Modulators of complex I

Abstract

The present invention describes compounds modulating the function of mitochondrial complex I (NADH-quinone oxidoreductase) having formula (I) ##STR00001##

Claims

1. A compound having formula (I) or a salt thereof ##STR00632## wherein: R1 is C.sub.1-4-alkyl unsubstituted or substituted with MeO; tetrahydropyranyl, tetrahydrofuranyl, oxetanyl, dioxepanyl, pyrrolidinyl or piperidinyl; or pyrrolidinyl or piperidinyl with the nitrogen substituted by methyl, isopropyl, oxetanyl, ethoxycarbonyl, acetyl, or trifluoroacetyl; R2 is a 5-, 6- or 7-membered ring containing 1 or 2 heteroatoms selected from O or N bound in formula (I) by a C═C double bond, in which one or both N-atoms can be substituted by methyl, isopropyl, acetyl, benzyloxycarbonyl, phenyl, oxetanyl or tetrahydropyranyl, and in which one or more C-atoms can be substituted by -methyl or —OH; R3 and R4 are independently from one another hydrogen; C.sub.1-6-alkyl, unsubstituted or substituted with one or more F, methoxy, C.sub.3-8-cycloalkyl unsubstituted or substituted with one or more F; aryl; heteroaryl consisting of 5 to 6 ring atoms; or heterocyclyl selected from the group consisting of oxetanyl, tetrahydropyranyl and pyrrolidinyl, said heterocyclyl being unsubstituted or substituted with C.sub.1-6-alkyl, acetyl, tetrahydrofuranyl, oxetanyl or hydroxyethylacetyl; or R3 and R4 together with the attached N form a heterocyclyl ring selected from the group consisting of morpholinyl and pyrrolidinyl both unsubstituted or substituted with C.sub.1-6-alkyl, F, or hydroxyl; or R3 and R4 together represent one of the following groups: ##STR00633##

2. A compound selected from the group consisting of the following compounds 1 to 175, or a salt thereof: TABLE-US-00036 # Structure  1  2  3  4  5  6  7  8  9  10  11  12  13  14  15  16  17  18  19  20  21  22  23  24  25  26  27  28  29  30  31  32  33  34  35  36  37  38  39  40  41  42  43  44  45  46  47  48  49  50  51  52  53  54  55  56  57  58  59  60  61  62  63  64  65  66  67  68  69  70  71  72  73  74  75  76  77  78  79  80  81  82  83  84  85  86  87  88  89  90  91  92  93  94  95  96  97  98  99 100 101 102 103 104 105 106 107 108 109 110 111 112 113 114 115 116 117 118 119 120 121 122 123 124 125 126 127 128 129 130 131 132 133 134 135 136 137 138 139 140 141 142 143 144 145 146 147 148 149 150 151 152 153 154 155 156 157 158 159 160 161 162 163 164 165 166 167 168 169 170 171 172 173 174 175

3. A compound selected from the group consisting of the following compounds 1001-1178, or a salt thereof: TABLE-US-00037 # Structure 1001 1002 1003 1004 1005 1006 1007 1008 1009 1010 1011 1012 1013 1014 1015 1016 1017 1018 1019 1020 1021 1022 1023 1024 1025 1026 1027 1028 1029 1030 1031 1032 1033 1034 1035 1036 1037 1038 1039 1040 1041 1042 1043 1044 1045 1046 1047 1048 1049 1050 1051 1052 1053 1054 1055 1056 1057 1058 1059 1060 1061 1062 1063 1064 1065 1066 1067 1068 1069 1070 1071 1072 1073 1074 1075 1076 1077 1078 1079 1080 1081 1082 1083 1084 1085 1086 1087 1088 1089 1090 1091 1092 1093 1094 1095 1096 1097 1098 1099 1100 1101 1102 1103 1104 1105 1106 1107 1108 1109 1110 1111 1112 1113 1114 1115 1116 1117 1118 1119 1120 1121 1122 1123 1124 1125 1126 1127 1128 1129 1130 1131 1132 1133 1134 1135 1136 1137 1138 1139 1140 1141 1142 1143 1144 1145 1146 1147 1148 1149 1150 1151 1152 1153 1154 1155 1156 1157 1158 1159 1160 1161 1162 1163 1164 1165 1166 1167 1168 1169 1170 1171 1172 1173 1174 1175 1176 1177 1178

4. The compound 26 according to claim 2 having the following formula or a salt thereof: ##STR00987##

5. The compound 26 according to claim 2 having the following formula: ##STR00988##

6. The compound 1024 according to claim 3 having the following formula or a salt thereof: ##STR00989##

7. The compound 1024 according to claim 3 having the following formula: ##STR00990##

Description

EXAMPLES

Example 1

HPLC/UPLC Methods

(1) Method A

(2) TABLE-US-00002 Device: Waters Alliance with DAD and MS detector Column: Waters XBridge C18, 4.6 × 30 mm, 3.5 μm Time % Solvent A % Solvent B Flow rate Temperature [min] [H.sub.2O, 0.1% NH.sub.3] [MeOH] [mL/min] [° C.] 0.00 95   5 4.0 60 0.20 95   5 4.0 60 1.50  0 100 4.0 60 1.75  0 100 4.0 60

(3) Method B

(4) TABLE-US-00003 Device: Waters Alliance with DAD and MS detector Column: Waters SunFire C18, 4.6 × 30 mm, 3.5 μm Time % Solvent A % Solvent B Flow rate Temperature [min] [H.sub.2O, 0.1% TFA] [MeOH] [mL/min] [° C.] 0.00 95 5 4.0 60 1.60 0 100 4.0 60 1.85 0 100 4.0 60 1.90 95 5 4.0 60

(5) Method C

(6) TABLE-US-00004 Device: Waters Alliance with DAD and MS detector Column: Waters XBridge C18, 4.6 × 30 mm, 3.5 μm Time % Solvent A % Solvent B Flow rate Temperature [min] [H.sub.2O, 0.1% TFA] [MeOH] [mL/min] [° C.] 0.00 95 5 4.8 60 1.60 0 100 4.8 60 1.85 0 100 4.8 60 1.90 95 5 4.8 60

(7) Method D

(8) TABLE-US-00005 Device: Waters Acquity with DAD and MS detector Column: Waters SunFire C18, 2.1 × 20 mm, 2.5 μm Time % Solvent A % Solvent B Flow rate Temperature [min] [H.sub.2O, 0.1% TFA] [MeOH] [mL/min] [° C.] 0.00 99 1 1.3 60 0.15 99 1 1.3 60 1.10 0 100 1.3 60 1.25 0 100 1.3 60

(9) Method E

(10) TABLE-US-00006 Device: Waters Acquity with DAD and MS detector Column: Supelco Ascentis Express C18, 2.1 × 50 mm, 2.7 μm Time % Solvent A % Solvent B Flow rate Temperature [min] [H.sub.2O, 0.1% TFA] [ACN, 0.08% TFA] [mL/min] [° C.] 0.00 95 5 1.5 60 0.70 1 99 1.5 60 0.80 1 99 1.5 60 0.81 95 5 1.5 60

(11) Method F

(12) TABLE-US-00007 Device: Waters Alliance with DAD and MS detector Column: Waters XBridge C18, 4.6 × 30 mm, 3.5 μm Time % Solvent A % Solvent B Flow rate Temperature [min] [H.sub.2O, 0.1% TFA] [MeOH, 0.1% TFA] [mL/min] [° C.] 0.00 95 5 4.0 60 0.20 95 5 4.0 60 1.50 0 100 4.0 60 1.75 0 100 4.0 60 1.85 95 5 4.0 60

(13) Method G

(14) TABLE-US-00008 Device: Waters Alliance with DAD and MS detector Column: Waters XBridge C18, 4.6 × 30 mm, 3.5 μm Time % Solvent A % Solvent B Flow rate Temperature [min] [H.sub.2O, 0.1% TFA] [MeOH] [mL/min] [° C.] 0.00 95 5 4.0 60 1.60 0 100 4.0 60 1.85 0 100 4.0 60 1.90 95 5 4.0 60

(15) Method H

(16) TABLE-US-00009 Device: Waters Alliance with 2695 with PDA detector 2996 and micromass ZQ 2000 Column: Microsorb C18, 4.6 × 20 mm, 5 μm Time % Solvent A % Solvent B Flow rate Temperature [min] [H.sub.2O, 0.15% TFA] [MeOH] [mL/min] [° C.] 0.00 95 5 5.2 rt 0.25 95 5 5.2 rt 1.90 0 100 5.2 rt 2.05 0 100 5.2 rt 2.15 95 5 5.2 rt 2.25 95 5 5.2 rt 2.30 95 5 0.1 rt

