Supporting Information Small molecule inhibitors that discriminate between protein arginine - methyltransferases PRMT1 and CARM1 James Dowden,* a Richard A. Pike, a Richard V. Parry, b Wei Hong, a Usama A. Muhsen, a and Stephen G. Ward b a School of Chemistry, University Park, University of ottingham, ottingham, G7 2RD, UK. Fax: +44 () 115 95113564; Tel: +44 () 115 9513566; E-mail: james.dowden@nottingham.ac.uk b Department of Pharmacy and Pharmacology, University of Bath, Claverton Down, Bath, BA2 7AY, UK. Chemistry: Intermediates not explicitly described within the manuscript are detailed with characterisation in the same order below. These protected compounds are numbered P-o., where o. corresponds to the number of the denuded compound in the manuscript. General procedure for first reductive amination: method 1 5'-Amino-5'-deoxy-2',3'-,-(1-methylethylidene)adenosine 8 1 (1.2 mol eq.) and aldehyde 5, 2 6, 3 7 4 or 12 5 (1. mol eq.) were suspended in ClCH 2 CH 2 Cl (5 ml) at room temperature under Ar and the mixture gently heated with vigorous stirring to obtain a solution. abh(ac) 3 (1.4 mol eq.) was slowly added portionwise and the reaction left for 2-4 hrs at room temperature under Ar. The reaction was then quenched by addition of saturated aqueous a 2 C 3 solution (5 ml). The mixture was extracted with CH 2 Cl 2 (3 x 5 ml) and the combined organic layers were dried over MgS 4 and concentrated in vacuo to yield the crude product. The product was purified by column chromatography on silica gel eluting with Me: CH 2 Cl 2, 5:95, unless specified otherwise. General procedure for second reductive amination: method 2 Secondary amines 9, 3 1 or 11 (1. mol eq.) and aldehydes 12, 5 13, 14, or 15 (1.1 mol eq.) with MgS 4 (1 mol eq.) were used, then as method 1 above. General procedure for deprotection: method 3 Intermediates from method 2 above were dissolved in TFA (4 ml) and water (.1 ml) and stirred overnight. The mixture was concentrated in vacuo, dissolved in water (5 ml) and washed with EtAc (2 x 5 ml). The aqueous layer was concentrated in vacuo to ~ 1mL then applied to a column of Amberlite IRA- 4 (Cl - ) ion exchange resin, eluted with water, and fractions containing product freeze dried to give the hydrochloride salts 16-18, 3 19-24.
General procedure for Swern oxidations xalyl chloride (1.2 mol eq) was added into a solution of dry DMS (2.4 mol eq.) in dry CH 2 Cl 2 (15 ml) at -78 C and stirred for 3 mins under Ar. A solution of alcohol (1 mol eq.) in dry CH 2 Cl 2 (15 ml) was slowly added via cannula and the reaction stirred for 3 min. Dry Et 3 (5 mol eq.) was slowly added and the reaction stirred for 3 mins then slowly warmed to r.t. The reaction was quenched by the addition of H 2 (15 ml) and the mixture extracted with CH 2 Cl 2 (3 x 25 ml). The combined organic extracts were dried over MgS 4, filtered and concentrated. The product was purified by column chromatography on silica gel. 2,3-di(tert-Butoxycarbonyl)-1-(4-oxobutyl)guanidine (13) Swern oxidation of 2,3-di(tert-butoxycarbonyl)-1-(4-hydroxybutyl)-guanidine. 6 Elution with EtAc: petroleum ether (4/6) 2:8 gave the product as a clear oil (1.32 g, 88%), max / cm -1 3328 (w), 3285 (w), 37 (w), 2984 (w), 2935 (w), 1722 (s), 1636 (s), 1617 (s), 1417 (m), 1394 (w), 1369 (s), 1332 (s), 1289 (w), 1253 (w), 124 (w), 1158 (s), 1136 (s), 152 (w), 127 (w); 1 H MR H (4 MHz, CDCl 3 ) 9.8 (1H, s, CH), 8.37 (1H, br s, C()HCH 2 ), 3.45 (2H, app. q, J = 7.1, HCH 2 ), 2.54 (2H, t, J = 7., CH 2 CH), 1.92 (2H, quintet, J = 7.1, CH 2 CH 2 CH 2 ), 1.51 (18H, s, C(CH 3 ) 3 ); 13 C MR C (1 MHz, CDCl 3 ) 21.2 (CH), 163.5 (C), 156.3 (C), 153.3 (C), 83.2 (C), 79.3 (C), 41.1 (CH 2 ), 4. (CH 2 ), 28.3 (CH 3 ), 28.2 (CH 3 ), 21.7 (CH 2 ); m/z (ESI) 362.2 (1%, M + Me + H + ), 33.2 (24%, M + H + ) Found 33.214 C 15 H 28 3 5 requires 33.223. 2,3-di(tert-Butoxycarbonyl)-1-(5-oxopentyl)-guanidine (14) Swern oxidation of 2,3-di(tert-butoxycarbonyl)-1-(5-hydroxypentyl)-guanidine. 6 Elution with EtAc: petroleum ether (4/6) 3:7 gave the product as a white solid (.98 g, 81%), mp 14-16 C; max / cm -1 3329 (w), 2986 (w), 2945 (w), 1722 (s), 1681 (m), 1635 (s), 1476 (w), 1455 (w), 1417 (s), 1394 (m), 1368 (s), 1334 (s), 128 (m), 1254 (m), 116 (s), 1136 (s), 154 (w), 989 (w); 1 H MR H (4 MHz, CDCl 3 ) 9.78 (1H, d, J = 1.4, CH), 8.34 (1H, br s, C()HCH 2 ), 3.45 (2H, app. q, J = 7., HCH 2 ), 2.5 (2H, td, J = 7.2, 1.4, CH 2 CH), 1.75-1.6 (4H, m, CH 2 CH 2 CH 2 CH 2 ), 1.51 (9H, s, C(CH 3 ) 3 ), 1.5 (9H, s, C(CH 3 ) 3 ); 13 C MR C (1 MHz, CDCl 3 ) 22. (CH), 163.6 (C), 156.2 (C), 153.3 (C), 83.1 (C), 79.3 (C), 43.4 (CH 2 ), 4.4 (CH 2 ), 28.5 (CH 2 ), 28.3 (CH 3 ), 28.1 (CH 3 ), 19.3 (CH 2 ); m/z (ESI) 376.2 (39%, M + Me + H + ), 344.2 (1%, M + H + ) Found 344.2181 C 16 H 3 3 5 requires 344.218.
