
 was complete , reaction mixture was cooled to rt , then transferred 
 to a separatory funnel , and ea and nahco3 were added and 
 the layers separated .
the aqueous layer was extracted with ea , and 
 the combined organic layers washed with brine then dried over mgso4 and purified using silica gel chromatography to yield the 
 title compound 2c as a clear , colorless oil ( 120 mg , 
 85% ) .
h nmr ( 500 mhz , cdcl3 )  7.86 ( d , j = 0.9 , 1h ) , 7.39 ( br s , 1h ) , 7.35 ( dd , j = 8.2 , 2.2 , 1h ) , 7.29 ( t , j = 7.4 , 2h ) , 7.237.16 
 ( m , 3h ) , 4.20 ( t , j = 6.2 , 2h ) , 3.98 ( s , 2h ) , 2.75 
 ( t , j = 6.2 , 2h ) , 2.58 ( q , j = 7.4 , 
 2h ) , 1.20 ( t , j = 7.4 , 3h ) .
c nmr ( 126 
 mhz , cdcl3 )  194.27 , 173.03 , 142.21 , 140.13 , 138.78 , 
 134.63 , 128.90 , 128.71 , 127.62 , 126.49 , 126.11 , 124.44 , 43.88 , 41.29 , 
 39.62 , 27.98 , 9.91 .
2d was synthesized 
 according to a modified general procedure a using 1 ( 0.18 
 g , 0.78 mmol , 1.0 equiv ) and excess butyric anhydride ( 5 ml ) and pyridine 
 ( 4 ml ) .
after reaction was complete ( 24 h ) , the reaction was quenched 
 with h2o .
dcm was added , and the aqueous layer was extracted 
 with dcm . combined organic extracts were washed with 2 m naoh and 
 dried with mgso4 .
solvents were filtered and removed under 
 reduced pressure , and the crude residue was purified via silica gel 
 chromatography to yield the title compound 2d as an oil 
 ( 0.029 g , 12% ) .
h nmr ( 400 mhz , cdcl3 )  
 7.87 ( t , j = 1.3 , 1h ) , 7.397.34 ( m , 2h ) , 
 7.337.25 ( m , 2h ) , 7.257.16 ( m , 3h ) , 4.21 ( t , j = 6.2 , 2h ) , 3.99 ( s , 2h ) , 2.76 ( t , j = 
 6.2 , 2h ) , 2.54 ( t , j = 8.0 , 2h ) , 1.791.66 
 ( m , 2h ) , 0.95 ( t , j = 7.4 , 3h ) .
c nmr 
 ( 101 mhz , cdcl3 )  194.37 , 172.35 , 142.31 , 140.18 , 
 138.87 , 134.71 , 128.99 , 128.80 , 127.77 , 126.58 , 126.18 , 124.55 , 43.97 , 
 41.38 , 39.82 , 36.57 , 19.25 , 13.94 .
2e was synthesized 
 according to general procedure b starting from 1 ( 100 
 mg , 0.42 mmol , 1.0 equiv ) to yield the title compound 2e as a clear , colorless oil ( 105 mg , 80.8% ) .
h nmr ( 500 
 mhz , cdcl3 )  7.87 ( d , j = 2.0 hz , 
 1h ) , 7.377.25 ( m , 4h ) , 7.257.17 ( m , 3h ) , 4.21 ( t , j = 6.3 hz , 2h ) , 3.99 ( s , 2h ) , 3.14 ( hept , j = 6.7 hz , 1h ) , 2.75 ( t , j = 6.2 hz , 2h ) , 1.18 ( d , j = 6.7 hz , 6h ) .
c nmr ( 126 mhz , cdcl3 )  194.33 , 176.97 , 142.29 , 140.05 , 138.84 , 134.59 , 128.90 , 
 128.87 , 128.68 , 127.64 , 126.46 , 126.15 , 124.22 , 43.90 , 41.25 , 39.79 , 
 31.13 , 19.88 .
2f was synthesized 
 following a modified general procedure b starting with 1 ( 0.15 g , 0.65 mmol , 1.0 equiv ) and also using et3n ( 0.18 
 ml , 1.29 mmol , 2.0 equiv ) .
the reaction stirred for 3 h , after which 
 time satd nahco3 was added .
solvents were filtered and removed under reduced 
 pressure , and crude residue was purified via silica gel chromatography 
 to yield the title compound 2f as a colorless oil ( 0.15 
 g , 78% ) .
h nmr ( 500 mhz , cdcl3 )  7.88 
 ( d , j = 2.2 , 1h ) , 7.46 ( d , j = 8.3 , 
 1h ) , 7.35 ( dd , j = 8.4 , 2.2 , 1h ) , 7.317.26 
 ( m , 2h ) , 7.227.17 ( m , 3h ) , 4.26 ( t , j = 6.3 , 
 2h ) , 3.98 ( s , 2h ) , 2.76 ( t , j = 6.3 , 2h ) , 2.061.96 
 ( m , 1h ) , 1.211.15 ( m , 2h ) , 0.910.82 ( m , 2h ) .
c nmr ( 126 mhz , cdcl3 )  194.41 , 173.00 , 
 142.45 , 140.13 , 138.53 , 134.49 , 128.89 , 128.71 , 127.74 , 126.49 , 125.85 , 
 124.09 , 43.53 , 41.28 , 39.70 , 13.74 , 9.77 .
2 g was synthesized 
 following general procdure b starting with 1 ( 0.15 g , 
 0.62 mmol , 1.0 equiv ) .
the reaction stirred for 3 h , after which time 
 satd nahco3 was added .
solvents were filtered and removed under reduced 
 pressure , and crude residue was purified via silica gel chromatography 
 to yield the title compound 2 g as a white solid ( 0.20 
 g , 99% ) .
h nmr ( 500 mhz , cdcl3 )  7.85 
 ( d , j = 1.3 , 1h ) , 7.487.37 ( br s , 1h ) , 7.35 
 ( dd , j = 8.2 , 2.2 , 1h ) , 7.29 ( t , j = 7.5 , 2h ) , 7.247.13 ( m , 3h ) , 4.12 ( t , j = 6.2 , 2h ) , 3.98 ( s , 2h ) , 3.52 ( p , j = 8.4 , 1h ) , 
 2.72 ( t , j = 6.2 , 2h ) , 2.43 ( dq , j = 11.8 , 9.2 , 2h ) , 2.13 ( q , j = 9.9 , 2h ) , 2.021.88 
 ( m , 2h ) .
c nmr ( 126 mhz , cdcl3 )  194.09 , 
 174.02 , 142.14 , 140.14 , 138.52 , 134.63 , 128.85 , 128.64 , 127.49 , 126.41 , 
 125.75 , 123.84 , 43.78 , 41.24 , 39.57 , 37.95 , 25.72 , 17.83 .
2h was synthesized following general procedure b starting from the 1 ( 150 mg , 0.63 mmol , 1.0 equiv ) to yield the title compound 2h as a clear , colorless oil ( 111 mg , 59.4% ) .
h nmr ( 500 mhz , cdcl3 )  7.86 ( d , j = 2.1 hz , 1h ) , 7.71 ( d , j = 8.6 hz , 1h ) , 7.34 ( dd , j = 8.6 , 2.2 hz , 1h ) , 7.29 ( t , j = 7.5 
 hz , 2h ) , 7.237.16 ( m , 3h ) , 4.18 ( t , j = 6.3 
 hz , 2h ) , 3.97 ( s , 2h ) , 3.84 ( s , 3h ) , 2.76 ( t , j = 
 6.3 hz , 2h ) .
c nmr ( 126 mhz , cdcl3 )  
 193.79 , 154.23 , 141.74 , 140.24 , 137.32 , 134.79 , 128.74 , 128.51 , 127.13 , 
 126.25 , 124.79 , 123.58 , 53.30 , 44.40 , 41.09 , 38.81 .
2i was synthesized 
 following general procedure b starting from the 1 ( 150 
 mg , 0.63 mmol , 1.0 equiv ) to yield the title compound 2i as a white solid ( 176 mg , 89.8% ) .
h nmr ( 500 mhz , cdcl3 )  7.87 ( d , j = 2.2 hz , 1h ) , 7.37 
 ( dd , j = 8.4 , 2.2 hz , 1h ) , 7.28 ( t , j = 7.6 hz , 2h ) , 7.237.15 ( m , 3h ) , 4.26 ( s , 2h ) , 4.17 ( t , j = 6.3 hz , 2h ) , 3.98 ( s , 2h ) , 3.45 ( s , 3h ) , 2.78 ( t , j = 6.2 hz , 2h ) .
c nmr ( 126 mhz , cdcl3 )  193.62 , 168.07 , 141.25 , 139.91 , 139.02 , 134.72 , 128.77 , 
 128.58 , 128.55 , 127.56 , 126.37 , 125.78 , 123.66 , 76.74 , 71.84 , 59.24 , 
 43.95 , 41.17 , 39.41 .
