Conformationally constrained, fully synthetic macrocyclic compounds
09695191 · 2017-07-04
Assignee
Inventors
- Daniel Obrecht (Bättwil, CH)
- Philipp Ermert (Allschwil, CH)
- Said Oumouch (Mulhouse, FR)
- Franck Lach (Les Grandes Loges, FR)
- Anatol Luther (Binzen, DE)
- Karsten Marx (Basel, CH)
- Kerstin MÖHLE (Wettswil, CH)
Cpc classification
A61P29/00
HUMAN NECESSITIES
A61P25/18
HUMAN NECESSITIES
A61K31/437
HUMAN NECESSITIES
A61K31/407
HUMAN NECESSITIES
A61P43/00
HUMAN NECESSITIES
A61P1/06
HUMAN NECESSITIES
A61P1/00
HUMAN NECESSITIES
A61P25/28
HUMAN NECESSITIES
A61K31/4985
HUMAN NECESSITIES
International classification
C07D273/02
CHEMISTRY; METALLURGY
A61K31/4985
HUMAN NECESSITIES
C07D273/00
CHEMISTRY; METALLURGY
A61K31/407
HUMAN NECESSITIES
Abstract
Conformationally restricted, spatially defined 12-30 membered macrocyclic ring systems of type (I) are constituted by three distinct building blocks: an aromatic template a, a conformation modulator b and a spacer moiety c as detailed in the description and the claims. Macrocycles of type (I) are readily manufactured by parallel synthesis or combinatorial chemistry. They are designed to interact with specific biological targets. In particular, they show agonistic or antagonistic activity on the motilin receptor (MR receptor), on the serotonin receptor of subtype 5-HT.sub.2B (5-HT.sub.2B receptor), and on the prostaglandin F2receptor (FP receptor). They are thus potentially useful for the treatment of hypomotility disorders of the gastrointestinal tract such as diabetic gastroparesis and constipation type irritable bowl syndrome; of CNS related diseases like migraine, schizophrenia, psychosis or depression; of ocular hypertension such as associated with glaucoma and preterm labour. ##STR00001##
Claims
1. Compounds of the general formula I incorporating the building blocks A, B and C ##STR02704## wherein the encircled moiteties a, b and c in building blocks A, B and C are selected from Tables 1, 2 and 3, respectively, and are appropriately and independently substituted as defined below: TABLE-US-00082 TABLE 1
2. Compounds according to claim 1 wherein A, B and C are selected from Tables 10, 2 and 12, and are appropriately and independently substituted as defined below: TABLE-US-00091 TABLE 10 Radicals A1(a1)-A626(a25)
3. Compounds according to claim 2 wherein A is A1(a1); A2(a1); A3(a1); A4(a1); A5(a1); A6(a1); A7(a1); A9(a1); A10(a1); A73(a2); A170(a4); A209(a7); A240(a10); A272(a10); A532(a18); A609(a24); A612(a24) and A614(a24) as shown in Table 13, below; TABLE-US-00094 TABLE 13 Building Blocks of Type A
4. Compounds according to claim 3 wherein the building blocks of type A are A1(a1); A2(a1); A3(a1); A4(a1); A5(a1); A6(a1); A7(a1); A9(a1); A10(a1); A73(a2); A170(a4); A209(a7); A240(a10); A272(a10); A532(a18); A614(a24) as shown in Table 16, below; TABLE-US-00097 TABLE 16 Building Blocks of Type A
5. Compounds according to claim 1 wherein readily accessible substances that define possible subunits of the linker C are those listed in Table 19, below; TABLE-US-00100 TABLE 19 Code Chemical Name Ala L-Alanine .sup.DAla D-Alanine Arg L-Arginine .sup.DArg D-Arginine Asn L-Asparagine .sup.DAsn D-Asparagine Asp L-Aspartic acid .sup.DAsp D-Aspartic acid Cys L-Cysteine .sup.DCys D-Cysteine Glu L-Glutamic acid .sup.DGlu D-Glutamic acid Gln L-Glutamine .sup.DGln D-Glutamine Gly Glycine His L-Histidine .sup.DHis D-Histidine Ile L-Isoleucine .sup.DIle D-Isoleucine Leu L-Leucine .sup.DLeu D-Leucine Lys L-Lysine .sup.DLys D-Lysine Met L-Methionine .sup.DMet D-Methionine Phe L-Phenylalanine .sup.DPhe D-Phenylalanine Pro L-Proline .sup.DPro D-Proline Ser L-Serine .sup.DSer D-Serine Thr L-Threonine .sup.DThr D-Threonine Trp L-Tryptophan .sup.DTrp D-Tryptophan Tyr L-Tyrosine .sup.DTyr D-Tyrosine Val L-Valine .sup.DVal D-Valine Apa 3-Amino-propanoic acid H-.sup.3-HAla-OH (3S)-3-Amino-butyric acid H-.sup.3-HVal-OH (3R)-3-Amino-4-methyl-valeric acid H-.sup.3-HIle-OH (3R,4S)-3-Amino-4-methyl-hexanoic acid H-.sup.3-HLeu-OH (3S)-3-Amino-5-methyl-hexanoic acid H-.sup.3-HMet-OH (3S)-3-Amino-5-methylthio pentanoic acid H-.sup.3-HTyr-OH (3S)-3-Amino-4-(4-hydroxyphenyl)-butyric acid H-.sup.3-HHis-OH (3S)-3-Amino-4-(imidazole-4-yl)-butyric acid H-.sup.3-HPhe-OH (3S)-3-Amino-4-phenyl butyric acid H-.sup.3-HTrp-OH (3S)-3-Amino-4-(indol-3-yl)-butyric acid H-.sup.3-HSer-OH (3R)-3-Amino-4-hydroxy-butyric acid H-.sup.3-HAsp-OH 3-Amino-pentanedioic acid H-.sup.3-HGlu-OH (3S)-3-Amino-hexanedioic acid H-.sup.3-HLys-OH (3S)-3,7-Diamino-heptanoic acid H-.sup.3-HArg-OH (3S)-3-Amino-6-guanidino-hexanoic-acid H-.sup.3-HCys-OH (3R)-3-Amino-4-mercapto-butyric acid H-.sup.3-HAsn-OH (3S)-3-Amino-4-carbamoyl-butyric acid H-.sup.3-HGln-OH (3S)-3-Amino-5-carbamoyl-pentanoic acid H-.sup.3-HThr-OH (3R,4R)-3-Amino-4-hydroxy-pentanoic acid Gaba 4-Amino-butyric acid H-.sup.4-DiHAla-OH (4S)-4-Amino-pentanoic acid H-.sup.4-DiHVal-OH (4R)-4-Amino-5-methyl-hexanoic acid H-.sup.4-DiHIle-OH (4R,5S)-4-Amino-5-methyl-heptanoic acid H-.sup.4-DiHLeu-OH (4R)-4-Amino-6-methyl-heptanoic acid H-.sup.4-DiHMet-OH (4R)-4-Amino-6-methylthio-hexanoic acid H-.sup.4-DiHTyr-OH (4R)-4-Amino-5-(4-hydroxyphenyl)- pentanoic acid H-.sup.4-DiHHis-OH (4R)-4-Amino-5-(imidazole-4-yl)- pentanoic acid H-.sup.4-DiHPhe-OH (4R)-4-Amino-5-phenyl-pentanoic acid H-.sup.4-DiHTrp-OH (4R)-4-Amino-5-(indol-3-yl)-pentanoic acid H-.sup.4-DiHSer-OH (4R)-4-Amino-5-hydroxy-pentanoic acid H-.sup.4-DiHAsp-OH (4R)-4-Amino-hexanedioic acid H-.sup.4-DiHGlu-OH 4-Amino-heptanedioic acid H-.sup.4-DiHLys-OH (4S)-4,8-Diamino-octanoic acid H-.sup.4-DiHArg-OH (4S)-4-Amino-7-guanidino-heptanoic-acid H-.sup.4-DiHCys-OH (4R)-4-Amino-5-mercapto-pentanoic acid H-.sup.4-DiHAsn-OH (4R)-4-Amino-5-carbamoyl-pentanoic acid H-.sup.4-DiHGln-OH (3S)-3-Amino-5-carbamoyl-hexanoic acid H-.sup.4-DiHThr-OH (4R,5R)-4-Amino-5-hydroxy-hexanoic acid Cit L-Citrulline .sup.DCit D-Citrulline Orn L-Ornithine .sup.DOrn D-Ornithine tBuA L-t-Butylalanine .sup.DtBuA D-t-Butylalanine Sar Sarcosine Pen L-Penicillamine .sup.DPen D-Penicillamine tBuG L-tert.-Butylglycine .sup.DtBuG D-tert.-Butylglycine 4AmPhe L-para-Aminophenylalanine .sup.D4AmPhe D-para-Aminophenylalanine 3AmPhe L-meta-Aminophenylalanine .sup.D3AmPne D-meta-Aminophenylalanine 2AmPhe L-ortho-Aminophenylalanine .sup.D2AmPhe D-ortho-Aminophenylalanine Phe(mC(NH.sub.2)NH) L-meta-Amidinophenylalanine .sup.DPhe(mC(NH.sub.2)NH) D-meta-Amidinophenylalanine Phe(pC(NH.sub.2)NH) L-para-Amidinophenylalanine .sup.DPhe(pC(NH.sub.2)NH) D-para-Amidinophenylalanine Phe(mNHC(NH.sub.2)NH) L-meta-Guanidinophenylalanine .sup.DPhe(mNHC(NH.sub.2)NH) D-meta-Guanidinophenylalanine Phe(pNHC(NH.sub.2)NH) L-para-Guanidinophenylalanine .sup.DPhe(pNHC(NH.sub.2)NH) D-para-Guanidinophenylalanine 2Pal (2S)-2-Amino-3-(pyridine-2-yl)-propionic acid .sup.D2Pal (2R)-2-Amino-3-(pyridine-2-yl)-propionic acid 4Pal (2S)-2-Amino-3-(pyridine-4-yl)-propionic acid .sup.D4Pal (2R)-2-Amino-3-(pyridine-4-yl)-propionic acid Phg L-Phenylglycine .sup.DPhg D-Phenylglycine Cha L-Cyclohexylalanine .sup.DCha D-Cyclohexylalanine C.sub.4al L-3-Cyclobutylalanine .sup.DC.sub.4al D-3-Cyclobutylalanine C.sub.5al L-3-Cyclopentylalanine .sup.DC.sub.5al D-3-Cyclopentylalanine Nle L-Norleucine .sup.DNle D-Norleucine 2-Nal L-2-Naphthylalanine .sup.D2Nal D-2-Naphthylalanine 1-Nal L-1-Naphthylalanine .sup.D1Nal D-1-Naphthylalanine 4ClPhe L-4-Chlorophenylalanine .sup.D4ClPhe D-4-Chlorophenylalanine 3ClPhe L-3-Chlorophenylalanine .sup.D3ClPhe D-3-Chlorophenylalanine 2ClPhe L-2-Chlorophenylalanine .sup.D2ClPhe D-2-Chlorophenylalanine 3,4Cl.sub.2Phe L-3,4-Dichlorophenylalanine .sup.D3,4Cl.sub.2Phe D-3,4-Dichlorophenylalanine 4FPhe L-4-Fluorophenylalanine .sup.D4FPhe D-4-Fluorophenylalanine 3FPhe L-3-Fluorophenylalanine .sup.D3FPhe D-3-Fluorophenylalanine 2FPhe L-2-Fluorophenylalanine .sup.D2FPhe D-2-Fluorophenylalanine Thi L--2-Thienylalanine .sup.DThi D--2-Thienylalanine Tza L-2-Thiazolylalanine .sup.DTza D-2-Thiazolylalanine Mso L-Methionine sulfoxide .sup.DMso D-Methionine sulfoxide AcLys N-Acetyllysine .sup.DAcLys N-Acetyl-D-lysine Dap 2,3-Diaminopropionic acid .sup.DDap D-2,3-Diaminopropionic acid Dab 2,4-Diaminobutyric acid .sup.DDab (2R)-2,4-Diaminobutyric acid Dbu (2S)-2,3-Diamino-butyric acid .sup.DDbu (2R)-2,3-Diamino-butyric acid Abu -Aminobutyric acid (GABA) Aha -Aminohexanoic acid Aib -Aminoisobutyric acid Cyp 1-Amino cyclopentane carboxylic acid Y(Bzl) L-O-Benzyltyrosine .sup.DY(Bzl) D-O-Benzyltyrosine H(Bzl) (3S)-2-Amino-3-(1-benzylimidazole-4- yl)-propionic acid .sup.DH(Bzl) (3R)-2-Amino-3-(1-benzylimidazole-4- yl)-propionic acid Bip L-(4-phenyl)phenylalanine .sup.DBip D-(4-phenyl)phenylalanine S(Bzl) L-O-Benzylserine .sup.DS(Bzl) D-O-Benzylserine T(Bzl) L-O-Benzylthreonine .sup.DT(Bzl) D-O-Benzylthreonine alloT (2S,3S)-2-Amino-3-hydroxy-butyric acid .sup.DalloT (2R,3S)-2-Amino-3-hydroxy-butyric acid Leu3OH (2S,3R)-2-Amino-3-hydroxy-4-methyl- pentanoic acid .sup.DLeu3OH (2R,3R)-2-Amino-3-hydroxy-4-methyl- pentanoic acid hAla L-Homo-alanine .sup.DhAla D-Homo-alanine hArg L-Homo-arginine .sup.DhArg D-Homo-arginine hCys L-Homo-cysteine .sup.DhCys D-Homo-cysteine hGlu L-Homo-glutamic acid .sup.DhGlu D-glutamic acid hGln L-Homo-glutamine .sup.DhGln D-Homo-glutamine hHis L-Homo-histidine .sup.DhHis D-Homo-histidine hIle L-Homo-isoleucine .sup.DhIle D-Homo-isoleucine hLeu L-Homo-leucine .sup.DhLeu D-Homo-leucine hNle L-Homo-norleucine .sup.DhNle D-Homo-norleucine hLys L-Homo-lysine .sup.DhLys D-Homo-lysine hMet L-Homo-Methionine .sup.DhMet D-Homo-Methionine hPhe L-Homo-phenylalanine .sup.DhPhe D-Homo-phenylalanine hSer L-Homo-serine .sup.DhSer D-Homo-serine hThr L-Homo-threonine .sup.DhThr D-Homo-threonine hTrp L-Homo-tryptophan .sup.DhTrp D-Homo-tryptophan hTyr L-Homo-tyrosine .sup.DhTyr D-Homo-tyrosine hVal L-Homo-valine .sup.DhVal D-Homo-valine hCha L-Homo-cyclohexylalanine .sup.DhCha D-Homo-cyclohexylalanine Bpa L-4-Benzoylphenylalanine .sup.DBpa D-4-Benzoylphenylalanine OctG L-Octylglycine .sup.DOctG D-Octylglycine Tic (3S)-1,2,3,4-Tetrahydroisoquinoline-3- carboxylic acid .sup.DTic (3R)-1,2,3,4-Tetrahydroisoquinoline-3- carboxylic acid Tiq (1S)-1,2,3,4-Tetrahydroisoquinoline-1- carboxylic acid .sup.DTiq (1R)-1,2,3,4-Tetrahydroisoquinoline-1- carboxylic acid Oic (2S,3aS,7aS)-1-Octahydro-1H-indole-2- carboxylic acid .sup.DOic (2R,3aS,7aS)-1-Octahydro-1H-indole-2- carboxylic acid 4AmPyrr1 (2S,4S)-4-Amino-pyrrolidine-2-carboxylic acid .sup.D4AmPyrr1 (2R,4S)-4-Amino-pyrrolidine-2-carboxylic acid 4AmPyrr2 (2S,4R)-4-Amino-pyrrolidine-2-carboxylic acid .sup.D4AmPyrr2 (2R,4R)-4-Amino-pyrrolidine-2-carboxylic acid 4PhePyrr1 (2S,4R)-4-Phenyl-pyrrolidine-2- carboxylic acid .sup.D4PhePyrr1 (2R,4R)-4-Phenyl-pyrrolidine-2- carboxylic acid 4PhePyrr2 (2S,4S)-4-Phenyl-pyrrolidine-2- carboxylic acid .sup.D4PhePyrr2 (2R,4S)-4-Phenyl-pyrrolidine-2- carboxylic acid 5PhePyrr1 (2S,5R)-5-Phenyl-pyrrolidine-2- carboxylic acid .sup.D5PhePyrr1 (2R,5R)-5-Phenyl-pyrrolidine-2- carboxylic acid 5PhePyrr2 (2S,5S)-5-Phenyl-pyrrolidine-2- carboxylic acid .sup.D5PhePyrr2 (2R,5S)-5-Phenyl-pyrrolidine-2- carboxylic acid 4Hyp1 (4S)-L-Hydroxyproline .sup.D4Hyp1 (4S)-D-Hydroxyproline 4Hyp2 (4R)-L-Hydroxyproline .sup.D4Hyp2 (4R)-D-Hydroxyproline 4Mp1 (4S)-L-Mercaptoproline .sup.D4Mp1 (4S)-D-Mercaptoproline 4Mp2 (4R)-L-Mercaptoproline .sup.D4Mp2 (4R)-D-Mercaptoproline Pip L-Pipecolic acid .sup.DPip D-Pipecolic acid H-.sup.3-HCit-OH (3S)-3-Amino-6-carbamidyl-hexanoic acid H-.sup.3-HOrn-OH (3S)-3,6-Diamino-hexanoic acid H-.sup.3-HtBuA-OH (3S)-3-Amino-5,5-dimethyl-hexanoic acid H-.sup.3-HSar-OH N-Methyl-3-amino-propionic acid H-.sup.3-HPen-OH (3R)-3-Amino-4-methyl-4-mercapto- pentanoic acid H-.sup.3-HtBuG-OH (3R)-3-Amino-4,4-dimethyl-pentanoic acid H-.sup.3-H4AmPhe-OH (3S)-3-Amino-4-(4-aminophenyl)-butyric acid H-.sup.3-H3AmPhe-OH (3S)-3-Amino-4-(3-aminophenyl)-butyric acid H-.sup.3-H2AmPhe-OH (3S)-3-Amino-4-(2-aminophenyl)-butyric acid H-.sup.3- (3S)-3-Amino-4-(3-amidinophenyl)-butyric HPhe(mC(NH.sub.2)NH)OH acid H-.sup.3- (3S)-3-Amino-4-(4-amidinophenyl)-butyric