Glucose Sensitivity of Boronic Acid Block Copolymers at Physiological ph

Size: px

Start display at page:

Download "Glucose Sensitivity of Boronic Acid Block Copolymers at Physiological ph"

Terence Fisher
5 years ago
Views:

1 Eltroni Supporting Informtion for Gluos Snsitivity of Boroni Ai Blok Copolymrs t Physiologil p Dshish Roy n Brnt S. Sumrlin* * Dprtmnt of Chmistry n th Cntr for Drug Disovry, Dsign, n Dlivry Southrn Mthoist Univrsity 3215 Dnil Avnu, Dlls, Txs , USA Mtrils. 2-Doylsulfnylthioronylsulfnyl-2-mthylpropioni i (DMP) hin trnsfr gnt (CTA) ws prpr s prviously rport. 1,-Dimthylrylmi (DMA, Fluk, 98%) ws pss through smll olumn of si lumin prior to polymriztion. 2,2 - Azoisisoutyronitril (AIB, Sigm, 98%) ws rrystlliz from thnol. Anhyrous 1,2- iminothn (TCI), i-trt-utyliront (Alf Asr, 97+%), 4-roxynznoroni i (Boron Molulr), pinol (Aros rgnis, 99%), ryloyl hlori (Alf Asr, 96%), oxlylhlori (Alf Asr), trifluroti i (EMD), 1,3,5-trioxn (Aros rgnis, 99.5%),,-imthylformmi (DMF) (Alrih 99.9%), ttrhyrofurn (TF) (Aros rgnis, 99.9%), ithyl thr (Fishr), 1,4-ioxn (Alf Asr, 99+%, st. with ppm BT), soium hyroxi (Alf Asr, 97%), hyrohlori i (Alrih 37%), polystyrn-support oroni i rsin (Lnstr), molulr sivs (4 Å, Mllinkrot), trithylmin

2 (Mllinkrot), thyl tt (Fishr), soium hlori (EMD), nhyrous soium sulft (EMD), nhyrous soium ront, (99%, Pfltz & Bur), soium hlori (Mllinkrot), ihloromthn (Fluk, ri ovr 4 ngstroms molulr sivs for us), D-gluos (nhyrous, grnulr) (Mllinkrot), imthylsulfoxi- 6 (DMS- 6, Cmrig Isotop, 99.9% D), mthnol- 4 (Cmrig Isotop, 99.8% D) n CDCl 3 (Cmrig Isotop, 99% D) wr us s riv, unlss othrwis not. All othr rgnts wr otin from VWR n us without furthr purifition, unlss othrwis not. Instrumnts n nlyss. Siz xlusion hromtogrphy (SEC) ws onut in 0.05 M LiBr in DMF t 55 C with flow rt of 1.0 ml/min (Visotk VE 2001 GPCmx; Columns: gur + two Polymr Lortoris PolrGl-M mix olumns: molulr wight rng 0 60 x 10 3 n x 10 3 g/mol, rsptivly). Dttion onsist of Visotk VE 3580 rfrtiv inx ttor oprting t 660 nm n Visotk Mol 270 Sris Pltform, onsisting of lsr light sttring ttor (oprting t 3 mw, 670 nm with ttion ngls of 7 n 90 ) n four pillry visomtr. Molulr wights wr trmin y th tripl ttion mtho. Th n/ of th lok opolymrs ws trmin in situ y ssuming 100% mss rovry. 1 MR sptrosopy ws onut with Brukr Avn 400 sptromtr oprting t 400 Mz or JEL Dlt 500 oprting t 500 Mz. Dynmi light sttring (DLS) ws onut with Mlvrn Ztsizr no-s quipp with 4 mw, 633 nm - lsr n n Avlnh photoio ttor t n ngl of 173.

3 Monomr synthsis 2 C 2 1,4-ioxn rt, 48 h 2 Cl nhyrous 2 C 3 DCM, 0-5 o C, 3 h rt, 16 h B DCM rt, 48 h i) TFA, DCM, rt, 2 h ii) vport TFA/DCM, TF, Et 3, 2 h C Cl Cl CCl 2 B DCM rt, 16 h B istill TF Et o C, 2 h rt, 22 h B (1) Shm S1 Synthsis of monomr -(2-ryloylmino-thyl)-4-(4,4,5,5-ttrmthyl- [1,3,2]ioxoroln-2-yl)-nzmi (AEBB-Pin, 1) Synthsis of trt-utyl--(2-minothyl)ront. 1,2-Diminothn (28 g, 31 ml, 0.46 mol) ws issolv in 500 ml thr-nk roun-ottom flsk ontining 150 ml 1,4- ioxn. To this stirr solution, i-trt-utyliront (13.2 g, mol), pr-issolv in 1,4-ioxn (150 ml), ws slowly t room tmprtur ovr prio of 5 h. Aftr 48 h, th solution ws filtr to rmov th whit pripitt form uring th rtion. Exss 1,2- iminothn n 1,4-ioxn ws vport unr ru prssur. Aition of wtr (250 ml) l to th pripittion of is(, -t-utyloxyronyl)-1,2-iminothn, whih ws