2j was synthesized 
 following general procedure b starting from the 1 ( 100 
 mg , 0.42 mmol , 1.0 equiv ) to yield the title compound 2j as a clear , slightly yellow oil ( 139 mg , 96.5% ) .
h nmr 
 ( 500 mhz , cdcl3 )  7.78 ( d , j = 
 2.1 hz , 1h ) , 7.427.34 ( m , 3h ) , 7.317.25 ( m , 2h ) , 7.227.17 
 ( m , 2h ) , 7.147.10 ( m , 1h ) , 7.087.04 ( m , 2h ) , 7.01 
 ( dd , j = 8.5 , 2.2 hz , 1h ) , 6.81 ( d , j = 8.4 hz , 1h ) , 4.21 ( t , j = 6.3 hz , 2h ) , 3.85 ( s , 
 2h ) , 2.77 ( t , j = 6.3 hz , 2h ) .
c nmr 
 ( 126 mhz , cdcl3 )  193.73 , 170.04 , 142.60 , 139.99 , 
 138.06 , 135.06 , 134.31 , 130.99 , 128.78 , 128.58 , 128.47 , 128.39 , 127.38 , 
 126.36 , 124.82 , 124.63 , 45.30 , 41.10 , 39.50 .
3c was synthesized following 
 general procedure c using 2c ( 120 mg , 0.40 mmol , 1.0 
 equiv ) , ( r)-2-methylpropane-2-sulfinamide ( 150 mg , 
 1.2 mmol , 3.0 equiv ) , and ti(oet)4 ( 0.63 ml , 2.4 mmol , 
 6.0 equiv ) to form the ( r)-tert - butanesulfinyl 
 imine intermediate in situ . once sufficient ketone was converted into 
 imine intermediate ( after 17 h ) , the reaction mixture was transferred 
 via cannula to a round - bottom flask containing nabh4 ( 90 
 mg , 2.4 mmol , 6.0 equiv ) and 20 ml of thf in a xylenes dry ice bath ; 
 after addition , the solution was stirred at room temperature for 3 
 h before being quenched with meoh .
once resultant solid was removed , 
 crude residue was purified using silica gel chromatography to yield 
 the title compound 3c as a clear , colorless oil ( 120 
 mg , 78% ) .
h nmr ( 500 mhz , cdcl3 )  7.69 
 ( d , j = 8.2 hz , 1h ) , 7.297.21 ( m , 2h ) , 7.207.15 
 ( qd , j = 6.5 , 5.4 , 1.6 hz , 4h ) , 7.06 ( dd , j = 8.5 , 2.3 hz , 1h ) , 4.52 ( q , j = 3.4 
 hz , 1h ) , 3.973.87 ( m , 3h ) , 3.57 ( tdd , j = 
 12.9 , 4.2 , 1.7 hz , 1h ) , 3.32 ( bs , 1h ) , 2.202.13 ( m , 1h ) , 1.991.90 
 ( m , 1h ) , 1.50 ( s , 9h ) , 1.19 ( s , 9h ) .
c nmr ( 126 mhz , 
 cdcl3 )  153.43 , 140.77 , 136.51 , 136.42 , 128.88 , 
 128.71 , 128.55 , 128.47 , 128.34 , 125.97 , 123.89 , 80.97 , 55.52 , 50.36 , 
 41.08 , 40.00 , 29.41 , 28.22 , 22.48 .
3d was synthesized following 
 general procedure c using 2d ( 0.029 g , 0.094 mmol , 1.0 
 equiv ) to yield the title compound 3d as a colorless 
 oil ( 0.010 g , 26% ) .
h nmr ( 400 mhz , cdcl3 ) 
  7.337.27 ( m , 2h ) , 7.247.18 ( m , 3h ) , 7.10 ( dd , j = 8.3 , 2.1 , 1h ) , 4.53 ( t , j = 4.6 , 1h ) , 
 3.95 ( s , 2h ) , 3.943.85 ( m , 1h ) , 3.803.68 ( m , 1h ) , 
 3.34 ( br s , 1h ) , 2.512.41 ( m , 2h ) , 2.272.15 ( m , 1h ) , 
 2.081.97 ( m , 1h ) , 1.741.63 ( m , 2h ) , 1.20 ( s , 9h ) , 
 0.92 ( t , j = 7.4 , 3h ) .
3e was synthesized following 
 general procedure c using 2e ( 105 mg , 0.34 mmol , 1.0 
 equiv ) to yield the title compound 3e as a clear , colorless 
 oil ( 70 mg , 50% ) .
h nmr ( 500 mhz , cdcl3 )  
 7.347.26 ( m , 3h ) , 7.247.19 ( m , 3h ) , 7.10 ( dd , j = 8.2 , 2.2 hz , 1h ) , 4.53 ( q , j = 3.9 
 hz , 1h ) , 3.96 ( s , 2h ) , 3.943.89 ( m , 1h ) , 3.753.67 
 ( m , 1h ) , 3.30 ( bs , 1h ) , 3.12 ( p , j = 6.9 hz , 1h ) , 
 2.242.15 ( m , 1h ) , 2.081.98 ( m , 1h ) , 1.231.16 
 ( s , 9h ) , 1.13 ( dd , j = 9.0 , 6.7 hz , 6h ) .
c nmr ( 126 mhz , cdcl3 )  177.35 , 140.53 , 138.70 , 
 136.67 , 128.92 , 128.67 , 128.58 , 128.54 , 126.28 , 124.60 , 55.76 , 51.04 , 
 41.37 , 40.00 , 31.09 , 30.96 , 22.55 , 20.03 , 20.01 .
3f was synthesized following 
 general procedure c using 2f ( 0.16 g , 0.51 mmol , 1.0 
 equiv ) to yield the title compound 3f as a colorless 
 oil ( 0.12 g , 55% ) .
h nmr ( 400 mhz , cdcl3 )  
 7.36 ( d , j = 8.2 , 1h ) , 7.327.26 ( m , 3h ) , 
 7.237.18 ( m , 3h ) , 7.09 ( dd , j = 8.3 , 2.0 , 
 1h ) , 4.55 ( q , j = 4.3 , 1h ) , 4.013.93 ( m , 
 3h ) , 3.75 ( ddd , j = 12.9 , 9.3 , 5.6 , 1h ) , 3.33 ( d , j = 3.6 , 1h ) , 2.23 ( dq , j = 14.9 , 5.1 , 
 1h ) , 2.101.89 ( m , 3h ) , 1.20 ( s , 9h ) , 1.141.08 ( m , 
 2h ) , 0.78 ( dd , j = 7.9 , 2.5 , 2h ) .
c 
 nmr ( 101 mhz , cdcl3 )  173.36 , 140.65 , 138.53 , 136.83 , 
 131.83 , 128.98 , 128.71 , 128.58 , 128.51 , 126.36 , 124.97 , 55.85 , 51.18 , 
 41.47 , 39.85 , 30.74 , 22.61 , 13.65 , 9.28 .
3 g was synthesized following 
 general procedure c using 2 g ( 0.20 g , 0.62 mmol , 1.0 
 equiv ) to yield the title compound 3 g as a colorless 
 oil ( 0.16 g , 60% ) .
h nmr ( 500 mhz , cdcl3 )  
 7.317.25 ( m , 2h ) , 7.237.17 ( m , 2h ) , 7.09 ( dd , j = 8.3 , 2.1 , 1h ) , 4.51 ( q , j = 4.3 , 1h ) , 
 3.95 ( s , 2h ) , 3.863.77 ( m , 1h ) , 3.743.63 ( m , 1h ) , 
 3.503.39(m , 2h ) , 2.442.30 ( m 2h ) , 2.19 ( dq , j = 14.3 , 4.9 , 1h ) , 2.121.97 ( m , 3h ) , 1.971.82 
 ( m , 2h ) , 1.21 ( s , 9h ) .
c nmr ( 126 mhz , cdcl3 )  174.44 , 140.63 , 138.39 , 136.58 , 128.89 , 128.73 , 128.56 , 
 128.50 , 126.23 , 124.20 , 55.77 , 50.96 , 41.37 , 39.90 , 38.22 , 30.72 , 
 25.80 , 22.60 , 17.84 .