1-[4-(formyl)phenyl]-2,3-bis-(tert-butoxycarbonyl)guanidine (15) Swern oxidation of 1-[4-(hydroxymethyl)phenyl]([2,3-bis-(tertbutoxycarbonyl)guanidine. 7 Elution with EtAc: petroleum ether (4/6) 1:9 gave the product as a white solid (1.545 g, 5%), mp 133 C (dec.) (CH 2 Cl 2 ); max / cm -1 3147 (w), 2986 (w), 2936 (w), 283 (w), 2744 (w), 1722 (m), 1698 (m), 1637 (s), 159 (s), 1558 (s), 1476 (w), 1455 (w), 1413 (s), 137 (m), 1342 (m), 137 (s); 1 H MR H (4 MHz, CDCl 3 ) 11.75 (1H, br s, H), 1.67 (1H, s, H), 9.94 (1H, s, CH), 7.82-7.9 (4H, m, ArH), 1.55 (18H, s, C(CH 3 ) 3 ); 13 C MR C (1 MHz, CDCl 3 ) 191. (C), 153.2 (C), 142.6 (Cx2), 132.4 (C), 13.9 (CH), 121.5 (CH), 84.3 (C), 8.2 (C), 28.1 (CH 3 ); m/z (ESI) 749.3 (1%, 2M + a + ), 386.2 (39%, M + a + ), 364.2 (38%, M + H + ) Found 364.1867 C 18 H 26 3 5 requires 364.1867. 5'-[[(3S)-4-(tert-butoxy)-3-[[(tert-butoxy)carbonyl]amino]-4-oxobutyl]amino]- 5'-deoxy-2',3'--(1-methylethylidene)-adenosine (9) 2 2 Method 1. Column chromatography on silica gel, elution with EtAc:Me, 25:1, gave secondary amine 9 as a white solid (.9 g, 73%), mp 87-89 C; [ ] 23 D -34.6 (c 1.7 in CHCl 3 ); max / cm -1 3525 (w), 3414 (w), 298 (m), 2933 (m), 177 (s), 1632 (s), 1589 (s), 1328 (s), 1151 (s), 177 (m); 1 H MR H (4 MHz; CDCl 3 ) 8.33 (1H, s, ArH), 7.93 (1H, s, ArH), 6.6 (2H, s, adenosine-h 2 ), 6. (1H, d, J = 8.4, HBoc), 5.98 (1H, d, J = 3.6, 1'-H), 5.5 (1H, dd, J = 6.2, 3.6, 2'-H), 5.8 (1H, dd, J = 6.2, 3.2, 3'-H), 4.41-4.37 (1H, m, 4'-H), 4.34-4.25 (1H, m, CHCH 2 CH 2 ), 2.96 (1H, dd, J = 12.7, 3.8, 5'-CH a H b ), 2.8 (1H, dd, J = 12.7, 5.2, 5'-CH a H b ), 2.77-2.72 (1H, m, CH a H b H), 2.7-2.59 (1H, m, CH a H b H), 1.9-1.77 (2H, m, CHCH 2 CH 2 ) 1.62 (3H, s, CH 3 ), 1.51 (9H, s, C(CH 3 ) 3 ), 1.4 (3H, s, CH 3 ), 1.38 (s, 9H, C(CH 3 ) 3 ); 13 C MR C (1 MHz; CDCl 3 ) 171.9 (C), 155.7 (C), 155.6 (C), 153.1 (CH), 149.5 (C), 139.9 (CH), 12.4 (C), 114.6 (C), 91.1 (CH), 85. (CH), 83. (CH), 82.2 (CH), 81.7 (C), 79.4 (C), 53. (CH), 51.4 (CH 2 ), 46.3 (CH 2 ), 32.3 (CH 2 ), 28.3 (CH 3 ), 28. (CH 3 ), 27.4 (CH 3 ), 25.5 (CH 3 ); m/z (ESI) 564.3 (1%, M + H + ) Found 564.3111 C 26 H 42 7 7 requires 564.3146.
5'-[3-[2,3-di-(tert-butoxycarbonyl)guanidino]propyl]amino]-5'-deoxy-2',3'-- (1-methylethylidene)-adenosine Method 1. Elution Me: CH 2 Cl 2, from 1:9 to 15:85 gave the product as a white solid (.516 g, 43%), mp 13-14 C (CH 2 Cl 2 ); [ ] 19 D -21.1 (c 1. in CHCl 3 ); max / cm -1 2986 (m), 2936 (w), 1721 (m), 1632 (s), 1588 (s), 1473 (w), 1456 (w), 1417 (m), 1393 (w), 1369 (s), 133 (s); 1 H MR H (4 MHz; CDCl 3 ) 8.46 (1H, br s, H), 8.32 (1H, s, ArH), 7.95 (1H, s, ArH), 6.1 (1H, d, J = 3.1, 1'- H), 5.98 (2H, br s, adenosine-h 2 ), 5.47 (1H, dd, J = 6.4, 3.1, 2'-H), 5.2 (1H, dd, J = 6.4, 3.3, 3'-H), 4.42-4.36 (1H, m, 4'-H), 3.51-3.44 (2H, m, C()HCH 2 ), 2.94-2.81 (2H, m, 5'-CH 2 ), 2.72-2.59 (2H, m, CH 2 H), 1.7 (2H, quin, J = 6.8, CH 2 CH 2 H), 1.6 (3H, s, CH 3 ), 1.48 (18H, s, C(CH 3 ) 3 ), 1.38 (3H, s, CH 3 ); 13 C MR C (1 MHz; CDCl 3 ) 163.5 (C), 156.2 (C), 155.6 (C), 153.2 (C), 153.1 (CH), 149.4 (C), 139.9 (CH), 12.3 (C), 114.5 (C), 9.8 (CH), 58.6 (CH), 83.5 (CH), 82.9 (C), 82.4 (CH), 79.2 (C), 51.3 (CH 2 ), 47.1 (CH 2 ), 38.9 (CH 2 ), 29.1 (CH 2 ), 28.2 (CH 3 ), 28. (CH 3 ), 27.2 (CH 3 ), 25.4 (CH 3 ); m/z (ESI) 66.3 (M + H +,1%) Found 66.336 C 27 H 44 9 7 requires 66.3358; Calc. for C 27 H 43 9 7.