4 rmov vi filtrtion. Th quous solution ws sturt with soium hlori, n th rsulting turi solution ws rptly xtrt with ihloromthn (100 6 ml). Th rsulting orgni lyr ws ri ovr nhyrous soium sulft n filtr. Susqunt rmovl of solvnt rotry vportion provi olorlss oil in 89% (8.6 g) yil. 1 MR (CDCl 3, 400 Mz, ppm): 5.22 (s, 1, Bo), (m, 2, C 2 ), (m, 2, C 2 2 ), 1.34 (s, 9, 3C 3 ), 1.18 (s, 2, 2 ). 13 C MR (CDCl 3, 125 Mz, ppm): 156 (C=), 78.5 (CM 3 ), 43 (C 2 ), 41.6 (C 2 2 ), 28 (C 3 ). Synthsis of -trt-utyloxyronyl- -ryl-1,2-iminothn. trt-butyl--(2- minothyl)ront (8.5 g, mol) ws issolv in ihloromthn (100 ml), n th solution ws ool to 0 C using n i-slt th. Anhyrous soium ront (5.6 g, mol) ws, n th solution ws stirr for n hour. Aryloyl hlori (6.0 g, mol), pr-issolv in ihloromthn ( 30 ml), ws rop wis t 0 C ovr prio of 3 h. Th rtion ws furthr ontinu for 16 h t room tmprtur. DCM (50 ml) ws to th rtion mixtur, n th soli mtrils wr filtr. Th orgni lyr ws xtrt with sturt soium iront (50 2 ml), wtr (50 3 ml), n sturt rin solution (50 3 ml). Anhyrous soium sulft ws to ry th orgni lyr, n th solvnt ws rmov rotry vportion. A soli whit prout ws otin in 77% (8.7 g) yil. 1 MR (CDCl 3, 400 Mz, ppm): 6.62 (s, 1, C), 5.1 (s, 1, Bo), 6.19 (, J= z, 1, C 2 =), 5.56 (, J= z, 1, C 2 =), 6.05 (, J=10.2, z, C=), (m, 2, CC 2 ), (m, 2, C 2 Bo), 1.36 (s, 9, 3C 3 ). 13 C MR (CDCl 3, 125 Mz): δ (C=), (C=, Bo), (=C), (C 2 =), (t-c), (CC 2 ), (C 2 Bo), (3C 3 ).

5 Synthsis of -(2-minothyl)rylmi: To stirr solution of -trt-utyloxyronyl- - ryl-1,2-iminothn (2.0 g, 9.3 mmol) in 10 ml DCM, n xss of trifluroti i (TFA, 30 ml) ws. Th rtion ws ontinu for 2 h t room tmprtur. Th solvnt ws rmov y rotry vportion to giv th TFA slt of -(2-minothyl)rylmi. Th prout ws kpt unr nitrogn for ing to th nxt rtion. Synthsis of 4-roxyphnyloroni i, pinol str (CPBAE): 4-Croxyphnyl oroni i (CPBA) (8.0 g, mol) ws suspn into DCM (100 ml) in 250 ml roun ottom flsk. Exss pinol (6.8 g, mol) n molulr sivs wr to th flsk. Th rtion ws ontinu t room tmprtur for 48 h. Th rtion solution ws filtr to rmov molulr sivs n ll soli impuritis. Th solvnt ws rmov in vuo to giv soli powr tht ws furthr purifi vi rpt wshing with hxn (5 50 ml) to rmov ny xss pinol. Th prout ws otin in 48% yil (5.7 g) ftr rying unr vuum. 1 MR (DMS- 6, 400 Mz, ppm): (s, 1, C), 7.95 (, J=8.2 z, 2, C 6 4 ), (, J=8.2 z, 2, C 6 4 ), 1.30 (s, 12, 4C 3 ). Synthsis of 4-(hloroformyl) phnyloroni i, pinol str: To solution of CPBAE (2.2 g, 8.9 mmol) in DCM (35 ml) in roun ottom flsk fitt with rying tu of lium hlori ws n xss of oxlyl hlori (5.6 g, 3.8 ml, mol). Th rtion ontnts wr llow to stir t room tmprtur for 16 h. Th solvnt n xss oxlyl hlori wr rmov in vuo n kpt unr nitrogn in n i-slt th for using irtly for th nxt rtion with -(2-minothyl)rylmi.