3h was synthesized following general procedure c using 2h ( 111 mg , 0.376 mmol , 1.0 equiv ) to yield the title compound 3h as a clear , colorless oil ( 124 mg , 82.1% ) .
h nmr ( 500 mhz , cdcl3 )  7.73 ( d , j = 8.7 hz , 1h ) , 7.27 ( t , j = 7.8 hz , 2h ) , 7.237.16 
 ( m , 4h ) , 7.117.06 ( m , 1h ) , 4.54 ( q , j = 3.6 
 hz , 1h ) , 4.01 ( dt , j = 13.0 , 4.6 hz , 1h ) , 3.93 ( s , 
 2h ) , 3.78 ( d , j = 1.0 hz , 3h ) , 3.64 ( ddd , j = 12.9 , 11.1 , 3.8 hz , 1h ) , 3.26 ( d , j = 2.7 hz , 1h ) , 2.242.14 ( m , 1h ) , 2.031.94 ( m , 1h ) , 
 1.20 ( d , j = 1.0 hz , 9h ) .
c nmr ( 126 
 mhz , cdcl3 )  154.94 , 140.75 , 137.11 , 136.11 , 129.11 , 
 128.79 , 128.77 , 128.57 , 128.44 , 126.09 , 126.08 , 123.68 , 55.59 , 52.94 , 
 50.24 , 41.15 , 40.23 , 29.43 , 22.51 .
3i was synthesized following 
 general procedure c using 2i ( 176 mg , 0.60 mmol , 1.0 
 equiv ) to yield the title compound 3i as a clear , colorless 
 oil ( 148 mg , 62.7% ) .
h nmr ( 500 mhz , cdcl3 ) 
  7.307.24 ( m , 3h ) , 7.18 ( d , j = 7.6 
 hz , 3h ) , 7.09 ( dd , j = 8.3 , 2.0 hz , 1h ) , 4.52 ( q , j = 4.3 hz , 1h ) , 4.17 ( s , 2h ) , 3.93 ( s , 2h ) , 3.79 ( d , j = 40.2 hz , 11h ) , 3.44 ( d , j = 3.6 hz , 
 1h ) , 3.39 ( s , 3h ) , 2.20 ( dq , j = 14.5 , 4.9 hz , 1h ) , 
 2.04 ( qd , j = 9.2 , 8.7 , 5.0 hz , 1h ) , 1.20 ( s , 35h ) , 
 1.17 ( s , 10h ) .
c nmr ( 126 mhz , cdcl3 )  
 168.45 , 140.40 , 135.69 , 128.97 , 128.82 , 128.68 , 128.50 , 128.46 , 128.21 , 
 126.20 , 126.14 , 123.95 , 71.69 , 59.16 , 55.72 , 55.22 , 50.80 , 41.38 , 
 41.29 , 39.90 , 30.47 , 29.60 , 22.52 , 22.21 , 22.07 .
3j was synthesized following 
 general procedure c using 2j ( 139 mg , 0.41 mmol , 1.0 
 equiv ) to yield the title compound 3j as an off - white 
 solid ( 127 mg , 69.8% ) .
h nmr ( 500 mhz , cdcl3 )  7.31 ( td , j = 8.0 , 1.4 hz , 3h ) , 7.277.15 
 ( m , 5h ) , 7.147.09 ( m , 1h ) , 7.097.04 ( m , 2h ) , 6.78 
 ( t , j = 9.0 hz , 2h ) , 4.54 ( q , j = 
 3.9 hz , 1h ) , 3.93 ( dt , j = 12.5 , 5.1 hz , 1h ) , 3.82 
 ( s , 2h ) , 3.783.68 ( m , 1h ) , 3.32 ( d , j = 3.1 
 hz , 1h ) , 2.20 ( dq , j = 14.0 , 4.7 hz , 1h ) , 2.02 ( ddt , j = 14.5 , 9.9 , 5.0 hz , 1h ) , 1.13 ( d , j = 
 1.5 hz , 11h ) .
c nmr ( 126 mhz , cdcl3 )  
 170.22 , 140.50 , 138.06 , 136.69 , 135.96 , 130.33 , 129.87 , 128.79 , 128.68 , 
 128.46 , 128.34 , 128.32 , 128.23 , 126.15 , 125.31 , 60.32 , 55.72 , 50.61 , 
 41.47 , 41.19 , 30.24 , 22.53 , 20.99 , 14.15 .
4c was synthesized following 
 general procedure d using 3c ( 120 mg , 0.31 mmol , 1.0 
 equiv ) and conc hcl ( 0.1 ml , 1.0 mmol , 6.0 equiv ) . after removing 
 solvent ,
after washing the solid 3 with fresh et2o , the 
 remaining et2o was decanted off , yielding a white solid 
 amine hydrochloride salt ( 27 mg ) .
the ( r)-amine intermediate 
 ( 27 mg , 0.082 mmol , 1.0 equiv ) and diboc - dmt ( 33 mg , 0.082 mmol , 1.0 
 equiv ) and the coupling reagents hatu ( 31 mg , 0.082 mmol , 1.0 equiv ) 
 and hobt - cl ( 14 mg , 0.082 mmol , 1.0 equiv ) were dissolved in dmf ( 1015 
 ml ) , followed by the addition of the and dipea ( 14 ml , 0.82 mmol , 
 10.0 equiv ) .
after concentration under reduced pressure , the crude residue was 
 dissolved in a 1:1 mixture of dcm and tfa ( 10 ml ) and stirred for 
 1 h. the mixture was concentrated and purified by semipreparative 
 hplc to yield the title compound as a white solid .
note that not all 
 crude product was purified , so a yield was not calculated .
h nmr ( 500 mhz , cd3od )  8.15 ( d , j = 8.2 , 1h ) , 7.42 ( br s , 1h ) , 7.277.19 ( m , 2h ) , 7.187.11 
 ( m , 4h ) , 7.05 ( dd , j = 8.4 , 2.1 , 1h ) , 6.51 ( s , 2h ) , 
 4.964.91 ( m , 1h ) , 3.91 ( s , 2h ) , 3.86 ( dd , j = 11.5 , 5.0 , 1h ) , 3.78 ( br s , 1h ) , 3.26 ( dd , j = 
 13.6 , 11.5 , 1h ) , 3.183.10 ( m , 1h ) , 3.03 ( dd , j = 13.7 , 5.1 , 1h ) , 2.582.39 ( m , 2h ) , 2.27 ( s , 6h ) , 1.901.80 
 ( m , 1h ) , 1.481.39 ( m , 1h ) , 1.11 ( t , j = 7.4 , 
 3h ) .
4d was synthesized following 
 general procedure d using 3d ( 10 mg , 0.024 mmol , 1.0 
 equiv ) with the coupling reagent pybop to yield the tfa salt of the 
 title compound , 4d , as a white solid .
h nmr ( 500 mhz , cd3od )  7.39 ( br s , 1h ) , 7.277.20 ( m , 2h ) , 7.187.11 
 ( m , 4h ) , 7.06 ( dd , j = 8.4 , 2.1 , 1h ) , 6.50 ( s , 2h ) , 
 4.93 ( t , j = 6.5 , 1h ) , 3.91 ( s , 2h ) , 3.883.75 
 ( m , 2h ) , 3.25 ( dd , j = 13.6 , 11.5 , 1h ) , 3.153.11 
 ( m , 1h ) , 3.02 ( dd , j = 13.7 , 5.1 , 1h ) , 2.542.39 
 ( m , 2h ) , 2.27 ( s , 6h ) , 1.911.81 ( m , 1h ) , 1.691.58 
 ( m , 2h ) , 1.441.40 ( m , 1h ) , 0.93 ( t , j = 7.2 , 
 3h ) .
4e was synthesized following 
 general procedure d using 3e ( 70 mg , 0.17 mmol , 1.0 equiv ) 
 with the coupling reagent pybop to yield the tfa salt of the title 
 compound , 4e , as a white solid ( 42 mg , 40.7% ) .
h nmr ( 500 mhz , cd3od )  8.18 ( d , j = 8.2 hz , 0h ) , 7.19 
 ( dt , j = 35.4 , 7.7 hz , 6h ) , 7.04 ( d , j = 8.3 hz , 3h ) , 6.51 ( s , 2h ) , 3.92 ( d , j = 14.2 
 hz , 3h ) , 3.26 ( t , j = 12.6 hz , 1h ) , 3.12 ( ddt , j = 41.3 , 13.4 , 5.3 hz , 4h ) , 2.27 ( s , 7h ) , 1.86 ( dq , j = 12.6 , 6.2 hz , 1h ) , 1.37 ( dd , j = 12.8 , 
 6.4 hz , 1h ) , 1.12 ( d , j = 6.6 hz , 5h ) , 1.05 ( d , j = 6.9 hz , 3h ) .
c nmr ( 126 mhz , cd3od )  179.32 , 169.29 , 162.72 , 157.37 , 142.26 , 140.05 , 137.53 , 
 129.77 , 129.45 , 129.20 , 127.13 , 125.65 , 123.29 , 116.40 , 53.46 , 46.89 , 
 42.12 , 32.38 , 31.92 , 31.83 , 20.44 , 20.17 , 19.90 .
esi - ms 500.3 [ m + h ] and 522.3 
 [ m + na ] .