5'-deoxy-5'-[[(3S)-4-(tert-butoxy)-3-[[(1,1 dimethylethoxy)carbonyl]amino]-4- oxobutyl] [(3-(tert-butoxycarbonyl)aminopropyl]amino]-2',3'--(1- methylethylidene)-adenosine (P19) t 2 2 Method 2. gave a white solid (.447 g, 71%), mp 86-87 C (CH 2 Cl 2 ); [ ] 23 D -19.5 (c 1.5 in CHCl 3 ); max / cm -1 311 (w), 2984 (m), 2936 (w), 1712 (s), 1633 (s), 164 (w), 155 (m), 1369 (s), 1248 (m), 1158 (s); 1 H MR H (4 MHz; CDCl 3 ) 8.33 (1H, s, ArH), 7.93 (1H, s, ArH), 6.7 (1H, d, J = 1.5, 1'-H), 5.87 (2H, br s, H 2 ), 5.48-5.56 (2H, m, 2'-H and H), 5.-5.7 (1H, br m, 3'-H), 4.32-4.36 (1H, m, 4'-H), 4.15-4.25 (1H, br m, CHCH 2 ), 3.5-3.21 (2H, m, HCH 2 ), 2.74 (1H, dd, J = 13., 7.5, 5'-CH a H b ), 2.54-2.63 (1H, m, CHCH 2 CH a H b ), 2.44-2.54 (2H, m, 5'- CH a H b and HCH 2 CH 2 CH a H b ), 2.31-2.42 (2H, m, CHCH 2 CH a H b and HCH 2 CH 2 CH a H b ), 1.89-2. (1H, br m, CHCH a H b ), 1.65-1.72 (1H, m, CHCH a H b ), 1.63 (3H, s, CH 3 ), 1.5-1.58 (2H, m, HCH 2 CH 2 CH 2 ), 1.45 (9H, s, C(CH 3 ) 3 ), 1.44 (18H, s, C(CH 3 ) 3 ), 1.41 (3H, s, CH 3 ); 13 C MR C (1 MHz; CDCl 3 ) 171.9 (C), 156.2 (C), 155.6 (C), 153. (CH), 149.1 (C), 14.2 (CH), 12.3 (C), 114.4 (C), 9.9 (CH), 85.7 (CH), 84. (CH), 83.5 (CH), 81.7 (C), 79.6 (C), 78.8 (C), 77.2 (C), 56.1 (CH 2 ), 52.8 (CH), 51.6 (CH 2 ), 5.5 (CH 2 ), 38.2 (CH 2 ), 29.3 (CH 2 ), 28.5 (CH 3 ), 28.4 (CH 3 ), 28. (CH 3 ), 27.1 (CH 3 ), 26.9 (CH 2 ), 25.5 (CH 3 ); m/z (ESI) 721.4 (1%, M + H + ) Found 721.4247 C 34 H 57 8 9 requires 721.4243;
5'-deoxy-5'-[[(3S)-4-(tert-butoxy)-3-[[(1,1 dimethylethoxy)carbonyl]amino]-4- oxobutyl][[4-[2,3-bis-(tert-butoxycarbonyl)]guanidinophenyl]methyl]amino]- 2',3'--(1-methylethylidene)-adenosine (P2) Method 2 gave the product as a white solid (.57 g, 71%), mp 119-121 C (CH 2 Cl 2 ); [ ] 26 D -1.3 (c 1. in CHCl 3 ); max / cm -1 312 (s), 2985 (m), 2937 (m), 1714 (s), 1633 (s), 166 (s), 1564 (m), 156 (m), 1476 (m), 1456 (m), 1416 (s), 1394 (s), 137 (m), 134 (s), 133 (m), 124 (s), 1154 (s); 1 H MR H (4 MHz; CDCl 3 ) 11.65 (1H, s, H), 1.26 (1H, s, H), 8.24 (1H, s, ArH), 7.8 (1H, s, ArH), 7.46 (2H, d, J = 7.8, ArH), 7.16 (2H, d, J = 7.8, ArH), 6.5 (1H, br m, 1'-H), 5.69 (2H, s, adenosine-h 2 ), 5.62 (1H, br m, H), 5.37 (1H, d, J = 5.4, 2'-H), 4.79-4.87 (1H, br m, 3'-H), 4.3-4.43 (1H, br m, 4'-H), 4.1-4.25 (1H, br m, CHCH 2 ), 3.62 (1H, d, J = 13.4, ArCH a H b ), 3.43 (1H, d, J = 13.4, ArCH a H b ), 2.46-2.74 (4H, m, 5'-CH 2 and CHCH 2 CH 2 ), 1.93-2.7 (1H, m, CHCH a H b ), 1.68-1.87 (1H, m, CHCH a H b ), 1.61 (3H, s, CH 3 ), 1.55 (9H, s, (CH 3 ) 3 ), 1.49 (9H, s, (CH 3 ) 3 ), 1.45 (9H, s, C(CH 3 ) 3 ), 1.43 (9H, s, C(CH 3 ) 3 ), 1.38 (3H, s, CH 3 ); 13 C MR C (1 MHz; CDCl 3 ) 171.7 (C), 163.6 (C), 155.4 (C), 153.5 (C x 2), 153.3 (C), 153.1 (CH), 149.2 (C), 139.9 (CH), 135.7 (C), 134.9 (C), 129.4 (CH), 122.2 (CH), 12.3 (C), 114.3 (C), 9.7 (CH), 85.3 (CH), 83.8 (CH), 83.7 (C), 83.4 (CH), 81.6 (C), 79.6 (C), 79.4 (C), 58.3 (CH 2 ), 55.5 (CH 2 ), 53. (CH), 5.7 (CH 2 ), 29.3 (CH 2 ), 28.4 (CH 3 ), 28.2 (CH 3 ), 28.1 (CH 3 ), 28. (CH 3 ), 27.1 (CH 3 ), 25.4 (CH 3 ); m/z (ESI) 911.5 (1%, M + H + ) Found 911.4965 C 44 H 67 1 11 requires 911.4985;
5'-deoxy-5'-[[(4-(tert-butoxy)-4-oxobutyl][3-[2,3-di-(tert- butoxycarbonyl)guanidino]propyl]amino]-2',3'--(1-methylethylidene)- adenosine (P21) Method 2. White solid (.468 g, 79%), mp 81-83 C (CH 2 Cl 2 ); [ ] 26 D -13.6 (c 1. in CHCl 3 ); max / cm -1 2984 (m), 172 (s), 1633 (s), 1588 (s), 1475 (m), 1456 (w), 1418 (m), 1369 (s), 1331 (s), 129 (m), 124 (m), 1157 (s), 1135 (s); 1 H MR H (4 MHz; CDCl 3 ) 11.5 (1H, s, H), 8.44 (1H, br m, H), 8.36 (1H, s, ArH), 7.97 (1H, s, ArH), 6.5 (1H, d, J = 2.1, 1'-H), 5.73 (2H, s, adenosine-h 2 ), 5.52 (1H, dd, J = 6.4, 2.1, 2'-H), 4.97 (1H, dd, J = 6.4, 3.2, 3'-H), 4.34 (1H, app. td, J = 6.8, 3.2, 4'-H), 3.33-3.5 (2H, m, HCH 2 ), 2.74 (1H, dd, J = 13.6, 6.8, 5'-CH a H b ), 2.58 (1H, dd, J = 13.6, 6.8, 5'-CH a H b ), 2.39-2.55 (4H, m, CCH 2 CH 2 CH 2 and HCH 2 CH 2 CH 2 ), 2.16-2.23 (2H, m, CCH 2 CH 2 CH 2 ), 1.61-1.73 (4H, m, CCH 2 CH 2 CH 2 and HCH 2 CH 2 CH 2 ), 1.6 (3H, s, CH 3 ), 1.5 (18H, s, C(CH 3 ) 3 ), 1.43 (9H, s, C(CH 3 ) 3 ), 1.39 (3H, s, CH 3 ); 13 C MR C (1 MHz; CDCl 3 ) 172.8 (C), 163.6 (C), 156.1 (C), 155.5 (C), 153.2 (C), 153.1 (CH), 149.3 (C), 14.2 (CH), 12.3 (C), 114.3 (C), 91. (CH), 85.6 (CH), 83.7 (CH), 83.4 (CH), 82.9 (C), 8.1 (C), 79.2 (C), 56.1 (CH 2 ), 53.5 (CH 2 ), 52. (CH 2 ), 39.3 (CH 2 ), 33.1 (CH 2 ), 28.3 (CH 3 ), 28.1 (CH 3 ), 28.8 (CH 3 ), 27.1 (CH 3 ), 26.5 (CH 2 ), 25.3 (CH 3 ), 22. (CH 2 ); m/z (ESI) 748.4 (1%, M + H + ) Found 748.4351 C 35 H 58 9 9 requires 748.4352; Calc. for C 35 H 57 9 9.