HPhe(pC(NH.sub.2)NH)OH acid H-.sup.3- (3S)-3-Amino-4-(3-guanidinophenyl)- HPhe(mNHC(NH.sub.2)NH)OH butyric acid H-.sup.3- (3S)-3-Amino-4-(4-guanidino-phenyl)- HPhe(pNHC(NH.sub.2)NH)OH butyric acid H-.sup.3-H2Pal-OH (3S)-3-Amino-4-(pyridine-2-yl)-butyric acid H-.sup.3-H4Pal-OH (3S)-3-Amino-4-(pyridine-4-yl)-butyric acid H-.sup.3-HPhg-OH (3R)-3-Amino-3-phenyl-propionic acid H-.sup.3-HCha-OH (3S)-3-Amino-4-cyclohexyl-butyric acid H-.sup.3-HC.sub.4al-OH (3S)-3-Amino-4-cyclobutyl-butyric acid H-.sup.3-HC.sub.5al-OH (3S)-3-Amino-4-cyclopentyl-butyric acid H-.sup.3-HNle-OH (3S)-3-Amino-heptanoic acid H-.sup.3-H2Nal-OH (3S)-3-Amino-4-(2-naphthyl)-butyric acid H-.sup.3-H1Nal-OH (3S)-3-Amino-4-(1-naphthyl)-butyric acid H-.sup.3-H4ClPhe-OH (3S)-3-Amino-4-(4-chlorophenyl)-butyric acid H-.sup.3-H3ClPhe-OH (3S)-3-Amino-4-(3-chlorophenyl)-butyric acid H-.sup.3-H2ClPhe-OH (3S)-3-Amino-4-(2-chlorophenyl)-butyric acid H-.sup.3-H3,4Cl.sub.2Phe-OH (3S)-3-Amino-4-(3,4-dichlorophenyl)- butyric acid H-.sup.3-H4FPhe-OH (3S)-3-Amino-4-(4-fluorophenyl)-butyric acid H-.sup.3-H3FPhe-OH (3S)-3-Amino-4-(3-fluorophenyl)-butyric acid H-.sup.3-H2FPhe-OH (3S)-3-Amino-4-(2-fluorophenyl)-butyric acid H-.sup.3-HThi-OH (3R)-3-Amino-4-(2-thienyl)-butyric acid H-.sup.3-HTza-OH (3R)-3-Amino-4-(2-thiazolyl)-butyric acid H-.sup.3-HMso-OH (3R)-3-Amino-4-methylsulfoxyl-butyric acid Code Chemical Name H-.sup.3-HAcLys-OH (3S)-7-Acetylamino-3-amino-heptanoic acid H-.sup.3-HDpr-OH (3R)-3,4-diamino-butyric acid H-.sup.3-HA.sub.2BuOH (3S)-3,5-Diamino-pentanoic acid H-.sup.3-HDbu-OH (3R)-3,4-Diamino-pentanoic acid H-.sup.3-HAib-OH Amino-dimethyl acetic acid H-.sup.3-HCyp-OH 1-Amino-cyclopentane-1-yl-acetic acid H-.sup.3-HY(Bzl)-OH (3S)-3-Amino-4-(4-benzyloxyphenyl)- butyric acid H-.sup.3-HH(Bzl)-OH (3S)-3-Amino-4-(1-benzylimidazole-4- yl)-butyric acid H-.sup.3-HBip-OH (3S)-3-Amino-4-biphenylyl-butyric acid H-.sup.3-HS(Bzl)-OH (3S)-3-Amino-4-(benzyloxy)-butyric acid H-.sup.3-HT(Bzl)-OH (3R,4R)-3-Amino-4-benzyloxy-pentanoic acid H-.sup.3-HalloT-OH (3R,4S)-3-Amino-4-hydroxy-pentanoic acid H-.sup.3-HLeu3OHOH (3R,4R)-3-Amino-4-hydroxy-5-methyl- hexanoic acid H-.sup.3-HhAla-OH (3S)-3-Amino-pentanoic acid H-.sup.3-HhArg-OH (3S)-3-Amino-7-guanidino-heptanoic acid H-.sup.3-HhCys-OH (3R)-Amino-5-mercapto-pentanoic acid H-.sup.3-HhGlu-OH (3S)-3-Amino-heptanedioic acid H-.sup.3-HhGln-OH (3S)-3-Amino-6-carbamoyl hexanoic acid H-.sup.3-HhHis-OH (3S)-3-Amino-5-(imidazole-4-yl)- pentanoic acid H-.sup.3-HhIle-OH (3S,5S)-3-Amino-5-methyl-heptanoic acid H-.sup.3-HhLeu-OH (3S)-3-Amino-6-methyl-heptanoic acid H-.sup.3-HhNle-OH (3S)-3-Amino-octanoic acid H-.sup.3-DiAoc-OH (3S)-3,8-Diamino-octanoic acid H-.sup.3-HhMet-OH (3S)-3-Amino-6-methylthio-hexanoic acid H-.sup.3-HhPe-OH (3S)-3-Amino-5-phenyl-pentanoic acid H-.sup.3-HhSer-OH (3S)-3-Amino-5-hydroxy-pentanoic acid H-.sup.3-HhThr-OH (3S,5R)-3-Amino-5-hydroxy-hexanoic acid H-.sup.3-HhTrp-OH (3S)-3-Amino-5-(indol-3-yl)-pentanoic acid H-.sup.3-HhThr-OH (3S)-3-Amino-5-(4-hydroxyphenyl)- pentanoic acid H-.sup.3-HhCha-OH (3S)-3-Amino-5-cyclohexyl-pentanoic acid H-.sup.3-HBpa-OH (3S)-3-Amino-4-(4-benzoylphenyl)-butyric acid H-.sup.3-HOctG-OH (3S)-3-Amino-undecanoic acid H-.sup.3-HNle-OH (3S)-3-Amino-heptanoic acid H-.sup.3-HTic-OH (3S)-1,2,3,4-Tetrahydroisoquinoline-3-yl- acetic acid H-.sup.3-HTiq-OH (1S)-1,2,3,4-Tetrahydroisoquinoline-1- acetic acid H-.sup.3-HOic-OH (2S,3aS,7aS)-1-Octahydro-1H-indole-2- yl-acetic acid H-.sup.3-H4AmPyrr1-OH (2S,4S)-4-Amino-pyrrolidine-2-acetic acid H-.sup.3-H4AmPyrr2-OH (2S,4R)-4-Amino-pyrrolidine-2-acetic acid H-.sup.3-H4PhePyrr1-OH (2S,4R)-4-Phenyl-pyrrolidine-2-acetic acid H-.sup.3-H4PhePyrr2-OH (2S,4S)-4-Phenyl-pyrrolidine-2-acetic acid H-.sup.3-H5PhePyrr1-OH (2S,5R)-5-Phenyl-pyrrolidine-2-acetic acid H-.sup.3-H5PhePyrr2-OH (2S,5S)-5-Phenyl-pyrrolidine-2-acetic acid H-.sup.3-H4Hyp1-OH (2S,4S)-4-Hydroxy-pyrrolidine-2-acetic acid H-.sup.3-H4Hyp2-OH (2S,4R)-4-Hydroxy-pyrrolidine-2-acetic acid H-.sup.3-H4Mp1-OH (2R,4S)-4-Mercapto-pyrrolidine-2-acetic acid H-.sup.3-H4Mp2-OH (2R,4R)-4-Mercapto-pyrrolidine-2-acetic acid H-.sup.3-HPip-OH (2S)-piperidine-2-acetic acid H-.sup.3-HPro-OH (2S)-pyrrolidine-2-acetic acid H-.sup.3-H.sup.DPro-OH (2R)-pyrrolidine-2-acetic acid Ahb 4-Amino-2-hydroxy butyric acid H-.sup.4-DiHCit-OH (4S)-4-Amino-7-carbamidyl-heptanoic acid H-.sup.4-DiHOrn-OH (4S)-4,7-Diamino-heptanoic acid H-.sup.4-DiHtBuA-OH (4R)-4-Amino-6,6-dimethyl-heptanoic acid H-.sup.4-DiHSar-OH N-Methyl-4-amino-butyric acid H-.sup.4-DiHPen-OH (4R)-4-Amino-5-methyl-5-mercapto-hexanoic acid H-.sup.4-DiHtBuG-OH (4R)-4-Amino-5,5-dimethyl-hexanoic acid H-.sup.4-DiH4AmPhe-OH (4R)-4-Amino-5-(4-aminophenyl)-pentanoic acid H-.sup.4-DiH3AmPhe-OH (4R)-4-Amino-5-(3-aminophenyl)-pentanoic acid H-.sup.4-DiH2AmPhe-OH (4R)-4-Amino-5-(2-aminophenyl)-pentanoic acid H-.sup.4- (4R)-4-Amino-5-(3-amidinophenyl)- DiHPhe(mC(NH.sub.2)NH)OH pentanoic acid H-.sup.4- (4R)-4-Amino-5-(4-amidinophenyl)- DiHPhe(pC(NH.sub.2)NH)OH pentanoic acid H-.sup.4- (4R)-4-Amino-5-(3-guanidino-phenyl)- DiHPhe(mNHC(NH.sub.2)NH)OH pentanoic acid H-.sup.4- (4R)-4-Amino-5-(4-guanidino-phenyl)- DiHPhe(pNHC(NH.sub.2)NH)OH pentanoic acid H-.sup.4-DiH2Pal-OH (4R)-4-Amino-5-(pyridine-4-yl)-pentanoic acid H-.sup.4-DiH4Pal-OH (4R)-4-Amino-5-(pyridine-4-yl)-pentanoic acid H-.sup.4-DiHPhg-OH (4R)-4-Amino-4-phenyl-butyric acid H-.sup.4-DiHCha-OH (4R)-4-Amino-5-cyclohexyl-pentanoic acid H-.sup.4-DiHC.sub.4al-OH (4R)-4-Amino-5-cyclobutyl-pentanoic acid H-.sup.4-DiHC.sub.5al-OH (4R)-4-Amino-5-cyclopentyl-pentanoic acid H-.sup.4-DiHNle-OH (4S)-4-Amino-octanoic acid H-.sup.4-DiH2Nal-OH (4S)-4-Amino-5-(2-naphthyl)-pentanoic acid H-.sup.4-DiH1Nal-OH (4S)-4-Amino-5-(1-naphthyl)-pentanoic acid H-.sup.4-DiH4ClPhe-OH (4R)-4-Amino-5-(4-chlorophenyl)- pentanoic acid H-.sup.4-DiH3ClPhe-OH (4R)-4-Amino-5-(3-chlorophenyl)- pentanoic acid H-.sup.4-DiH2ClPhe-OH (4R)-4-Amino-5-(2-chlorophenyl)- pentanoic acid H-.sup.4-DiH3,4Cl.sub.2Phe-OH (4R)-4-Amino-5-(3,4-dichloro-phenyl)- pentanoic acid H-.sup.4-DiH4FPhe-OH (4R)-4-Amino-5-(4-fluorophenyl)- pentanoic acid H-.sup.4-DiH3FPhe-OH (4R)-4-Amino-5-(3-fluorophenyl)- pentanoic acid H-.sup.4-DiH2FPhe-OH (4R)-4-Amino-5-(2-fluorophenyl)- pentanoic acid H-.sup.4-DiHThi-OH (4R)-4-Amino-5-(2-thienyl)-pentanoic acid H-.sup.4-DiHTza-OH (4R)-4-Amino-5-(2-thiazolyl)-pentanoic acid H-.sup.4-DiHMso-OH (4R)-4-Amino-5-methylsulfoxyl-pentanoic acid H-.sup.4-DiHAcLys-OH (4S)-8-Acetylamino-4-amino-ocatanoic acid H-.sup.4-DiHDpr-OH (4R)-4,5-diamino-pentanoic acid H-.sup.4-DiHA.sub.2BuOH (4R)-4,5-Diamino-hexanoic acid H-.sup.4-DiHDbu-OH (4R)-4,5-Diamion-hexanoic acid H-.sup.4-DiHAib-OH 3-Amino-3,3-dimethyl propionic acid H-.sup.4-DiHCyp-OH (1-Amino-cyclopentane-1-yl)-3-propionic acid H-.sup.4-DiHY(Bzl)-OH (4R)-4-Amino-5-(4-benzyloxyphenyl)- pentanoic acid H-.sup.4-DiHH(Bzl)-OH (4R)-4-Amino-5-(1-benzylimidazole-4- yl)-pentanoic acid H-.sup.4-DiHBip-OH (4R)-4-Amino-5-biphenylyl-pentanoic acid H-.sup.4-DiHS(Bzl)-OH (4S)-4-Amino-5-(benzyloxy)-pentanoic acid H-.sup.4-DiHT(Bzl)-OH (4R,5R)-4-Amino-5-benzyloxy-hexanoic acid H-.sup.4-DiHalloT-OH (4R,5S)-4-Amino-5-hydroxy-hexanoic acid H-.sup.4-DiHLeu3OHOH (4R,5R)-4-Amino-5-hydroxy-6-methyl- heptanoic acid H-.sup.4-DiHhAla-OH (4S)-4-Amino-hexanoic acid H-.sup.4-DiHhArg-OH (4S)-4-Amino-8-guanidino-octanoic acid H-.sup.4-DiHhCys-OH (4R)-Amino-6-mercapto-hexanoic acid H-.sup.4-DiHhGlu-OH (4S)-4-Amino-ocatanedioic acid H-.sup.4-DiHhGln-OH (4S)-4-Amino-7-carbamoyl-heptanoic acid H-.sup.4-DiHhHis-OH (4S)-4-Amino-6-(imidazole-4-yl)-hexanoic acid H-.sup.4-DiHhIle-OH (4S,6S)-4-Amino-6-methyl-octanoic acid H-.sup.4-DiHhLeu-OH (4S)-4-Amino-7-methyl-ocatanoic acid H-.sup.4-DiHhNle-OH (4S)-4-Amino-nonanoic acid H-.sup.4-DiHhLys-OH (4S)-4,9-Diamino-nonanoic acid H-.sup.4-DiHhMet-OH (4R)-4-Amino-7-methylthioheptanoic acid H-.sup.4-DiHhPhe-OH (4S)-4-Amino-6-phenyl-hexanoic acid H-.sup.4-DiHhSer-OH (4R)-4-Amino-6-hydroxy-hexanoic acid H-.sup.4-DiHhThr-OH (4R,6R)-4-Amino-6-hydroxy-heptanoic acid H-.sup.4-DiHhTrp-OH (4S)-4-Amino-6-(indol-3-yl)-hexanoicacid H-.sup.4-DiHhTyr-OH (4S)-4-Amino-6-(4-hydroxyphenyl)- hexanoic acid H-.sup.4-DiHhCha-OH (4R)-4-Amino-5-cyclohexyl-pentanoic acid H-.sup.4-DihBpa-OH (4R)-4-Amino-5-(4-benzoylphenyl)- pentanoic acid H-.sup.4-DiHOctG-OH (4S)-4-Amino-dodecanoic acid H-.sup.4-DiHNle-OH (4S)-4-Amino-octanoic acid H-.sup.4-DiHTic-OH (3R)-1,2,3,4-Tetrahydroisoquinoline- 3-yl-3-propionic acid H-.sup.4-DiHTiq-OH (1R)-1,2,3,4-Tetrahydroisoquinoline- 1-yl-3-propionic acid H-.sup.4-DiHOic-OH (2S,3aS,7aS)-1-Octahydro-1H-indole- 2-yl-3-propionic acid H-.sup.4-DiH4AmPyrr1-OH (2R,4S)-4-Amino-pyrrolidine-2-yl-3- propionic acid H-.sup.4-DiH4AmPyrr2-OH (2R,4R)-4-Amino-pyrrolidine-2-yl-3- propionic acid H-.sup.4-DiH4PhePyrr1-OH (2R,4R)-4-Phenyl-pyrrolidine-2-yl-3- propionic acid H-.sup.4-DiH4PhePyrr2-OH (2R,4S)-4-Phenyl-pyrrolidine-2-yl-3- propionic acid H-.sup.4-DiH5PhePyrr1-OH (2S,5R)-5-Phenyl-pyrrolidine-2-yl-3- propionic acid H-.sup.4-DiH5PhePyrr2-OH (2S,5S)-5-Phenyl-pyrrolidine-2-yl-3- propionic acid H-.sup.4-DiH4Hyp1-OH (2R,4S)-4-Hydroxy-pyrrolidine-2-yl- 2-propionic acid H-.sup.4-DiH4Hyp2-OH (2R,4R)-4-Hydroxy-pyrrolidine-2-yl- 3-propionic acid H-.sup.4-DiH4Mp1-OH (2R,4S)-4-Mercapto-pyrrolidine-2-yl- 3-propionic acid H-.sup.4-DiH4Mp2-OH (2R,4R)-4-Mercapto-pyrrolidine-2-yl- 3-propionic acid H-.sup.4-DiHPip-OH (2S)-Piperidine-2-yl-3-propionic acid H-.sup.4-DiHPro-OH (2S)-Pyrrolidine-2-yl-3-propionic acid (AEt)G N-(2-Aminoethyl)glycine (APr)G N-(3-Amino-n-propyl)glycine (ABu)G N-(4-Amino-n-butyl)glycine (APe)G N-(5-Amino-n-pentyl)glycine (GuEt)G N-(2-Guanidinoethyl)glycine (GuPr)G N-(3-Guanidino-n-propyl)glycine (GuBu)G N-(4-Guanidino-n-butyl)glycine (GuPe)G N-(5-Guanidino-n-pentyl)glycine (PEG.sub.3-NH.sub.2)G N[H.sub.2N(CH.sub.2).sub.3(OCH.sub.2CH.sub.2).sub.2O(CH.sub.2).sub.3]glycine (Me)G N-Methylglycine (Et)G N-Ethylglycine (Bu)G N-Butylglycine (Pe)G N-Pentylglycine (Ip)G N-Isopropylglycine (2MePr)G N-(2-Methylpropyl)glycine (3MeBu)G N-(3-Methylbutyl)glycine (1MePr)G (1S)-N-(1-Methylpropyl)glycine (2MeBu)G (2S)-N-(2-Methylbutyl)glycine (MthEt)G N-(Methylthioethyl)glycine (MthPr)G N-(Methylthiopropyl)glycine (Ben)G N-(Benzyl)glycine (PhEt)G N-(2-Phenylethyl)glycine (HphMe)G N-([4-hydroxyphenyl]methyl)glycine (HphEt)G N-(2-[4-hydroxyphenyl]ethyl)glycine (ImMe)G N-(Imidazol-5-yl-methyl)glycine (ImEt)G N-(2-(Imidazol-5-yl)ethyl)glycine (InMe)G N-(Indol-2-yl-methyl)glycine (InEt)G N-(2-(Indol-2-yl)ethyl)glycine (CboMe)G N-(Carboxymethyl)glycine (CboEt)G N-(2-Carboxyethyl)glycine (CboPr)G N-(3-Carboxypropyl)glycine (CbaMe)G N-(Carbamoylmethyl)glycine (CbaEt)G N-(2-Carbamoylethyl)glycine (CbaPr)G N-(3-Carbamoylpropyl)glycine (HyEt)G N-(2-Hydroxyethyl)glycine (HyPr)G (2R)-N-(2-Hydroxypropyl)glycine (Mcet)G N-(2-Mercaptoethyl)glycine NMeAla L-N-Methylalanine NMe.sup.DAla D-N-Methylalanine NMeVal L-N-Methylvaline NMe.sup.DVal D-N-Methylvaline NMeIle L-N-Methylisoleucine NMe.sup.DIle D-N-Methylisoleucine NMeLeu L-N-Methylleucine NMe.sup.DLeu D-N-Methylleucine NMeNle L-N-Methylnorleucine NMe.sup.DNle D-N-Methylnorleucine NMeMet L-N-Methylmethionine NMe.sup.DMet D-N-Methylmethionine NMeTyr L-N-Methyltyrosine NMe.sup.DTyr D-N-Methyltyrosine NMeHis L-N-Methylhistidine NMe.sup.DHis D-N-Methylhistidine NMePhe L-N-Methylphenylalanine NMe.sup.DPhe D-N-Methylphenylalanine NMeTrp L-N-Methyltryptophane NMe.sup.DTrp D-N-Methyltryptophane NMeSer L-N-Methylserine NMe.sup.DSer D-N-Methylserine NMeAsp L-N-Methylaspartic acid NMe.sup.DAsp D-N-Methylaspartic acid NMeGlu L-N-Methylglutamic acid NMe.sup.DGlu D-N-Methylglutamic acid NMeLys L-N-Methyllysine NMe.sup.DLys D-N-Methyllysine NMeArg L-N-Methylarginine NMe.sup.DArg D-N-Methylarginine NMeDab L-N-Methyl-2,4-diamino butyric acid NMe.sup.DDab D-N-Methyl-2,4-diamino butyric acid NMeCys L-N-Methylcysteine NMe.sup.DCys D-N-Methylcysteine NMeAsn L-N-Methylasparagine NMe.sup.DAsn D-N-Methylasparagine NMeGln L-N-Methylglutamine NMe.sup.DGln D-N-Methylglutamine NMeThr L-N-Methylthreonine NMe.sup.DThr D-N-Methylthreonine.