(17) Method I

(18) TABLE-US-00010 Device: Waters Acquity with DAD and MS detector Column: Waters SunFire C18, 2.1 × 30 mm, 2.5 μm Time % Solvent A % Solvent B Flow rate Temperature [min] [H.sub.2O, 0.13% TFA] [MeOH, 0.05% TFA] [mL/min] [° C.] 0.00 99 1 1.2 60 0.15 99 1 1.2 60 1.10 0 100 1.2 60 1.25 0 100 1.2 60

(19) Method J

(20) TABLE-US-00011 Device: Waters Alliance with DAD and MS detector Column: Waters XBridge C18, 4.6 × 30 mm, 3.5 μm Time % Solvent A % Solvent B Flow rate Temperature [min] [H.sub.2O, 0.1% TFA] [MeOH] [mL/min] [° C.] 0.00 95 5 4.0 60 0.20 95 5 4.0 60 1.50 0 100 4.0 60 1.90 0 100 4.0 60 2.00 95 5 4.0 60

(21) Method K

(22) TABLE-US-00012 Device: Waters Alliance with DAD and MS detector Column: Waters XBridge C18, 4.6 × 30 mm, 3.5 μm Time % Solvent A % Solvent B Flow rate Temperature [min] [H.sub.2O, 0.1% TFA] [MeOH] [mL/min] [° C.] 0.00 95 5 4.0 60 0.20 95 5 4.0 60 1.50 0 100 4.0 60 1.75 0 100 4.0 60 1.85 95 5 4.0 60

(23) Method L

(24) TABLE-US-00013 Device: Waters Alliance with DAD and MS detector Column: Waters XBridge C18, 4.6 × 30 mm, 3.5 μm Time % Solvent A % Solvent B Flow rate Temperature [min] [H.sub.2O, 0.1% TFA] [MeOH] [mL/min] [° C.] 0.00 95 5 4.9 60 1.60 0 100 4.9 60 2.20 95 5 4.9 60

(25) Method M

(26) TABLE-US-00014 Device: Waters Alliance with DAD and MS detector Column: Waters XBridge C18, 4.6 × 30 mm, 3.5 μm Time % Solvent A % Solvent B Flow rate Temperature [min] [H.sub.2O, 0.1% NH.sub.3] [MeOH, 0.1% NH.sub.3] [mL/min] [° C.] 0.00 95 5 4.0 60 0.20 95 5 4.0 60 1.50 0 100 4.0 60 1.75 0 100 4.0 60

(27) Method N

(28) TABLE-US-00015 Device: Waters Alliance with DAD and MS detector Column: Waters SunFire C18, 4.6 × 30 mm, 3.5 μm Time % Solvent A % Solvent B Flow rate Temperature [min] [H.sub.2O, 0.1% TFA] [MeOH] [mL/min] [° C.] 0.00 95 5 4.0 60 0.20 95 5 4.0 60 1.50 0 100 4.0 60 1.75 0 100 4.0 60 1.85 95 5 4.0 60

(29) Method O

(30) TABLE-US-00016 Device: Waters Acquity with DAD and MS detector Column: Waters XBridge C18, 2.1 × 20 mm, 2.5 μm Time % Solvent A % Solvent B Flow rate Temperature [min] [H.sub.2O, 0.1% TFA] [MeOH] [mL/min] [° C.] 0.00 95 5 1.4 60 0.05 95 5 1.4 60 1.00 0 100 1.4 60 1.10 0 100 1.4 60

(31) Method P

(32) TABLE-US-00017 Device: Waters Acquity with DAD and MS detector Column: Waters XBridge BEH C18, 2.1 × 30 mm, 1.7 μm Time % Solvent A % Solvent B Flow rate Temperature [min] [H.sub.2O, 0.13% TFA] [MeOH, 0.08% TFA] [mL/min] [° C.] 0.00 99 1 1.3 60 0.05 99 1 1.3 60 0.35 0 100 1.3 60 0.50 0 100 1.3 60

Example 2

Synthesis of intermediates A1, A2, A4, A5, A7 to A9 and A11

(33) Intermediate A1:

(34) ##STR00196##

(35) (2-Fluoro-5-nitro-phenyl)-acetic acid (4.60 g; 23.10 mmol) and tetrahydro-furan-3-ylamine (10.0 g; 114.78 mmol) in DMSO (20 mL) are stirred at 45° C. over night. HCl (aq. solution; 2M; 92.4 mL; 184.80 mmol) is added. After stirring for 1.5 h at 45° C. the resulting precipitate is filtered off, washed with water and dried.

(36) MS (ESI.sup.+): m/z=249 [M+H].sup.+

(37) HPLC (Method B): R.sub.t=1.0 min

(38) The following intermediates were prepared in an analogous manner to intermediate A1:

(39) TABLE-US-00018 Mass R.sub.f value or Structure Educt 1 Educt 2 signal(s) R.sub.t No. Comment A2 (2- Fluoro- 5-nitro- phenyl)- acetic acid Tetrahydro- pyran-4- ylamine (M + H).sup.+ = 263 1.03 min (Method B) A4 (2- Fluoro- 5-nitro- phenyl)- acetic acid Isopropylamine (M + H).sup.+ = 221 HCl (4M, aq. solution) is used instead of HCl (2M, aq. solution) A5 (2- Fluoro- 5-nitro- phenyl)- acetic acid 1,4-Dioxepan- 6-amine (M + H).sup.+ = 279 1.08 min (Method A) Purification by MPLC (DCM/MeOH; 0/0 -> 99/1) A7 00 (2- Fluoro- 5-nitro- phenyl)- acetic acid (S)-3-Amino- pyrrolidine-1- carboxylic acid tert-butyl ester (M + H).sup.+ = 346 1.34 min (Method A) A8 01 (2- Fluoro- 5-nitro- phenyl)- acetic acid tert-Butylamine (M + H).sup.+ = 235 0.77 min (Method O) Additional amine (6 eq.) is added. Mixture is stirred at 100° C. over night A9 02 (2- Fluoro- 5-nitro- phenyl)- acetic acid 4-Amino- piperidine-1- carboxylic acid ethyl ester (M + H).sup.+ = 334 1.24 min (Method B) A11 03 (2- Fluoro- 5-nitro- phenyl)- acetic acid 2-Methoxy-1- methyl- ethylamine (M + H).sup.+ = 251 1.08 min (Method B)

Example 3

Synthesis of Intermediate A3

(40) ##STR00204##

(41) Step 1:

(42) (2-Fluoro-5-nitro-phenyl)-acetic acid (500 mg; 2.51 mmol) and oxetan-3-ylamine (936 mg; 12.81 mmol) in DMSO (2 mL) are stirred at 45° C. over night. The mixture is purified by preparative HPLC (eluent A. water+0.15% conc. ammonia, eluent B: MeOH).

(43) MS (ESI.sup.+): m/z=251 [M−H].sup.−

(44) (Method A): R.sub.t=0.66 min

(45) Step 2:

(46) Intermediate A3 Step 1 (462 mg; 0.92 mmol) and TBTU (0.71 g; 2.20 mmol) in DMF (8 mL) are stirred at room temperature over night. The resulting precipitate is filtered off and dried.

(47) MS (ESI.sup.+): m/z=235 [M+H].sup.+

(48) HPLC (Method A): R.sub.t=0.95 min

Example 4

Synthesis of Intermediate A6

(49) ##STR00205##

(50) Step 1:

(51) 1,3-Dihydro-1-(piperidin-4-yl)-(2H)-indol-2-one (5.51 g; 25.49 mmol) and TEA (7.16 mL; 50.99 mmol) in DCM (30 mL) are cooled in an ice bath. Trifluoroacetic anhydride (4.25 mL; 30.59 mmol) is added drop wise. The mixture is stirred at room temperature for 2 h. The mixture is diluted with NaHCO.sub.3 (aq. solution; 9%; 20 mL). After the gas development has stopped, the mixture is further diluted with DCM and water. The organic layer is separated, dried and evaporated. The residue is purified by MPLC (DCM/MeOH; 1/0.fwdarw.97/3).

(52) MS (ESI.sup.+): m/z=313 [M+H].sup.+

(53) HPLC (Method A): R.sub.t=1.25 min

(54) Step 2:

(55) Intermediate A6 Step 1 (8.02 g; 25.68 mmol) is solved in conc. sulphuric acid (45 mL) and cooled to −5° C. A cooled mixture of conc. sulphuric acid (15 mL) and conc. nitric acid (1.80 mL; 28.25 mmol) is added drop wise. After stirring for 1 h at −5° C. the mixture is poured on ice water. The resulting precipitate is filtered off and dried. The residue is taken up in DCM. The organic layer is washed with NaHCO.sub.3 (aq. solution; 9%), separated, dried and evaporated.