4f was synthesized following 
 general procedure d using 3f ( 120 mg , 0.28 mmol , 1.0 
 equiv ) with the coupling reagent pybop to yield the tfa salt of the 
 title compound , 4f , as a white solid .
note that not all 
 crude product was purified , so a yield was not calculated .
h nmr ( 500 mhz , cd3od )  7.39 ( d , j = 8.3 , 1h ) , 7.267.20 ( m , 2h ) , 7.197.12 ( m , 4h ) , 
 7.07 ( dd , j = 8.4 , 2.1 , 1h ) , 6.50 ( s , 2h ) , 4.96 ( t , j = 6.3 , 1h ) , 3.92 ( s , 2h ) , 3.913.80 ( m , 2h ) , 3.293.20 
 ( m , 2h ) , 3.072.99 ( m , 1h ) , 2.28 ( s , 6h ) , 1.981.92 
 ( m , 1h ) , 1.891.81 ( m , 1h ) , 1.471.38 ( m , 1h ) , 1.071.01 
 ( m , 1h ) , 0.990.92 ( m , 1h ) , 0.910.79 ( m , 2h ) .
4 g was synthesized following 
 general procedure d using 3 g ( 160 mg , 0.37 mmol , 1.0 
 equiv ) with the coupling reagent pybop to yield the tfa salt of the 
 title compound , 4 g , as a white solid . note that not all 
 crude product was purified , so a yield was not calculated .
h nmr ( 500 mhz , cd3od )  7.39 ( br s , 1h ) , 7.267.21 
 ( m , 2h ) , 7.187.11 ( m , 4h ) , 7.05 ( dd , j = 
 8.3 , 2.1 , 1h ) , 6.50 ( s , 2h ) , 4.984.89 ( m , 1h ) , 3.91 ( s , 2h ) , 
 3.85 ( dd , j = 11.5 , 5.1 , 1h ) , 3.68 ( br s , 1h ) , 3.50 
 ( p , j = 8.5 , 1h ) , 3.25 ( dd , j = 
 13.6 , 11.6 , 1h ) , 3.123.01 ( m , 2h ) , 2.351.94 ( m , 11h ) , 
 1.891.77 ( m , 2h ) , 1.431.32 ( m , 1h ) .
4h was synthesized following general procedure d using 3h ( 124 mg , 0.31 mmol , 1.0 equiv ) and the coupling reagent pybop to 
 yield the tfa salt of the title compound , 4h , as a white 
 solid ( 50 mg , 69.5% ) .
h nmr ( 400 mhz , cd3od ) 
  7.65 ( d , j = 8.6 hz , 1h ) , 7.267.20 
 ( m , 2h ) , 7.177.12 ( m , 3h ) , 7.07 ( d , j = 2.2 
 hz , 1h ) , 7.03 ( dd , j = 8.8 , 1.9 hz , 1h ) , 6.50 ( s , 
 2h ) , 4.97 ( t , j = 5.2 hz , 1h ) , 3.87 ( s , 2h ) , 3.863.81 
 ( m , 1h ) , 3.75 ( d , j = 1.4 hz , 4h ) , 3.25 ( ddd , j = 13.1 , 11.5 , 1.4 hz , 1h ) , 3.16 ( tdd , j = 6.7 , 3.8 , 1.4 hz , 12h ) , 3.052.94 ( m , 2h ) , 2.27 ( d , j = 1.4 hz , 7h ) , 1.901.82 ( m , 12h ) , 1.77 ( tt , j = 9.7 , 4.8 hz , 1h ) , 1.541.45 ( m , 1h ) .
esi - ms 488.3 [ m + h ] and 510.3 [ m + na ] .
4i was synthesized following 
 general procedure d using 3i ( 148 mg , 0.36 mmol , 1.0 
 equiv ) and the coupling reagent pybop to yield the tfa salt of the 
 title compound , 4i , as a white solid .
note that not all 
 crude product was purified , so a yield was not calculated .
h nmr ( 400 mhz , cd3od )  7.23 ( t , j = 7.8 hz , 2h ) , 7.14 ( dd , j = 9.3 , 2.6 hz , 4h ) , 
 7.07 ( dd , j = 8.4 , 1.9 hz , 1h ) , 6.51 ( s , 2h ) , 4.96 
 ( t , j = 5.7 hz , 1h ) , 4.25 ( dd , j = 14.6 , 1.1 hz , 1h ) , 4.14 ( d , j = 14.5 hz , 1h ) , 
 3.90 ( s , 2h ) , 3.84 ( dd , j = 11.5 , 5.0 hz , 1h ) , 3.40 
 ( s , 3h ) , 3.25 ( t , j = 12.6 hz , 1h ) , 3.15 ( tdd , j = 6.7 , 3.7 , 1.2 hz , 7h ) , 3.02 ( dd , j = 
 13.7 , 5.0 hz , 1h ) , 2.27 ( s , 6h ) , 1.881.81 ( m , 8h ) , 1.48 ( s , 
 1h ) .
esi - ms 502.3 [ m + h ] and 524.3 [ m + na ] .
4j was synthesized following 
 general procedure d using 3j ( 182 mg , 0.41 mmol , 1.0 
 equiv ) and the coupling reagent pybop to yield the tfa salt of the 
 title compound , 4j , as a white solid .
note that not all 
 crude product was purified , so a yield was not calculated .
h nmr ( 400 mhz , cd3od )  7.477.42 ( m , 1h ) , 
 7.37 ( d , j = 6.8 hz , 4h ) , 7.257.20 ( m , 2h ) , 
 7.15 ( d , j = 7.6 hz , 1h ) , 7.11 ( d , j = 7.6 hz , 2h ) , 6.876.76 ( m , 2h ) , 6.49 ( s , 2h ) , 5.04 ( t , j = 6.0 hz , 1h ) , 3.923.81 ( m , 4h ) , 3.25 ( d , j = 12.3 hz , 1h ) , 3.06 ( dd , j = 13.8 , 5.1 
 hz , 1h ) , 2.28 ( s , 6h ) , 1.94 ( t , j = 10.4 hz , 1h ) , 
 1.48 ( dd , j = 13.0 , 6.3 hz , 1h ) . c nmr 
 ( 126 mhz , cd3od )  172.42 , 169.14 , 157.49 , 142.26 , 
 140.04 , 139.67 , 137.75 , 137.19 , 131.74 , 130.37 , 129.71 , 129.50 , 129.45 , 
 129.39 , 128.99 , 127.15 , 126.27 , 123.18 , 116.45 , 53.53 , 49.00 , 46.85 , 
 42.06 , 31.98 , 31.42 , 20.44 .
esi - ms 534.3 [ m + h ] and 556.2 [ m + na ] .
after removal of 
 solvent under reduced pressure , the crude product was purified by 
 semipreparative hplc and lyophilized to yield the tfa salt of title 
 compound , 9k , as a white powder ( 2 mg , 18% ) .
h nmr ( 500 mhz , cd3od )  8.33 ( d , j = 7.8 hz , 1h ) , 7.19 ( t , j = 7.4 hz , 2h ) , 
 7.147.07 ( m , 3h ) , 6.92 ( d , j = 8.5 hz , 1h ) , 
 6.86 ( s , 1h ) , 6.46 ( s , 2h ) , 6.38 ( d , j = 8.5 hz , 
 1h ) , 3.91 ( d , j = 17.8 hz , 1h ) , 3.80 ( dd , j = 11.8 , 5.1 hz , 1h ) , 3.76 ( s , 2h ) , 3.663.59 ( m , 
 1h ) , 3.25 ( t , j = 12.4 hz , 1h ) , 3.00 ( dd , j = 13.6 , 5.1 hz , 1h ) , 2.86 ( d , j = 12.1 
 hz , 1h ) , 2.38 ( t , j = 12.1 hz , 1h ) , 1.81 ( t , j = 13.1 hz , 1h ) , 1.52 ( d , j = 13.4 hz , 
 1h ) , 1.29 ( s , 1h ) .
c nmr ( 126 mhz , cd3od ) 
  175.88 , 157.43 , 144.84 , 140.04 , 131.51 , 131.21 , 130.86 , 129.63 , 
 129.29 , 126.85 , 123.21 , 121.80 , 116.48 , 112.92 , 66.62 , 55.53 , 53.45 , 
 49.00 , 47.37 , 47.27 , 46.62 , 41.77 , 31.89 , 28.93 , 20.46 .
esi - ms = 487.3.3 [ m + h ] and 
 509.3 [ m + na ] .
