MEDICIAL CEMISTY Design Synthesis and SA of on-peptidic Urotensin II Agonists C Val Cys Tyr Lys Trp 2 Glu Thr Pro Asp Cys Phe uman urotensin II (S) pec 50 7.49 () pec 50 5.84 FL-104 AC-7954 Fredrik Lehmann 081203
MEDICIAL CEMISTY Where to start? AC7954 identified as the first non-peptidic Agonist for Urotensin II
MEDICIAL CEMISTY The mission: Try to improve the potency and efficacy relative to AC7954 ow? (step 1) different heterocycles different amines various substitution change "spacer" length various substitution
etrosynthetic analysis MEDICIAL CEMISTY Li Li + + + + Br
Synthesis MEDICIAL CEMISTY Br n-buli C 2 S 2 Me 2 90-99% 80-99% Dioxane mw 180 + + o, 6 min TF, rt + 18-78% 95-99% + n-buli 10-75%
SA (selection) MEDICIAL CEMISTY FL66 Compound pec50 Efficacy AC7954 4-5.95 ± 0.12 133 ± 8 FL60 4-F 5.60 ± 0.02 147 ± 40 FL47 4-CF 3 6.05 ± 0.07 109 ± 10 FL52 4-Me 5.01 ± 0.01 51 ± 1 FL56 3-Me 5.33 ± 0.08 135 ± 51 FL54 2-Me 4.97 ± 0.04 33 ± 4 FL62 3-5.59 ± 0.03 105 ± 18 FL66-6.20 ± 0.01 138 ± 21
Combinations? MEDICIAL CEMISTY 1 2 Compound 1 2 pec50 Efficacy AC7954 4-5.95 ± 0.12 133 ± 8 FL97 3,4-Di- 6.18 181 FL98 6,7-DiMe 2-apthyl 6.91 126 FL99 6,7-DiMe 3-Me 5.58 154 FL101 6,7-DiMe 3,5-Di-Me 5.28 163 Lehmann et al. J Bioorg Med Chem, 2005
Small changes can be vital! MEDICIAL CEMISTY AC-7954 pec 50 = 5.95 Inactive!
Stereochemistry? MEDICIAL CEMISTY AC-7954 16 (+) 16 pec 50 = 7.11 (-) 16 pec 50 = 5.78
Summary of SA MEDICIAL CEMISTY naphthyl tolerated methyl good, phenyl too large not necessery for activity can be exhanged for -alkyl chirality important two carbons most favorable substitution forbidden larger rings tolerated but not smaller dimethyl optimal, larger systems deterimental for activity Ph tolerated, Ph or CCPh too sterically demanding substitution pattern important, but difficult to predict
MEDICIAL CEMISTY The mission: Try to improve the potency and efficacy relative to AC7954 ow? (step 2) + pec 50 = 5.95 pec 50 = 5.77
MEDICIAL CEMISTY i 1 ii 3v, w iii iv, v vi 2 4a - e vii 2 viii ix 6a - u S 5a - s 8d, e, v 7a - u i) LA, TF, 91%. ii) -C 2 -X, PS-DIPEA, C 2 2, 48 and 78%. iii) -C, EDC, DMAP, C 2 2, 43-98%. iv) C 3 C, 2 S 4. v) 6M (aq), >99%. vi) -C, Et 3, TF, 29-91%. vii) -C, S 2, Et 3, TF, 42-98% or -C, EDC, DMAP, C 2 2, 60-67%. viii) -S 2 -, Et 3, TF, quant. ix) -C, Et 3, TF, 38-98% Lehmann et al. J Med Chem 2006
SA (selected) MEDICIAL CEMISTY 3v, w 4a - e 5a - e cmpd pec50 Efficacy n 3v 4-MePh A c 2 3w 2-Me-Ph A c 2 4a 2-MePh 5.77 ± 0.01 126 ± 34 2 4b 4-Me-Ph 5.66 ± 0.2 84 ± 28 3 4c 4-CF 3 -Ph 5.76 ± 0.15 47 ± 18 3 4d 4-Ph-Ph 6.28 ± 0.14 53 ± 27 2 4e 2-aphthyl 5.68 ± 0.04 49 ± 28 3 5a 2-MePh 5.45 ± 0.04 175 ± 15 2 5b 4-Me-Ph 5.87 ± 0.2 162 ± 38 5 5c 4-CF 3 -Ph 7.11 ± 0.1 115 ± 14 3 5d 4-Ph-Ph 7.11 ± 0.01 116 ± 11 2 (-) 5d 4-Ph-Ph 5.84 ± 0.1 96 ± 16 5 (+) 5d 4-Ph-Ph 7.49 ± 0.33 116 ± 18 5 5e 2-aphthyl 6.39 ± 0.19 109 ± 18 2
SA (selected) MEDICIAL CEMISTY S 6a - u 7a - u 8d, e, v cmpd pec50 Efficacy n 6a 2-MePh 5.41 ± 0.07 176 ± 28 4 6b 4-Me-Ph 5.78 ± 0.21 170 ± 48 3 6c 4-CF 3 -Ph 6.60 ± 0.13 94 ± 22 3 6d 4-Ph-Ph 6.57 ± 0.12 58 ± 16 3 6e 2-aphthyl 6.28 ± 0.26 89 ± 19 3 7a 2-MePh 5.64 ± 0.15 175 ± 23 4 7b 4-Me-Ph 5.70 ± 0.06 195 ± 31 3 7c 4-CF 3 -Ph 6.93 ± 0.11 105 ± 14 3 7d 4-Ph-Ph 6.84 ± 0.11 91 ± 18 3 7e 2-aphthyl 6.54 ± 0.1 106 ± 21 3 8d 4-Ph-Ph A 3 8e 2-aphthyl A 3 8v 4-MePh 5.19 ± 0.09 74 ± 10 4