(56) MS (ESI.sup.+): m/z=358 [M+H].sup.+

(57) HPLC (Method A): R.sub.t=1.22 min

Example 5

Synthesis of Intermediate A10

(58) ##STR00206##

(59) To a cooled mixture of sodium nitrate (12.71 g; 149.48 mmol) and conc. sulphuric acid (22.9 mL; 407.68 mmol) additional conc. sulphuric acid (40 mL) is added drop wise. 1-Methyl-1,3-dihydro-indol-2-one (20.0 g; 135.89 mmol) is taken up in conc. sulphuric acid (120 mL) and added drop wise to the cooled nitrosulphuric acid. The mixture is allowed to warm up to room temperature over night. The mixture is poured on ice water. The resulting precipitate is filtered off, washed with water and dried. The residue is taken up in DCM, washed with water and brine, separated, dried and evaporated.

(60) MS (ESI.sup.+): m/z=193 [M+H].sup.+

(61) HPLC (Method H): R.sub.t=0.90 min

Example 6

Synthesis of Intermediates B1, B2, B4, B6, B7, B9 and B10

(62) The following intermediates are prepared according to the given references or are commercially available:

(63) TABLE-US-00019 Name Structure Reference B1 07 WO2006/72350 B2 08 WO2005/111029 B4 09 WO2010/68520 B6 0 WO2010/68520 B7 EP1790641 B9 WO2008/76356 B10 WO2004/60376

Example 7

Synthesis of Intermediates B3, B5 and B8

(64) Intermediate B3:

(65) ##STR00214##

(66) 2-Oxa-6-aza-spiro[3.4]octan-7-one (1.00 g; 7.87 mmol) and trimethyloxonium tetrafluoroborate (1.28 g; 8.65 mmol) in DCM (120 mL) are stirred at room temperature over night. The mixture is diluted with saturated NaHCO.sub.3 solution until gas development stops. The organic layer is separated, dried and evaporated.

(67) MS (ESI.sup.+): m/z=142 [M+H].sup.+

(68) The following intermediates are prepared in an analogous manner to intermediate B3:

(69) TABLE-US-00020 Mass R.sub.f value or Structure Educt 1 Educt 2 signal(s) R.sub.t No. Comment B5 3-Oxo-1-oxa- 4,9-diaza- spiro[5.5]un- decane-9- carboxylic acid tert- butyl ester Trimethyl- oxonium tertafluoro- borate (M + H).sup.+ = 285 B8 5-Methyl- morpholin-3- one Trimethyl- oxonium tertafluoro- borate

Example 8

Synthesis of intermediates C1-C3, C5-C8, C10-C12, C14, C16, C18, C20-C25, C27-C29, C32-C36

(70) Intermediate C1:

(71) ##STR00217##

(72) Intermediate A1 (700 mg; 2.82 mmol) and Intermediate B1 (714 mg; 6.20 mmol) are stirred at 130° C. for 20 min in a microwave. The resulting precipitate is suspended in MeOH, filtered off and dried.

(73) MS (ESI.sup.+): m/z=332 [M+H].sup.+

(74) HPLC (Method B): R.sub.t=1.29 min

(75) The following intermediates were prepared in an analogous manner to intermediate C1:

(76) TABLE-US-00021 R.sub.f value Mass or Structure Educt 1 Educt 2 signal(s) R.sub.t No. Comment C2 A2 B2 (M + H).sup.+ = 360 1.29 min (Method B) C3 A3 B1 (M + H).sup.+ = 318 1.19 min (Method A) Combined reaction time in microwave 2.5 h at 130° C. C5 0 A2 B1 (M + H).sup.+ = 346 0.84 min (Method D) After 20 min additional B1 is added and the mixture is stirred for 20 min at 130° C. C6 A4 B2 (M + H).sup.+ = 318 1.31 min (Method N) C7 A2 B3 (M + H).sup.+ = 372 1.22 min (Method B) The mixture is stirred for 40 min at 130° C. and for 20 min at 140° C. C8 1- Methyl- 5-nitro- 1,3- dihydro- indol-2- one B1 (M + H).sup.+ = 276 0.58 min (Method O) C10 A4 B1 (M + H).sup.+ = 304 1.24 min (Method C) The mixture is stirred for 40 min at 130° C. and for 20 min at 140° C. C11 A5 B1 (M + H).sup.+ = 362 1.27 min (Method A) Combined reaction time in microwave 165 min at 130° C. C12 A4 B4 (M + H).sup.+ = 302 1.45 min (Method A) C14 A6 B1 (M + H).sup.+ = 441 1.41 min (Method B) Combined reaction time in microwave 40 min at 130° C. C16 A7 B1 (M + H).sup.+ = 431 1.46 min (Method A) C18 A4 B5 (M + H).sup.+ = 473 1.65 min (Method B) C20 0 A8 B1 (M + H).sup.+ = 318 1.45 min (Method A) Combined reaction time in microwave 4.5 h at 130° C. Purification by MPLC (CH/EA = 4/1) C21 A2 B6 (M + H).sup.+ = 360 1.31 min (Method B) C22 A9 B1 (M + H).sup.+ = 417 1.43 min (Method B) C23 A10 B7 (M + H).sup.+ = 423 1.64 min (Method H) C24 A2 B7 (M + H).sup.+ = 493 1.5 min (Method B) C25 A10 B2 (M + H).sup.+ = 290 1.29 min (Method H) C27 A2 B8 (M + H).sup.+ = 260 1.36 min (Method B) Additional reaction time in microwave 30 min at 140° C. Purification by preparative HPLC (eluent A. water + 0.1% conc. ammonia, eluent B: MeOH) C28 A11 B1 (M + H).sup.+ = 334 1.32 min (Method B) Combined reaction time in microwave 40 min at 130° C. C29 A6 B9 (M + H).sup.+ = 574 0.98 min (Method B) Combined reaction time in microwave 40 min at 130° C. C32 A10 B9 (M + H).sup.+ = 409 1.45 min (Method A) C33 0 A2 B9 (M + H).sup.+ = 479 1.47 min (Method A) C34 A4 B9 (M + H).sup.+ = 437 1.54 min (Method A) C35 A2 B10 (M + H).sup.+ = 330 1.16 min (Method L) C36 A6 B10 (M + H).sup.+ = 425 1.28 min (Method K)

Example 9

Synthesis of intermediate C4 and C31

(77) Intermediate C4:

(78) ##STR00244##

(79) Intermediate A4 (1.50 g; 6.81 mmol) and tetrahydro-4H-pyran-4-one (13.0 mL; 140.76 mmol) in piperidine (1.36 mL; 13.62 mmol) are stirred at 100° C. for 15 min in a microwave. The solvent is evaporated. The residue is stirred in TBME. The precipitate is filtered off and dried.

(80) MS (ESI.sup.+): m/z=303 [M+H].sup.+

(81) HPLC (Method A): R.sub.t=1.40 min

(82) The following intermediates were prepared in an analogous manner to intermediate C4:

(83) TABLE-US-00022 R.sub.f Structure Educt Educt Mass value or 1 2 signal(s) R.sub.t No. Comment C31 A2 Tetra- hydro- 4H- pyran- 4-one M.sup.+ = 344 1.32 min (Method A)

Example 10

Synthesis of Intermediate C9 and C13

(84) Intermediate C9:

(85) ##STR00246##

(86) The following reaction is performed under a nitrogen atmosphere.

(87) Intermediate A2 (225 mg; 1.24 mmol) and 2,2-dimethoxy-1-methyl-pyrrolidine (450 mg; 3.10 mmol) in chloroform (2.5 mL) are stirred at reflux for 3 h. Additional 2,2-dimethoxy-1-methyl-pyrrolidine (1.2 eq.) is added and the mixture is stirred at 65° C. over night. The mixture is washed with sat. NaHCO.sub.3 solution. The organic layer is separated, washed with brine, dried and evaporated. The residue is purified by preparative HPLC (eluent A. water+0.15% conc. ammonia, eluent B: MeOH).

(88) MS (ESI.sup.+): m/z=344 [M+H].sup.+

(89) HPLC (Method A): R.sub.t=1.26 min

(90) The following intermediates were prepared in an analogous manner to intermediate C9:

(91) TABLE-US-00023 R.sub.f value Mass or Structure Educt 1 Educt 2 signal(s) or R.sub.t No. Comment C13 A4 2,2- dimethoxy- 1-methyl- pyrrolidine (M + H).sup.+ = 302 1.34 min (Method A)

Example 11

Synthesis of Intermediate C15 and C37

(92) Intermediate C15:

(93) ##STR00248##

(94) Intermediate C14 (81 mg; 0.16 mmol) in THF (5 mL) and potassium carbonate (51 mg; 0.37 mmol) in water (1.5 mL) are stirred at 40° C. for 2 h. The mixture is diluted with brine and EA. The organic layer is separated, dried and evaporated.