6. Compounds according to claim 1, selected from: (2S,11S,19aS)-2-(acetylamino)-15-fluoro-N-[2-(1H-indol-3-yl)ethyl]-7,12-dimethyl-5,8,13-trioxo-2,3,6,7,8,9,10,11,12,13,19,19a-dodecahydro-1H,5H-pyrrolo[2,1-c][1,4,7,12]benzoxatriazacyclopentadecine-11-carboxamide, (2S,11S,19aS)N-[2-(dimethylamino)ethyl]-15-fluoro-2-{[2-(1H-indol-3-yl)acetyl]amino}-7,12-dimethyl-5,8,13-trioxo-2,3,6,7,8,9,10,11,12,13,19,19a-dodecahydro-1H,5H-pyrrolo[2,1-c][1,4,7,12]benzoxatriazacyclopentadecine-11-carboxamide; (2S,11S,19aS)-15-fluoro-2-{[2-(1H-indol-3-yl)acetyl]amino}-N-[2-(1H-indol-3-yl)ethyl]-7,12-dimethyl-5,8,13-trioxo-2,3,6,7,8,9,10,11,12,13,19,19a-dodecahydro-1H,5H-pyrrolo[2,1-c][1,4,7,12]benzoxatriazacyclopentadecine-11-carboxamide; (2S,11S,19aS)-2-{[2-(dimethylamino)acetyl]amino}-15-fluoro-N-[2-(1H-indol-3-yl)ethyl]-7,12-dimethyl-5,8,13-trioxo-2,3,6,7,8,9,10,11,12,13,19,19a-dodecahydro-1H,5H-pyrrolo[2,1-c][1,4,7,12]benzoxatriazacyclopentadecine-11-carboxamide; tert-butyl N-[(2S,11S,19aS)-15-fluoro-11-({[2-(1H-indol-3-yl)ethyl]amino}carbonyl)-7,12-dimethyl-5,8,13-trioxo-2,3,6,7,8,9,10,11,12,13,19,19a-dodecahydro-1H,5H-pyrrolo[2,1-c][1,4,7,12]benzoxatriazacyclopentadecin-2-yl]carbamate; (2S,11S,19aS)N-[2-(dimethylamino)ethyl]-15-fluoro-7,12-dimethyl-2-{[2-(1-naphthyl)acetyl]amino}-5,8,13-trioxo-2,3,6,7,8,9,10,11,12,13,19,19a-dodecahydro-1H,5H-pyrrolo[2,1-c][1,4,7,12]benzoxatriazacyclopentadecine-11-carboxamide; benzyl N-[(4S,6S,10S)-14-methyl-6-{[2-(2-naphthyl)acetyl]amino}-9,15-dioxo-2-oxa-8,14-diazatricyclo[14.3.1.04,8]icosa-1(20),16,18-trien-10-yl]carbamate; benzyl N-[(4S,6S,13S)-6-{[2-(1H-indol-3-yl)acetyl]amino}-11,15-dimethyl-9,12,16-trioxo-2-oxa-8,11,15-triazatricyclo[15.3.1.04,8]henicosa-[(21),17,19-trien-13-yl]carbamat; N-[(4S,6S,13S)-6-{[2-(1H-indol-3-yl)acetyl]amino}-11,15-dimethyl-9,12,16-trioxo-2-oxa-8,11,15-triazatricyclo[15.3.1.04,8]henicosa-[(21),17,19-trien-13-yl]decanamide.
7. A composition having the compound according to any one of claims 2, 3, 5, 6 or 1 in a therapeutically active amount and having agonistic activity on the motilin receptor (MR receptor), on the serotonin receptor of subtype 5-HT.sub.2B (5-HT.sub.2B receptor), and on the prostaglandin F2 receptor (FP receptor).
8. A pharmaceutical composition containing a compound according to any one of claims 2, 3, 5, 6 or 1 and a therapeutically inert carrier.
9. The composition according to claim 8 having agonistic or antagonistic activity on the motilin receptor (MR receptor), on the serotonin receptor of subtype 5-HT.sub.2B (5-HT.sub.2B receptor), and on the prostaglandin F2 receptor (FP receptor).
10. The composition according to claim 9 in a form suitable for oral, topical, transdermal, injection, buccal, transmucosal, pulmonary or inhalation administration.
11. The composition according to claim 10 in form of tablet, degree, capsule, solution, liquid, gel, plaster, scream, ointment, syrup, slurry, suspension, spray, nebuliser or suppository.
12. A medicament comprising the compound according to claim 1 having agonistic or antagonistic activity on the motilin receptor (MR receptor), on the serotonin receptor of subtype 5-HT.sub.2B (5-HT.sub.2B receptor), and on the prostaglandin F2 receptor (FP receptor).
13. A method of treating hypomotility disorders of the gastrointestinal tract selected from the group consisting of diabetic gastroparesis and constipation type irritable bowl syndrome; CNS diseases selected from the group consisting of migraine, schizophrenia, psychosis and depression; ocular hypertension associated with glaucoma or preterm labour; said method comprising: administering the compound of claim 1 to a patient in need thereof.
Description
EXAMPLES
(1) The following Examples illustrate the invention in more detail but are not intended to limit its scope in any way. The following abbreviations are used in these Examples: ADDP: azodicarboxylic dipiperidide All: allyl Alloc: allyloxycarbonyl AllocCl: allyl chloroformate AllocOSu: allyloxycarbonyl-N-hydroxysuccinimide AM-resin: aminomethyl resin aq.: aqueous arom.: aromatic BnBr: benzyl bromide Boc: tert-butoxycarbonyl br.: broad Cbz: benzyloxycarbonyl CbzOSu: N-(benzyloxycarbonyloxy)succinimide Cl-HOBt: 6-chloro-1-hydroxybenzotriazole CMBP: cyanomethylenetributyl-phosphorane m-CPBA: 3-chloroperbenzoic acid d: day(s) or doublet (spectral) DBU: 1,8-diazabicyclo[5.4.0]undec-7-ene DCE: 1,2-dichloroethane DEAD: diethyl azodicarboxylate DFPE polystyrene: 2-(3,5-dimethoxy-4-formylphenoxy)ethyl polystyrene DIAD: diisopropyl azodicarboxylate DIC: N,N-diisopropylcarbodiimide DMF: dimethylformamide DMSO: dimethyl sulfoxide DPPA: diphenyl phosphoryl azide DVB: divinylbenzene EDC: 1-[3-(dimethylamino)propyl-3-ethylcarbodiimide equiv.: equivalent Et.sub.3N: triethylamine EtOAc: ethyl acetate FC: flash chromatography FDPP: pentafluorophenyl diphenylphosphinate Fmoc: 9-fluorenylmethoxycarbonyl h: hour(s) HATU: O-(7-azobenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate HBTU: O-(benortriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate HCTU: O-(1H-6-chlorobenortriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate HOAt: 1-hydroxy-7-azabenzotriazole HOBt.H.sub.2O: 1-hydroxybenzotriazole hydrate HMPA: hexamethylphosphoramide i.v.: in vacuo m: multiplet (spectral) MeOH: methanol NMP: 1-methyl-2-pyrrolidinone Pd(PPh.sub.3).sub.4: Tetrakis(triphenylphosphine)palladium(0) PEG PS resin: polyethyleneglycol coated polystyrene resin PG: protective group PPh.sub.3: triphenylphosphine prep.: preparative i-Pr.sub.2NEt: N-ethyl-N,N-diisopropylamine PyBOP: (Benzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate PyBroP: Bromotripyrrolidinophosphonium hexafluorophosphate q: quartet (spectral) quant.: quantitative sat.: saturated soln: solution t: triplet (spectral) TBAF: tetrabutylammonium fluoride Teoc: 2-(trimethylsilyl)ethoxycarbonyl TeocONp: 2-(trimethylsilyl)ethyl 4-nitrophenyl carbonate TFA: trifluoroacetic acid THF: tetrahydrofuran tlc: thin layer chromatography T3P: propanphosphonic acid cyclic anhydride p-TsOH: p-toluenesulfonic acid
General Methods
(2) TLC: Merck (silica gel 60 F254, 0.25 mm).
(3) Flash chromatography (FC): Fluka silica gel 60 (0.04-0.063 mm) and Interchim Puriflash IR 60 silica gel (0.04-0.063 mm).
(4) I. Analytical HPLC-MS Methods:
(5) R.sub.t in min (purity at 220 nm in %), m/z [M+H].sup.+
(6) Volume of injection: 5 L for all methods
(7) Method 1a and 1b
(8) TABLE-US-00020 Column: XBridge C18 2.5 m, 2.1 50 mm (186003085 - Waters) Mobile Phases A: 0.1% TFA in Water B: 0.085% TFA in Acetonitrile Column oven temp. 45 C. Time Flow (min.) (l/min) % A % B Gradient 0 500 97 3 0.1 500 97 3 3 500 3 97 3.6 500 3 97 3.7 500 97 3 4.3 500 97 3 Method 1, cont. UV Wavelenght: 220 nm 254 nm MS scan Range: Method 1a 100-800 Da Method 1b Centroid 300-2000 Da mode Scan Time: 1 sec. Ionization type: Electrospray
Method 2
(9) TABLE-US-00021 Column: Gemini NX C18 3 m, 2.1 50 mm (00B-4453-B0 - Phenomenex) Mobile Phases A: 0.1% TFA in Water B: 0.085% TFA in Acetonitrile Column oven temp. 45 C. Time Flow (min.) (l/min) % A % B Gradient 0 800 97 3 0.1 800 97 3 2.2 800 3 97 2.5 800 3 97 2.55 1000 97 3 2.75 1000 97 3 2.8 800 97 3 UV Wavelenght: 220 nm 254 nm MS scan Range: 100-2000 Da Centroid mode Scan Time: 1 sec. Ionization type: Electrospray
Method 3
(10) TABLE-US-00022 Column: Gemini NX C18 3 m, 2.1 50 mm (00B-4453-B0 - Phenomenex) 1 mM ammonium Mobile Phases A: bicarbonate pH 10 B: Acetonitrile Column oven temp. 45 C. Time Flow (min.) (l/min) % A % B Gradient 0 800 97 3 0.1 800 97 3 2.2 800 3 97 2.5 800 3 97 2.55 1000 97 3 2.75 1000 97 3 2.8 800 97 3 Method 3, cont. UV Wavelenght: 220 nm 254 nm MS scan Range: 100-2000 Da Centroid mode Scan Time: 1 sec. Ionization type: Electrospray
Method 4a-4-b
(11) TABLE-US-00023 Column: Gemini NX C18 3 m, 2.1 50 mm (00B-4453-B0 - Phenomenex) Mobile Phases A: 0.1% TFA in Water B: 0.085% TFA in Acetonitrile Column oven temp. 45 C. Time Flow (min.) (l/min) % A % B Gradient 0 800 97 3 0.1 800 97 3 2.7 800 3 97 3 800 3 97 3.05 1000 97 3 3.25 1000 97 3 3.3 800 97 3 UV Wavelenght: 220 nm 254 nm MS scan Range: Method 4a Centroid 100-2000 Da mode Method 4b Profile 350-2000 Da mode Scan Time: 1 sec. Ionization type: Electrospray
Method 5a-5b
(12) TABLE-US-00024 Column: Gemini NX C18 3 m, 2.1 50 mm (00B-4453-B0 - Phenomenex) 1 mM ammonium Mobile Phases A: bicarbonate pH 10 B: Acetonitrile Column oven temp. 45 C. Time Flow (min.) (l/min) % A % B Gradient 0 800 97 3 0.1 800 97 3 2.7 800 3 97 3 800 3 97 3.05 1000 97 3 3.25 1000 97 3 3.3 800 97 3 Method 5, cont. UV Wavelenght: 220 nm 254 nm MS scan Range: Method 5a Centroid 100-2000 Da mode Method 5b Profile 350-2000 Da mode Scan Time: 1 sec. Ionization type: Electrospray
Method 6
(13) TABLE-US-00025 Column: Acquity UPLC BEH C18 1.7 m, 2.1 50 mm (cod. 186002350 - Waters) Mobile Phases A: 0.1% TFA in Water B: 0.085% TFA in Acetonitrile Column oven temp. 55 C. Time Flow (min.) (l/min) % A % B Gradient 0 1250 97 3 0.05 1250 97 3 1.65 1250 3 97 1.95 1250 3 97 2.00 1250 97 3 2.30 1250 97 3 UV Wavelenght: 220 nm 254 nm MS scan Range: 100-1650 Da Centroid mode Scan Time: 0.5 sec. Ionization type: Electrospray
Method 7
(14) TABLE-US-00026 Column: Acquity UPLC BEH C18 1.7 m, 2.1 50 mm (cod. 186002350 - Waters) Mobile Phases A: 0.1% TFA in Water B: 0.085% TFA in Acetonitrile Column oven temp. 55 C. Time Flow (min.) (l/min) % A % B Gradient 0 1250 97 3 0.05 1250 97 3 1.65 1250 3 97 1.95 1250 3 97 2.00 1250 97 3 2.30 1250 97 3 Method 7, cont. UV Wavelenght: 220 nm 254 nm MS scan Range: 100-1650 Da Profile mode Scan Time: 0.5 sec. Ionization type: Electrospray
Method 8
(15) TABLE-US-00027 Column: Acquity UPLC BEH C18 1.7 m, 2.1 50 mm (cod. 186002350 - Waters) 1 mM ammonium Mobile Phases A: bicarbonate pH 10 B: Acetonitrile Column oven temp. 55 C. Volume of 5 l injection: Time Flow (min.) (l/min) % A % B Gradient 0 1250 97 3 0.05 1250 97 3 1.65 1250 3 97 1.95 1250 3 97 2.00 1250 97 3 2.30 1250 97 3 UV Wavelenght: 220 nm 254 nm MS scan Range: 100-1650 Da Profile mode Scan Time: 0.5 sec. Ionization type: Electrospray
Method 9a-9c
(16) TABLE-US-00028 Column: Acquity UPLC BEH C18 1.7 m, 2.1 100 mm (cod. 186002352 - Waters) Mobile Phases A: 0.1% TFA in Water/ Acetonitrile B: 95/5 v/v 0.085% TFA in Acetonitrile Column oven temp. 55 C. Method 9, cont. Time Flow (min.) (l/min) % A % B Gradient 0 700 99 1 0.2 700 99 1 2.5 700 3 97 2.85 700 3 97 2.86 700 99 1 3.20 700 99 1 UV Wavelenght: 220 nm MS scan Range: Method 9a: 100-800 Da; Method 9b: 100-1200 Da; Method 9c: 200-1400 Da Profile mode Scan Time: 1 sec. Ionization type: Electrospray
(17) Analytical HPLC (x % CH.sub.3CN): R.sub.t in min (purity at 220 nm in %) Column: Develosil RPAq 5 m, 4.650 mm; Flow rate: 1.5 ml/min 0.0-0.5 min (x % CH.sub.3CN, 100-x % H.sub.2O containing 0.1% TFA); 0.5-5.0 min (x % CH.sub.3CN, 100-x % H.sub.2O containing 0.1% TFA to 100% CH.sub.3CN) 5.0-6.2 min (100% CH.sub.3CN)
II. Preparative HPLC Methods:
1. Reverse PhaseAcidic Conditions Column: XBridge C18 5 m, 30150 mm (Waters)
Mobile Phases: A: 0.1% TFA in Water/Acetonitrile 95/5 v/v 1B: 0.1% TFA in Water/Acetonitrile 5/95 v/v
2. Reverse PhaseBasic Conditions Column: XBridge C18 5 m, 30150 mm (Waters) Mobile Phases: A: 10 mM Ammonium Bicarbonate pH 10/Acetonitrile 95/5 v/v B: Acetonitrile
3. Normal Phase
(18) TABLE-US-00029 Column: VP 100/21 NUCLEOSIL 50-10, 21 100 mm (Macherey-Nagel) Mobile phases: A: Hexane B: Ethylacetate C: Methanol
(19) NMR Spectroscopy: Bruker Avance 300, .sup.1H-NMR (300 MHz) in the indicated solvent at ambient temperature. Chemical shifts in ppm, coupling constants J in Hz.