4l was synthesized by treating crude 9l with 3 n hcl to 
 remove the remaining boc - groups and yield the crude product which 
 was purified by semipreparative hplc and lyophilized to yield the 
 title compound as the tfa salt of 4l , as a white powder 
 ( 44 mg , 41.1% ) .
h nmr ( 500 mhz , cd3od )  
 8.17 ( d , j = 7.6 hz , 1h ) , 7.227.17 ( m , 2h ) , 
 7.147.07 ( m , 3h ) , 6.926.85 ( m , 2h ) , 6.43 ( s , 2h ) , 
 6.39 ( d , j = 8.4 hz , 1h ) , 4.87 ( s , 6h ) , 4.24 ( s , 
 0h ) , 4.234.15 ( m , 2h ) , 3.81 ( q , j = 5.3 hz , 
 1h ) , 3.76 ( d , j = 8.6 hz , 2h ) , 3.31 ( s , 2h ) , 3.25 
 ( dd , j = 13.5 , 11.8 hz , 1h ) , 3.00 ( dd , j = 13.6 , 5.0 hz , 1h ) , 2.85 ( dt , j = 11.8 , 4.1 hz , 
 1h ) , 2.42 ( td , j = 12.3 , 3.2 hz , 1h ) , 2.27 ( s , 6h ) , 
 1.75 ( tt , j = 12.8 , 4.1 hz , 1h ) , 1.57 ( dq , j = 13.3 , 3.5 hz , 1h ) , 1.29 ( td , j = 7.1 , 
 1.0 hz , 3h ) .
c nmr ( 126 mhz , cd3od )  
 173.03 , 168.05 , 157.42 , 144.44 , 143.28 , 140.00 , 131.60 , 131.03 , 130.84 , 
 129.64 , 129.29 , 126.84 , 123.20 , 121.53 , 116.47 , 112.40 , 62.35 , 53.49 , 
 53.20 , 49.51 , 49.34 , 49.17 , 49.00 , 48.83 , 48.66 , 48.49 , 47.56 , 46.18 , 
 41.75 , 31.87 , 28.78 , 20.48 , 14.51 .
esi - ms 516.3 [ m + h ] and 538.3 [ m + na ] .
4 m was formed by treating the crude residue of 9 m with 
 1:1 tfa : dcm to form crude product which was purified by semipreparative 
 hplc and lyophilized to yield the tfa salt of the title compound , 4 m ( 1.6 mg , 2.2% ) .
note that purification was difficult and 
 not all crude product was purified , so a yield was not calculated . 

esi - ms 488.3 [ m + h ] and 510.3 [ m + na ] .
no h or c nmr data acquired . instead , formation of final product 4 m verified by mass spectrometry .
6 was 
 synthesized following general procedure c using 5 ( 1.1 
 g , 4.8 mmol , 1.0 equiv ) , ( r)-2-methylpropane-2-sulfinamide 
 ( 175 mg , 1.44 mmol , 3.0 equiv ) , and ti(oet)4 ( 0.604 ml , 
 2.88 mmol , 6.0 equiv ) to form the ( r)-tert - butanesulfinyl imine intermediate in situ . once sufficient ketone 
 was converted into imine intermediate ( after 48 h ) , the reaction mixture 
 was transferred via cannula to a round - bottom flask containing nabh4 ( 109 mg , 2.44 mmol , 6.0 equiv ) and 20 ml of thf in a xylenes 
 dry ice bath ; after addition , the solution was stirred at room temperature 
 for 3 h before being quenched with meoh .
once resultant solid was 
 removed , crude residue was purified using silica gel chromatography 
 to yield the title compound 6 as a clear , colorless oil 
 ( 175 mg , 82.2% ) .
h nmr ( 500 mhz , cdcl3 )  
 7.69 ( d , j = 8.2 hz , 1h ) , 7.297.21 ( m , 2h ) , 
 7.207.15 ( qd , j = 6.5 , 5.4 , 1.6 hz , 4h ) , 
 7.06 ( dd , j = 8.5 , 2.3 hz , 1h ) , 4.52 ( q , j = 3.4 hz , 1h ) , 3.973.87 ( m , 3h ) , 3.57 ( tdd , j = 12.9 , 4.2 , 1.7 hz , 1h ) , 3.32 ( bs , 1h ) , 2.202.13 
 ( m , 1h ) , 1.991.90 ( m , 1h ) , 1.50 ( s , 9h ) , 1.19 ( s , 9h ) .
c nmr ( 126 mhz , cdcl3 )  153.43 , 140.77 , 
 136.51 , 136.42 , 128.88 , 128.71 , 128.55 , 128.47 , 128.34 , 125.97 , 123.89 , 
 80.97 , 55.52 , 50.36 , 41.08 , 40.00 , 29.41 , 28.22 , 22.48 .
7 was synthesized by first 
 treating 6 ( 400 mg , 0.904 mml , 1.0 equiv ) with excess 
 1:1 tfa : dcm .
next , the ellman auxiliary was cleaved using conc hcl ( 0.133 ml , 5.43 
 mmol , 6.0 equiv ) . after removing solvent ,
after washing the solid 
 3 with fresh et2o , the remaining et2o 
 was decanted off , yielding the title 7 compound as tan 
 solid ( 110 mg , 44.4% from 7 ) .
h nmr ( 500 
 mhz , cd3od )  7.68 ( d , j = 1.8 hz , 
 1h ) , 7.44 ( dd , j = 8.3 , 1.8 hz , 1h ) , 7.37 ( d , j = 8.3 hz , 1h ) , 7.317.24 ( m , 4h ) , 7.227.17 
 ( m , 1h ) , 4.77 ( t , j = 6.0 hz , 1h ) , 4.06 ( s , 2h ) , 
 3.73 ( ddd , j = 12.4 , 9.0 , 3.1 hz , 1h ) , 3.65 ( ddd , j = 13.0 , 7.8 , 3.3 hz , 1h ) , 2.61 ( dddd , j14.8 , 9.0 , 5.7 , 
 3.3 hz , 1h ) , 2.38 ( dddd , j = 14.8 , 7.8 , 6.4 , 3.1 
 hz , 1h ) .
8 was synthesized following 
 a modified version of general procedure d using 7 ( 110 
 mg , 0.40 mmol , 1.0 equiv ) . after coupling to diboc - dmt ,
residue was 
 not deprotected with tfa . instead , the crude residue was purified 
 by semipreparative hplc and lyophilized to yield the title compound 8 ( 287 mg , quant ) .
esi - ms 630.3 [ m + h ] and esi - ms 652.3 [ m + na ] .
first acetone was degassed for 30 min , then flooded with ar for 
 an additional 30 min . to a flame - dried round - bottom flask equipped 
 with a stir bar with ar atmosphere
was added sodium iodide ( 25 mg , 
 0.16 mmol , 2.0 equiv ) and 2-chloroacetamide ( 15 mg , 0.16 mmol , 2.0 
 equiv ) , which was then placed back under vacuum .
next , 10 ml of degassed 
 acetone was added to the reaction vessel via cannula and reaction 
 stirred for 10 min . meanwhile , to another flame - dried round - bottom 
 flask equipped with a stirbar was added 8 , degassed acetone , 
 followed by dipea ( 0.021 ml , 0.12 mmol , 1.5 equiv ) , and the solution 
 stirred for 10 min .
next , the contents of the round - bottom flask containing 
 the sodium iodide and 2- chloroacetamide was transferred to the flask 
 containing 8 via cannula .
the reaction stirred stirred 
 at room temp for 24 h. the next day , the solvent was removed under 
 reduced pressure to yield the crude product ( 13 mg ) that was taken 
 ahead to next step ( formation of 4k ) without further 
 purification , isolation , or characterization .
esi - ms 687.3 [ m + h ] and esi - ms 709.3 [ m 
 + na ] .
no h or c nmr data acquired . to synthesize 91 , 8 ( 214 mg
, 0.34 mmol , 1.0 equiv ) and k2co3 ( 94 mg , 0.68 mmol , 2.0 equiv ) were placed in 
 a round - bottom flask and the atmosphere was evacuated then the reaction 
 vessel was flooded with ar .
next , bromo ethyl acetate 
 ( 0.377 ml , 3.4 mmol , 10 equiv ) was added to the reaction vessel , which 
 was equipped with a condenser and placed in an 80 c oil bath . 
 after 6 h ,
after 24 h , the solvent was removed under reduced pressure and 
 crude residue was resuspended in dcm and di h2o and the 
 layers separated .
the organic layer was washed with 5% citric acid 
 solution ( 1  20 ml ) and then with brine ( 1  20 ml ) and 
 dried over mgso4 to yield the crude residue of 91 .
esi - ms 716.3 [ m + h ] and esi - ms 738.3 [ m + na ] .
the residue was carried 
 forward ( to synthesize 4l and 4 m ) without 
 further purification , isolation , or characterization .
9 m was synthesized by treating crude 9l ( 107 mg , 0.15 
 mmol , 1.0 equiv ) with lioh ( excess ) in etoh at 60 c for 2.5 
 h. reaction was monitored for loss of starting material by analytical 
 hplc ( gradient a ) : starting material retention time = 72.4 ) .
the residue was 
 carried forward ( 4 m ) without further purification , isolation , 
 or characterization .