(95) MS (ESI.sup.+): m/z=345 [M+H].sup.+

(96) HPLC (Method B): R.sub.t=0.9 min

(97) The following intermediates were prepared in an analogous manner to intermediate C15:

(98) TABLE-US-00024 Educt Mass R.sub.f value or Structure 1 signal(s) R.sub.t No. Comment C37 C36 (M + H).sup.+ = 329 1.07 min (Method N) The residue is taken up in DCM/water. The precipitate is filtered off and dried.

Example 12

Synthesis of Intermediate C17

(99) ##STR00250##

(100) Intermediate C16 (800 mg; 1.86 mmol) in DCM/TFA (1/1; 15 mL) are stirred at room temperature for 1 h. The solvent is evaporated. The residue is taken up in DCM and washed with NaOH (aq. solution; 1M). The organic layer is separated, dried and evaporated.

(101) MS (ESI.sup.+): m/z=331 [M+H].sup.+

(102) HPLC (Method B): R.sub.t=0.91 min

Example 13

Synthesis of Intermediate C19

(103) ##STR00251##

(104) Step 1:

(105) Intermediate C18 (2.70 g; 5.71 mmol) in DCM/TFA (1/1; 30 mL) are stirred at room temperature for 1 h. The solvent is evaporated.

(106) MS (ESI.sup.+): m/z=373 [M+H].sup.+

(107) HPLC (Method B): R.sub.t=1.15 min

(108) Step 2:

(109) Intermediate C19 Step 1 (600 mg; 1.23 mmol), acetone (448 μL; 6.17 mmol) and glacial acetic acid (182 μL; 3.33 mmol) in MeOH (20 mL) are stirred at room temperature for 1 h. Sodium cyanoborohydride (155 mg; 2.47 mmol) is stirred at room temperature for 2 h. Additional acetone (2 mL) is added. After stirring over night the mixture is diluted with sat. NaHCO.sub.3 solution. The organic layer is separated, dried and evaporated.

(110) MS (ESI.sup.+): m/z=415 [M+H].sup.+

(111) HPLC (Method B): R.sub.t=1.12 min

Example 14

Synthesis of Intermediate C26

(112) ##STR00252##

(113) Step 1:

(114) Intermediate C18 (2.70 g; 5.71 mmol) in DCM/TFA (1/1; 30 mL) are stirred at room temperature for 1 h. The solvent is evaporated.

(115) MS (ESI.sup.+): m/z=373 [M+H].sup.+

(116) HPLC (Method B): R.sub.t=1.15 min

(117) Step 2:

(118) Intermediate C26 Step 1 (300 mg; 0.62 mmol), acetic anhydride (87 μL; 0.93 mmol) and TEA (316 μL; 1.85 mmol) in DCM (7 mL) are stirred at room temperature for 1 h. The mixture is diluted with sat. NaHCO.sub.3 solution and DCM. The organic layer is separated, dried and evaporated.

(119) MS (ESI.sup.+): m/z=415 [M+H].sup.+

(120) HPLC (Method B): R.sub.t=1.40 min

Example 15

Synthesis of Intermediate C30

(121) ##STR00253##

(122) Step 1:

(123) Intermediate C29 (500 mg; 0.87 mmol) in THF (17 mL) and potassium carbonate (157 mg; 1.13 mmol) in water (12 mL) are stirred at 40° C. for 5 h. The mixture is diluted with NaHCO.sub.3 (aq. solution; 9%) and extracted with EA. The organic layer is washed with brine, separated, dried and evaporated. The residue is purified by preparative HPLC (eluent A. water+0.15% conc. ammonia, eluent B: MeOH).

(124) MS (ESI.sup.+): m/z=478 [M+H]+

(125) HPLC (Method A): R.sub.t=1.46 min

(126) Step 2:

(127) Intermediate C30 Step 1 (118 mg; 0.25 mmol), acetone (90 μL; 1.24 mmol) and glacial acetic acid (36 μL; 0.67 mmol) in MeOH (8 mL) are stirred at room temperature for 2 h. Sodium cyanoborohydride (31 mg; 0.49 mmol) is added and the mixture stirred is stirred at 40° C. for 2 days. The mixture is diluted with NaHCO.sub.3 (aq. solution; 9%) and DCM. The organic layer is separated, dried and evaporated. The residue is stirred in MeOH, filtered off and dried. The residue is purified by preparative HPLC (eluent A. water+0.15% conc. ammonia, eluent B: MeOH).

(128) MS (ESI.sup.+): m/z=520 [M+H].sup.+

(129) HPLC (Method A): R.sub.t=1.58 min

Example 16

Synthesis of Intermediate C38

(130) ##STR00254##

(131) Intermediate C29 (203 mg; 0.35 mmol) in THF (7 mL) and potassium carbonate (64 mg; 0.46 mmol) in water (5 mL) are stirred at 40° C. for 3 days. The mixture is diluted with NaHCO.sub.3 (aq. solution; 9%) and extracted with EA. The organic layer is washed with brine, separated, dried and evaporated. The residue is purified by preparative HPLC (eluent A. water+0.15% conc. ammonia, eluent B: MeOH).

(132) MS (ESI.sup.+): m/z=494 [M+H].sup.+

(133) HPLC (Method A): R.sub.t=1.38 min

Example 17

Synthesis of intermediates D1-D12, D15-D26, and D28-D31

(134) Intermediate D1:

(135) ##STR00255##

(136) Intermediate C1 (862 mg; 2.60 mmol) and Raney-Nickel (150 mg) in MeOH (25 mL) and THF (50 mL) are hydrogenated in a Parr apparatus (rt; 50 psi; 4.5 h). The catalyst is filtered off and the solvent is evaporated.

(137) MS (ESI.sup.+): m/z=302 [M+H].sup.+

(138) HPLC (Method B): R.sub.t=0.67 min

(139) The following intermediates were prepared in an analogous manner to intermediate D1:

(140) TABLE-US-00025 Mass R.sub.f value or Structure Educt 1 signal(s) R.sub.t No. Comment D2 C2 (M + H).sup.+ = 330 0.73 min (Method B) D3 C3 (M + H).sup.+ = 288 0.89 min (Method A) D4 C6 (M + H).sup.+ = 288 0.65 min (Method M) D5 C7 (M + H).sup.+ = 342 0.69 min (Method B) D6 0 C8 (M + H).sup.+ = 246 0.39 min (Method O) D7 C9 (M + H).sup.+ = 314 0.96 min (Method A) D8 C10 (M + H).sup.+ = 274 0.71 min (Method C) D9 C11 (M + H).sup.+ = 332 0.95 min (Method A) D10 C12 (M + H).sup.+ = 272 1.14 min (Method A) D11 C13 (M + H).sup.+ = 272 0.71 min (Method G) D12 C15 (M + H).sup.+ = 315 D15 C19 (M + H).sup.+ = 385 0.71 min (Method B) D16 C20 (M + H).sup.+ = 288 0.91 min (Method G) D17 C21 (M + H).sup.+ = 330 0.76 min (Method B) D18 0 C22 (M + H).sup.+ = 387 0.92 min (Method B) Purification by preparative HPLC ((eluent A. water + 0.1% conc. ammonia, eluent B: MeOH) D19 C23 (M + H).sup.+ = 393 1.15 min (Method H) Purification by preparative HPLC (eluent A. water + 0.1% conc. ammonia, eluent B: MeOH) D20 C24 (M + H).sup.+ = 329 1.5 min (Method B) D21 C25 (M + H).sup.+ = 260 0.78 min (Method F) D22 C26 (M + H).sup.+ = 385 0.84 min (Method B) D23 C27 (M + H).sup.+ = 330 0.79 min (Method B) D24 C17 (M + H).sup.+ = 301 0.36 min (Method B) D25 C28 (M + H).sup.+ = 304 0.69 min (Method B) D26 C30 (M + H).sup.+ = 356 1.14 min (Method A) D28 C32 (M + H).sup.+ = 379 1.25 min (Method A) D29 0 C33 (M + H).sup.+ = 449 1.28 min (Method A) D30 C24 (M + H).sup.+ = 463 1.05 min (Method B) D31 C34 (M + H).sup.+ = 407 1.34 min (Method a)

Example 18

Synthesis of Intermediate D13

(141) ##STR00283##

(142) Step 1:

(143) Intermediate C17 (270 mg; 0.82 mmol), acetic anhydride (85 μL; 0.90 mmol) and DIPEA (423 μL; 2.46 mmol) in DCM (5 mL) are stirred at room temperature for 15 min. The solvent is evaporated.

(144) MS (ESI.sup.+): m/z=373 [M+H].sup.+

(145) HPLC (Method A): R.sub.t=1.17 min

(146) Step 2:

(147) Intermediate D13 Step 1 (430 mg; 0.58 mmol) and Raney-Nickel (50 mg) in MeOH (5 mL) and THF (10 mL) are hydrogenated in a Parr apparatus (rt; 50 psi; 17 h). The catalyst is filtered off and the solvent is evaporated. The residue is purified by preparative HPLC (eluent A. water+0.1% conc. ammonia, eluent B: MeOH).