6 Synthsis of -(2-ryloylmino-thyl)-4-(4,4,5,5-ttrmthyl-[1,3,2]ioxoroln-2-yl)- nzmi (1): Th TFA slt of -(2-minothyl)rylmi ws issolv in istill TF (15 ml). Trithylmin (15 ml) ws to this solution t 0 C to otin th fr s ryloyl min monomr, -(2-minothyl)rylmi. Th solution ws stirr for n hour for ommning rop wis ition of ol solution of 4-(hloroformyl) phnyloroni i, pinol str (pr-issolv in 15 ml istill TF). Th solution ws stirr t 0 C for th first 2 h n t room tmprtur for th nxt 22 h. Th solvnt ws rmov unr vuum to giv soli prout tht ws susquntly suspn in istill wtr (100 ml). A fw rops of Cl (2 ) wr until th p of th solution ws 4.0. Th rsulting solution ws xtrt with thyl tt (5 100 ml). Th thyl tt lyr ws furthr wsh ( 4) with istill wtr n susquntly ri ovr nhyrous soium sulft. Th solvnt ws rmov rotry vportion to giv soli whit prout (1). Th ru prout ws purifi vi rrystlliztion from thyl tt (1.9 g, Yil= 59%). 1 MR (DMS- 6, 400 Mz, ppm): 8.62 (s, 1, C 2 CC 6 4 ), 8.25 (s, 1, CC 2 ), 7.85 (, J=8.24 z, 2, C 6 4 ), 7.74 (, J=8.24 z, 2, C 6 4 ), 6.21 (, J=9.98, z, C=), 6.08 (, J= z, 1, C 2 =), 5.59 (, J= 9.96 z, 1, C 2 =), 1.31 (s, 12, 4C 3 ). 13 C MR (DMS- 6, 125 Mz, ppm): , (2 C=, C), (2C, C 6 4 ), (2C, C 6 4 ), (=C), (C 2 =), (C (C 3 ) 2 ), (C 2 CC 6 4 ), (CC 2 ), (4C 3 ). RAFT homopolymriztions of 1. An xmpl RAFT polymriztion of 1 ws s follows. 1 (0.5 g, 1.46 mmol), DMP (5.3 mg, mmol), AIB (1.2 mg, 7.30 µmol), DMF (5 ml) n trioxn (44 mg) (s n intrnl stnr) wr sl in 20 ml vil. Th ontnts wr

7 oxygnt y uling with nitrogn for pproximtly 30 min for th vil ws pl in prht rtion lok t 70 C. Smpls wr rmov prioilly y syring to trmin molulr wight n polyisprsity inx y SEC n monomr onvrsion y 1 MR sptrosopy. Synthsis of PDMA mrocta (4). An xmpl RAFT polymriztion of DMA ws s follows. DMA (2.2 g, mol), DMP (54 mg, 0.15 mmol), AIB (1.2 mg, 7.3 µmol), DMF (8.8 ml), n trioxn (100 mg, intrnl stnr) wr sl in vil quipp with mgnti stir r n purg with nitrogn for 30 min. Th rtion vil ws pl in prht rtion lok t 70 C, n th polymriztion ws qunh ftr 90 min y rmoving th polymriztion from ht n xposing th rtion solution to ir. Aftr onntrtion unr vuum, th polymr ws pripitt into ol thr ( 2) n susquntly ilys ginst istill wtr for 48 h using 3,500 molulr wight ut off mmrn. Th polymr (4) ws lyophiliz n ri unr vuum t 40 C for 6 h to yil yllow powr (M n = 8,900 g/mol, M w /M n = 1.16). RAFT lok opolymriztion of 1 with PDMA mrocta. An xmpl RAFT lok opolymriztion of th oroni i monomr 1 with PDMA mrocta ws onut s follows. 1 (0.21 g, 0.60 mmol), PDMA mrocta (M n = 8,900 g/mol, M w /M n = 1.16) (54 mg, 6.0 µmol), trioxn (10 mg, intrnl stnr), AIB [0.49 mg, 3.0 µmol), n DMF (2 ml) wr sl in vil n purg with nitrogn for 30 min. Th rtion vil ws pl in prht rtion lok t 70 C, n th polymriztion ws qunh ftr 14 h y rmoving th polymriztion from ht n xposing th rtion solution to ir. Th rsulting PDMA--