5'-[3-[2,3-di-(tert-butoxycarbonyl)guanidino]propyl][(3-(tert- butoxycarbonyl)aminopropyl]amino]-5'-deoxy-2',3'--(1-methylethylidene)- adenosine (P22) 2 Method 2. Column chromatography on silica gel (Me:CH 2 Cl 2 5:95 to 1:9) gave the product as a white solid (.187 g, 35%), mp 87-88 C (CH 2 Cl 2 ); [ ] 23 D - 8.2 (c 1.5 in CHCl 3 ); max / cm -1 3414 (w), 3328 (w), 2984 (m), 1712 (s), 1632 (s), 156 (w), 1473 (w), 1417 (m), 1393 (w), 1368 (s), 133 (m), 1289 (m), 1248 (m), 1158 (s), 1136 (s); 1 H MR H (4 MHz; CDCl 3 ) 11.49 (1H, br s, H), 8.35 (1H, br s, H), 8.31 (1H, s, ArH), 7.95 (1H, s, ArH), 6.9 (2H, br s, H 2 ), 6.5 (1H, d, J = 1.9, 1'-H), 5.92 (1H, br m, 2'-H), 5.55 (1H, br m, H), 5.5 (1H, dd, 6.4, 3.2, 3'-H), 4.82 (1H, m, 4'-H), 3.41 (2H, td, J = 6.9, 5.4, CCH 2 ), 2.99-3.17 (2H, m, C()HCH 2 ), 2.65 (1H, dd, J = 13.3, 7., 5'-CH a H b ), 2.43-2.55 (2H, m, 5'-CH a H b and CCH 2 CH 2 CH 2 ), 2.34-2.43 (2H, m, C()HCH 2 CH 2 CH 2 ), 1.59 (3H, s, CH 3 ), 1.47 (18H, s, C(CH 3 ) 3 x 2), 1.43-1.45 (4H, m, CCH 2 CH 2 CH 2 and C()HCH 2 CH 2 CH 2 ), 1.41 (9H, s, C(CH 3 ) 3 ), 1.38 (3H, s, CH 3 ); 13 C MR C (1 MHz; CDCl 3 ) 163.5 (C), 156. (C), 155.7 (C), 153.2 (C), 153. (CH), 149.1 (C), 14.1 (CH), 12.2 (C), 114.2 (C), 9.8 (CH), 85.7 (CH), 83.8 (CH), 83.4 (CH), 82.9 (C), 79.1 (C x 2), 78.7 (C), 56.1 (CH 2 ), 53.4 (CH 2 ), 51.5 (CH 2 ), 39. (CH 2 ), 38.5 (CH 2 ), 28.4 (CH 3 ), 28.2 (CH 3 ), 28. (CH 3 ), 27. (CH 3 ), 26.5 (CH 2 ), 26.2 (CH 2 ), 25.3 (CH 3 ); m/z (ESI) 763 (1%, M + H + ) Found 763.445 C 35 H 59 1 9 requires 763.4461.
5'-[di(3-(tert-butoxycarbonyl)aminopropyl]amino]-5'-deoxy-2',3'--(1- methylethylidene)-adenosine (P23) Method 2. White solid (.421 g, 75%), mp 74-75 C (CH 2 Cl 2 ); [ ] 21 D -11.8 (c 1. in CH 2 Cl 2 ); max / cm -1 3413 (m), 2982 (s), 2819 (w), 176 (s), 1631 (s), 1588 (m), 158 (s), 1473 (m), 1423 (w), 1392 (w), 1368 (s), 133 (w), 1248 (m), 1167 (s); 1 H MR H (4 MHz; CDCl 3 ) 8.32 (1H, s, ArH), 7.94 (1H, s, ArH), 6.8 (1H, d, J = 1.6, 1'-H), 6.2 (2H, s, adenosine-h 2 ), 5.49-5.57 (1H, m, 2'-H), 5.16-5.23 (2H, s, H), 5.1-5.8 (1H, m, 3'-H), 4.31-4.41 (1H, m, 4'-H), 3.2-3.18 (4H, m, HCH 2 CH 2 CH 2 ), 2.58-2.69 (1H, m, 5'-CH a H b ), 2.5-2.58 (1H, m, 5'-CH a H b ), 2.34-2.5 (4H, m, HCH 2 CH 2 CH 2 ), 1.62 (3H, s, CH 3 ), 1.47-1.59 (4H, m, HCH 2 CH 2 CH 2 ), 1.43 (18H, s, C(CH 3 ) 3 ), 1.41 (3H, s, CH 3 ); 13 C MR C (1 MHz; CDCl 3 ) 156.1 (C), 155.7 (C), 153. (CH), 149.2 (C), 14.3 (CH), 12.3 (C), 114.3 (C), 9.9 (CH), 85.9 (CH), 84. (CH), 83.4 (CH), 78.9 (C), 56.2 (CH 2 ), 51.6 (CH 2 ), 38.6 (CH 2 ), 28.7 (CH 3 ), 28.5 (CH 3 ), 27. (CH 3 ), 26.8 (CH 2 ), 25.4 (CH 3 ); m/z (ESI) 621.4 (1%, M + H + ) Found 621.3726 C 29 H 49 8 7 requires 621.3719.