11 was synthesized by first degassing a solution of 3:1 acetone : di 
 h2o for 1 h , then ar was bubbled through solution for 1 
 h to ensure removal and displacement of ambient oxygen .
intermediate 10 ( 1.0 equiv ) , 2-naphthylboronic acid ( 2.0 equiv ) , k2co3 ( 3.0 equiv ) , and pd(dppf)cl2 ( 0.1 
 equiv ) were added to a microwave tube , and the tube was placed under 
 vacuum for 15 min , then flooded with ar .
roughly 12 ml of 
 the 3:1 acetone : di h2o solution was added to a tube via 
 syringe , then the tube was placed in microwave for 3060 min 
 with a maximum power of 300 w and a maximum temperature of 100 c 
 with the  powermax  option enabled . once the microwave 
 reaction was complete , reaction mixture was filtered through a pad 
 of celite to remove palladium and solvents were removed under reduced 
 pressure to yield a crude brown residue which was purified using silica 
 gel chromatography to obtain the title compound 11 as 
 an off - white solid .
h nmr ( 500 mhz , cdcl3 )  7.90 ( d , j = 2.2 hz , 1h ) , 7.817.73 
 ( m , 3h ) , 7.69 ( d , j = 8.7 hz , 1h ) , 7.63 ( s , 1h ) , 
 7.497.40 ( m , 2h ) , 7.35 ( dt , j = 8.6 , 1.8 
 hz , 1h ) , 7.30 ( dt , j = 8.4 , 1.5 hz , 1h ) , 4.164.10 
 ( m , 4h ) , 2.782.71 ( m , 2h ) , 1.54 ( d , j = 1.3 
 hz , 9h ) .
c nmr ( 126 mhz , cdcl3 )  194.26 , 
 152.75 , 142.47 , 137.89 , 136.72 , 134.71 , 133.58 , 132.13 , 128.24 , 127.61 , 
 127.54 , 127.35 , 127.31 , 127.25 , 127.08 , 126.78 , 126.05 , 125.46 , 124.82 , 
 123.87 , 82.12 , 44.28 , 41.33 , 38.99 , 28.29 .
12 was synthesized 
 by deprotecting 11 with a 1:1 mixture of tfa : dcm .
after 
 stirring at rt for 1 h , solvent was removed under reduced pressure 
 to yield the crude final product , which was then purified using silica 
 gel chromatography to yield the title compound 12 as 
 a yellow solid .
note that not all crude product was purified , so a 
 yield was not calculated .
h nmr ( 500 mhz , cdcl3 )  7.847.72 ( m , 4h ) , 7.62 ( s , 1h ) , 7.497.38 
 ( m , 2h ) , 7.337.24 ( m , 1h ) , 7.16 ( ddd , j = 
 8.3 , 4.8 , 1.9 hz , 1h ) , 6.65 ( ddd , j = 15.4 , 8.3 , 
 2.4 hz , 1h ) , 4.03 ( s , 1h ) , 3.593.53 ( m , 2h ) , 2.772.66 
 ( m , 2h ) , 2.25 ( s , 1h ) .
c nmr ( 126 mhz , cdcl3 )  193.66 , 138.64 , 136.34 , 136.11 , 133.56 , 132.05 , 130.95 , 
 128.11 , 127.58 , 127.51 , 127.40 , 127.36 , 127.20 , 126.87 , 125.95 , 125.31 , 
 119.67 , 119.42 , 116.41 , 116.22 , 42.57 , 42.41 , 41.14 , 38.08 .
13c was 
 synthesized according to general procedure a using 12 ( 150 mg , 0.52 mmol , 1.0 equiv ) and excess propionic anhydride .
once 
 complete , excess anhydride was removed and the crude residue was purified 
 using silica gel chromatography to yield the title compound 13c as a clear , pale - orange oil ( 156 mg , 87.2% ) .
h nmr ( 500 mhz , cdcl3 )  7.91 ( s , 1h ) , 7.817.73 
 ( m , 3h ) , 7.63 ( s , 1h ) , 7.477.32 ( m , 4h ) , 7.28 ( dd , j = 8.4 , 1.9 hz , 1h ) , 4.224.15 ( m , 2h ) , 4.12 ( s , 
 2h ) , 2.74 ( td , j = 6.3 , 1.9 hz , 2h ) , 2.612.52 
 ( m , 2h ) , 1.21 ( t , 3h ) .
c nmr ( 126 mhz , cdcl3 )  194.08 , 172.83 , 142.08 , 138.43 , 137.45 , 134.74 , 134.52 , 
 133.42 , 132.01 , 128.23 , 127.51 , 127.49 , 127.41 , 127.17 , 127.03 , 126.03 , 
 125.92 , 125.45 , 124.30 , 123.97 , 43.69 , 41.27 , 39.43 , 27.80 , 9.71 .
13d was 
 synthesized according to general procedure a using 12 ( 155 mg , 0.54 mmol , 1.0 equiv ) and excess butyric anhydride .
once 
 complete , excess anhydride was removed and the crude residue was purified 
 using silica gel chromatography to yield the title compound 13d as a clear , slightly dark - red oil ( 112 mg , 58.0% ) .
h nmr ( 500 mhz , cdcl3 )  7.94 ( s , 1h ) , 7.857.76 
 ( m , 3h ) , 7.66 ( s , 1h ) , 7.517.39 ( m , 4h ) , 7.32 ( d , j = 8.5 hz , 1h ) , 4.20 ( t , j = 6.3 hz , 2h ) , 
 4.15 ( s , 2h ) , 2.76 ( t , j = 5.9 hz , 2h ) , 2.602.49 
 ( m , 2h ) , 1.75 ( h , j = 7.7 hz , 2h ) , 0.97 ( t , j = 7.6 hz , 3h ) .
c nmr ( 126 mhz , cdcl3 )  194.00 , 171.99 , 142.10 , 138.38 , 137.42 , 134.69 , 134.47 , 
 133.42 , 132.01 , 128.21 , 127.49 , 127.47 , 127.39 , 127.16 , 127.02 , 126.00 , 
 125.90 , 125.42 , 124.32 , 124.00 , 43.71 , 41.25 , 39.50 , 36.28 , 18.94 , 
 13.66 .
13e was synthesized according to general procedure b using 12 ( 150 mg , 0.52 mmol , 1.0 equiv ) and methylchloroformate ( 0.200 ml , 
 2.3 mmol , 2.3 equiv ) .
the reaction stirred overnight ; once complete , 
 solvent was removed and the crude residue was purified using silica 
 gel chromatography to yield the title compound 109 as 
 a clear oil ( 65 mg , 36.1% ) .
additionally , not all product 
 was purified as the rf values for the starting material and product 
 were similar and separation was difficult .
h nmr ( 500 
 mhz , cdcl3 )  7.92 ( d , j = 2.2 hz , 
 1h ) , 7.827.74 ( m , 3h ) , 7.72 ( d , j = 8.6 hz , 
 1h ) , 7.64 ( s , 1h ) , 7.487.41 ( m , 2h ) , 7.38 ( dd , j = 8.6 , 2.3 hz , 1h ) , 7.30 ( dd , j = 8.4 , 1.8 hz , 
 1h ) , 4.18 ( t , j = 6.3 hz , 2h ) , 4.13 ( s , 2h ) , 3.84 
 ( s , 3h ) , 2.77 ( t , j = 6.3 hz , 2h ) .
c 
 nmr ( 126 mhz , cdcl3 )  193.81 , 154.23 , 141.81 , 137.73 , 
 137.18 , 134.88 , 133.49 , 132.05 , 128.21 , 127.55 , 127.47 , 127.26 , 127.23 , 
 127.03 , 126.01 , 125.42 , 124.81 , 123.63 , 53.32 , 44.39 , 41.26 , 38.81 .
14c was synthesized according 
 to general procedure c using 13c ( 156 mg , 0.45 mmol , 
 1.0 equiv ) , ( r)-2-methylpropane-2-sulfinamide ( 165 
 mg , 1.36 mmol , 3.0 equiv ) , and ti(oet)4 ( 0.57 ml , 2.7 mmol , 
 6.0 equiv ) to form the ( r)-tert - butanesulfinyl 
 imine intermediate in situ .
once sufficient ketone was converted into 
 imine intermediate ( after 48 h ) , the reaction mixture was transferred 
 via cannula to a round - bottom flask containing nabh4 ( 103 
 mg , 2.7 mmol , 6.0 equiv ) and 20 ml of thf in a xylenes dry ice bath , 
 after addition , the solution was stirred at room temperature for 3 
 h before being quenched with meoh .
once resultant solid was removed , 
 crude residue was purified using silica gel chromatography to yield 
 the title compound 14c as a clear , colorless oil ( 148 
 mg , 72.5% ) .
no h or c nmr data acquired . 
 instead , product taken ahead to next reaction ( formation of 15c ) without any further isolation , purification , or characterization .