(148) MS (ESI.sup.+): m/z=343 [M+H].sup.+

Example 19

Synthesis of Intermediate D14

(149) ##STR00284##

(150) Step 1:

(151) Intermediate C17 (489 mg; 1.48 mmol), oxetan-3-one (160 mg; 2.22 mmol) and glacial acetic acid (218 μL; 4.00 mmol) in MeOH (20 mL) are stirred at room temperature for 1 h. Sodium cyanoborohydride (186 mg; 2.96 mmol) is added and the mixture is stirred at room temperature for 1 h. THF (5 mL) is added. After stirring over night additional sodium cyanoborohydride (186 mg; 2.96 mmol) is added. The mixture is diluted with water. The organic solvent is evaporated and the aqueous layer is extracted with DCM. The organic layer is separated, dried and evaporated. The residue is stirred in MeOH, filtered off and dried. The residue is purified by preparative HPLC (eluent A. water+0.1% conc. ammonia, eluent B: MeOH).

(152) MS (ESI.sup.+): m/z=387 [M+H].sup.+

(153) HPLC (Method A): R.sub.t=1.24 min

(154) Step 2:

(155) Intermediate D14 Step 1 (164 mg; 0.42 mmol) and Raney-Nickel (50 mg) in MeOH (5 mL) and THF (10 mL) are hydrogenated in a Parr apparatus (rt; 50 psi; 4 h). The catalyst is filtered off and the solvent is evaporated.

(156) MS (ESI.sup.+): m/z=357 [M+H].sup.+

(157) HPLC (Method B): R.sub.t=0.43 min

Example 20

Synthesis of Intermediate D27

(158) ##STR00285##

(159) Intermediate C31 (448 mg; 1.30 mmol) and powdered iron (392 mg; 7.02 mmol) in water (14 mL) and ethanol (29 mL) are heated to reflux. Glacial acetic acid (0.79 mL; 13.78 mmol) is added drop wise and the mixture is stirred for 1 h. The organic solvent is evaporated and the residue is taken up in DCM and water. The mixture is alkalised with NaOH (aq. solution; 5 mL). The mixture is filtered through celite. The organic layer is separated, dried and evaporated. The residue is stirred in MeOH/ACN, filtered off and dried.

(160) MS (ESI.sup.+): m/z=315 [M+H].sup.+

(161) HPLC (Method A): R.sub.t=1.07 min

Example 21

Synthesis of Intermediate E1

(162) ##STR00286##

(163) Intermediate A10 (11.0 g; 57.24 mmol) and Pd/C (10%; 1.0 g) in DCM (200 mL) are hydrogenated in a Parr apparatus (rt; 50 psi; 5 h). Additional Pd/C (10%; 1.0 g) and MeOH (100 mL) is added and the mixture is hydrogenated for 3 h. Formaldehyde (aq. solution; 37%; 22.70 mL; 304.90 mmol) is added and the mixture is stirred for 10 min without H.sub.2-pressure and for further 3 h with H.sub.2-pressure. The catalyst is filtered off and the solvent is evaporated. The residue is taken up in NaOH (aq. solution; 1M) and extracted with DCM. The organic layer is separated, dried and evaporated. The residue is purified by MPLC (DCM/MeOH=98/2).

(164) MS (ESI.sup.+): m/z=191 [M+H].sup.+

(165) HPLC (Method P): R.sub.t=0.25 min

Example 22

Synthesis of Intermediates F1, F2 and F4

(166) Intermediate F1:

(167) ##STR00287##

(168) Step 1:

(169) Intermediate D19 (430 mg; 1.10 mmol), 1,4-diiodo-butane (145 μL; 1.10 mmol) and potassium carbonate (303 mg; 2.19 mmol) in DMF (12 mL) are stirred at 70° C. for 2 h. After stirring over night at room temperature the mixture is diluted with NaHCO.sub.3 (aq. solution; 9%) and extracted with EA. The organic layer is washed with brine, separated, dried and evaporated. The residue is purified by preparative HPLC (eluent A. water+0.15% conc. ammonia, eluent B: MeOH).

(170) MS (ESI.sup.+): m/z=447 [M+H].sup.+

(171) HPLC (Method A): R.sub.t=1.57 min

(172) Step 2:

(173) Intermediate F1 Step 1 (203 mg; 0.46 mmol) and Pd/C (10%; 20 mg) in MeOH (20 mL) and THF (10 mL) are hydrogenated in a Parr apparatus (rt; 50 psi; 1 h). Additional Pd/C (10%) is added and the mixture is hydrogenated. The catalyst is filtered off and the solvent is evaporated.

(174) MS (ESI.sup.+): m/z=313 [M+H].sup.+

(175) HPLC (Method A): R.sub.t=1.33 min

(176) The following intermediates were prepared in an analogous manner to intermediate F1:

(177) TABLE-US-00026 Mass R.sub.f value or Structure Educt 1 Educt 2 signal(s) R.sub.t No. Comment F2 D28 1,4- Diiodo- butane (M + H).sup.+ = 299 F4 D30 1,4- Diiodo- butane (M + H).sup.+ = 383 0.65 min (Method B)

Example 23

Synthesis of Intermediates F3 and F5

(178) Intermediate F3:

(179) ##STR00290##

(180) Step 1:

(181) Intermediate D29 (0.78 g; 1.74 mmol), acetone (631 μL; 8.70 mmol) and glacial acetic acid (256 μL; 4.70 mmol) in MeOH (20 mL) are stirred for 2 h at room temperature. Sodium cyanoborohydride (219 mg; 3.48 mmol) is added and the mixture is stirred at room temperature over night. The mixture is diluted with NaHCO.sub.3 (aq. solution; 9%) and extracted with DCM. The organic layer is separated, dried and evaporated. The residue is purified by preparative HPLC (eluent A. water+0.15% conc. ammonia, eluent B: MeOH).

(182) MS (ESI.sup.+): m/z=491 [M+H].sup.+

(183) HPLC (Method A): R.sub.t=1.49 min

(184) Step 2:

(185) Intermediate F3 Step 1 (348 mg; 0.71 mmol) and Pd/C (10%; 35 mg) in MeOH (14 mL) and THF (10 mL) are hydrogenated in a Parr apparatus (rt; 50 psi; 1.25 h). The catalyst is filtered off and the solvent is evaporated. The residue is purified by preparative HPLC (eluent A. water+0.15% conc. ammonia, eluent B: MeOH).

(186) MS (ESI.sup.+): m/z=357 [M+H].sup.+

(187) HPLC (Method A): R.sub.t=1.19 min

(188) The following intermediate was prepared in an analogous manner to intermediate F3:

(189) TABLE-US-00027 Mass R.sub.f value or Structure Educt 1 Educt 2 signal(s) R.sub.t No. Comment F5 D31 Acetone (M + H).sup.+ = 315 1.29 min (Method A)

Example 24

Synthesis of Intermediate G1

(190) ##STR00292##

(191) The following reaction is performed under an argon atmosphere.

(192) Intermediate E1 (700 mg; 3.68 mmol) and carbon disulfide (0.24 mL; 4.05 mmol) in DMF (15 mL) are cooled in an ice bath. Sodium hydride (55% in mineral oil; 0.32 g; 7.36 mmol) is added and the mixture is stirred for 20 min. The mixture is allowed to warm up to room temperature. After 1 h of stirring at room temperature the mixture is poured on ice water. The resulting precipitate is filtered off, washed with water and dried.

(193) MS (ESI.sup.+): m/z=295 [M+H].sup.+

(194) HPLC (Method H): R.sub.t=1.05 min

Example 25

Synthesis of Compounds 1001 to 1123

(195) Compound 1001:

(196) ##STR00293##

(197) Intermediate D1 (150 mg; 0.50 mmol), dihydro-furan-3-one (58 μL; 0.75 mmol) and glacial acetic acid (73 μL; 1.34 mmol) in MeOH (3 mL) are stirred for 1 h at room temperature. Sodium cyanoborohydride (63 mg; 1.00 mmol) is added and the mixture is stirred for 1 h at room temperature. The mixture is purified by preparative HPLC (eluent A. water+0.1% conc. ammonia, eluent B: MeOH).