5'-5'-deoxy-[di[2,3-di-(tert-butoxycarbonyl)guanidino]propyl]amino]-2',3'-- (1-methylethylidene)-adenosine (P24) 2 Method 2. Elution with Me:CH 2 Cl 2 5:95 to 1:9 gave the product as a white solid (.181 g, 3%), mp 117-119 C (CH 2 Cl 2 ); [ ] 21 D -7.62 (c 1. in CH 2 Cl 2 ); max / cm -1 3414 (w), 2985 (m), 2938 (m), 1721 (s), 1632 (s), 1587 (s), 1474 (m), 1455 (m), 1417 (s), 1394 (m), 1369 (s), 133 (s), 1157 (s), 1135 (s),; 1 H MR H (4 MHz; CDCl 3 ) 11.49 (2H, s, (C()HC) 2 ), 8.42 (2H, br m, (C()HCH 2 ) 2 ), 8.34 (1H, s, ArH), 7.99 (1H, s, ArH), 6.5 (1H, d, J = 2.1, 1'-H), 5.76 (2H, br s, adenosine-h 2 ), 5.54 (1H, dd, J = 6.4, 2.1, 2'-H), 4.97 (1H, dd, J = 6.4, 3.3, 3'-H), 4.34 (1H, app. td, J = 6.7, 3.3, 4'-H), 3.35-3.46 (4H, m, (HCH 2 CH 2 CH 2 ) 2 ), 2.72 (1H, dd, J = 13.6, 6.7, 5'-CH a H b ), 2.43-2.61 (5H, m, 5'-CH a H b and (HCH 2 CH 2 CH 2 ) 2 ), 1.6-1.72 (4H, m, (HCH 2 CH 2 CH 2 ) 2 ), 1.59 (3H, s, CH 3 ), 1.49 (36H, s, C(CH 3 ) 3 x 4), 1.38 (3H, s, CH 3 ); 13 C MR C (1 MHz; CDCl 3 ) 163.6 (C x 2), 156.1 (C), 155.5 (C), 153.2 (CH), 149.3 (C), 14.3 (CH), 12.3 (C), 114.3 (C), 9.9 (CH), 85.5 (CH), 83.6 (CH), 83.4 (CH), 82.9 (C), 79.1 (C), 56. (CH 2 ), 52. (CH 2 ), 39.3 (CH 2 ), 28.3 (CH 3 ), 28. (CH 3 ), 27. (CH 3 ), 26. (CH 2 ), 25.2 (CH 3 ); m/z (ESI) 95.5 (M + H +,1%), Found 95.5196 C 41 H 69 12 11 requires 95.523; References 1. A. M. Reeve, C. A. Townsend, Tetrahedron 1998, 54, 15959. 2. R. M. Werner,.Shokek and J. T. Davis, J. rg. Chem. 1997, 62, 8243. 3. M. Kolb, J. Barth, J.-G. Heydt and M. J. Jung, J. Med. Chem. 1987, 3, 267-72. 4. S. Hourcade, A. Ferdenzi, P. Retailleau, S. Mons and C. Marazano, Eur. J. rg. Chem., 25, 132. 5.. S. Freeman,Y. Tal-Gan, S. Klein, A. Levitzki and C. Gilon, J. rg. Chem. 211, 76, 378. 6. eubert B. J.; Snider B. B. rg. Lett. 23, 5, 765-8. 7. Thuring J. W.; Li H.; Porter. A. Biochemistry 22, 41, 22-13.
H 2 H 2 H H H H 2 1..9.8.7.6.5.4.3.2.1 1.98.2.2 1.3.1.13 2.15 1.26 9.42.1 4.52.4 9.5 9. 8.5 8. 7.5 7. 6.5 6. 5.5 5. 4.5 4. 3.5 3. 2.5 2. 1.5 1..5
H 2 H 2 H H H H 2 1. 71.54 73.5.9.8.7 144.4 143.49 9.22 38.1 24.41.6.5 17.79 148. 149.91 119.29 54.97 5.64.4.3 156.67.2 78.4 5.91 22.58.1 23 22 21 2 19 18 17 16 15 14 13 12 11 1 9 8 7 6 5 4 3 2 1-1 -2
@7 Celcius H 2 H 2 H H H H 2 1..9 144.22 145.29 9.92 78.83 73.78 72.32 38.94 23.54 25.29.8.7 52.28 56.17 52. 51.72.6.5.4.3 171.38.2 157.66 148.85 15.73 119.97.1 22 21 2 19 18 17 16 15 14 13 12 11 1 9 8 7 6 5 4 3 2 1-1 -2-3
This Print journal Date: is 25 The Feb Royal 21 Society 15:46:52 of Chemistry 211 MS Data Review All Plots - 2/25/21 3:46 PM File: c:\varianws\data\dmt\18129\rp116,8 _2-5-37 pm.xms Sample: RP116,8 Scan Range: 1-899 Time Range:.6-3.32 min. kcounts 2.:6.> 1 perator: dmt Date: 12/18/29 2:5 PM RP116,8 _2-5-37 PM.xms TIC 75 5 25 uv 3 2.:6.> 1A RP116,8 _2-5-37 PM.xms uv 2 1-1 -2 1% 5 1 15 2 25 Spectrum 1A BP: 467 (3136=1%), rp116,8 _2-5-37 pm.xms minutes 3.366 min, Scans: 98-1, 2.:6.>, Ion: 1 us, RIC: 99216 467 3136 75% 5% 25% 2 8243 21 4159 23 1657 332 3213 468 649 47 178 % 2 3 4 5 6 m/z
H 2 H 2 H H H 2 H 1..9.8.7.6.5.4 M1(d).3 6.11 6.1 4.44.2.1 4.47 4.4 3.95 3.75 3.69 3.66 3.63 3.46 3.42 3.27 3.26 3.25 3.23 3.7 2.38 2.32 2.28 2.18 1.7 1.53 8.4 2.1 1.11 2.39 1.8 2.35 2.42 2.25 1.12 2.11 9.5 9. 8.5 8. 7.5 7. 6.5 6. 5.5 5. 4.5 4. 3.5 3. 2.5 2. 1.5 1..5
H 2 H 2 H H H H 2 73.1 1..9.8 144.45 9. 71.55 4.4.7 24.5 24.91.6.5.4.3 171.4 156.7 148.9 149.95 143.43 119.25 78.9 51.26 2.33.2 55..1 23 22 21 2 19 18 17 16 15 14 13 12 11 1 9 8 7 6 5 4 3 2 1-1 -2
@7 Celcius H 2 H 2 H H H H 2 73.75 72.35 1..9 144.16 145.39 9.78 78.93 41.22 25.71 25.39 21.24.8 52.33.7 56.7.6.5 54.34 52.25.4.3.2 171.98 148.91 15.82 119.94.1 157.64 22 21 2 19 18 17 16 15 14 13 12 11 1 9 8 7 6 5 4 3 2 1-1 -2-3