14d was synthesized according 
 to general procedure c using 13d ( 112 mg , 0.31 mmol , 
 1.0 equiv ) , ( r)-2-methylpropane-2-sulfinamide ( 114 
 mg , 0.94 mmol , 3.0 equiv ) , and ti(oet)4 ( 0.39 ml , 1.9 mmol , 
 6.0 equiv ) to form the ( r)-tert - butanesulfinyl 
 imine intermediate in situ . once sufficient ketone was converted into 
 imine intermediate ( after 48 h ) , the reaction mixture was transferred 
 via cannula to a round - bottom flask containing nabh4 ( 71 
 mg , 1.9 mmol , 6.0 equiv ) and 20 ml of thf in a xylenes a dry ice bath ; 
 after addition , the solution was stirred at room temperature for 3 
 h before being quenched with meoh .
once resultant solid was removed , 
 crude residue was purified using silica gel chromatography to yield 
 the title compound 14d as a clear , colorless oil ( 54 
 mg , 37.2% ) .
h nmr ( 500 mhz , cdcl3 )  
 7.78 ( q , j = 7.7 hz , 3h ) , 7.66 ( s , 1h ) , 7.44 ( pd , j = 7.1 , 3.4 hz , 2h ) , 7.33 ( dd , j = 6.4 , 
 2.1 hz , 2h ) , 7.14 ( dt , j = 8.2 , 2.0 hz , 1h ) , 4.52 
 ( t , j = 4.4 hz , 1h ) , 4.11 ( s , 2h ) , 3.89 ( dt , j = 12.2 , 5.5 hz , 1h ) , 3.74 ( ddd , j = 13.6 , 
 9.8 , 6.2 hz , 1h ) , 3.32 ( d , j = 3.3 hz , 1h ) , 2.522.39 
 ( m , 2h ) , 2.20 ( dh , j = 15.3 , 7.4 , 5.6 hz , 1h ) , 2.04 
 ( dq , j = 12.7 , 4.6 , 3.9 hz , 1h ) , 1.741.61 
 ( m , 2h ) , 1.17 ( d , j = 1.9 hz , 9h ) , 0.91 ( t , j = 7.5 hz , 3h ) .
c nmr ( 126 mhz , cdcl3 )  187.72 , 172.68 , 138.03 , 136.64 , 133.57 , 132.10 , 128.79 , 
 128.72 , 128.57 , 128.21 , 127.58 , 127.52 , 127.46 , 127.10 , 126.03 , 125.43 , 
 124.85 , 124.60 , 55.73 , 50.96 , 41.52 , 39.95 , 36.69 , 30.68 , 22.51 , 19.14 , 
 13.88 .
14e was synthesized according to general procedure c using 13e ( 65 mg , 0.19 mmol , 1.0 equiv ) , ( r)-2-methylpropane-2-sulfinamide 
 ( 68 mg , 0.57 mmol , 3.0 equiv ) , and ti(oet)4 ( 0.24 ml , 1.1 
 mmol , 6.0 equiv ) to form the ( r)-tert - butanesulfinyl imine intermediate in situ . once sufficient ketone 
 was converted into imine intermediate ( after 48 h ) , the reaction mixture 
 was transferred via cannula to a round - bottom flask containing nabh4 ( 43 mg , 1.1 mmol , 6.0 equiv ) and 20 ml of thf in a xylenes 
 dry ice bath , after addition , the solution was stirred at room temperature 
 for 3 h before being quenched with meoh .
once resultant solid was 
 removed , crude residue was purified using silica gel chromatography 
 to yield the title compound 14e as a clear , colorless 
 oil ( 34 mg , 40.0% ) .
h nmr ( 500 mhz , cdcl3 ) 
  7.817.71 ( m , 4h ) , 7.64 ( d , j = 5.2 
 hz , 1h ) , 7.487.39 ( m , 2h ) , 7.347.26 ( m , 2h ) , 7.13 
 ( ddd , j = 8.5 , 6.0 , 2.2 hz , 1h ) , 4.55 ( h , j = 3.7 hz , 1h ) , 4.09 ( s , 2h ) , 4.053.96 ( m , 1h ) , 
 3.78 ( s , 3h ) , 3.693.59 ( m , 1h ) , 3.33 ( q , j = 2.7 hz , 1h ) , 2.18 ( ddt , j = 12.8 , 10.4 , 4.3 hz , 
 1h ) , 2.061.93 ( m , 1h ) , 1.18 ( s , 9h ) .
c nmr ( 126 
 mhz , cdcl3 )  154.94 , 138.25 , 136.95 , 136.16 , 133.53 , 
 132.02 , 129.23 , 128.84 , 128.65 , 128.08 , 127.52 , 127.48 , 127.43 , 126.96 , 
 125.92 , 125.30 , 123.70 , 55.60 , 52.95 , 50.35 , 41.33 , 40.28 , 29.49 , 
 22.49 .
15c was synthesized following 
 general procedure d using 14c ( 148 mg , 0.33 mmol , 1.0 
 equiv ) and conc hcl ( 6 drops ) . after removing solvent ,
after washing 
 the solid 3 with fresh et2o , the remaining et2o was decanted off , yielding a white solid amine hydrochloride 
 salt ( 126 mg ) .
the synthesis was completed by following general procedure 
 f with newly formed ( r ) amine intermediate ( 60 mg , 
 0.16 mmol , 1.0 equiv ) to yield crude product which was purified by 
 semipreparative hplc and lyophilized to yield the title compound 15c as a tfa salt ( 15 mg , 14.7% ) . note that not all crude 
 product was purified .
h nmr ( 500 mhz , cdcl3 )  7.76 ( ddd , j = 21.9 , 8.2 , 3.1 hz , 3h ) , 
 7.60 ( d , j = 3.2 hz , 1h ) , 7.477.37 ( m , 2h ) , 
 7.28 ( dt , j = 8.6 , 2.2 hz , 1h ) , 7.19 ( d , j = 3.5 hz , 1h ) , 7.157.09 ( m , 1h ) , 6.51 ( d , j = 3.4 hz , 2h ) , 4.93 ( p , j = 5.2 hz , 1h ) , 
 4.08 ( d , j = 3.4 hz , 2h ) , 3.86 ( dt , j = 11.2 , 4.3 hz , 1h ) , 3.78 ( s , 1h ) , 3.24 ( td , j = 
 12.6 , 11.2 , 3.5 hz , 1h ) , 3.15 ( s , 1h ) , 3.03 ( dt , j = 13.9 , 4.5 hz , 1h ) , 2.582.41 ( m , 2h ) , 2.26 ( d , j = 3.6 hz , 6h ) , 1.85 ( dtd , j = 13.5 , 8.7 , 
 4.7 hz , 1h ) , 1.45 ( s , 1h ) , 1.11 ( td , j = 7.1 , 3.2 
 hz , 3h ) .
c nmr ( 126 mhz , cd3od )  175.85 , 
 169.20 , 157.47 , 140.07 , 139.86 , 137.62 , 135.07 , 133.59 , 129.47 , 129.07 , 
 128.58 , 128.46 , 128.38 , 127.87 , 127.09 , 126.46 , 125.87 , 123.24 , 116.45 , 
 53.49 , 47.11 , 42.27 , 31.95 , 31.29 , 28.92 , 20.42 , 10.06 .
esi - ms 536.3 [ m + h ] and 558.3 
 [ m + na ] .
15d was synthesized following 
 general procedure d using 14d ( 34 mg , 0.076 mmol , 1.0 
 equiv ) and conc hcl ( 3 drops ) . after removing solvent ,
after washing 
 the solid 3 with fresh et2o , the remaining et2o was decanted off , yielding a white solid amine hydrochloride 
 salt ( 46 mg ) .
the synthesis was completed by following general procedure 
 f with newly formed ( r ) amine intermediate ( 46 mg , 
 0.12 mmol , 1.0 equiv ) to yield crude product which was purified by 
 semipreparative hplc and lyophilized to yield the title compound 15d as a tfa salt ( 12 mg , 15.6% ) . note that not all crude 
 product was purified .
esi - ms 550.3 [ m + h ] and 572.3 [ m + na ] .