(198) MS (ESI.sup.+): m/z=372 [M+H].sup.+

(199) HPLC (Method A): R.sub.t=1.11 min

(200) In analogy to the preparation of compound 1001 the following compounds are obtained:

(201) TABLE-US-00028 Mass Nr. Structure Educt 1 Educt 2 signal(s) R.sub.t 1002 D1 (M + H).sup.+ = 374 0.98 min (Method A) 1003 D1 (M + H).sup.+ = 386 1.13 min (Method A) 1004 D1 (M + H).sup.+ = 374 1.21 min (Method A) 1005 00 D1 01 (M + H).sup.+ = 344 0.85 min (Method B) 1006 02 D2 03 (M + H).sup.+ = 398 1.4 min (Method A) 1007 04 D2 05 (M + H).sup.+ = 372 0.9 min (Method B) 1008 06 D2 07 (M + H).sup.+ = 384 1.33 min (Method A) 1009 08 D2 09 (M + H).sup.+ = 400 1.15 min (Method A) 1010 0 D17 (M + H).sup.+ = 372 0.92 min (Method B) 1011 D17 (M + H).sup.+ = 400 0.85 min (Method B) 1012 D3 (M + H).sup.+ = 342 1.27 min (Method A) 1013 D3 (M + H).sup.+ = 358 1.41 min (Method A) 1014 D3 (M + H).sup.+ = 356 1.35 min (Method A) 1015 0 D3 (M + H).sup.+ = 356 1.33 min (Method A) 1016 D3 (M + H).sup.+ = 330 1.22 min (Method A) 1017 D3 (M + H).sup.+ = 358 1.42 min (Method A) 1018 D3 (M + H).sup.+ = 360 1.18 min (Method A) 1019 Example 176 (M + H).sup.+ = 343 1.30 min (Method A) 1020 0 Example 176 (M + H).sup.+ = 341 1.53 min (Method A) 1021 Example 176 (M + H).sup.+ = 315 1.44 min (Method A) 1022 Example 176 (M + H).sup.+ = 327 1.48 min (Method A) 1023 Example 173 (M + H).sup.+ = 398 1.53 min (Method A) 1024 Example 173 (M + H).sup.+ = 358 0.83 min (Method C) 1025 0 Example 173 (M + H).sup.+ = 384 1.38 min (Method A) 1026 Example 173 (M + H).sup.+ = 370 1.33 min (Method A) 1027 D4 (M + H).sup.+ = 370 0.74 min (Method D) 1028 D4 (M + H).sup.+ = 330 1.37 min (Method A) 1029 D4 (M + H).sup.+ = 342 1.42 min (Method A) 1030 0 D5 (M + H).sup.+ = 412 0.78 min (Method B) 1031 D5 (M + H).sup.+ = 384 0.84 min (Method B) 1032 D6 (M + H).sup.+ = 369 1033 D6 (M + H).sup.+ = 300 1.29 min (Method A) 1034 D6 (M + H).sup.+ = 288 1.23 min (Method A) 1035 0 D7 (M + H).sup.+ = 368 1.31 min (Method A) 1036 D7 (M + H).sup.+ = 384 1.12 min (Method A) 1037 D7 (M + H).sup.+ = 356 1.26 min (Method A) 1038 D8 (M + H).sup.+ = 316 0.9 min (Method C) 1039 D9 (M + H).sup.+ = 374 1.28 min (Method A) 1040 0 D9 (M + H).sup.+ = 402 1.45 min (Method A) 1041 D9 (M + H).sup.+ = 400 1.39 min (Method A) 1042 D10 (M + H).sup.+ = 314 1.42 min (Method A) 1043 D10 (M + H).sup.+ = 380 1.66 min (Method A) 1044 D11 (M + H).sup.+ = 314 0.67 min (Method A) 1045 0 D11 (M + H).sup.+ = 326 0.81 min (Method A) 1046 D12 (M + H).sup.+ = 399 0.71 min (Method B) 7 eq. of acetone and 4 eq. of borohydride were used, due to double alkylation 1047 D12 (M + H).sup.+ = 441 0.74 min (Method B) 7 eq. of acetone and 4 eq. of borohydride were used, due to triple alkylation 1048 D13 (M + H).sup.+ = 385 1.21 min (Method A) 1049 D14 (M + H).sup.+ = 399 0.63 min (Method B) 1050 0 D15 (M + H).sup.+ = 427 1.46 min (Method A) 1051 D16 (M + H).sup.+ = 330 1.45 min (Method A) 1052 D18 (M + H).sup.+ = 415 0.94 min (Method B) 1053 D19 (M + H).sup.+ = 421 1.45 min (Method F) 1054 D20 (M + H).sup.+ = 413 1.45 min (Method B) 5 eq. of acetone and 2 eq. of borohydride were used, due to double alkylation 1055 00 D21 01 (M + H).sup.+ = 288 0.86 min (Method F) 1056 02 D22 03 (M + H).sup.+ = 427 0.97 min (Method B) 1057 04 D23 05 (M + H).sup.+ = 372 0.98 min (Method B) 1058 06 D24 07 (M + H).sup.+ = 385 0.7 min (Method B) 5 eq. of acetone and 2 eq. of borohydride were used, due to double alkylation 1059 08 D25 09 (M + H).sup.+ = 346 0.85 min (Method B) 1060 0 D18 (M + H).sup.+ = 429 1.01 min (Method B) 1061 D26 (M + H).sup.+ = 440 1.51 min (Method A) 1062 D27 (M + H).sup.+ = 440 0.97 min (Method G) 1063 D8 (M + H).sup.+ = 379 0.29 min (Method E) 1064 D8 (M + H).sup.+ = 388 0.30 min (Method E) 1065 0 D8 (M + H).sup.+ = 381 0.30 min (Method E) 1066 D8 (M + H).sup.+ = 358 0.30 min (Method E) 1067 D8 (M + H).sup.+ = 356 0.37 min (Method E) 1068 D8 (M + H).sup.+ = 371 0.24 min (Method E) 1069 D8 (M + H).sup.+ = 358 0.30 min (Method E) 1070 0 D8 (M + H).sup.+ = 330 0.34 min (Method E) 1071 D8 (M + H).sup.+ = 358 0.38 min (Method E) 1072 D8 (M + H).sup.+ = 370 0.39 min (Method E) 1073 D8 (M + H).sup.+ = 380 0.31 min (Method E) 1074 D8 (M + H).sup.+ = 328 0.33 min (Method E) 1075 0 D8 (M + H).sup.+ = 379 0.32 min (Method E) 1076 D8 (M + H).sup.+ = 392 0.36 min (Method E) 1077 D8 (M + H).sup.+ = 376 0.30 min (Method E) 1078 D8 (M + H).sup.+ = 354 0.28 min (Method E) 1079 D8 (M + H).sup.+ = 372 0.31 min (Method E) 1080 0 D8 (M + H).sup.+ = 342 0.35 min (Method E) 1081 D8 (M + H).sup.+ = 344 0.36 min (Method E) 1082 D8 (M + H).sup.+ = 399 0.26 min (Method E) 1083 D8 (M + H).sup.+ = 342 0.34 min (Method E) 1084 D8 (M + H).sup.+ = 330 0.31 min (Method E) 1085 0 D8 (M + H).sup.+ = 356 0.34 min (Method E) 1086 D8 (M + H).sup.+ = 358 0.31 min (Method E) 1087 D8 (M + H).sup.+ = 380 0.30 min (Method E) 1088 D8 (M + H).sup.+ = 358 0.30 min (Method E) 1089 D8 (M + H).sup.+ = 379 0.30 min (Method E) 1090 0 D8 (M + H).sup.+ = 385 0.36 min (Method E) 1091 D8 (M + H).sup.+ = 344 0.37 min (Method E) 1092 D8 (M + H).sup.+ = 344 0.36 min (Method E) 1093 D8 (M + H).sup.+ = 399 0.29 min (Method E) 1094 D8 (M + H).sup.+ = 330 0.35 min (Method E) 1095 0 Example 173 (M + H).sup.+ = 441 0.24 min (Method E) 1096 Example 173 (M + H).sup.+ = 400 0.28 min (Method E) 1097 Example 173 (M + H).sup.+ = 384 0.32 min (Method E) 1098 Example 173 (M + H).sup.+ = 396 0.26 min (Method E) 1099 Example 173 (M + H).sup.+ = 372 0.31 min (Method E) 1100 0 Example 173 (M + H).sup.+ = 384 0.32 min (Method E) 1101 Example 173 (M + H).sup.+ = 412 0.37 min (Method E) 1102 Example 173 (M + H).sup.+ = 386 0.33 min (Method E) 1103 Example 173 (M + H).sup.+ = 421 0.29 min (Method E) 1104 Example 173 (M + H).sup.+ = 434 0.34 min (Method E) 1105 00 Example 173 01 (M + H).sup.+ = 424 0.28 min (Method E) 1106 02 Example 173 03 (M + H).sup.+ = 418 0.28 min (Method E) 1107 04 Example 173 05 (M + H).sup.+ = 414 0.29 min (Method E) 1108 06 Example 173 07 (M + H).sup.+ = 388 0.29 min (Method E) 1109 08 Example 173 09 (M + H).sup.+ = 421 0.27 min (Method E) 1110 0 Example 173 (M + H).sup.+ = 439 0.27 min (Method E) 1111 Example 173 (M + H).sup.+ = 398 0.34 min (Method E) 1112 Example 173 (M + H).sup.+ = 370 0.30 min (Method E) 1113 Example 173 (M + H).sup.+ = 422 0.29 min (Method E) 1114 Example 173 (M + H).sup.+ = 422 0.28 min (Method E) 1115 0 Example 173 (M + H).sup.+ = 398 0.34 min (Method E) 1116 Example 173 (M + H).sup.+ = 430 0.28 min (Method E) 1117 Example 173 (M + H).sup.+ = 413 0.23 min (Method E) 1118 Example 173 (M + H).sup.+ = 400 0.28 min (Method E) 1119 Example 173 (M + H).sup.+ = 400 0.29 min (Method E) 1120 0 Example 173 (M + H).sup.+ = 372 0.28 min (Method E) 1121 D14 (M + H).sup.+ = 399 0.63 min (Method B) 1122 D6 (M + H).sup.+ = 328 1.52 min (Method A) 1123 D8 (M + H).sup.+ = 397 0.29 min (Method E)