r another way MEDICIAL CEMISTY 1 2 + 2 i 2 1 A - C 1-10 {A1 - C10} i) PS-DCC 2 eqv, PS-DMAP 0.2 eqv, C 2 2, rt, 96h, 46-98% Works even in small scale... Cmpd Scale (mg) Yield Purity Scale (mg) Yield Purity {A1} 25 94 100 5 92 100 {A7} 25 83 100 5 97 100 Lehmann et al. Eur J Med Chem 2007
2 2 2 A B C MEDICIAL CEMISTY 1. = 2. = CF 3 3. =Me 4. = 5. = CF 3 6. =Me 7. = 8. = CF 3 9. = Me cmpd A B C 10 Extraction Yield Purity Ion ex Yield Purity Extraction Yield Purity Ion ex Yield Purity Extraction Yield Purity Ion ex Yield Purity 1 92 100 - - 93 100 - - 67 100 - - 2 96 100 - - 98 100 - - 88 100 - - 3 96 100 - - 93 100 - - 79 100 - - 4 92 100 - - 96 100 - - 66 100 - - 5 83 100 - - 78 100 - - 84 100 - - 6 91 100 - - 98 100 - - 94 100 - - 7 87 85 62 100 132 93 86 100 76 100 - - 8 97 100 - - 56 100 - - 200 51 59 100 9 83 97 68 100 87 100 - - 118 88 47 100 10 147 88 46 100 120 75 60 100 102 76 64 100
2 2 2 A B C MEDICIAL CEMISTY 1. = 2. = CF 3 3. = Me 4. = 5. = CF 3 6. =Me 7. = 8. = CF 3 9. = Me 10 cmpd A B C P EC 50 a Efficacy b P EC 50 a Efficacy b P EC 50 a Efficacy b 1 5.73 ± 0.47 76 ± 5 5.38 ± 0.07 83 ± 11 5.76 ± 0.17 121 ± 31 2 6.06 ± 0.23 53 ± 7 5.95 ± 0.03 25 ± 4 6.24 ± 0.06 40 ± 3 3 5.82 ± 0.21 65 ± 18 5.23 ± 0.05 62 ± 5 A c A c 4 5.78 ± 0.10 139 ± 5 5.34 ± 0.14 73 ± 9 6.22 ± 0.20 107 ± 22 5 5.95 ± 0.31 73 ± 10 5.64 ± 0.09 60 ± 6 6.43 ± 0.07 75 ± 1 6 5.53 ± 0.03 101 ± 12 5.27 ± 0.02 67 ± 14 5.85 ± 0.11 113 ± 9 7 6.37 ± 0.12 128 ± 10 5.81 ± 0.07 97 ± 6 6.23 ± 0.18 116 ± 3 8 6.89 ± 0.06 133 ± 3 6.36 ± 0.08 96 ± 1 6.87 ± 0 117 ± 1 9 6.29 ± 0.04 99 ± 4 5.70 ± 0.08 79 ± 14 6.42 ± 0.06 106 ± 16 10 6.40 ± 0.04 124 ± 6 5.51 ± 0.07 131 ± 0 6.34 ± 0.05 115 ± 14
2 2 2 A B C MEDICIAL CEMISTY 1. = 2. = CF 3 3. = Me 4. = 5. = CF 3 6. =Me 7. = 8. = CF 3 9. = Me 10 140 6,7 120 Efficacy 100 80 60 40 Aliphatic Conjugated pec50 6,2 5,7 A B C 20 5,2 5,7 6,2 6,7 pec50 5,2 1 2 3 4 5 6 7 8 9 10 Acid
Determination of the absolute configuration MEDICIAL CEMISTY 2 Ph Me (,S) + Ph Me (,) C20 1 C9 C10 C8 C11 C7 2 C6 C1 1 C2 21 13 2 C14 C13 C12 1 C15 C3 C4 1 C19 C16 C18 C17 2 4-Ph-Ph-C 2 4-Ph-Ph-C C5 2 X-ray structure of the (,)-methoxy phenyl acet amide (S)-FL104 pec 50 7.49 ()-FL104 pec 50 5.84
MEDICIAL CEMISTY Molecular modelling proposed a pharmacophore for UII-agonists A1 6.70 Å A2 A3 10.8 Å 8.4 Å 4.8 Å 4.8 Å
Focus on Mannich reaction App 300 reaction in 2 mmol scale were run in for various time in seven solvents (water, acetic acid, dioxane, TF, DMF, ethanol, and methanol) at three different temperatures. At 150 o C, pressure jumped one bar => release of formaldehyde
Focus on Mannich reaction MEDICIAL CEMISTY Table 3. esults from synthesis of Mannich bases using various secondary amines. + + 180 o C, 300s dioxane Product Yield a Product Yield a 19 Diethylamine 78 21 Thiomorpholine 46 20 Morpholine 44 22 Piperidine 61 a) All yields are LC yields given as averages of at least two experiments
Focus on Mannich reaction MEDICIAL CEMISTY Lehmann et al, Mol Div 2003
Mannich eaction MEDICIAL CEMISTY a) Values for small scale reactions (2 mmol), values within parenthesis refers to large (40 mmol) scale Lehmann et al, Mol Div 2003
Mannich eaction Lehmann et al, Mol Div 2003