15e was synthesized following general procedure d using 14e ( 34 mg , 0.076 mmol , 1.0 equiv ) and conc hcl ( 3 drops ) . after removing 
 solvent ,
after washing the solid 3 with fresh et2o , the 
 remaining et2o was decanted off , yielding a white solid 
 amine hydrochloride salt ( 19 mg ) .
the synthesis was completed by following 
 general procedure f with newly formed ( r ) amine intermediate 
 ( 19 mg , 0.0.16 mmol , 1.0 equiv ) to yield crude product which was purified 
 by semipreparative hplc and lyophilized to yield the title compound 15e as a tfa salt ( 13 mg , 40.6% ) .
h nmr ( 500 mhz , cd3od ) 
  7.78 ( d , j = 7.9 hz , 1h ) , 7.73 ( dd , j = 8.3 , 2.9 hz , 2h ) , 7.67 ( d , j = 8.5 
 hz , 1h ) , 7.59 ( s , 1h ) , 7.42 ( dt , j = 8.9 , 6.2 hz , 
 2h ) , 7.27 ( dt , j = 8.4 , 1.7 hz , 1h ) , 7.12 ( s , 1h ) , 
 7.09 ( dt , j = 8.6 , 1.8 hz , 1h ) , 6.50 ( s , 2h ) , 4.96 
 ( t , j = 5.1 hz , 1h ) , 4.04 ( s , 2h ) , 3.873.80 
 ( m , 1h ) , 3.75 ( d , j = 1.5 hz , 4h ) , 3.283.19 
 ( m , 1h ) , 3.00 ( dt , j = 12.6 , 8.3 hz , 2h ) , 2.26 ( d , j = 1.5 hz , 7h ) , 1.78 ( ddt , j = 14.7 , 9.9 , 
 4.7 hz , 1h ) , 1.50 ( ddd , j = 13.4 , 8.6 , 4.8 hz , 1h ) .
c nmr ( 126 mhz , cd3od )  168.96 , 157.48 , 
 156.61 , 139.99 , 138.03 , 137.48 , 135.07 , 133.57 , 130.31 , 129.72 , 129.02 , 
 128.78 , 128.57 , 128.46 , 128.39 , 127.80 , 127.05 , 126.41 , 124.63 , 123.19 , 
 116.43 , 111.40 , 53.54 , 53.41 , 47.06 , 42.32 , 42.14 , 31.92 , 30.30 , 20.44 . 

esi - ms 538.3 [ m + h ] and 560.3 [ m + na ] .
all tissue culture reagents were purchased from 
 gibco life sciences ( grand island , ny , u.s . ) .
c6-rat glioma cells 
 stably transfected with a rat  ( c6-mor ) or rat  ( c6-dor ) 
 opioid receptor and chinese hamster ovary 
 ( cho ) cells stably expressing a human  ( cho - kor ) opioid receptor were used for all in vitro assays .
cells were 
 grown to confluence at 37 c in 5% co2 in dulbecco s 
 modified eagle medium ( dmem ) containing 10% fetal bovine serum and 
 5% penicillin / streptomycin .
membranes were prepared by washing confluent 
 cells three times with ice - cold phosphate buffered saline ( 0.9% nacl , 
 0.61 mm na2hpo4 , 0.38 mm kh2po4 , ph 7.4 ) .
cells were detached from the plates by incubation 
 in warm harvesting buffer ( 20 mm hepes , 150 mm nacl , 0.68 mm edta , 
 ph 7.4 ) and pelleted by centrifugation at 1600 rpm for 3 min .
the 
 cell pellet was suspended in ice - cold 50 mm tris - hcl buffer , ph 7.4 , 
 and homogenized with a tissue tearor ( biospec products , inc . ,
bartlesville , 
 ok , u.s . ) for 20 s. the homogenate was centrifuged at 15000 rpm for 
 20 min at 4 c .
the pellet was rehomogenized in 50 mm tris - hcl 
 with a tissue tearor for 10 s , followed by recentrifugation .
the final 
 pellet was resuspended in 50 mm tris - hcl and frozen in aliquots at 
 80 c .
protein concentration was determined via a bca protein 
 assay ( thermo scientific pierce , waltham , ma , u.s . ) using bovine serum 
 albumin as the standard .
radiolabeled 
 compounds were purchased from perkinelmer ( waltham , ma , u.s . ) .
opioid 
 ligand binding assays were performed by competitive displacement of 
 0.2 nm [ h]diprenorphine ( 250 ci , 1.85 tbq / mmol ) 
 by the peptidomimetic from membrane preparations containing opioid 
 receptors as described above .
the assay mixture , containing membranes 
 ( 20 g protein / tube ) in 50 mm tris - hcl buffer ( ph 7.4 ) , [ h]diprenorphine , and various concentrations of test peptidomimetic , 
 was incubated at room temperature for 1 h to allow binding to reach 
 equilibrium .
the samples were rapidly filtered through whatman gf / c 
 filters using a brandel harvester ( brandel , gaithersburg , md , u.s . ) 
 and washed five times with 50 mm tris - hcl buffer . bound radioactivity 
 on dried filters
was determined by liquid scintillation counting , 
 after saturation with ecolume liquid scintillation cocktail , in a 
 wallac 1450 microbeta ( perkinelmer , waltham , ma , u.s . ) .
the results presented 
 are the mean  standard error ( se\m ) from at least three separate 
 assays performed in duplicate .
ki ( nm ) 
 values were calculated using nonlinear regression analysis to fit 
 a logistic equation to the competition data using graphpad prism , 
 version 6.0c , for mac os x ( graphpad software inc . , la jolla , ca ) .
agonist 
 stimulation of [ s]guanosine 5-o-[-thio]triphosphate ( [ 5s]gtps , 1250 ci , 
 46.2 tbq / mmol ) binding to g - protein was measured as described previously .
briefly , membranes ( 1020 g of 
 protein / tube ) were incubated 1 h at 25 c in gtps buffer 
 ( 50 mm tris - hcl , 100 mm nacl , 5 mm mgcl2 , ph 7.4 ) containing 
 0.1 nm [ s]gtps , 30 m guanosine diphosphate 
 ( gdp ) , and varying concentrations of test peptidomimetic .
g - protein 
 activation following receptor stimulation of [ s]gtps 
 ( % stimulation ) with peptidomimetic was compared with 10 m 
 of the standard compounds [ d - ala2,n - mephe4,gly - ol]enkephalin 
 ( damgo ) at mor , d - pen2,5-enkephalin ( dpdpe ) at dor , or u69,593 
 at kor .
the reaction was terminated by vacuum filtration of gf / c filters 
 that were washed 10 times with gtps buffer . bound radioactivity 
 was measured as described above .
the results are presented as the 
 mean  standard error ( sem ) from at least three separate assays 
 performed in duplicate ; potency ( ec50 ( nm ) ) and % stimulation 
 were determined using nonlinear regression analysis with graphpad 
 prism , as above .
adult 
 male c57bl/6 mice weighing between 20 and 30 g at 816 weeks 
 old purchased from harlan ( indianapolis , in ) .
mice were group - housed 
 and had free access to food and water at all times .
experiments were 
 conducted in the housing room , which was maintained on a 12 h light / dark 
 cycle ( with lights on at 0700 ) ; experiments were conducted during 
 the light cycle . each mouse was used only once .
studies were performed 
 in accordance with the university of michigan committee on the use 
 and care of animals and the guide for the care and use of laboratory 
 animals ( national research council , 2011 publication ) .
three - dimensional ( 3d ) models of opioid receptors 
 in the inactive conformation were produced as previously described using x - ray structures of the mouse mor ( pdb 
 i d : 4dkl ) , the human dor ( pdb i d : 4n6h ) , and the human kor ( pdb i d : 4djh ) as structural 
 templates .
the recently obtained crystal structure of mouse mor in 
 the active conformation ( pdb i d : 5c1 m ) was used 
 as a template for homology modeling of active conformations of dor 
 and kor .
structures of receptor loops in the active state were kept 
 similar to those in the crystal structures of corresponding receptors 
 in the inactive state .
n - termini of dor ( residues 3345 ) and 
 kor ( residues 4557 ) were modeled using the structure of mor 
 n - terminus in the active conformation with a few adjustments that 
 transformed the central bulge ( residues 5357 ) into the extended 
 conformation of the polypeptide chain . such modifications increased 
 the size of the binding pocket and allowed docking of relatively bulky 
 tetra- and pentapeptides and peptidomimetics without hindrances with 
 the n - terminus .
structures of peptidomimetic ligands were generated 
 using the 3d - builder application of quanta ( accelrys , inc . ) , followed 
 by conformational search included in the program package .
ligand conformations 
 that demonstrated the best superposition of aromatic substituents 
 of the thq core with the pharmacophore elements ( tyr and 
 phe ) of receptor - bound conformations of cyclic tetrapeptides were selected and minimized with charmm implemented in quanta ( adopted - basis 
 newton  raphson method , 100 steps ,  = 10 ) .
low energy 
 conformations ( within 2 kcal / mol ) were manually positioned inside 
 the receptor binding cavity to reproduce the binding modes of cyclic 
 tetrapeptides .
the docking pose of each ligand was subsequently refined 
 using the solid docking module of quanta .
the receptor  ligand 
 complexes were then minimized with charmm ( adopted - basis newton  raphson 
 method , 30 steps ,  = 10 ) . models of opioid ligand 
 n - acetylation of the tetrahydroquinoline ( thq ) core of a 
 series of -opioid receptor ( mor ) agonist/-opioid receptor 
 ( dor ) antagonist ligands increases dor affinity , resulting in ligands 
 with balanced mor and dor affinities . 
 we report a series of n - substituted thq analogues that incorporate various carbonyl - containing 
 moieties to maintain dor affinity and define the steric and electronic 
 requirements of the binding pocket across the opioid receptors . 
 4h produced in vivo antinociception ( ip ) for 1 h at 10 mg / kg . 
Introduction
Results
Discussion and Conclusions
Experimental
Section