Example 26

Synthesis of Compounds 1124 to 1147

(202) Compound 1124:

(203) ##STR00538##

(204) Compound 1173 (70 mg; 0.22 mmol), 2-iodo-2-(-2-iodo-ethoxy)-ethane (32 μL; 0.22 mmol) and potassium carbonate (61 mg; 0.44 mmol) in DMF (3 mL) are stirred for 2 h at 70° C. Additional 2-iodo-2-(2-thoxy)-ethane (0.22 mmol) and potassium carbonate (0.44 mmol) are added. The mixture is stirred at 70° C. for 1.5 h and at room temperature for 3 days. The mixture is diluted with NaHCO.sub.3 (aq. solution; 9%) and extracted with EA. The organic layer is washed with brine, separated, dried and evaporated. The residue is purified by preparative HPLC (eluent A: water+0.1% conc. Ammonia, eluent B: MeOH).

(205) MS (ESI.sup.+): m/z=386 [M+H].sup.+

(206) HPLC (Method A): R.sub.t=1.19 min

(207) In analogy to the preparation of example 1124 the following compounds are obtained:

(208) TABLE-US-00029 Mass Nr. Structure Educt 1 Educt 2 signal(s) R.sub.t 1125 Example 1173 0 (M + H).sup.+ = 398 1.51 min (Method A) 1126 Example 1173 (M + H).sup.+ = 384 1.43 min (Method A) 1127 Example 1173 (M + H).sup.+ = 370 0.83 min (Method G) 1128 D23 (M + H).sup.+ = 412 1.58 min (Method A) 1129 D2 (M + H).sup.+ = 412 0.91 min (Method B) 1130 D27 0 (M + H).sup.+ = 369 1.51 min (Method A) 1131 D17 (M + H).sup.+ = 412 1.54 min (Method A) 1132 D8 (M + H).sup.+ = 342 1.29 min (Method A) 1133 D8 (M + H).sup.+ = 328 0.83 min (Method C) 1134 D15 (M + H).sup.+ = 467 1.71 min (Method A) 1135 D15 0 (M + H).sup.+ = 439 1.67 min (Method A) 1136 D22 (M + H).sup.+ = 439 0.93 min (Method B) 1137 D4 (M + H).sup.+ = 342 0.72 min (Method D) 1138 D10 (M + H).sup.+ = 342 1.36 min (Method A) 1139 Example 1177 (M + H).sup.+ = 327 1.58 min (Method A) 1140 D25 0 (M + H).sup.+ = 358 0.81 min (Method B) 1141 D25 (M + H).sup.+ = 374 0.8 min (Method B) 1142 D18 (M + H).sup.+ = 442 1.05 min (Method B) 1143 D18 (M + H).sup.+ = 441 0.99 min (Method B) 1144 D6 (M + H).sup.+ = 328 1.52 min (Method A) 1145 D21 0 (M + H).sup.+ = 314 0.92 min (Method F) 1146 D19 (M + H).sup.+ = 447 1.17 min (Method F) 1147 D1 (M + H).sup.+ = 384 0.87 min (Method B)

Example 27

Synthesis of Compounds 1148 to 1151

(209) Compound 1148:

(210) ##STR00585##

(211) Intermediate E1 (200 mg; 1.05 mmol) and 7-methoxy-3,4,5,6-tetrahydro-2H-azepine (134 mg; 1.05 mmol) are stirred for 20 minutes in a microwave at 170° C. The residue is purified by preparative HPLC (eluent A: water+0.1% conc. Ammonia, eluent B: MeOH).

(212) MS (ESI.sup.+): m/z=286 [M+H].sup.+

(213) HPLC (Method H): R.sub.t=1.05 min

(214) In analogy to the preparation of example 27 the following compounds are obtained:

(215) TABLE-US-00030 Mass Nr. Structure Educt 1 Educt 2 signal(s) R.sub.t 1149 E1 (M + H).sup.+ = 272 0.98 min (Method H) 1150 E1 (M + H).sup.+ = 274 0.43 min (Method I) 1151 0 E1 (M + H).sup.+ = 258 0.89 min (Method H)

Example 28

Synthesis of Compounds 1152 to 1163

(216) Compound 1152:

(217) ##STR00592##

(218) Intermediate F1 (30 mg; 0.10 mmol), 3-(2H)-furanone dihydrochloride (9 μL; 0.12 mmol) and glacial acetic acid (13 μL; 0.24 mmol) in MeOH (2 mL) are stirred at 50° C. for 1 h. Sodium cyanoborohydride (12 mg; 0.19 mmol) is added and stirred at room temperature over night. The mixture is diluted with NaHCO.sub.3 (aq. solution; 9%) and extracted with DCM. The organic layer is separated, dried and evaporated. The residue is purified by preparative HPLC (eluent A: water+0.1% conc. Ammonia, eluent B: MeOH).

(219) MS (ESI.sup.+): m/z=383 [M+H].sup.+

(220) HPLC (Method A): R.sub.t=1.42 min

(221) In analogy to the preparation of example 1152 the following compounds are obtained:

(222) TABLE-US-00031 Mass Nr. Structure Educt 1 Educt 2 signal(s) R.sub.t 1153 F1 (M + H).sup.+ = 355 1.54 min (Method A) 1154 F1 (M + H).sup.+ = 369 1.38 min (Method A) 1155 F1 (M + H).sup.+ = 327 1.42 min (Method A) 1156 F2 00 (M + H).sup.+ = 341 1.53 min (Method A) 1157 01 F3 02 (M + H).sup.+ = 413 1.24 min (Method A) 1158 03 F3 04 (M + H).sup.+ = 441 1.30 min (Method A) 1159 05 F3 06 (M + H).sup.+ = 399 1.38 min (Method A) 1160 07 F4 08 (M + H).sup.+ = 439 0.67 min (Method A) 1161 09 F5 0 (M + H).sup.+ = 357 1.45 min (Method A) 1162 F5 (M + H).sup.+ = 371 1.34 min (Method A) 1163 F5 (M + H).sup.+ = 399 1.38 min (Method A)

Example 29

Synthesis of Compounds 1164

(223) ##STR00615##

(224) Intermediate F4 (100 mg; 0.26 mmol), acetic anhydride (37 μL; 0.39 mmol) and DIPEA (134 μL; 0.78 mmol) in DCM (3 mL) are stirred at room temperature for 1 h. The mixture is diluted with NaHCO.sub.3 (aq. solution; 9%) and extracted with DCM. The organic layer is separated, dried and evaporated. The residue is taken up in MeOH and the precipitate is filtered off. The precipitate is purified by preparative HPLC (eluent A: water+0.1% conc. ammonia, eluent B: MeOH).

(225) MS (ESI.sup.+): m/z=425 [M+H].sup.+

(226) HPLC (Method B): R.sub.t=0.84 min

Example 30

Synthesis of Compound 1165 to 1166

(227) Compound 1165:

(228) ##STR00616##

(229) Intermediate F5 (30 mg; 0.10 mmol), acetyl chloride (6 μL; 0.09 mmol) and TEA (20 μL; 0.14 mmol) in THF (3 mL) are stirred at room temperature for 10 minutes. Additional acetyl chloride is added. The mixture is diluted with water and extracted with DCM. The organic layer is separated, dried and evaporated. The residue is purified by preparative HPLC (eluent A: water+0.1% TFA, eluent B: MeOH).

(230) MS (ESI.sup.+): m/z=357 [M+H].sup.+

(231) HPLC (Method D): R.sub.t=0.62 min

(232) In analogy to the preparation of compound 1165 the following compound is obtained:

(233) TABLE-US-00032 Mass Nr. Structure Educt 1 Educt 2 signal(s) R.sub.t 1166 F3 (M + H).sup.+ = 399 0.88 min (Method J)

Example 31

Synthesis of Compounds 1167 to 1168

(234) Compound 1167:

(235) ##STR00619##

(236) Intermediate G1 (90 mg; 0.31 mmol) and N1-phenyl-ethane-1,2-diamine (42 mg; 0.31 mmol) in n-butanol (2 mL) are stirred for 15 min in a microwave at 220° C. The solvent is evaporated and the residue is purified by preparative HPLC (eluent A: water+0.1% conc. ammonia, eluent B: MeOH).