This Print journal Date: is 25 The Feb Royal 21 Society 16:45:54 of Chemistry 211 MS Data Review All Plots - 2/25/21 4:45 PM File: c:\varianws\data\dmt\22129\rp124,8 _9-5-32 am.xms Sample: RP124,8 Scan Range: 1-897 Time Range:.7-3.26 min. kcounts 1 2.:6.> perator: dmt Date: 12/22/29 9:5 AM RP124,8 _9-5-32 AM.xms TIC 75 5 25 uv 3 2.:6.> 1A RP124,8 _9-5-32 AM.xms uv 2 1-1 -2 1% 5 1 15 2 25 Spectrum 1A BP: 481 (384=1%), rp124,8 _9-5-32 am.xms minutes 3.171 min, Scans: 92-94, 2.:6.>, Ion: 1 us, RIC: 9341 481 384 75% 5% 25% 214 7227 482 7767 233 2414 483 1929 % 2 3 4 5 6 m/z
H 2 H 2 H H H H 2 1..9.8.7.6.5.4 M1(d) 6.11 6.1 4.44.3.2.1 4.47 4.41 3.96 3.75 3.69 3.61 3.65 3.43 3.23 3.24 3.37 3.2 3.2 2.32 2.19 1.65 8.4 1.45 1.29 2. 1.1 2.39 1.24 2.45 2.422.312.15 1.15 2.33 2.18 9.5 9. 8.5 8. 7.5 7. 6.5 6. 5.5 5. 4.5 4. 3.5 3. 2.5 2. 1.5 1..5
H 2 H 2 H H H H 2 73.1 1..9.8.7 144.45 143.5 9.5 78.15 71.59 4.73 27.32 24.49.6.5 148.8 149.95 22.77.4 171.3 119.25.3.2 156.64 54.94 51.18 22.46.1 23 22 21 2 19 18 17 16 15 14 13 12 11 1 9 8 7 6 5 4 3 2 1-1 -2
@7 Celcius H 2 H 2 H H H H 2 1..9 144.21 145.39 9.81 78.98 73.74 72.38 41.58 28.1 23.53 23.36.8.7 55.98 52.21.6.5.4 54.7 52.9.3.2 171.87 148.89 15.79 119.94.1 22 21 2 19 18 17 16 15 14 13 12 11 1 9 8 7 6 5 4 3 2 1-1 -2-3
This Print journal Date: is 25 The Feb Royal 21 Society 16:52:11 of Chemistry 211 MS Data Review All Plots - 2/25/21 4:52 PM File: c:\varianws\data\dmt\22129\rp125,8 _9-5-44 am.xms Sample: RP125,8 Scan Range: 1-898 Time Range:.5-3.28 min. kcounts 1 2.:6.> perator: dmt Date: 12/22/29 9:5 AM RP125,8 _9-5-44 AM.xms TIC 75 5 25 uv 3 2.:6.> 1A RP125,8 _9-5-44 AM.xms uv 2 1-1 -2 1% 5 1 15 2 25 Spectrum 1A BP: 495 (14617=1%), rp125,8 _9-5-44 am.xms minutes 3.261 min, Scans: 95-97, 2.:6.>, Ion: 1 us, RIC: 9633 495 14617 75% 228 9817 5% 25% 212 3749 211 2375 213 918 24 3627 241 174 453 2176 496 4458 % 2 3 4 5 6 m/z
H 2 H 2 H H 2 1..9.8.7.6.5.4.3.2.1 M1(d) 6.11 6.1 4.76 4.45 4.46 4.43 3.92 3.78 3.91 3.56 3.66 3.69 3.48 3.34 3.33 2.99 3.4 2.97 2.8 2.32 2.17 2.37 8.4 2.7 2.5 1.67.87 2. 2.22 1.93 2.35 1.6 1.93 9.5 9. 8.5 8. 7.5 7. 6.5 6. 5.5 5. 4.5 4. 3.5 3. 2.5 2. 1.5 1..5 -.5
72.99 71.51 1..95 H 2.9.85.8.75 H H 2 H 2 144.42 89.99.7 36.41.65.6.55.5 143.48 24.48 21.47.45.4.35.3 51.5.25.2.15.1 171.64 148.1 149.94 119.27 78.1 55.28.5 23 22 21 2 19 18 17 16 15 14 13 12 11 1 9 8 7 6 5 4 3 2 1-1 -2
@7 Celcius H 2 H 2 H H 2 1..9 9.77 78.85 73.75 72.3 22.37 25.44.8.7.6.5 145.43 144.12 56.3 52.85 52.75 51.64 37.32.4.3.2.1 22 21 2 19 18 17 16 15 14 13 12 11 1 9 8 7 6 5 4 3 2 1-1 -2-3
This Print journal Date: is 25 The Feb Royal 21 Society 16:55:29 of Chemistry 211 MS Data Review All Plots - 2/25/21 4:55 PM File: c:\varianws\data\dmt\2911\rp135d,1v _11-26-46 am.xms Sample: RP135d,1v Scan Range: 1-898 Time Range:.5-3.27 min. kcounts 17.5 2.:65.> perator: dmt Date: 1/29/21 11:26 AM RP135d,1v _11-26-46 AM.xms TIC 15. 12.5 1. 7.5 5. 2.5 nv 2.:65.> 1A RP135d,1v _11-26-46 AM.xms uv 4 3 2 1 1% 5 1 15 2 25 Spectrum 1A BP: 29 (2294=1%), rp135d,1v _11-26-46 am.xms 29 2294 minutes 8.25 min, Scans: 242-244, 2.:65.>, Ion: us, RIC: 1366 75% 425 1574 5% 25% 217 485 213 2 254 471 252 265 31 318 326 144 352 367 383 127 455 242 492 147 % 2 3 4 5 6 m/z
H 2 H 2 H H + H 2 H 2 1..9.8 M3(d) 7.46.7 7.48 M4(d).6.5.4 7.25 7.23 M2(d) 6.1 6.1 4.47 M1(dd) 3.65 3.66.3.2 8.32 4.5 4.41 3.91 3.9 3.88 3.67 8.35 3.52 2.37 2.25.1 3.47 2.43.78 2.1.85 1.1 3.99.99 2.23 1.7 9.5 9. 8.5 8. 7.5 7. 6.5 6. 5.5 5. 4.5 4. 3.5 3. 2.5 2. 1.5 1..5
H 2 H 2 H H + H 2 H 2 1..9.8.7.6.5.4.3 127.62 125.35 119.14 9.24 77.95 132.65 171.48 155.88 147.92 149.88 144.31 143.42 136.15 71.55 73.2 54.68 51.41 51.32 24.55.2.1 57.56 23 22 21 2 19 18 17 16 15 14 13 12 11 1 9 8 7 6 5 4 3 2 1-1 -2
@ 7 Celcius H 2 H 2 H H + H 2 H 2 1..9.8.7 52.56.6.5.4 9.93 78.87 73.84 72.38 126.33 128.66 133.38 144.16 145.33 55.8 58.53 25.48.3.2.1 172.11 156.86 148.82 15.79 136.83 119.89 22 21 2 19 18 17 16 15 14 13 12 11 1 9 8 7 6 5 4 3 2 1-1 -2-3