(20) The term isomers comprises in the present invention species of identical chemical formula, constitution and thus molecular mass, such as but not limited to amide cis/trans isomers, rotamers, conformers, diastereomers.
Examples
Starting Materials
Building Blocks of Type A (Scheme 1)
(21) 2-Acetoxy-5-fluoro benzoic acid (2) was prepared according to the method of C. M. Suter and A. W. Weston, J. Am. Chem. Soc. 1939, 61, 2317-2318.
(22) 3-Acetoxybenzoic acid (3) is commercially available.
(23) 4-Acetoxybenzoic acid (4) is commercially available.
(24) 5-Hydroxy nicotinic acid (5) is commercially available.
(25) 8-Acetoxyquinoline-2-carboxylic acid (8) was prepared according to the method of R. W. Hay, C. R. Clark, J. Chem. Soc. Dalton 1977, 1993-1998.
(26) (S)-2-tert-Butoxycarbonylamino-8-hydroxy-1,2,3,4-tetrahydro-naphthalene-2-carboxylic acid (10) was prepared according to the method of M. M. Altorfer, Dissertation Universitt Zrich, 1996.
(27) 3-Mercaptobenzoic acid (11) is commercially available
Building Blocks of Type B (Scheme 2)
(28) tert-Butyl (3S,5S)-5-(hydroxymethyl)pyrrolidin-3-ylcarbamate (13) as well as the corresponding HCl salt (13.HCl) are commercially available.
(29) tert-Butyl (3R,5S)-5-(hydroxymethyl)pyrrolidin-3-ylcarbamate (17) as well as the corresponding HCl salt (17.HCl) are commercially available.
(30) (S)-tert-Butyl 3-(hydroxymethyl)piperazine-1-carboxylate hydrochloride (21.HCl) is commercially available.
(31) (R)-tert-Butyl 3-(hydroxymethyl)piperazine-1-carboxylate hydrochloride (83.HCl) (Scheme 5) is commercially available.
(32) (2S,4S)-Allyl 2-(hydroxymethyl)-4-((2-(trimethylsilyl)ethoxy)carbonylamino)pyrrolidine-1-carboxylate (16) was prepared in three steps (1. Alloc protection of the secondary amino group with allyloxycarbonyl-N-hydroxysuccinimide (AllocOSu) in CH.sub.2Cl.sub.2, 2. cleavage of the Boc group with dioxane-HCl;
(33) 3. Teoc protection of the primary amino group with 2-(trimethylsilyl)ethyl 4-nitrophenyl carbonate (Teoc-ONp) in CH.sub.2Cl.sub.2 in the presence of Et.sub.3N) from amino alcohol 13, applying standard conditions; as leading references cf. T. W. Greene, P. G. M. Wuts, Protective Groups in Organic Synthesis, 3rd edition, John Wiley & Sons, 1999; P. J. Kocienski, Protecting Groups, 3rd edition, Georg Thieme Verlag, 2005.
(34) Data of 16: C.sub.15H.sub.28N.sub.2O.sub.5Si (344.5): Flow injection MS (APCI): 689 ([2M+H].sup.+), 345 ([M+H].sup.+). .sup.1H-NMR (DMSO-d.sub.6): 7.28 (d, J=6.1, 1 H), 5.90 (m, 1 H), 5.25 (qd, J=1.7, 17.2, 1 H), 5.16 (qd, J=1.5, 10.5, 1 H), 4.90 (br. t, 1 H), 4.54-4.42 (m, 2 H), 4.04-3.97 (m, 2 H), 3.90 (q, J=6.8, 1 H), 3.80-3.66 (br. m and dd, 2 H), 3.57-3.43 (br. m, 2 H), 2.96 (br. m, 1 H), 2.19 (br. m, 1 H), 1.78 (br. m, 1 H), 0.89 (t, J ca 8.3, 2 H), 0.00 (s, 9 H)
(35) (2S,4R)-Allyl 2-(hydroxymethyl)-4-((2-(trimethylsilyl)ethoxy)carbonylamino)pyrrolidine-1-carboxylate (20) was prepared from amino alcohol hydrochloride 17.HCl, applying the same transformations as described for the synthesis of diastereomer 16 with the exception of the Alloc protection step which was performed using allyl chloroformate in CH.sub.2Cl.sub.2 in the presence of aqueous NaHCO.sub.3 solution.
(36) Data of 20: C.sub.15H.sub.28N.sub.2O.sub.5Si (344.5): LC-MS (method 9a): R.sub.t=1.98, 345 ([M+H].sup.+); 317; 259. .sup.1H-NMR (DMSO-d.sub.6): 7.26 (d, J=6.6, 1 H), 5.89 (m, 1 H), 5.25 (br. d, J=17.0, 1 H), 5.15 (br. d, J=10.2, 1 H), 4.75 (m, 1 H), 4.48 (m, 2 H), 4.16-3.98 (m, 3 H), 3.82 (br. m, 1 H), 3.48-3.30 (m, 3 H), 3.21 (m, 1 H), 2.01 (m, 1 H), 1.80 (m, 1 H), 0.89 (t, J=8.3, 2H), 0.00 (s, 9 H).
(37) (S)-1-Allyl 4-tert-butyl 2-(hydroxymethyl)piperazine-1,4-dicarboxylate (22) was prepared from amino alcohol hydrochloride 21.HCl, applying allyl chloroformate in CH.sub.2Cl.sub.2 in the presence of aqueous NaHCO.sub.3 solution; as leading references cf. T. W. Greene, P. G. M. Wuts, Protective Groups in Organic Synthesis, 3rd edition, John Wiley & Sons, 1999; P. J. Kocienski, Protecting Groups, 3rd edition, Georg Thieme Verlag, 2005.
(38) Data of 22: C.sub.14H.sub.24N.sub.2O.sub.5 (300.4): LC-MS (method 9a): R.sub.t=1.70, 201 ([M+H].sup.+). .sup.1H-NMR (DMSO-d.sub.6): 5.90 (m, 1 H), 5.29 (qd, J=1.7, 17.3, 1 H), 5.18 (qd, J=1.5, 10.5, 1 H), 4.81 (t, J=4.9, 1 H), 4.53 (d-like m, J ca. 5.1, 2 H), 4.04-3.75 (br. m, 4 H), 3.39 (m, 2 H), 2.95-2.70 (br. m, 3 H), 1.40 (s, 9 H).
Building Blocks of Type C (Scheme 3)
(S)-5-Allyl 1-benzyl 2-(methylamino)pentanedioate hydrochloride (27.HCl)
(39) A mixture of Boc-L-Glu(OAll)OH (23; 33 g, 115 mmol) and NaHCO.sub.3 (27 g, 322 mmol) in DMF (500 mL) was stirred for 1 h at room temperature followed by the slow addition of benzyl bromide (35 mL, 299 mmol) in DMF (15 mL). Stirring was continued for 16 h followed by aqueous workup (diethyl ether, sat. aq. NaHCO.sub.3 soln, sat aq. NaCl soln) and purification by FC(CH.sub.2Cl.sub.2/MeOH 100:0 to 98:2) to give the corresponding benzyl ester (34.4 g, 79%), which was dissolved in dioxane (40 mL) and treated with 4 M HCl-dioxane (400 mL) for 1 h. The volatiles were evaporated. The residue was crystallized from diethyl ether to afford 24.HCl (23.8 g, 83%).
(40) 4-Nitrobenzenesulfonyl chloride (39 g, 178 mmol) was added at 0 C. to a solution of 24.HCl (46.5 g, 148 mmol) and pyridine (42 mL, 519 mmol) in CH.sub.2Cl.sub.2 (700 mL). The mixture was stirred for 15 h followed by aqueous workup (CH.sub.2Cl.sub.2, 1 M aq. HCl soln) and purification of the crude by FC (hexane/EtOAc 80:20 to 75:25) to yield 25 (55.54 g, 81%).
(41) A solution of 25 (41.3 g, 89 mmol) in dry DMF (200 mL) was cooled to 0 C. Methyliodide (5.8 mL, 94 mmol) in DMF (100 mL) was slowly added, followed by a solution of DBU (14 mL, 94 mmol) in DMF (100 mL). The mixture was stirred for 4 h at room temperature followed by aqueous workup (EtOAc, 1 M aq. HCl soln., H.sub.2O, sat. aq. NaHCO.sub.3 soln, sat. aq. NaCl soln) to afford 26 (42.8 g, 99%).
(42) A solution of 26 (17.4 g, 37 mmol) in dry, degassed CH.sub.3CN (270 mL) was treated with thiophenol (6.7 mL, 66 mmol) and Cs.sub.2CO.sub.3 (39 g, 121 mmol) at room temperature for 16 h. The mixture was filtered and the residue was washed with diethyl ether. The filtrate was carefully concentrated (bath temperature 20 C.) and immediately purified by FC (hexane/EtOAc 80:20 to 50:50). The combined product fractions were carefully concentrated, immediately treated with 4 M HCl-dioxane (20 mL) for 5 min and concentrated to give 27.HCl (8.62 g, 72%).
(43) Data of 27.HCl: C.sub.16H.sub.21NO.sub.4.HCl (291.3, free base). LC-MS (method 9b): R.sub.t=1.44, 292 ([M+H].sup.+). .sup.1H-NMR (DMSO-d.sub.6): 9.57 (br. s, NH.sub.2.sup.+), 7.45-7.34 (m, 5 arom. H), 5.88 (M, 1 H), 5.32-5.19 (m, 4 H), 4.53 (td, J=1.3, 5.4, 1 H), 4.13 (br. t, J ca. 6.0, 1 H), 2.69-2.40 (m, 2 H), 2.56 (s, 3 H), 2.30-2.05 (m, 2 H).
(44) (R)-5-Allyl 1-benzyl 2-(methylamino)pentanedioate hydrochlorided (29.HCl) was prepared from Boc-D-Glu(OAll)OH (28) applying the methods described above for the synthesis of the enantiomer (27.HCl).
(45) Data of 29.HCl: C.sub.16H.sub.21NO.sub.4.HCl (291.3, free base). LC-MS (method 9b): R.sub.t=1.44, 292 ([M+H].sup.+). .sup.1H-NMR (DMSO-d.sub.6): 9.92 (br. s, NH.sup.+), 9.54 (br. s, NH.sup.+), 7.45-7.34 (m, 5 arom. H), 5.88 (M, 1 H), 5.32-5.19 (m, 4 H), 4.53 (td, J=1.3, 5.4, 1 H), 4.13 (br. t, J ca. 6.0, 1 H), 2.69-2.40 (m, 2 H), 2.56 (s, 3 H), 2.30-2.05 (m, 2 H).
(S)-Allyl 2-(benzyloxycarbonylamino)-3-(methylamino)propanoate hydrochloride (32.HCl)
(46) Cbz-L-SerOH (30) was converted into amino acid 31 by -lactone formation and opening with HNCH.sub.3Si(CH.sub.3).sub.3 (see J. Kim, S. G. Bott, D. M. Hoffman Inorg. Chem. 1998, 37, 3835-3841), following the procedures of J. K. Kretsinger and J. P. Schneider, J. Am. Chem. Soc. 2003, 125, 7907-7913 and E. S. Ratemi and J. C. Vederas, Tetrahedron Lett. 1994, 35, 7605-7608.
(47) A solution of 31.HCl (2.2 g, 7.6 mmol) in allyl alcohol (55 mL) was treated with thionyl chloride (1.7 mL, 23 mmol) for 15 min at room temperature and for 1.5 h at 70 C. The volatiles were evaporated. The crude product was dissolved in CH.sub.2Cl.sub.2 and washed with aq. NaHCO.sub.3 solution. The aqueous layers were extracted with CH.sub.2Cl.sub.2 and with EtOAc. The combined organic phase was dried (Na.sub.2SO.sub.4), filtered, and concentrated. The resulting oil (2.18 g) was dissolved in CH.sub.2Cl.sub.2 (80 mL), treated with 4 M HCl-dioxane (20 mL), stirred for 5 min and concentrated to afford 32.HCl (2.5 g, quantitative).
(48) Data of 32.HCl: C.sub.15H.sub.20N.sub.2O.sub.4.HCl (292.3, free base). LC-MS (method 9a): R.sub.t=1.26, 293 ([M+H].sup.+). .sup.1H-NMR (DMSO-d.sub.6):
(49) 9.20 (br. s, NH.sup.+), 9.03 (br. s, NH.sup.+), 8.02 (d, J=8.2, NH), 7.38-7.30 (m, 5 arom. H), 5.89 (m, 1 H), 5.33 (d, J=17.3, 1 H), 5.23 (d, J=10.5, 1 H), 5.08 (s, 2 H), 4.63 (d, J=5.3, 2 H), 4.56 (m, 1 H), 3.35 (br. m, 1 H), 3.25 (br. m, 1 H), 2.56 (br. s, 3 H).
(50) As an alternative, 32.HCl was prepared from Cbz-L-DapOH applying the method described below for the synthesis of the enantiomer 36.HCl.
(R)-Allyl 2-(benzyloxycarbonylamino)-3-(methylamino)propanoate hydrochloride (36.HCl)
(51) Cbz-D-DapOH was converted into the allylester-pTsOH salt 33 pTsOH according to the procedure of T. M. Kamenecka and S. J. Danishefsky, Chem. Eur. J. 2001, 7, 41-63, describing the synthesis of D-threonine allyl ester.
(52) The amino ester 33 pTsOH was converted into the free base by extraction (CH.sub.2Cl.sub.2, sat. aq. NaHCO.sub.3 soln) and treated with 4-nitrobenzenesulfonyl chloride (1.05 equiv.) in CH.sub.2Cl.sub.2 in the presence of pyridine (3.0 equiv.) to give the p-nitrophenyl sulfonamide 34.
(53) At 0 C., a solution of methyl iodide (2.3 mL, 37 mmol) in DMF (80 mL) was added to a solution of 34 (16.4 g, 35 mmol) in DMF (80 mL). A solution of DBU (5.6 mL, 37 mmol) in DMF (80 mL) was slowly added over 2 h. The mixture was stirred at room temperature for 1.5 h, followed by an aqueous workup (EtOAC, 1 M HCl soln, H.sub.2O, sat. aq. NaHCO.sub.3 soln, sat. aq. NaCl soln) to afford 35 (17.07 g, quant.).
(54) At 0 C., thiophenol (3.02 mL, 29.6 mmol) was added (dropwise, rapidly) to a mixture of 35 (7.85 g, 16.5 mmol) and K.sub.2CO.sub.3 (7.95 g, 57.5 mmol) in DMF (78 mL). The mixture was stirred for 2.5 h at 0-10 C. The mixture was diluted with EtOAc and washed with H.sub.2O and sat. aq. NaCl soln. The organic layer was extracted with ice-cold 1 M aqueous HCl soln. The aqueous phase (base extract) was poured onto aqueous Na.sub.2CO.sub.3 soln to reach pH ca 7; 2 M aq. NaOH soln. was added to reach pH ca 10, followed by extraction with EtOAc. The organic phase was dried (Na.sub.2SO.sub.4) and concentrated. The remaining oil (2.72 g) was dissolved in CH.sub.2Cl.sub.2 (30 mL) and treated with 4 M HCl-dioxane (10 mL) to afford after evaporation of the volatiles 36.HCl (3.34 g, 62%).
(55) Data of 36.HCl: C.sub.25H.sub.20N.sub.2O.sub.4.HCl (292.3, free base). LC-MS (method 7): R.sub.t=0.88, 293 ([M+H].sup.+). .sup.1H-NMR (DMSO-d.sub.6): 9.06 (br. s, NH.sup.+), 8.94 (br. s, NH.sup.+), 8.00 (d, J=8.3, NH), 7.38-7.30 (m, 5 arom. H), 5.88 (m, 1 H), 5.33 (d, J=17.3, 1 H), 5.23 (d, J=10.5, 1 H), 5.08 (s, 2 H), 4.63 (d, J=5.3, 2 H), 4.56 (m, 1 H), 3.35 (br. m, 1 H), 3.20 (br. m, 1 H), 2.57 (br. s, 3 H).
(S)-Allyl 2-(benzyloxycarbonylamino)-4-(methylamino)butanoate hydrochloride (40.HCl)
(56) Cbz-L-DabOH (37) was converted into the allylester-pTsOH salt 38.pTsOH according to the procedure of T. M. Kamenecka and S. J. Danishefsky, Chem. Eur. J. 2001, 7, 41-63, describing the synthesis of D-threonine allyl ester.