(237) MS (ESI.sup.+): m/z=335 [M+H].sup.+

(238) HPLC (Method H): R.sub.t=1.02 min

(239) In analogy to the preparation of example 1167 the following compounds are obtained:

(240) TABLE-US-00033 Mass Nr. Structure Educt 1 Educt 2 signal(s) R.sub.t 1168 0 G1 (M + H).sup.+ = 341 1.16 min (Method J)

Example 32

Synthesis of Compound 1169 to 1171

(241) Compound 1169:

(242) ##STR00622##

(243) Intermediate C35 (100 mg; 0.26 mmol), formaldehyde (aq. solution; 37%; 0.19 mL; 2.58 mmol) and Raney-Nickel (50 mg) in MeOH (10 mL) are hydrogenated in a Parr apparatus (rt; 1.1 bar; 8 h). The catalyst is filtered off and the solvent is evaporated. The residue is stirred in MeOH, filtered off and dried.

(244) MS (ESI.sup.+): m/z=328 [M+H]+

(245) HPLC (Method ?): R.sub.t=0.72 min

(246) In analogy to the preparation of example 1169 the following compounds are obtained:

(247) TABLE-US-00034 Educt Mass Nr. Structure 1 signal(s) R.sub.t 1170 C37 (M + H).sup.+ = 341 0.75 min (Method K) 15 eq. of formaldehyde used, due triple alkylation 1171 C36 (M + H).sup.+ = 423 1.06 min (Method K)

Example 33

Synthesis of Compound 1172 to 1174

(248) Compound 1172:

(249) ##STR00625##

(250) Intermediate C35 (200 mg; 0.52 mmol) and Raney-Nickel (50 mg) in MeOH (10 mL) are hydrogenated in a Parr apparatus (rt; 1.1 bar; 8 h). The catalyst is filtered off and the solvent is evaporated. The residue is purified by preparative HPLC (eluent A: water+0.1% conc. ammonia, eluent B: MeOH).

(251) MS (ESI.sup.+): m/z=300 [M+H].sup.+

(252) HPLC (Method L): R.sub.t=0.63 min

(253) In analogy to the preparation of example 1172 the following compounds are obtained:

(254) TABLE-US-00035 Educt Mass Nr. Structure 1 signal(s) R.sub.t 1173 C5 (M + H).sup.+ = 316 0.69 min (Method B) 1174 C38 (M + H).sup.+ = 464 1.24 min (Method A)

Example 34

Synthesis of Compound 1175

(255) ##STR00628##

(256) Intermediate C35 (200 mg; 0.52 mmol) and Raney-Nickel (50 mg) in MeOH (10 mL) are hydrogenated in a Parr apparatus (rt; 1.1 bar; 8 h). The catalyst is filtered off and the solvent is evaporated. The residue is purified by preparative HPLC (eluent A: water+0.1% conc. ammonia, eluent B: MeOH).

(257) MS (ESI.sup.+): m/z=314 [M+H].sup.+

(258) HPLC (Method M): R.sub.t=1.12 min

Example 35

Synthesis of Compound 1176

(259) ##STR00629##

(260) Intermediate C4 (1.78 g; 5.90 mmol) and powdered iron (1.78 g; 31.87 mmol) in water (56.5 mL) and ethanol (116 mL) are stirred at 80° C. Glacial acetic acid (3.58 mL; 62.55 mmol) are added drop wise and the mixture is stirred for 1 h at 80° C. The organic solvent is evaporated and the aq. layer is alkalised with NaOH (7 mL) and extracted with DCM. Iron is filtered off through celite. The organic layer is separated, dried and evaporated. The residue is purified by HPLC (eluent A: water+0.15% conc. ammonia, eluent B: MeOH).

(261) MS (ESI.sup.+): m/z=273 [M+H].sup.+

(262) HPLC (Method A): R.sub.t=1.16 min

Example 36

Synthesis of Compound 1177

(263) ##STR00630##

(264) Intermediate D1 (150 mg; 0.50 mmol), bromo-cyclobutane (101 mg; 0.75 mmol) and potassium carbonate (138 mg; 1.00 mmol) in DMF (3 mL) are stirred at 70° C. for 4 h. Additional bromo-cyclobutane (101 mg; 0.75 mmol) is added and the mixture is stirred at 80° C. over night. Additional bromo-cyclobutane (101 mg; 0.75 mmol) is added. After stirring for 4 h at 90° C. the mixture is purified by preparative HPLC (eluent A: water+0.15% conc. ammonia, eluent B: MeOH).

(265) MS (ESI.sup.+): m/z=356 [M+H].sup.+

(266) HPLC (Method B): R.sub.t=0.89 min

Example 37

Synthesis of Compound 1178

(267) ##STR00631##

(268) Example 146 (65 mg; 0.15 mmol) and Pd/C (10%; 10 mg) in MeOH (5 mL) is hydrogenated in a Parr apparatus (rt; 50 psi; 1 h). The catalyst is filtered off and the solvent is removed.

(269) The residue is purified by preparative HPLC (eluent A: water+0.15% conc. ammonia, eluent B: MeOH).

(270) MS (ESI.sup.+): m/z=313 [M+H].sup.+

(271) HPLC (Method A): R.sub.t=1.34 min

Example 38

Biological Assays

(272) The biological activity of compounds is determined by the following methods:

(273) Assay A: Determination of Complex I mediated ROS-inhibition (CI)

(274) Enzyme kinetic experiments permit the detection of ROS generated through Complex I. Herefore Complex I was purified from bovine heart (Sharpley et al. 2006 Biochemistry. 45(1):241-8. First a subcellular fractionation was conducted to obtain a crude mitochondria fraction, followed by a hypotonoc lysis and differential centrifugation, from which mitochondrial membranes were obtained. Solubilzation of mitochondrial membranes followed by an ion exchange chromatography and size exclusion chromatography resulted in enzyme preparations which contain Complex I with little contaminations of Complex IV. These preparations were used to study ROS-generation by Complex I, the substrate NADH (1 μM) and oxygen (ambient). The generated ROS are detected via the Oxidation of Amplex red in a coupled reaction containing Amplex Red and horse radish peroxidase

(275) IC50 of a compound of the invention was estimated by testing the compound using a 8 point concentration-response experiment.

(276) In 384-well microtiter plates 5 μl of test compound (final concentrations ranging from 0.01 nM to 30 μM, diluted in assay buffer and 1% DMSO final) or control was mixed with 5 μl of substrate mix (3 μM NADH, 10 μM AmplexRed, 1 mM Fructose 1,6 bis-phopshate and 1 mM AsO4). The enzymatic reaction was started by addition of 15 μl of enzyme mix (containing 20 μg/ml Complex I, 2 U/ml horse radish peroxidase, 1 U/ml Aldolase, 1 U/ml Trioseisomerase, 1 U/ml Glycerinaldehyd-3-phosphat-Dehydrogenase) and the generation of ROS was followed by measuring the increase in absorption at 557 nm every 53 seconds at room temperature for 12 minutes, followed by linear regression (slope analysis). To assess the potency of Compounds IC50 values are calculated as 50% activity of Complex I by nonlinear regression curve fitting, using a 4-parameter sigmoidal dose-response model.

(277) Assay B: Determination of Cellular Protection (HT22)

(278) To show selective pathway engagement in a cellular context, a mice neuroblastoma cells (HT-22) were depleted of the endogenous antioxidant glutathione resulting on oxidative stress on the mitochondrial and cellular level and cell death (Tan S, Sagara Y, Liu Y, Maher P, Schubert D. The regulation of reactive oxygen species production during programmed cell death. J Cell Biol. 1998; 141:1423-1432). By incubation with high concentrations of Glutamate (5 mM) these cells are depleted of intracellular glutathione do to a inhibition of cystine uptake, which results in an accumulation of mitochondrial derived ROS and ultimately cell death.

(279) In 384-well plates, 2000 HT-22 cells were seeded in 50 ul cell culture medium (DMEM containing 10% fetal calf serum and 1% penicillin/streptomycin) and were cultured for 24 hours, followed by the incubation with glutamate (to induce cell death) or vehicle (viable cells (100%) in presence of test compound (0.01-30 uM final) for 16 hours. Viability of the cells were assessed by adding 10% Alamar Blue reagent and incubation for 1 h at 37° C., followed by measuring Fluorescence (excitation 530 nm, emission 590 nm.

(280) To assess the potency of compounds, EC50 values were calculated by nonlinear regression curve fitting, using a 4-parameter sigmoidal dose-response model (see Complex I assay).