8 poly(-(2-ryloylmino-thyl)-4-(4,4,5,5-ttrmthyl-[1,3,2]ioxoroln-2-yl)-nzmi) (PDMA--PAEBB-Pin, 5) (38% onvrsion; M n = 19,400 g/mol; M w /M n =1.28) ws isolt y pripittion ( 3) from TF into thr n rying unr vuum. Dprottion of lok opolymr PDMA--PAEBB-Pin (5). An xmpl prottion ws onut s follows. Blok opolymr 5 (0.2 g, mmol, M n = 19,400 g/mol, M w /M n = 1.28) ws stirr with n xss mount of polystyrn-support oroni i rsin (2.9 g, ~3.0 mmol B() 2 /g) in 50 ml roun ottom flsk ontining 98% tonitril n 2 vol. % trifluoroti i (30 ml). Th rtion mixtur ws rflux for 24 h. Th solvnt ws rmov unr ru prssur, n 20 ml DMF/wtr (95%/5%, v/v) ws to th prout. Th solution ws filtr to rovr th polystyrn-support oroni i rsin, n th solvnt ws vport. Th rsulting soli mtril ws issolv in 2% (w/v) quous n ilyz ginst th sm si solution follow y ilysis ginst ioniz wtr. Th lyophiliztion of th solution provi whit powr of PDMA--poly(-(2- ryloylmino-thyl)-4-orono-nzmi) (PDMA--PAEBB) (6). Aquous solution stuis of PDMA--PAEBB. An xmpl milliztion prour for PDMA--PAEBB ws onut s follows. Dioniz wtr (6.2 ml) ws ropwis to ry smpl of PDMA--PAEBB (3.2 mg) to otin 0.05% (w/v) solution. For p-rspons stuis, th solution ws sonit for 30 min n titrt with 0.05 M solution filtr with 0.45 µm syring filtr. During titrtion, th mount of solution, th p, n th hyroynmi imtr (from DLS) wr ror. Aftr th formtion of unimr t high p (p=8.5), th p of th solution ws lowr y titrting with 1 Cl to vrify th rssmly

9 (unimr to mill formtion) of th PDMA--PAEBB. For gluos-rspons stuis, th solution ws sonit for 30 min, n th p of th solution ws just to 7.4 y ing solution of 0.05 M tht h n filtr with 0.45 µm syring filtr. An pproprit mount of gluos (60-95 mm) ws to th smpl using 2 M gluos stok solution filtr with 0.45 µm syring filtr. Upon ition of gluos, th solution m trnsprnt, n th hyroynmi imtrs wr ror for n ftr gluos ition ppm Figur S1. 1 MR sptrum of trt-utyl--(2-minothyl)ront.

10 g f f + g ppm) Figur S2. 1 MR sptrum of -trt-utyloxyronyl- -ryl-1,2-iminothn.

11 +f f 2 f CDCl 3 DMS (ppm) Figur S3. 1 MR sptrum of -(2-minothyl)rylmi.

12 C B DMS ppm Figur S4. 1 MR sptrum of 4-roxyphnyloroni i, pinol str.

13 h g f 2 + ( +) B DMS f h+g ppm Figur S5. 1 MR sptrum of -(2-ryloylmino-thyl)-4-(4,4,5,5-ttrmthyl- [1,3,2]ioxoroln-2-yl)-nzmi (1).

14 f B f ppm Figur S6. DEPT MR sptrum of -(2-ryloylmino-thyl)-4-(4,4,5,5-ttrmthyl- [1,3,2]ioxoroln-2-yl)-nzmi (1).

15 C S SC C n S B g h f 2 + (+) DMS +f (ppm) Figur S7. 1 MR sptrum of poly(-(2-ryloylmino-thyl)-4-(4,4,5,5-ttrmthyl- [1,3,2]ioxoroln-2-yl)-nzmi) (3).

16 C m B S SC n C S 2 mthnol ppm Figur S8. 1 MR sptrum of PDMA--PAEBB (6). (1) Li, J. T.; Fill, D.; Sh, R. Mromoluls 2002, 35,

Intramolecular Quaternization as Folding Strategy for the Synthesis of Catalytically Active Imidazolium-based Single Chain Nanoparticles

Intramolecular Quaternization as Folding Strategy for the Synthesis of Catalytically Active Imidazolium-based Single Chain Nanoparticles Supporting Informtion Intrmolulr Qutrniztion s Foling Strtgy for th Synthsis of Ctlytilly Ativ Imizolium-s Singl Chin Nnoprtils Romin Lmrt,, Ann-Lur Wirotius,, n Dnil Tton, * Lortoir Chimi s Polymèrs Orgniqus