(57) A mixture of 38.pTsOH (45 g, 97 mmol) in CH.sub.2Cl.sub.2 (600 mL) was cooled to 0 C. MeOH (60 mL) was added, followed by ethyl trifluoroacetate (23 mL, 194 mmol). Et.sub.3N (53 mL, 387 mmol) was added dropwise. The mixture was stirred at 0 C. for 15 min, then at room temperature for 4 h. The volatiles were evaporated. The residue was dissolved in EtOAc, washed (1 M aq. HCl soln, sat. aq. Na.sub.2CO.sub.3 soln), dried (Na.sub.2SO.sub.4), filtered and concentrated to afford the corresponding trifluoroacetamide (32 g, 84%). N-Methylation of the acetamide (21.78 g, 56 mmol; applying CH.sub.3I and K.sub.2CO.sub.3 in DMF) following the procedure described by Chu-Biao Xue et al. J. Med. Chem. 2001, 44, 2636-2660with the exception that the transformation was performed at room temperature for 4 hafforded 39 (25 g, ca 90%). Treatment of 39 (8.0 g, ca 18 mmol) in THF (80 mL) with Pd(PPh.sub.3).sub.4 (0.2 g) and morpholine (8.5 mL, 98 mmol) at room temperature for 3 h afforded after aqueous workup (EtOAc, 1 M aq HCl soln.) the corresponding trifluoroacetamido acid (7.3 g) which was treated with NH.sub.3 (25% in H.sub.2O; 50 mL) for 2 h and concentrated to give the corresponding aminoacid (8 g). This material was dissolved in allyl alcohol (150 mL) and treated at 0 C. with thionyl chloride (6.6 mL, 91 mmol). The mixture was stirred at 0 C. for 15 min and at room temperature for 3 h and concentrated to give 40.HCl (7.6 g, used in the next step without further purification)
(58) Data of 40.HCl: C.sub.16H.sub.22N.sub.2O.sub.4.HCl (306.3, free base). Flow injection MS (ESI, positive modus): 307 ([M+H].sup.+). .sup.1H-NMR (DMSO-d.sub.6): 8.97 (br. s, NH.sub.2.sup.+), 7.92 (d, J=7.8, NH), 7.40-7.25 (m, 5 arom. H), 5.88 (m, 1 H), 5.32 (d, J=17.2, 1 H), 5.22 (d, J=10.5, 1 H), 5.05 (s, 2 H), 4.60 (d, J=5.2, 2 H), 4.22 (m, 1 H), 2.94 (m, 2 H), 2.50 (s, 3 H, superimposed by DMSO-d signal), 2.10 (m, 1 H), 2.00 (m, 1 H).
(S)-Allyl 2-(benzyloxycarbonylamino)-5-(methylamino)pentanoate hydrochloride (44.HCl)
(59) Cbz-L-OrnOH (41) was converted into the allylester-pTsOH salt 42.pTsOH according to the procedure of T. M. Kamenecka and S. J. Danishefsky, Chem. Eur. J. 2001, 7, 41-63, describing the synthesis of D-threonine allyl ester.
(60) The ester 42.pTsOH (5.5 g, 11 mmol) was converted into 43 (3.97 g, 83%) applying the conditions described for the synthesis of 39, with the exception that the N-methylation was continued at room temperature for 8 h.
(61) The allyl ester group was then cleaved applying the conditions described for the treatment of 39. The saponification of the resulting trifluoroacetamido acid was performed according to the procedure of Chu-Biao Xue et al. J. Med. Chem. 2001, 44, 2636-2660, with the exception that 2 equiv. of LiOH were used. The resulting amino acid (3.80 g, containing LiCl ca. 9 mmol) was treated at room temperature with allyl alcohol (100 mL) and thionyl chloride (3.0 mL, 41 mmol). The mixture was heated for 2 h at 70 C. Stirring was continued at room temperature for 17 h. The volatiles were evaporated. The resulting solid was washed with CH.sub.2Cl.sub.2 to afford 44.HCl (3.62 g, ca 75% w/w; yield 83%, used without further purification).
(62) Data of 44.HCl: C.sub.17H.sub.24N.sub.2O.sub.4.HCl (320.4, free base). LC-MS (method 9b): R.sub.t=1.48, 321 ([M+H].sup.+). .sup.1H-NMR (DMSO-d.sub.6): 9.26 (br. s, NH.sub.2.sup.+), 7.86 (d, J=7.7, NH), 7.39-7.13 (m, 5 arom. H), 5.89 (m, 1 H), 5.31 (br. d, J=17.3, 1 H), 5.20 (br. d, J=10.4, 1 H), 5.04 (s, 2 H), 4.58 (d, J=5.2, 2 H), 4.05 (br. m, 1 H), 2.81 (br. m, 2 H), 2.44 (s, 3 H), 1.80-1.60 (br. m, 4 H), Sarcosine allyl ester (46) was prepared as p-TsOH salt applying the procedure of T. M. Kamenecka and S. J. Danishefsky, Chem. Eur. J. 2001, 7, 41-63, describing the synthesis of D-threonine allyl ester.
(63) 2-((Allyloxycarbonyl) (methyl)amino)acetic acid (47) was prepared according to the method of M. Mori, A. Somada, S. Oida, Chem. Pharm. Bull. 2000, 48, 716-728.
(64) 3-((Allyloxycarbonyl) (methyl)amino)propanoic acid (49) was prepared applying the method of M. Mori, A. Somada, S. Oida, Chem. Pharm. Bull. 2000, 48, 716-728, describing the synthesis of N-allyloxycarbonylsarcosine.
(65) (S)-2-(Benzyloxycarbonylamino)pent-4-enoic acid (51) was prepared from (S)-allylglycine by N-protection (CBzOSu, dioxane, aqueous Na.sub.2CO.sub.3) in analogy to the procedure of D. R. Ijzendoorn, P. N. M. Botman, R. H. Blaauw, Org. Lett 2006, 8, 239-242.
(66) Acid 51 was also described by Z-Y Sun, CH. Kwon, J. N. D. Wurpel, J. Med. Chem. 1994, 37, 2841-2845.
(67) General Procedures
(68) Synthesis of the A-c1 Fragment
(69) Procedure A
(70) A.1: Acid Chloride Formation
(71) Oxalyl chloride (3.5-5.0 equiv.) was added to a mixture of the acetoxyaryl carboxylic acid (Ac-A-OH) and dry diethyl ether or CH.sub.2Cl.sub.2. The resulting mixture was stirred at room temperature for 15 min followed by the addition of a few drops (ca 50-100 L) of dry DMF. Stirring was continued for 16 h. The mixture was filtered. The filtrate was concentrated and the residue dried i.v. to afford the crude acetoxyaryl carboxylic acid chloride (Ac-A-Cl), which was immediately used in the next step.
(72) A.2: Amide Coupling
(73) A mixture of the amino ester salt (H-c1-OAll.HCl), the crude acetoxyaryl carboxylic acid chloride (Ac-A-Cl, 1.1-1.5 equiv.) and dry CH.sub.2Cl.sub.2 or THF was cooled to 0 C. An auxiliary base (sym-collidine or i-Pr.sub.2NEt; 3.0 equiv.) was added dropwise. The mixture was stirred at room temperature for 16 h. The mixture was distributed between EtOAc and 1 M aq. HCl solution. The organic phase was washed (1 M aq. HCl soln., then sat. aq. NaHCO.sub.3 soln. or sat aq. NaCl soln.), dried (Na.sub.2SO.sub.4), filtered and concentrated. FC (hexane/EtOAc gradients) gave the acetoxyaryl amide (Ac-A-c1-OAll).
(74) A.3: Deacetylation
(75) A solution of acetoxyarylamide (Ac-A-c1-OAll) in dry THF was treated at 0 C. with 3-dimethylaminopropylamine (3.0-4.5 equiv.). The solution was stirred at room temperature for 1-5 h. The mixture was distributed between EtOAc and icecold 0.1 M or 1 M aq. HCl solution. The organic phase was washed (0.1 or 1 M aq. HCl soln., sat. aq. NaCl soln.), dried (Na.sub.2SO.sub.4), filtered and concentrated to afford the hydroxyaryl amide (H-A-c1-OAll).
(76) Synthesis of the Linear Cyclization Precursor HB-A-c1-OH
(77) Procedure B
(78) B.1.1: Mitsunobu Aryl Ether Synthesis Using PPh.sub.3/DEAD
(79) A mixture of the hydroxyaryl amide (H-A-c1-OAll) and PPh.sub.3 (1.5 equiv.) was dried i.v. for 15 min. Under argon a solution of alcohol (HOB-Alloc, 1.2 equiv.) in dry benzene was added and the resulting solution was cooled to 0 C. A solution DEAD (40% in toluene, 1.2 equiv.) in benzene was slowly added (by syringe pump). The mixture was stirred at room temperature for 18 h and concentrated. FC (hexane/EtOAc gradients) gave the protected amino acid (Alloc-B-A-c1-OAll, sometimes contaminated with byproducts such as e.g. triphenylphosphine oxide, however acceptable for the use in the next step without further purification).
(80) B.1.2: Mitsunobu Aryl Ether Synthesis Using CMBP
(81) A solution of the hydroxyaryl amide (HO-A-c1-OAll), the alcohol (HOB-Alloc, 1.2-1.3 equiv) and CMBP (2 equiv) was heated in dry toluene at reflux for 3-4 h. The solution was concentrated. FC (hexane/EtOAc gradients) afforded the protected amino acid (Alloc-B-A-c1-OAll).
(82) B.2: Cleavage of the Allyl/Alloc Protective Groups
(83) Pd(PPh.sub.3).sub.4 (0.05-0.1 equiv.) was added to a mixture of the protected amino acid (Alloc-B-A-c1-OAll) and 1,3-dimethylbarbituric acid (2.5 equiv.) in degassed EtOAc/CH.sub.2Cl.sub.2 (ca. 1:1). The resulting solution was stirred at room temperature for 1-3 h and concentrated. FC (EtOAC, CH.sub.2Cl.sub.2/EtOH, or CH.sub.2Cl.sub.2/MeOH gradients) afforded the free amino acid (HB-A-c1-OH)
(84) Synthesis of the Linear Cyclization Precursor HB-a-c1-c2-Oh
(85) Procedure C
(86) C.1: Alloc Carbamate Formation
(87) At 0 C., allylchloroformate (1.1 equiv.) was slowly added to a mixture of aminoacid (HB-A-c1-OH) and Na.sub.2CO.sub.3 (1.5-3 equiv.) in dioxane/H.sub.2O 1:1. The mixture was stirred at room temperature for 15 h. The mixture was diluted with EtOAc and treated with 1 M aq. HCl solution until pH ca 2 was reached. The organic phase was separated, washed (sat. aq. NaCl soln.), dried (Na.sub.2SO.sub.4), filtered, concentrated and dried i.v. to afford the alloc protected amino acid (Alloc-B-A-c1-OH).
(88) C.2: Amid Coupling
(89) i-Pr.sub.2NEt (5.0 equiv.) was slowly added to a mixture of the alloc protected amino acid (Alloc-B-A-c1-OH), the aminoacid ester salt (H-c2-0All.p-TsOH, 1.2 equiv.), HOAt (1.5 equiv.) and HATU (1.5 equiv.) in DMF. The mixture was stirred at room temperature for 20 h followed by distribution between EtOAc and ice-cold 0.5 M aq, HCl solution. The organic phase was washed (0.5 M aq. HCl soln., H.sub.2O, sat. aq. NaHCO.sub.3 soln., sat. aq. NaCl soln.), dried (Na.sub.2SO.sub.4), filtered and concentrated. FC (hexane/EtOAc gradients) afforded the protected amino acid (Alloc-B-A-c1-c2-OAll)
(90) C.3: Cleavage of the Allyl/Alloc Protective Groups
(91) Pd(PPh.sub.3).sub.4 (0.1 equiv.) was added to a mixture of the protected amino acid (Alloc-B-A-c1-c2-OAll) and 1,3-dimethylbarbituric acid (2.5 equiv.) in degassed EtOAc/CH.sub.2Cl.sub.2 1:1. The resulting solution was stirred at room temperature for 1-2 h and concentrated. FC (EtOAC, CH.sub.2Cl.sub.2/EtOH, or CH.sub.2Cl.sub.2/MeOH gradients) afforded the free amino acid (HB-A-c1-c2-OH).
(92) Synthesis of the c2-B Fragment
(93) Procedure D
(94) Synthesis in Two Steps, Via Amidoester and Subsequent Saponification
(95) i-Pr.sub.2NEt (5.0 equiv.) was slowly added to a mixture of the N-protected amino acid (Alloc-c2-OH, 2.2 equiv.), the aminoalcohol hydrochloride (HOBH HCl), Cl-HOBt (0.25 equiv.) and HCTU (2.5 equiv.) in DMF. The resulting solution was stirred at room temperature for 17 h, followed by distribution between EtOAc and sat. aq. Na.sub.2CO.sub.3 solution. The organic phase was washed (1 M aq. HCl soln, sat. aq. NaCl soln), dried (Na.sub.2SO.sub.4), filtered and concentrated. FC (hexane/EtOAc or CH.sub.2Cl.sub.2/MeOH gradients) afforded the corresponding amidoester, which was dissolved in THF/H.sub.2O 4:1 and treated with lithium hydroxide monohydrate (3.0 equiv.) for 2 h at room temperature. The mixture was concentrated to about 50% of the original volume, diluted with EtOAc and extracted with 1 M aq. NaOH solution. The organic phase was washed (H.sub.2O, sat. aq. NaCl soln), dried (Na.sub.2SO.sub.4), filtered and concentrated to afford the amidoalcohol (HOB-c2-Alloc).
(96) Synthesis of the Linear Cyclization Precursor H-c2-B-A-c1-OH
(97) Procedure E
(98) E.1.1: Mitsunobu Aryl Ether Synthesis Using PPh.sub.3/DEAD
(99) A mixture of the hydroxyaryl amide (HO-A-c1-OAll) and PPh.sub.3 (1.5-4.5 equiv.) was dissolved in benzene. The solution was concentrated and the residue was dried i.v. for 15-30 min. Under argon, a solution of the alcohol (HOB-c2-Alloc, 1.2-2.3 equiv.) in dry and degassed benzene was added and the resulting mixture was cooled to 0 C. A solution of DEAD (40% in toluene, 1.2-4.5 equiv.) was slowly added. The mixture was stirred at room temperature for 18 h. In case of incomplete consumption of the hydroxyaryl amide, additional triphenylphosphine (1.0-1.3 equiv.) and DEAD (40% in toluene, 1.0 equiv.) and alcohol (1.0 equiv.)if consumed according to tlcwere added and stirring was continued for 18 h. The mixture was concentrated.
(100) FC (hexane/EtOAc, CH.sub.2Cl.sub.2/EtOH, or CH.sub.2Cl.sub.2/MeOH gradients) afforded Alloc-c2-B-A-c1-OAll (possibly contaminated with byproducts such as e.g. triphenylphosphine oxide, however acceptable for the use in the next step without further purification).
(101) E.1.2: Mitsunobu Aryl Ether Synthesis Using CMBP
(102) CMBP (2-3 equiv.) was added to a mixture of the hydroxyaryl amide (H-A-c1-OAll) and the alcohol (HOB-c2-Alloc, 1.2-2.2 equiv.) in dry toluene. The mixture was heated at reflux for 16 h and concentrated. FC (hexane/EtOAc gradients) afforded the protected amino acid (Alloc-c2-B-A-c1-OAll).
(103) E.2: Cleavage of the Allyl/Alloc Protective Groups
(104) Pd(PPh.sub.3).sub.4 (0.05-0.1 equiv.) was added to a mixture of the protected amino acid (Alloc-c2-B-A-c1-OAll) and 1,3-dimethylbarbituric acid (2.4 equiv.) in degassed EtOAc/CH.sub.2Cl.sub.2 1:1. The resulting solution was stirred at room temperature for 1-3 h and concentrated. FC(EtOAC, CH.sub.2Cl.sub.2/EtOH, or CH.sub.2Cl.sub.2/MeOH gradients) afforded the free amino acid (H-c2-B-A-c1-OH).
(105) Synthesis of the Macrocycles Cyclo-(B-A-c1), and Cyclo-(c2-B-A-c1)
(106) Procedure F
(107) The macrolactamization was typically performed at final concentrations ranging from 0.01 M to 0.001 M
(108) F.1.1: T3P Mediated Lactam Formation
(109) A solution of the precursor (HB-A-c1-OH or H-c2-B-A-c1-OH or HB-A-c1-c2-OH, respectively) in dry CH.sub.2Cl.sub.2 was added within 2 h by syringe pump to a solution of T3P (50% in EtOAc, 2 equiv.) and i-Pr.sub.2NEt (4 equiv.) in dry CH.sub.2Cl.sub.2. The solution was stirred at room temperature for 20 h, extracted with sat. aq. Na.sub.2CO.sub.3 solution and with H.sub.2O, dried (Na.sub.2SO.sub.4), filtered and then concentrated. FC (hexane/EtOAc/MeOH or CH.sub.2Cl.sub.2/MeOH gradients) afforded the macrocyclic compound (cyclo-(B-A-c1) or cyclo-(c2-B-A-c1), respectively).
(110) F.1.2: FDPP Mediated Lactam Formation
(111) A solution of the precursor (HB-A-c1-OH or H-c2-B-A-c1-OH or HB-A-c1-c2-OH, respectively) in dry DMF was added within 2 h to a solution of FDPP (2.0 equiv.) in dry DMF. The solution was stirred at room temperature for 20 h. The volatiles were evaporated and the residue taken up in EtOAc and washed (sat. aq. NaHCO.sub.3 soln, H.sub.2O, sat. aq. NaCl soln). The organic phase was dried (Na.sub.2SO.sub.4), filtered and concentrated. FC (hexane/EtOAc/MeOH or CH.sub.2Cl.sub.2/MeOH gradients gradients) afforded the macrocyclic compound (cyclo-(B-A-c1) or cyclo-(c2-B-A-c1), respectively).