This Print journal Date: is 25 The Feb Royal 21 Society 17:25:23 of Chemistry 211 MS Data Review All Plots - 2/25/21 5:25 PM File: c:\varianws\data\dmt\121\rp183d,1v _3-35-52 pm.xms Sample: RP183d,1v Scan Range: 1-898 Time Range:.5-3.27 min. kcounts 2.:65.> 15. perator: dmt Date: 2/1/21 3:35 PM RP183d,1v _3-35-52 PM.xms TIC 12.5 1. 7.5 5. 2.5 nv 3 2.:65.> 1A RP183d,1v _3-35-52 PM.xms uv 25 2 15 1 5 1% 5 1 15 2 25 Spectrum 1A BP: 215 (62=1%), rp183d,1v _3-35-52 pm.xms 215 62 minutes 12.987 min, Scans: 384-386, 2.:65.>, Ion: us, RIC: 11474 515 551 75% 234 444 5% 25% 22 158 23 39 22 296 213 18 223 247 232 177 235 6 251 169 261 22 259 15 271 361 282 212 281 5 31 155 35 44 315 37 331 258 323 13 352 264 35 194 353 113 356 42 368 183 39 227 386 78 412 164 438 281 434 141 433 81 457 77 481 215 479 12 55 65 537 266 541 77 559 319 554 132 561 86 58 66 619 135 613 89 631 195 % 2 3 4 5 6 m/z
H 2 H H H H 2 1..9.8.7.6.5.4.3.2 M1(d) 6.12 6.11 4.47 4.49 4.44 3.79 3.75 3.61 3.65 3.26 3.31 3.19 3.24 2.38 8.4 8.41 1.93.1.86.95 2.1.99 6. 2.1 4.2 9.5 9. 8.5 8. 7.5 7. 6.5 6. 5.5 5. 4.5 4. 3.5 3. 2.5 2. 1.5 1..5
H 2 H H H H 2 1..9 144.68 9.11 78.19 71.66 38.1 3.23.8.7 72.91 54.71.6.5.4.3 176.52 156.7 15.14 148.1 143.46 119.33 18.44.2 5.75 22.55.1 23 22 21 2 19 18 17 16 15 14 13 12 11 1 9 8 7 6 5 4 3 2 1-1 -2
@7 Celcius H 2 H H H H 2 1..9 9.85 78.96 73.68 72.44 38.95 31.7 23.52 19.32.8 144.19 55.9 52.4.7 145.67 54.17.6.5.4.3 176.92.2.1 157.69 148.89 151.3 12.4 22 21 2 19 18 17 16 15 14 13 12 11 1 9 8 7 6 5 4 3 2 1-1 -2-3
This Print journal Date: is 25 The Feb Royal 21 Society 16:57:11 of Chemistry 211 MS Data Review All Plots - 2/25/21 4:57 PM File: c:\varianws\data\dmt\121\rp181d,1v _2-2-49 pm.xms Sample: RP181d,1v Scan Range: 1-898 Time Range:.4-3.26 min. kcounts 3 2.:65.> perator: dmt Date: 2/1/21 2:2 PM RP181d,1v _2-2-49 PM.xms TIC 25 2 15 1 5 7 nv 6 2.:65.> 1A RP181d,1v _2-2-49 PM.xms uv 5 4 3 2 1 1% 5 1 15 2 25 Spectrum 1A BP: 452 (1779=1%), rp181d,1v _2-2-49 pm.xms minutes 9.746 min, Scans: 288-29, 2.:65.>, Ion: 1 us, RIC: 23533 452 1779 75% 5% 25% 453 2432 215 665 317 166 % 2 3 4 5 6 m/z
H 2 H 2 H H H H 2.2.15.1.5 2.19.9.11 1.3 2.45. 1.81 4.42.13 9.5 9. 8.5 8. 7.5 7. 6.5 6. 5.5 5. 4.5 4. 3.5 3. 2.5 2. 1.5 1..5
@7 Celcius H 2 H 2 H H H H 2.16.15.14.13.12.11.1.9.8.7.6.5.4.3.2.1 1.94 1..5 1.5 2.15 2.6 8.68 4.49.11 9.5 9. 8.5 8. 7.5 7. 6.5 6. 5.5 5. 4.5 4. 3.5 3. 2.5 2. 1.5 1..5
H 2 H 2 H H H H 2 1..9.8.7.6.5.4.3.2.1 22 21 2 19 18 17 16 15 14 13 12 11 1 9 8 7 6 5 4 3 2 1
H 2 H 2 H H H H 2 1..9.8.7.6.5.4.3.2.1 22 21 2 19 18 17 16 15 14 13 12 11 1 9 8 7 6 5 4 3 2 1
This Print journal Date: is 25 The Feb Royal 21 Society 17:5:44 of Chemistry 211 MS Data Review All Plots - 2/25/21 5:5 PM File: c:\varianws\data\dmt\121\rp279,1v _11-7-5 am.xms Sample: RP279,1v Scan Range: 1-9 Time Range:.4-3.33 min. kcounts 2.:65.> 3 perator: dmt Date: 2/1/21 11:7 AM RP279,1v _11-7-5 AM.xms TIC 25 2 15 1 5 uv 1.5 1.25 2.:65.> 1A RP279,1v _11-7-5 AM.xms uv 1..75.5.25. 1% 5 1 15 2 25 Spectrum 1A BP: 423 (458=1%), rp279,1v _11-7-5 am.xms 423 458 minutes 9.473 min, Scans: 28-282, 2.:65.>, Ion: us, RIC: 19279 75% 5% 25% 26 467 236 343 27 341 288 698 352 345 374 279 424 393 468 286 % 2 3 4 5 6 m/z
This Print journal Date: is 25 The Feb Royal 21 Society 17:6:15 of Chemistry 211 MS Data Review All Plots - 2/25/21 5:6 PM File: c:\varianws\data\dmt\121\rp279,1v _11-7-5 am.xms Sample: RP279,1v Scan Range: 1-9 Time Range:.4-3.33 min. kcounts 2.:65.> 3 perator: dmt Date: 2/1/21 11:7 AM RP279,1v _11-7-5 AM.xms TIC 25 2 15 1 5 uv 1.5 2.:65.> 1A RP279,1v _11-7-5 AM.xms uv 1.25 1..75.5.25. 1% 5 1 15 2 25 Spectrum 1A BP: 233 (1536=1%), rp279,1v _11-7-5 am.xms 233 1536 minutes 3.4 min, Scans: 88-9, 2.:65.>, Ion: 1 us, RIC: 22474 75% 252 113 5% 27 78 243 562 269 55 311 56 43 495 25% 21 358 21 145 229 221 262 174 273 359 288 183 298 356 318 177 334 377 349 83 366 228 375 87 428 252 423 114 453 188 465 376 48 166 531 166 559 2 582 13 635 29 % 2 3 4 5 6 m/z
H 2 H 2 H H 2 1..9.8 M2(t) 3.2.7 3.37.6.5.4.3.2.1 M1(d) 6.15 6.14 4.51 4.5 4.42 4.45 3.73 3.74 3.8 3.68 3.39 3.35 8.41 8.42 3.3 3. 2.15 2.1 2.3 2.9.97 1. 2.22 2.12 4.15 4.1 4.14 9.5 9. 8.5 8. 7.5 7. 6.5 6. 5.5 5. 4.5 4. 3.5 3. 2.5 2. 1.5 1..5
H 2 H 2 H H 2 1..9.8 21.49.7 36.39.6.5.4 143.24 144.61 9.5 77.47 73.1 71.41.3 55.12.2.1 148.11 15.7 119.26 5.67 23 22 21 2 19 18 17 16 15 14 13 12 11 1 9 8 7 6 5 4 3 2 1-1 -2