(112) Attachment of Substituents to the Macrocyclic Core Structures: Synthesis of the Final Products
(113) Procedure H
(114) A solution of a macrocyclic benzylester in MeOH or MeOH/THF (ca 100 mL per g of starting material) was hydrogenated for 2 h at room temperature and at normal pressure in the presence of palladium hydroxide on activated charcoal (moistened with 50% H.sub.2O; 0.5 g per g of starting material). The mixture was filtered trough a pad of celite. The residue was washed (MeOH, MeOH/CH.sub.2Cl.sub.2 1:1, THF). The combined filtrate and washings were concentrated to obtain a macrocyclic acid.
(115) Procedure I
(116) I.1: Teoc Deprotection with Dioxane-HCl
(117) A solution of a macrocyclic Teoc-amine (1.5 mmol) in dioxane (18 mL) was treated with 4 M HCl in dioxane (18 mL) and stirred at room temperature for 4-16 h. The mixture was treated with diethyl ether and filtered. The solid was washed with diethyl ether and dried i.v. to give the macrocyclic amine hydrochloride.
(118) I.2: Teoc Deprotection with TBAF in THF
(119) A solution of TBAF (1 M in THF, 3 equiv.) was added at 0 C. to a solution of a macrocyclic Teoc-amine (1.3 mmol) in THF (34 mL). Stirring at 0 C. to room temperature was continued for 3 h. The solution was distributed between CH.sub.2Cl.sub.2 and H.sub.2O. The organic phase was washed (H.sub.2O), dried (Na.sub.2SO.sub.4), filtered and concentrated to provide after FC the macrocyclic amine.
(120) Procedure J
(121) A solution of a macrocyclic Boc-amine in dioxane (10 mL per g of starting material) was treated with 4 M HCl in dioxane (20 mL per g of starting material) and stirred at room temperature for 2 h. The mixture was filtered. The solid was washed with diethyl ether and dried i.v. to give the macrocyclic amine hydrochloride.
(122) Procedure K
(123) A solution of a macrocyclic benzylcarbamate (0.9 mmol) in MeCH (52 mL) was hydrogenated for 4 h at room temperature and at normal pressure in the presence of palladium hydroxide on activated charcoal (moistened with 50% H.sub.2O; 0.3 g). The mixture was filtered trough a pad of celite. The residue was washed (MeOH). The combined filtrate and washings were concentrated to obtain the macrocyclic amine.
(124) Procedure L
(125) Amide Coupling
(126) L.1.1: with Carboxylic Acid Anhydrides or Acylchlorides
(127) A Solution of an Amino macrocycle (free amine or hydrochloride; 0.09 mmol) in CH.sub.2Cl.sub.2 (1 mL) was at 0 C. subsequently treated with pyridine (10 equiv.) and the carboxylic acid anhydride (1.05-5 equiv.) or a carboxylic acid chloride (1.05-2.0 equiv.), respectively. The solution was stirred at room temperature for 15 h. After the addition of MeOH (0.1 mL) the solution was stirred for 10 min and concentrated. The resulting crude product was coevaporated with toluene and purified by chromatography (FC, normal phase or reversed phase prep. HPLC) to give an N-acylamino macrocycle.
(128) L.1.2: with Carboxylic Acid and Polymersupported Carbodiimide
(129) A solution of an amino macrocycle (free amine or hydrochloride; 0.09 mmol), a carboxylic acid (1.2 equiv.), HOBt.H.sub.2O (1.2 equiv.) in CH.sub.2Cl.sub.2 (1 mL) was treated with N-cyclohexyl-carbodiimide-N-methylpolystyrene (1.9 mmol/g; 1.5 equiv.) and i-Pr.sub.2NEt (3.0 equiv.). The mixture was stirred for 15 h at room temperature. (Polystyrylmethyl)trimethylammonium bicarbonate (3.5 mmol/g; 3 equiv.) was added and stirring was continued for 1 h. The mixture was diluted with CH.sub.2Cl.sub.2/MeOH 9:1 (2 mL) and filtered. The polymer was washed twice with CH.sub.2Cl.sub.2/MeOH 8:2 (5 mL). The combined filtrate and washings were concentrated. Purification of the crude product by chromatography (FC, normal phase or reversed phase prep. HPLC) afforded an N-acylamino macrocycle
(130) L.1.3: with a Carboxylic Acid and HATU
(131) A solution of an amino macrocycle (free amine or hydrochloride; 0.145 mmol), a carboxylic acid (2.0 equiv.), HATU (2.0 equiv.), HOAt (2.0 equiv.) in DMF (2 mL) was treated with i-Pr.sub.2NEt (4.0 equiv.). The mixture was stirred for 15 h at room temperature. The solvent was removed. The residue was distributed between CHCl.sub.3 and sat. aq. NaHCO.sub.3 solution. The organic phase was washed (H.sub.2O), dried (Na.sub.2SO.sub.4), filtered and concentrated. Purification of the crude product by chromatography (FC, normal phase or reversed phase prep. HPLC) afforded an N-acylamino macrocycle.
(132) L.2: with an Amine and HATU
(133) A solution of a macrocyclic carboxylic acid (0.78 mmol), an amine (2.0 equiv.), HATU (2.0 equiv.), HOAt (2.0 equiv.) in DMF (6 mL) was treated with i-Pr.sub.2NEt (4.0 equiv.). The mixture was stirred for 15 h at room temperature. The solvent was removed. The residue was distributed between CHCl.sub.3 and sat. aq. NaHCO.sub.3 solution. The organic phase was washed (H.sub.2O), dried (Na.sub.2SO.sub.4), filtered and concentrated. Purification of the crude product by chromatography (FC, normal phase or reversed phase prep. HPLC) afforded a macrocycle amide.
(134) Procedure M
(135) N,N-Diethylamino macrocycles by Reductive Amination
(136) At 0 C. NaBH(OAc).sub.3 (5 equiv.) and acetaldehyde (1 mL) were added to a solution of an the amino macrocycle (free amine or hydrochloride; 0.09 mmol) in THF (1 mL). The mixture was stirred at 0 C. to room temperature for 15 h. The mixture was diluted with CHCl.sub.3 and washed with sat. aq. NaHCO.sub.3 soln. The organic phase was dried (Na.sub.2SO.sub.4), filtered and concentrated. Purification of the crude product by chromatography (FC, normal phase or reversed phase prep. HPLC) afforded the diethylamino macocycle.
(137) Procedure N
(138) Methylester Cleavage
(139) A solution of the methylester (57 mol) in THF (1.5 mL) and MeOH (0.5 mL) was treated with H.sub.2O (0.5 mL) and lithium hydroxide monohydrate (3 equiv.) for 2 h at room temperature.
(140) The mixture was acidified by addition of aqueous 1 M HCl and concentrated. The crude product was purified by prep. HPLC.
Synthesis of A-c1 Fragments
1. Synthesis of (S)-5-allyl 1-benzyl 2-(5-fluoro-2-hydroxy-N-methylbenzamido)pentanedioate (54) (Scheme 4)
(141) Following procedure A (steps A.1-A.3), the reaction of 2-acetoxy-5-fluoro benzoic acid (2, 11.78 g, 59 mmol) and oxalylchloride (18 mL, 206 mmol) in dry CH.sub.2Cl.sub.2 (516 mL) in the presence of DMF (50 L) afforded 2-acetoxy-5-fluoro benzoyl chloride (52).
(142) Reaction of acid chloride 52 with (S)-5-allyl 1-benzyl 2-(methylamino)pentanedioate hydrochloride (27HCl, 15.0 g, 46 mmol) in THF (260 mL) in the presence of i-Pr.sub.2NEt (23 mL, 137 mmol) yielded the acetate 53 (19.35 g, 90%), which was treated with 3-dimethylamino-1-propylamine (23 mL, 185 mmol) in THF (200 mL) to afford after aqueous workup (EtOAc, 0.1 M aq. HCl soln, sat. aq. NaCl soln) and after FC (hexane/EtOAc 8:2 to 7:3) the phenol 54 (14.4 g, 81%).
(143) Data of 54: C.sub.23H.sub.24FNO.sub.6 (429.4). HPLC (30% CH.sub.3CN): R.sub.t=3.79 (87%). LC-MS (method 9a): R.sub.t=2.09, 430 ([M+H].sup.+).
2. Synthesis of (R)-5-allyl 1-benzyl 2-(5-fluoro-2-hydroxy-N-methylbenzamido)pentanedioate (56) (Scheme 4)
(144) Following procedure A (steps A.1-A.3), the reaction of 2-acetoxy-5-fluoro benzoic acid (2, 13.0 g, 67 mmol) and oxalylchloride (20 mL, 233 mmol) in dry CH.sub.2Cl.sub.2 (585 mL) in the presence of DMF (50 L) afforded 2-acetoxy-5-fluoro benzoyl chloride (52).
(145) Reaction of acid chloride 52 with (R)-5-allyl 1-benzyl 2-(methylamino)pentanedioate hydrochloride (29.HCl, 17.0 g, 52 mmol) in THF (280 mL) in the presence of i-Pr.sub.2NEt (27 mL, 156 mmol) yielded 55 (21.5 g, 88%), which was treated with 3-dimethylamino-1-propylamine (26 mL, 205 mmol) in THF (200 mL) to afford after aqueous workup (EtOAc, 0.1 M aq. HCl soln, sat. aq. NaCl soln) and after FC (hexane/EtOAc 8:2 to 7:3) the phenol 56 (14.8 g, 75%).
(146) Data of 56: C.sub.23H.sub.24FNO.sub.6 (429.4). HPLC (30% CH.sub.3CN): R.sub.t=3.79 (89). LC-MS (method 9c): R.sub.t=2.11, 430 ([M+H].sup.+).
3. Synthesis of (S)-allyl 2-(benzyloxycarbonylamino)-3-(3-hydroxy-N-methylbenzamido)propanoate (59) (Scheme 4)
(147) Following procedure A (steps A.1-A.3), the reaction of 3-acetoxybenzoic acid (3, 6.0 g, 33 mmol) and oxalylchloride (14 mL, 164 mmol) in dry diethyl ether (216 mL) in the presence of DMF (50 L) afforded 3-acetoxybenzoyl chloride (57, 7.0 g, quant.).
(148) Reaction of 57 (7.0 g, 35 mmol) with (S)-allyl 2-(benzyloxycarbonylamino)-3-(methylamino)propanoate hydrochloride (32.HCl, 10.5 g, 32 mmol) in CH.sub.2Cl.sub.2 (285 mL) in the presence of 2,4,6-collidine (12.8 mL, 96 mmol) yielded 58 (12.34 g, 82%).
(149) The acetate 58 (12.82 g, 28.2 mmol) was treated with 3-dimethylamino-1-propylamine (10.6 mL, 84.6 mmol) in THF (114 mL) to afford the phenol 59 (10.45 g, 90%).
(150) Data of 59: C.sub.22H.sub.24N.sub.2O.sub.6 (412.4). HPLC (10% CH.sub.3CN): R.sub.t=3.91 (96). LC-MS (method 9a): R.sub.t=1.77, 413 ([M+H].sup.+).
4. Synthesis of (R)-allyl 2-(benzyloxycarbonylamino)-3-(3-hydroxy-N-methylbenzamido)propanoate (61) (Scheme 4)
(151) Following procedure A (steps A.1-A.3), the reaction of 3-acetoxybenzoic acid (3, 5.82 g, 32.3 mmol) and oxalylchloride (11.1 mL, 129 mmol) in dry diethyl ether (210 mL) in the presence of DMF (50 L) afforded 3-acetoxybenzoyl chloride (57, 6.5 g, 100%).
(152) Reaction of 57 (6.5 g, 32.3 mmol) with (R)-allyl 2-(benzyloxycarbonylamino)-3-(methylamino)propanoate hydrochloride (36.HCl, 8.5 g, 26 mmol) in CH.sub.2Cl.sub.2 (220 mL) in the presence of 2,4,6-collidine (10.3 mL, 77.6 mmol) yielded 60 (10.73 g, 92%).
(153) The acetate 60 (15.46 g, 34 mmol) was treated with 3-dimethylamino-1-propylamine (12.8 mL, 102 mmol) in THF (140 mL) to afford the phenol 61 (12.92 g, 92%).
(154) Data of 61: C.sub.22H.sub.24N.sub.2O.sub.6 (412.4). LC-MS (method 2): R.sub.t=1.77 (98), 413 ([M+H].sup.+).
5. Synthesis of (S)-allyl 2-(benzyloxycarbonylamino)-4-(3-hydroxy-N-methylbenzamido)butanoate (63) (Scheme 4)
(155) Following procedure A (steps A.1-A.3), the reaction of 3-acetoxybenzoic acid (3, 7.65 g, 43 mmol) and oxalylchloride (18.2 mL, 213 mmol) in dry CH.sub.2Cl.sub.2 (140 mL) in the presence of DMF (300 L) afforded after 3 h at room temperature 3-acetoxybenzoyl chloride (57).
(156) Reaction of 57 thus obtained with (S)-allyl 2-(benzyloxycarbonylamino)-5-(methylamino)butanoate hydrochloride (40.HCl, 8.7 g, 28 mmol) in THF (140 mL) in the presence of i-Pr.sub.2NEt (15 mL, 85 mmol) yielded 62 (8.1 g, 61%).
(157) The acetate 62 (4.85 g, 10 mmol) was treated with 3-dimethylamino-1-propylamine (3.8 mL, 31 mmol) in THF (90 mL) to afford the phenol 63 (4.23 g, 95%).
(158) Data of 63: C.sub.23H.sub.26N.sub.2O.sub.6 (426.5). LC-MS: (method 6): R.sub.t=1.06 (99), 427 ([M+H].sup.+).
6. Synthesis of (S)-allyl 2-(benzyloxycarbonylamino)-5-(3-hydroxy-N-methylbenzamido)pentanoate (65) (Scheme 4)
(159) Following procedure A (steps A.1-A.3), the reaction of 3-acetoxybenzoic acid (3, 10 g, 58 mmol) and oxalylchloride (19 mL, 218 mmol) in dry CH.sub.2Cl.sub.2 (450 mL) in the presence of DMF (500 L) afforded 3-acetoxybenzoyl chloride (57).
(160) Reaction of 57 thus obtained with (S)-allyl 2-(benzyloxycarbonylamino)-5-(methylamino)pentanoate hydrochloride (44.HCl, 17.3 g, 48 mmol) in THF (200 mL) in the presence of i-Pr.sub.2NEt (25 mL, 145 mmol) yielded 64 (12.08 g, 51%), which was treated with 3-dimethylamino-1-propylamine (9.3 mL, 75 mmol) in THF (240 mL) to afford after aqueous workup (EtOAc, 1 M aq. HCl soln, sat. aq. NaHCO.sub.3 soln, sat.-aq. NaCl soln) the phenol 65 (10.84 g, 98%).
(161) Data of 65: C.sub.24H.sub.28N.sub.2O.sub.6 (440.5). LC-MS (method 6): R.sub.t=1.15 (91), 441 ([M+H].sup.+).
7. Synthesis of (S)-5-allyl 1-benzyl 2-(4-hydroxy-N-methylbenzamido)pentanedioate (68) (Scheme 4)
(162) Following procedure A (steps A.1-A.3), the reaction of 4-acetoxybenzoic acid (4, 10.7 g, 59.5 mmol) and oxalylchloride (17.7 mL, 206 mmol) in dry CH.sub.2Cl.sub.2 (350 mL) in the presence of DMF (50 L) afforded 4-acetoxybenzoyl chloride (66).
(163) Reaction of 66 with (S)-5-allyl 1-benzyl 2-(methylamino)pentanedioate hydrochloride (27.HCl, 15.0 g, 46 mmol) in THF (250 mL) in the presence of i-Pr.sub.2NEt (23.3 mL, 137 mmol) yielded 67 (16.24 g, 78%).
(164) The treatment of 67 (15.2 g, 33.5 mmol) with 3-dimethylamino-1-propylamine (12.6 mL, 101 mmol) in THF (140 mL) afforded the phenol 68 (14.86 g, quant.; the product was contaminated with 9% EtOAc).
(165) Data of 68: C.sub.23H.sub.25NO.sub.6 (411.4). LC-MS (method 9b): R.sub.t=1.96, 412 ([M+H].sup.+).
8. Synthesis of (S)-allyl 2-(benzyloxycarbonylamino)-3-(5-hydroxy-N-methylnicotinamido)propanoate (71) (Scheme 4)
(166) A mixture of 5-hydroxy nicotinic acid (5, 3.5 g, 25.1 mmol) and acetic anhydride (23 mL, 243 mmol) was heated at 95 C. for 45 min and cooled to room temperature. The mixture was filtered. The solid was washed (H.sub.2O, diethyl ether) and dried i.v. to give 5-acetoxynicotinic acid (6; 3.76 g, 82%) (Scheme 1) 5-Acetoxynicotinic acid (6; 5.7 g, 31.5 mmol) was suspended in CHCl.sub.3 (stabilized with amylene, 230 mL). Oxalylchloride (9.0 mL, 105 mmol) was added followed by DMF (ca. 50 l). The mixture was stirred at room temperature for 15 h, then concentrated, coevaporated with dry CH.sub.2Cl.sub.2 and dried i.v. to afford 5-acetoxynicotinoyl chloride (69). (S)-allyl 2-(benzyloxycarbonylamino)-3-(methylamino)propanoate hydrochloride (32, 8.6 g, 26.2 mmol) and THF (225 mL) were added. The mixture was cooled to 0 C. Et.sub.3N (13 mL, 92 mmol) was slowly added. The mixture was stirred at 0 C. to room temperature for 18 h. 3-dimethylamino-1-propylamine (9.9 mL, 78.6 mmol) was added and stirring at room temperature was continued for 2 h. The mixture was distributed between EtOAc and 1 M aq. NaH PO.sub.4 solution. The organic layer was separated, washed (sat. aq. NaCl soln), dried (Na.sub.2SO.sub.4), filtered and concentrated. FC(CH.sub.2Cl.sub.2/MeOH 19:1) afforded the phenol 71 (8.81 g, 81%).
(167) Data of 71: C.sub.21H.sub.23N.sub.3O.sub.6 (413.4). LC-MS (method 6): R.sub.t=0.94 (92), 414 ([M+H].sup.+).
9. Synthesis of allyl (2S)-2-[(benzyloxy)carbonyl]amino-3-[((2S)-2-[(tert-butoxycarbonyl)amino]-8-hydroxy-1,2,3,4-tetrahydro-2-naphthalenylcarbonyl) (methyl)amino]propanoate (72) (Scheme 4)
(168) A mixture of 10 (3.0 g, 9.76 mmol), HATU (5.57 g, 14.6 mmol), HOAt (1.99 g, 14.6 mmol) and 32.HCl (6.4 g, 19.5 mmol) were dissolved in DMF (113 mL). i-Pr.sub.2NEt (8.36 mL, 48.8 mmol) was added. The mixture was stirred at room temperature for 3 d. The mixture was distributed between H.sub.2O and EtOAc. The organic phase was dried (Na.sub.2SO.sub.4), filtered, and concentrated. FC (hexane/EtOAc 75:25 to 50:50) afforded 72 (2.58 g, 45%).
(169) Data of 72: C.sub.31H.sub.39N.sub.3O.sub.8 (581.3). LC-MS (method 7): R.sub.t=1.27 (97), 582 ([M+H].sup.+).
10. Synthesis of 5-allyl 1-benzyl (25)-2-[[(8-hydroxy-2-quinolinyl)carbonyl] (methyl)amino]pentanedioate (75) (Scheme 4)
(170) Following procedure A (steps A.1-A.3), the reaction of 8-Acetoxyquinoline-2-carboxylic acid (8, 2.22 g 9.6 mmol) and oxalylchloride (2.1 mL, 24 mmol) in dry CH.sub.2Cl.sub.2 (90 mL) (no addition of DMF) afforded after 2 h at room temperature acetoxyquinoline-2-carboxylic acid chloride (73).
(171) Reaction of 73 with (S)-5-allyl 1-benzyl 2-(methylamino)pentanedioate hydrochloride (27.HCl, 2.3 g, 8.0 mmol) in CH.sub.2Cl.sub.2 (200 mL) in the presence of i-Pr.sub.2NEt (5.5 mL, 32 mmol) yielded after 2.5 h at room temperature and purification by FC (hexane/EtOAc gradient) 74 (3.03 g, 74%), which was treated with 3-dimethylamino-1-propylamine (2.3 mL, 18 mmol) in THF (54 mL) to afford after aqueous workup (EtOAc, 1 M aq. HCl soln, sat. aq. NaHCO.sub.3 soln, sat.-aq. NaCl soln) the phenol 75 (2.79 g, 99%).
(172) Data of 75: C.sub.26H.sub.26N.sub.2O.sub.6 (462.5). LC-MS (method 7): R.sub.t=1.29 (94), 463 ([M+H].sup.+).
11. Synthesis of N-allyl-3-hydroxy-N-methylbenzamide (77) (Scheme 4)
(173) Following procedure A (steps A.1-A.3), the reaction of 3-acetoxybenzoic acid (3, 23.7 g, 132 mmol) and oxalylchloride (45.3 mL, 527 mmol) in dry diethyl ether (800 mL) in the presence of DMF (100 L) afforded 3-acetoxybenzoyl chloride (57).
(174) Reaction of 57 thus obtained with N-allylmethylamine (10.1 ml, 105 mmol) in CH.sub.2Cl.sub.2 (500 mL) in the presence of 2,4,6-collidine (42 mL, 316 mmol) yielded 76 (24 g, 98%).
(175) The acetate 76 (10.9 g, 46.7 mmol) was treated with 3-dimethylamino-1-propylamine (17.5 mL, 140 mmol) in THF (90 mL) to afford after aqueous workup (EtOAc, 1 M aq. HCl soln, sat. aq. NaCl soln) the phenol 77 (9.0 g, 100%).
(176) Data of 77: C.sub.11H.sub.13NO.sub.2 (191.2). LC-MS (method 2): R.sub.t=1.52 (99), 192 ([M+H].sup.+).
12. Synthesis of (S)-5-allyl-1-benzyl 2-(3-mercapto-N-methylbenzamido)pentanedioate (80) (Scheme 4)
(177) Acetic anhydride (0.46 mL, 4.86 mmol) was added at 0 C. to a solution of 3-mercaptobenzoic acid (11, 250 mg, 1.62 mmol) in 1 M aqueous NaOH solution (5.0 mL, 5.0 mmol). The mixture was stirred at 0 C. for 1 h. A precipitate was formed. The mixture was acidified by the addition of 1 M aqueous HCl solution and filtered. The solid was dried i.v. to afford 3-(acetylthio)benzoic acid (12; 280 mg, 88%).
(178) Oxalyl chloride (0.34 mL, 3.97 mmol) was added to a mixture of 12 (260 mg, 1.33 mmol) and CHCl.sub.3 (stabilized with amylene; 16 mL). DMF (7 L) was added. The mixture was stirred at room temperature for 2 h. The volatiles were evaporated to afford 3-(acetylthio)benzoyl chloride (78).
(179) (S)-5-allyl 1-benzyl 2-(methylamino)pentanedioate hydrochloride (27.HCl, 434 mg, 1.33 mmol) and dry THF (5 mL) were added. The mixture was cooled to 0 C., followed by the addition of i-Pr.sub.2NEt (0.79 mL, 4.6 mmol). The mixture was stirred at room temperature for 16 h and distributed between EtOAc and 1 M aqueous HCl solution. The organic phase was separated, dried (Na.sub.2SO.sub.4), filtered and concentrated. FC (hexane/EtOAc 2:1) afforded the acetate 79 (420 mg, 67%).
(180) At room temperature, a solution of 79 (246 mg, 0.52 mmol) in degassed THF (3.6 mL) was treated with 3-dimethylamino-1-propylamine (0.13 mL, 1.05 mmol) for 1 h. The mixture was distributed between EtOAc and 1 M aqueous HCl solution. The organic phase was separated, dried (Na.sub.2SO.sub.4), filtered and concentrated. FC (hexane/EtOAc 2:1) afforded 80 (153 mg, 68%).
(181) Data of 80: C.sub.23H.sub.25NO.sub.5S (427.5): LC-MS (method 7): R.sub.t=1.39 (84), 428 ([M+H].sup.+).
Synthesis of c2-B Fragments
1. Synthesis of allyl N-2-[(2S,4S)-4-[(tert-butoxycarbonyl)amino]-2-(hydroxymethyl)tetrahydro-1H-pyrrol-1-yl]-2-oxoethyl-N-methylcarbamate (81) (Scheme 5)
(182) A solution of (2-((allyloxycarbonyl) (methyl)amino)acetic acid (47, 8.0 g, 46 mmol) and aminoalcohol 13 (11.0 g, 51 mmol) in DMF (120 mL) was cooled to 0 C. 2,4,6-Collidine (11 mL, 82 mmol) was added followed by HATU (22 g, 58 mmol). The mixture was stirred for 1 h at 0 C. then for 16 h at room temperature followed by distribution between EtOAc and sat. aq. Na.sub.2CO.sub.3 solution. The organic phase was washed (1 M aq. HCl soln, sat. aq. NaCl soln), dried (Na.sub.2SO.sub.4), filtered and concentrated. FC (EtOAc/MeOH 100:0 to 95:5) afforded the amidoalcohol 81 (14.7 g, 86%).
(183) Data of 81: C.sub.17H.sub.29N.sub.3O.sub.6 (371.4). HPLC (20% CH.sub.3CN): R.sub.t=2.94 (97). LC-MS (method 9c): R.sub.t=1.55; 743 ([2M+H].sup.+), 372 ([M+H].sup.+).
2. Synthesis of allyl N-2-[(2S,4R)-4-[(tert-butoxycarbonyl)amino]-2-(hydroxymethyl)tetrahydro-1H-pyrrol-1-yl]-2-oxoethyl-N-methylcarbamate (82) (Scheme 5)
(184) Following procedure D, the reaction of the amnioalcohol 17.HCl (10.0 g, 39.6 mmol) and 2-((allyloxycarbonyl) (methyl)amino)acetic acid (47, 15.1 g, 87 mmol) in DMF (100 mL) in the presence of HCTU (40.9 g, 98.9 mmol), Cl-HOBt (1.68 g, 9.89 mmol) and i-Pr.sub.2NEt (33.6 mL, 198 mmol) afforded after FC (hexane/EtOAc 20:80 to 0:100) the corresponding amido ester intermediate (13.7 g) which was saponified with lithium hydroxide monohydrate (3.28 g, 78.1 mmol) in THF (350 mL) and H.sub.2O (90 mL) to yield the amidoalcohol 82 (8.89 g, 61%).
(185) Data of 82: C.sub.17H.sub.29N.sub.3O.sub.6 (371.4). LC-MS (method 9b): R.sub.t=1.57; 372 ([M+H].sup.+), 316, 272 ([M+H-Boc].sup.+), 156.
3. Synthesis of tert-butyl (3R)-4-{2-[[(allyloxy)carbonyl](methyl)amino]acetyl}-3-(hydroxymethyl)tetrahydro-1(2H)-pyrazinecarboxylate (84) (Scheme 5)
(186) Following procedure D, the reaction of (R)-tert-butyl 3-(hydroxymethyl)piperazine-1-carboxylate hydrochloride (83.HCl, 19.7 g, 78 mmol) and 3-((allyloxycarbonyl) (methyl)amino)acetic acid (47, 30 g, 172 mmol) in DMF (188 mL) in the presence of HCTU (81.0 g, 195 mmol), Cl-HOBt (3.3 g, 19 mmol) and i-Pr.sub.2NEt (67 mL, 390 mmol) afforded after FC (EtOAc) the corresponding amido ester intermediate (40 g) which was saponified with lithium hydroxide monohydrate (9.5 g, 228 mmol) in THF (1020 mL) and H.sub.2O (245 mL) to yield after FC (EtOAc) amidoalcohol 84; 22.8 g, 79%).
(187) Data of 84: C.sub.17H.sub.29N.sub.3O.sub.6 (371.4). LC-MS (method 7): R.sub.t=0.99 (93), 372 ([M+H].sup.+).
4. Synthesis of benzyl N-MS)-1-[(2S,4S)-4-[(tert-butoxycarbonyl)amino]-2-(hydroxymethyl)tetrahydro-1H-pyrrol-1-yl]carbonyl-3-butenyl)carbamate (85) (Scheme 5)
(188) Aminoalcohol-hydrochloride 13.HCl (3.7 g, 14.7 mmol) was added to a solution of acid 51 (5.22 g, 14.7 mmol) in DMF (80 ml). The mixture was cooled to 0 C. HATU (7.0 g, 18.4 mmol) and 2,4,6-collidine (3.51 ml, 26.4 mmol) were added. The solution was stirred at 0 C. to room temperature for 17 h, followed by distribution between EtOAc and sat. aq. Na.sub.2CO.sub.3 solution. The organic phase was washed (1 M aq. HCl soln, sat. aq. NaHCO.sub.3 soln, sat. aq. NaCl soln), dried (Na.sub.2SO.sub.4), filtered and concentrated. FC (hexane/EtOAc 30:70 to 20:80) afforded the amidoalcohol (85, 5.78 g, 88%)
(189) Data of 85: C.sub.23H.sub.33N.sub.3O.sub.6 (447.5). LC-MS (method 2): R.sub.t=1.92 (92), 448 ([M+H].sup.+).
5. Synthesis of allyl N-3-[(2S,4R)-4-[(tert-butoxycarbonyl)amino]-2-(hydroxymethyl)tetrahydro-1H-pyrrol-1-yl]-3-oxopropyl-N-methylcarbamate (86) (Scheme 5)
(190) Following procedure D, the reaction of aminoalcohol 17.HCl (7.5 g, 30 mmol) and 3-((allyloxycarbonyl) (methyl)amino)propanoic acid (49, 12.3 g, 66 mmol) in DMF (77 mL) in the presence of HCTU (31.0 g, 75.0 mmol), Cl-HOBt (1.27 g, 7.5 mmol) and i-Pr.sub.2NEt (25.6 mL, 150 mmol) afforded after FC(CH.sub.2Cl.sub.2/MeOH 100:0 to 97:3) the corresponding amido ester intermediate (17.1 g) which was saponified with lithium hydroxide monohydrate (3.8 g, 90 mmol) in THF (388 mL) and H.sub.2O (105 mL) to yield the amidoalcohol 86 (10.48 g, 86%).
(191) Data of 86: C.sub.18H.sub.31N.sub.3O.sub.6 (385.4). HPLC (10% CH.sub.3CN): R.sub.t=3.49 (88). LC-MS (method 9a): R.sub.t=1.62; 386 ([M+H].sup.+), 330 ([M+H-tBu].sup.+), 286 ([M+H-Boc].sup.+).
Core 01: Synthesis of Ex.1 (Scheme 6)
(192) Synthesis of the Mitsunobu Product 87
(193) To a solution of 54 (350 mg, 0.82 mmol), 16 (590 mg, 1.7 mmol) and PPh.sub.3 (1069 mg, 4.08 mmol) in dry degassed CHCl.sub.3 (11 mL) was added ADDP (1028 mg, 4.08 mmol) in one portion at 0 C., under a N.sub.2 atmosphere. The resulting mixture was stirred for 16 h at room temperature. The mixture was filtered and the slurry washed further with diethyl ether. The combined filtrates were concentrated in vacuo. The crude residue was purified by FC (CH.sub.2Cl.sub.2/EtOH 100:0 to 99:1) to afford 87 (1.05 g, contains triphenylphosphine oxide; used in the next step without further purification).
(194) Synthesis of the Amino Acid 88
(195) Following procedure B.2, the reaction of 87 (441 mg, contaminated with triphenylphosphine oxide, ca 0.5 mmol), 1,3-dimethylbarbituric acid (219 mg, 1.4 mmol) and Pd(PPh.sub.3).sub.4 (34 mg) in EtOAc/CH.sub.2Cl.sub.2 (55:45, 10 mL) yielded after 1.5 h and subsequent FC(CH.sub.2Cl.sub.2/MeOH 100:0 to 80:20) amino acid 88 (267 mg, 72%).
(196) Data of 88: C.sub.31H.sub.42FN.sub.3O.sub.8Si (631.7). LC-MS (method 9a):
(197) R.sub.t=2.02, 632 ([M+H].sup.+). HPLC (30% CH.sub.3CN): R.sub.t=3.41 (96).
(198) Synthesis of the Macrolactam Ex.1
(199) According to procedure F.1.1 the amino acid 88 (75 mg, 0.12 mmol) in dry CH.sub.2Cl.sub.2 (6 mL) was added within 4 h to T3P (50% in EtOAc, 0.21 mL, 0.36 mmol) and i-Pr.sub.2NEt (0.1 mL, 0.59 mmol) in dry CH.sub.2Cl.sub.2 (6 mL) to give after FC(CH.sub.2Cl.sub.2/MeOH 100:0 to 96:4) the macrolactam Ex.1 (45 mg, 61%).
(200) Data of Ex.1: C.sub.34H.sub.40FN.sub.3O.sub.7Si (613.7). LC-MS (method 7):
(201) R.sub.t=1.45 (41), 614 ([M+H].sup.+); 1.47 (44), 614 ([M+H].sup.+).
(202) .sup.1H-NMR (DMSO-d.sub.6): complex spectrum, several isomers; 7.45-7.01 (m, 8 H), 6.78-6.58 (2 m, 1 H), 5.42-5.06 (m, 3 H), 4.50-3.50 (several m, 7 H), 3.30-1.40 (several m, 7 H), 2.84, 2.70, 2.66 (s, 3 H), 0.97-0.82 (m, 2 H), 0.03, 0.02, 0.00 (s 9 H).
Core 02: Synthesis of Ex.2 (Scheme 11)
(203) Synthesis of the Protected Macrolactam Ex.2
(204) A solution of T3P (50% in EtOAc, 0.75 mL, 1.27 mmol) and i-Pr.sub.2NEt (0.36 mL, 2.2 mmol) in dry CH.sub.2Cl.sub.2 (20 mL) was added within 2 h to a solution of the amino acid 98 (250 mg, 0.43 mmol) in dry CH.sub.2Cl.sub.2 (730 mL). The solution was stirred at room temperature for 20 h, followed by extraction with sat. aq. Na.sub.2CO.sub.3 solution. The organic phase was dried (Na.sub.2SO.sub.4), filtered and concentrated. FC(CH.sub.2Cl.sub.2/MeOH 100:0 to 95:5) afforded Ex.2 (187 mg, 77%).
(205) Data of Ex.2: C.sub.30H.sub.36FN.sub.3O.sub.7 (569.6) LC-MS (method 7):
(206) R.sub.t=1.35 (62), 570 ([M+H].sup.+); 1.39 (15), 570 ([M+H].sup.+)
(207) .sup.1H-NMR (DMSO-d.sub.6): complex spectrum, several isomers; 7.46-7.30 (m, 5 H), 7.27-7.06 (m, 2 H), 6.98-6.67 (4 dd, 1 H), 5.54-5.06 (m, 3 H), 4.68-3.48 (m, 6 H), 3.05-1.98 (m 10 H; s at 2.82, 2.69, 2.64), 1.44-1.41 (3s, 9 H).
Core 03: Synthesis of Ex.3, Ex.4, and Ex.5 (Scheme 7)
(208) Synthesis of the Mitsunobu Product 89
(209) Following procedure E.1.1, the reaction of phenol 54 (7.8 g, 18 mmol), alcohol 81 (16 g, 43 mmol), DEAD (40% in toluene, 37 mL, 82 mmol), and PPh.sub.3 (21 g, 80 mmol) in dry benzene (250 mL) afforded after FC(CH.sub.2Cl.sub.2/EtOH 100:0 to 95:5) the protected amino acid 89 (15.9 g, contaminated with ca. 30% triphenylphosphine oxide; used in the next step without further purification).