@7 Celcius H 2 H 2 H H 2 22.31 1..9.8 51.85 37.3.7.6.5 143.9 145.81 9.77 78.33 73.79 72.22 56.17.4.3.2.1 148.94 151.8 119.94 22 21 2 19 18 17 16 15 14 13 12 11 1 9 8 7 6 5 4 3 2 1-1 -2-3
This Print journal Date: is 25 The Feb Royal 21 Society 17:17:3 of Chemistry 211 MS Data Review All Plots - 2/25/21 5:17 PM File: c:\varianws\data\dmt\121\rp278,1v _12-8-59 pm.xms Sample: RP278,1v Scan Range: 1-897 Time Range:.6-3.24 min. kcounts 35 2.:65.> perator: dmt Date: 2/1/21 12:9 PM RP278,1v _12-8-59 PM.xms TIC 3 25 2 15 1 5 uv 1.5 2.:65.> RP278,1v _12-8-59 PM.xms uv 1.25 1. 1A.75.5.25. 1% 5 1 15 2 25 Spectrum 1A BP: 246 (249=1%), rp278,1v _12-8-59 pm.xms 246 249 minutes 8.514 min, Scans: 251-253, 2.:65.>, Ion: us, RIC: 14446 75% 381 1824 5% 25% 211 32 236 384 247 62 283 36 357 212 382 576 412 131 423 496 443 254 485 133 53 183 565 154 % 2 3 4 5 6 m/z
This Print journal Date: is 25 The Feb Royal 21 Society 17:17:59 of Chemistry 211 MS Data Review All Plots - 2/25/21 5:17 PM File: c:\varianws\data\dmt\121\rp278,1v _12-8-59 pm.xms Sample: RP278,1v Scan Range: 1-897 Time Range:.6-3.24 min. kcounts 35 2.:65.> perator: dmt Date: 2/1/21 12:9 PM RP278,1v _12-8-59 PM.xms TIC 3 25 2 15 1 5 uv 1.5 2.:65.> 1A RP278,1v _12-8-59 PM.xms uv 1.25 1..75.5.25. 1% 5 1 15 2 25 Spectrum 1A BP: 423 (2939=1%), rp278,1v _12-8-59 pm.xms 423 2939 minutes 3.57 min, Scans: 89-91, 2.:65.>, Ion: 1 us, RIC: 2855 75% 5% 25% 28 66 25 394 261 572 233 358 26 299 295 458 35 382 293 211 374 23 4 527 422 28 461 24 485 353 523 385 562 225 6 321 648 155 % 2 3 4 5 6 m/z
H H 2 H H 2 H H H H 2 1. 8.43 8.44.9.8.7.6.5.4.3 M1(d) 6.14 6.13 4.5 M2(dd) 3.68 3.34 3.2 3.3 3.32 1.96.2 4.52 4.44 3.8 3.84 3.81 3.65.1.85.92 2.3 1.1 8. 4.11 9.5 9. 8.5 8. 7.5 7. 6.5 6. 5.5 5. 4.5 4. 3.5 3. 2.5 2. 1.5 1..5
H H 2 H H 2 H H H H 2 38.8 1..9.8.7.6.5.4.3 156.69 147.98 149.97 144.52 143.49 119.32 9.33 78.15 73.1 71.6 54.68 51.23.2.1 22.93 23 22 21 2 19 18 17 16 15 14 13 12 11 1 9 8 7 6 5 4 3 2 1-1 -2
@7 Celcius H H 2 H H 2 H H H H 2 1..9 23.63.8.7 38.98 52.35.6.5 144.18 145.51 9.96 78.85 73.8 72.36 55.88.4.3.2.1 157.66 148.85 15.87 119.99 22 21 2 19 18 17 16 15 14 13 12 11 1 9 8 7 6 5 4 3 2 1-1 -2-3
This Print journal Date: is 25 The Feb Royal 21 Society 17:9:16 of Chemistry 211 MS Data Review All Plots - 2/25/21 5:9 PM File: c:\varianws\data\dmt\2911\rp28,1v _2-46-1 pm.xms Sample: RP28,1v Scan Range: 1-899 Time Range:.4-3.29 min. kcounts 2.:65.> 5 perator: dmt Date: 1/29/21 2:46 PM RP28,1v _2-46-1 PM.xms TIC 4 3 2 1 uv 3 2.:65.> 1A RP28,1v _2-46-1 PM.xms uv 2 1-1 -2 1% 5 1 15 2 25 Spectrum 1A BP: 465 (11972=1%), rp28,1v _2-46-1 pm.xms minutes 3.16 min, Scans: 91-93, 2.:65.>, Ion: 1 us, RIC: 44874 465 11972 75% 5% 466 3876 25% 231 11 313 999 366 1337 423 1262 527 114 616 689 % 2 3 4 5 6 m/z
Information added after peer-review: Table 1: Distances (Å) measured between nearest oxygen of binding site residue and nearest nitrogen of guanidinium for flexibly docked ligand and rigid CARM1 receptor, or overlaid PRMT1 (bold). 16 17 18 2 Residue 1 2 3 1 2 3 1 2 3 1 2 3 E258 3.19 5.94 6.85 6.5 8.56 E267 3.41 3.16 2.98 3.3 2.86 3.69 3.65 3.15 Asn162 9.53 8.79 7.71 8.25 Glu47 6.4 4.4 3.15 4.5 19 Residue + H 3 E258 5.2 E267 2.73 Asn162 12.11 Glu47 7.71 Figure 1: Docked structure of 18 showing numbering of nitrogens in Table 1 above
Electronic Supplementary Material (ESI) for rganic & Biomolecular Chemistry Further images relevant to the Docking discussion: Figure 2: Result of docking inhibitor 2 (white backbone) with CARM1 binding site (2Y1X.pdb, selected residues green backbone), also AdoHcy 2 (yellow), substrate arginine channel (green mesh). PRMT1 residue superimposed, Glu47 (orange, bold in parentheses). Figure 3: Result of docking inhibitor 19 (white backbone) with CARM1 binding site (2Y1X.pdb, selected residues green backbone), also AdoHcy 2 (yellow), substrate arginine channel (green mesh). PRMT1 residue superimposed, Glu47 (orange, bold in parentheses).