(210) Synthesis of the Amino Acid 90
(211) Following procedure E.2, the reaction of 89 (9.6 g, contaminated with triphenylphosphine oxide, ca 9 mmol), 1,3-dimethylbarbituric acid (5.0 g, 32.0 mmol) and Pd(PPh.sub.3).sub.4 (0.4 g) in EtOAc/CH.sub.2Cl.sub.2 (55:45, 266 mL) yielded after 1.5 h and after FC(CH.sub.2Cl.sub.2/MeOH 90:10 to 50:50) amino acid 90 (4.34 g, 76%).
(212) Data of 90: C.sub.33H.sub.43FN.sub.4O.sub.9 (658.7). HPLC (10% CH.sub.3CN): R.sub.t=3.87 (99).
(213) LC-MS (method 9a): R.sub.t=1.77, 659 ([M+H].sup.+).
(214) Synthesis of the Protected Macrolactam Ex.3
(215) According to procedure F.1.2, the amino acid 90 (2.5 g, 3.80 mmol) in dry DMF (50 mL) was treated with FDPP (2.51 g, 6.53 mmol) in DMF (400 mL) to afford after FC (EtOAc/MeOH 100:0 to 95:5) the macrolactam Ex.3 (2.29 g, 94%).
(216) Data of Ex.3: C.sub.33H.sub.41FN.sub.4O.sub.8 (640.7). HPLC (30% CH.sub.3CN): R.sub.t=3.20 (96). LC-MS (method 9c): R.sub.t=2.06, 641 ([M+H].sup.+). .sup.1H-NMR (CDCl.sub.3): 7.45-7.32 (m, 5 H), 7.06 (m, 1 H), 6.94-6.88 (m, 2 H), 5.57 (dd, J=2.8, 12.6, 1 H), 5.42 (br. m, 1 H), 5.26 (d, J=12.2, 1 H), 5.15 (d, J=12.2, 1 H), 4.90 (dd, J=2.5, 11.0, 1 H), 4.34 (d, J=17.2, 1 H), 4.35-4.11 (m, 3 H), 3.82 (br. t, J ca. 8.5, 1 H), 3.65 (d, J=17.3, 1 H), 3.29 (t, J ca 8.8, 1 H), 3.14 (s, 3 H), 2.65 (s, 3 H), 2.51-1.98 (several m, 5 H), 1.76 (td, J=8.2, 12.7, 1 H), 1.36 (s, 9 H).
(217) Synthesis of the Acid Ex.4:
(218) According to procedure H, ester Ex.3 (2.0 g, 3.1 mmol) was hydrogenated in MeOH (120 mL)/THF (40 mL) in the presence of the catalyst (1 g) for 2 h to afford Ex.4 (1.68 g, 97%).
(219) Data of Ex.4: C.sub.26H.sub.35FN.sub.4O.sub.8 (550.6). HPLC (5% CH.sub.3CN): R.sub.t=3.60 (86). LC-MS: (method 9c): R.sub.t=1.53; 551 ([M+H].sup.+), 451 ([M+H-Boc].sup.+).
(220) Synthesis of the Amine Ex.5:
(221) According to procedure J, ester Ex.3 (100 mg, 0.16 mmol) in dioxane (3 mL) was treated with 4 M HCl-dioxane (3 mL) to afford Ex.5.HCl (100 mg, quant.).
(222) Data of Ex.5.HCl: C.sub.28H.sub.33FN.sub.4O.sub.6.HCl (540.6, free base). LC-MS: (method 9c): R.sub.t=1.44, 541 ([M+H].sup.+).
Core 04: Synthesis of Ex.56 and Ex.57 (Scheme 8)
(223) Synthesis of the Mitsunobu Product 91
(224) Following procedure E.1.1 the reaction of phenol 54 (8.0 g, 19 mmol), alcohol 82 (16.0 g, 43 mmol), DEAD (40% in toluene, 38 mL, 84 mmol), and PPh.sub.3 (22 g, 84 mmol) in dry benzene (260 mL) afforded after FC the protected amino acid 91 (33.5 g, contaminated with triphenylphosphine oxide. The material was used in the next step without further purification).
(225) Synthesis of the Amino Acid 92
(226) Following procedure E.2, the reaction of 91 (33.5 g, impure material), 1,3-dimethylbarbituric acid (16 g, 102 mmol) and Pd(PPh.sub.3).sub.4 (0.2 g) in EtOAc/CH.sub.2Cl.sub.2 (45:55, 340 mL) yielded after 3 h and after FC(CH.sub.2Cl.sub.2/EtOH 100:0 to 70:30 then CH.sub.2Cl.sub.2/MeOH 90:10 to 70:30) amino acid 92 (4.8 g, 39% over the two steps, based on phenol 54).
(227) Data of 92: C.sub.33H.sub.43FN.sub.4O.sub.9 (658.7). HPLC (10% CH.sub.3CN): R.sub.t=3.80 (95). LC-MS (method 9c): R.sub.t=1.81, 659 ([M+H].sup.+).
(228) Synthesis of the Protected Macrolactam Ex.56
(229) According to procedure F.1.1, amino acid 92 (3.8 g, 5.80 mmol) in dry CH.sub.2Cl.sub.2 (40 mL) was treated with T3P (50% in EtOAc, 6.8 mL, 12 mmol) and i-Pr.sub.2NEt (4.0 mL, 23 mmol) in dry CH.sub.2Cl.sub.2 (510 mL) to afford after FC (EtOAc/MeOH 100:0 to 95:5) the macrolactam Ex.56 (3.23 g, 87%).
(230) Data of Ex.56: C.sub.33H.sub.41FN.sub.4O.sub.8 (640.7). HPLC (30% CH.sub.3CN): R.sub.t=3.49 (88). LC-MS (method 9c): R.sub.t=2.02, 641 ([M+H].sup.+). .sup.1H-NMR (CDCl.sub.3): 7.41-7.32 (m, 5 H), 7.04 (m, 1 H), 6.94-6.83 (m, 2 H), 5.54 (dd, J=3.0, 12.7, 1 H), 5.25 (d, J=12.2, 1 H), 5.14 (d, J=12.2, 1 H), 4.89 (dd, J=2.1, 11.0, 1 H), 4.63 (br. m, 1 H), 4.39-4.10 (m, 4 H), 3.79-3.64 (m, 2 H), 3.49 (br. m, 1 H), 3.12 (s, 3 H), 2.64 (s, 3 H), 2.51-2.36 (m, 2 H), 2.23-1.98 (m, 4 H), 1.44 (s, 9 H).
(231) Synthesis of the Acid Ex.57:
(232) According to procedure H, the ester Ex.56 (2.25 g, 3.5 mmol) was hydrogenated in MeOH (120 mL)/THF (40 mL) in the presence of the catalyst (1.1 g) for 2 h to affordafter washing of the filtration residue with warm (50 C.) MeOH/THF 3:1the acid Ex.57 (1.9 g, 98%).
(233) Data of Ex.57: C.sub.26H.sub.35FN.sub.4O.sub.8 (550.6). HPLC LC-MS: (method 2): R.sub.t=1.54 (82), 551 ([M+H].sup.+).
Core 05: Synthesis of Ex.85 and Ex.86 (Scheme 9)
(234) Synthesis of the Mitsunobu Product 93
(235) Following procedure E.1.1, the reaction of phenol 56 (6.6 g, 15 mmol), alcohol 81 (13 g, 35 mmol), DEAD (40% in toluene, 32 mL, 69 mmol), and PPh.sub.3 (18 g, 69 mmol) in dry benzene (220 mL) afforded after FC(CH.sub.2Cl.sub.2/MeOH 100:0 to 94:6) the protected amino acid 93 (34.5 g, contaminated with triphenylphosphine oxide and diethyl hydrazine-1,2-dicarboxylate; acceptable for the use in the next step without further).
(236) Synthesis of the Amino Acid 94
(237) Following procedure E.2, the reaction of 93 (34.5 g, impure material), 1,3-dimethylbarbituric acid (17 g, 106 mmol) and Pd(PPh.sub.3).sub.4 (0.1 g) in EtOAc/CH.sub.2Cl.sub.2 (55:45, 350 mL) yielded after 3 h and after FC(CH.sub.2Cl.sub.2/EtOH 100:0 to 70:30 then CH.sub.2Cl.sub.2/MeOH 90:10 to 70:30) the amino acid 94 (5.6 g, 55% over the two steps, based on phenol 56).
(238) Data of 94: C.sub.33H.sub.43FN.sub.4O.sub.9 (658.7). HPLC (10% CH.sub.3CN): R.sub.t=3.79 (96). LC-MS (method 9c): R.sub.t=1.77, 659 ([M+H].sup.+).
(239) Synthesis of the Protected Macrolactam Ex.85
(240) According to procedure F.1.1, amino acid 94 (2.75 g, 4.2 mmol) in dry CH.sub.2Cl.sub.2 (35 mL) was treated with T3P (50% in EtOAc, 4.9 mL, 8.3 mmol) and i-Pr.sub.2NEt (2.9 mL, 17 mmol) in dry CH.sub.2Cl.sub.2 (355 mL) to yield after FC (EtOAc/MeOH 100:0 to 95:5) macrolactam Ex.85 (2.47 g, 92%).
(241) Data of Ex.85: C.sub.33H.sub.41FN.sub.4O.sub.8 (640.7). HPLC (30% CH.sub.3CN): R.sub.t=3.52 (96). LC-MS (method 9c): R.sub.t=2.06; 641 ([M+H].sup.+), 541 ([M+H-Boc].sup.+). .sup.1H-NMR (CDCl.sub.3): two isomers, ratio 85:15, 7.42-7.31 (m, 5 H), 7.08-6.77 (m, 3 H), 5.33 (d, J=8.3, 1 H), 5.23 (d, J=12.2, 1 H), 5.17 (d, J=12.1, 1 H), 4.84 (dd, J=2.9, 8.9, 1 H), 4.37-4.25 (m, 3 H), 4.11 (dd, J=4.2, 12.0, 1 H), 3.89 (t, J=8.3, 1 H), 3.80 (d, J=8.9, 1 H), 3.61 (d, J=17.1, 1 H), 3.16 (t, J=9.1, 1 H), 3.13 (s, 2.55 H, NCH.sub.3 of major isomer), 3.03 (s, 0.45 H, NCH.sub.3 of minor isomer), 2.98 (s, 2.55 H, NCH.sub.3 of major isomer), 2.87 (0.45 H, NCH.sub.3 of minor isomer), 2.64-2.41 (m, 2 H), 2.27-2.09 (m, 1 H), 1.98-1.83 (m, 2 H), 1.79-1.66 (m, 2 H), 1.45 (s, 7.65 H, Boc, major isomer), 1.35 (s, 1.35 H, Boc, minor isomer).
(242) Synthesis of the Acid Ex.86:
(243) According to procedure H, ester Ex.85 (2.0 g, 3.1 mmol) was hydrogenated in MeOH (120 mL)/THF (40 mL) in the presence of the catalyst (1 g) for 2 h to affordafter washing of the filtration residue with warm (50 C.) MeOH/TFH 3:1the acid Ex.86 (1.67 g, 97%).
(244) Data of Ex.86: C.sub.26H.sub.35FN.sub.4O.sub.8 (550.6). LC-MS: (method 3): R.sub.t=1.10 (83), 551 ([M+H].sup.+); 1.17 (15), 551 ([M+H].sup.+).
Core 06: Synthesis of Ex.104 and Ex.105 (Scheme 10)
(245) Synthesis of the Mitsunobu Product 95
(246) Following procedure E.1.2, the reaction of phenol 56 (13.1 g, 30.5 mmol), alcohol 82 (13.6 g, 36.6 mmol), and CMBP (14.7 g, 61 mmol) in dry toluene (500 mL) afforded after FC (hexane/EtOAc 50:50 to 30:70) the protected amino acid 95 (16 g, 67%).
(247) Synthesis of the Amino Acid 96
(248) Following procedure E.2, the reaction of 95 (16.0 g, 20 mmol), 1,3-dimethylbarbituric acid (8 g, 49 mmol) and Pd(PPh.sub.3).sub.4 (0.1 g) in EtOAc/CH.sub.2Cl.sub.2 (55:45, 220 mL) yielded after 3 h and after FC(CH.sub.2Cl.sub.2/EtOH 100:0 to 70:30 then CH.sub.2Cl.sub.2/MeOH 90:10 to 70:30) amino acid 96 (11 g, 81%).
(249) Data of 96: C.sub.33H.sub.43FN.sub.4O.sub.9 (658.7). LC-MS (method 2): R.sub.t=1.63 (97), 659 ([M+H].sup.+).
(250) Synthesis of the Protected Macrolactam Ex.104
(251) According to procedure F.1.1, amino acid 96 (4.0 g, 6.1 mmol) in dry CH.sub.2Cl.sub.2 (40 mL) was treated with T3P (50% in EtOAc, 7.2 mL, 12.1 mmol) and i-Pr.sub.2NEt (4.2 mL, 24.3 mmol) in dry CH.sub.2Cl.sub.2 (1160 mL) to give after FC(CH.sub.2Cl.sub.2/MeOH 100:0 to 95:5) macrolactam Ex.104 (2.32 g, 60%).
(252) Data of Ex.104: C.sub.33H.sub.41FN.sub.4O.sub.8 (640.7). LC-MS (method 7): R.sub.t=1.21 (47), 641 ([M+H].sup.+); 1.24 (53), 641 ([M+H]). .sup.1H-NMR (DMSO-d.sub.6): complex spectrum, mixture of isomers, 7.44-6.65 (m, 9 H), 5.32-5.05 (m, 2 H), 4.70-3.30 (several m, 9 H), 2.92 (s, NCH.sub.3 of major isomer), 2.84 (s, NCH.sub.3 of major isomer), 2.30-1.70 (several m, 6 H), 1.40, 1.38 (2 s, 9 H).
(253) Synthesis of the Acid Ex.105:
(254) According to procedure H, ester Ex.104 (2.15 g, 3.3 mmol) was hydrogenated in MeOH (215 mL) in the presence of the catalyst (1.07 g) for 4 h to afford acid Ex.105 (1.72 g, 93%).
(255) Data of Ex.105: C.sub.26H.sub.35FN.sub.4O.sub.8 (550.6). LC-MS: (method 7): R.sub.t=0.91 (45), 551 ([M+H].sup.+); 0.95 (38), 551 ([M+H].sup.+).
Core 07: Synthesis of Ex.115 and Ex.116 (Scheme 11)
(256) Synthesis of the Mitsunobu Product 97
(257) A mixture of the phenol 54 (6.42 g, 14.9 mmol), alcohol 22 (4.04 g, 13.5 mmol), and PPh.sub.3 (9.73 g, 37.1 mmol) was dried i.v. for 15 min. and dissolved in dry, degassed chloroform (130 mL). The solution was cooled to 0 C. A solution of ADDP (9.36 g, 37.1 mmol) in chloroform (20 mL) was slowly added. The mixture was stirred at room temperature for 3 h followed by the addition of more 22 (4.04 g, 13.5 mmol) and PPh.sub.3 (5.97 g, 22.8 mmol) in chloroform (20 mL). The mixture was cooled to 0 C. A solution of ADDP (5.74 g, 22.7 mmol) in chloroform (20 mL) was slowly added. The solution was stirred at room temperature for 16 h and concentrated. The residue was suspended in diethyl ether and filtered. The solid was washed with diethyl ether. The combined filtrate and washings were concentrated. FC (CH.sub.2Cl.sub.2/EtOAc 10:1) gave 97 (7.73 g, 73%).
(258) Synthesis of the Amino Acid 98
(259) Following procedure B.2, the reaction of 97 (7.72 g, 11 mmol), 1,3-dimethylbarbituric acid (4.1 g, 26.0 mmol) and Pd(PPh.sub.3).sub.4 (0.63 g) in EtOAc/CH.sub.2Cl.sub.2 (53:47, 190 mL) yielded after 2 h and after FC (EtOAc, then CH.sub.2Cl.sub.2/MeOH 95:5 to 90:10) amino acid 98 (4.31 g, 67%).
(260) Data of 98: C.sub.30H.sub.38FN.sub.3O.sub.8 (587.6). HPLC (10% CH.sub.3CN): R.sub.t=3.86 (84). LC-MS (method 9a): R.sub.t=1.76; 588 ([M+H].sup.+), 488 ([M+H-Boc].sup.+).
(261) Synthesis of the Alloc Protected Amino Acid 99
(262) Following procedure C.1, the reaction of the amino acid 98 (4.3 g, 7.3 mmol), allyl choroformate (0.86 mL, 8.0 mmol) and Na.sub.2CO.sub.3 (1.2 g, 11 mmol) in dioxane (62 mL) and H.sub.2O (60 mL) gave acid 99 (5.07 g, 100%).
(263) Synthesis of the Protected Diamide 100
(264) Following procedure C.2, the acid 99 (4.9 g, 7.3 mmol) was reacted with sarcosine allylester p-toluenesulfonate (46.p-TsOH, 2.6 g, 8.8 mmol), HOAt (1.5 g, 11 mmol), HATU (4.2 g, 11 mmol) and i-Pr.sub.2NEt (6.2 mL, 36 mmol) in DMF (75 mL) to afford the protected amino acid 100 (4.37 g, 76%).
(265) Data of 100: C.sub.40H.sub.51FN.sub.4O.sub.11 (782.8). HPLC (50% CH.sub.3CN): R.sub.t=3.56 (99). LC-MS (method 9a): R.sub.t=2.45; 783 ([M+H].sup.+), 683 ([M+H-Boc].sup.+).
(266) Synthesis of the Deprotected Amino Acid 101
(267) Following procedure C.3, the reaction of the protected amino acid 100 (4.36 g, 5.6 mmol), 1,3-dimethylbarbituric acid (2.1 g, 13 mmol) and Pd(PPh.sub.3).sub.4 (0.32 g) in EtOAc/CH.sub.2Cl.sub.2 (45:55, 106 mL) yielded amino acid 101 (3.46 g, 93%).
(268) Data of 101: C.sub.33H.sub.43FN.sub.4O.sub.9 (658.7). LC-MS (method 9b): R.sub.t=1.74; 659 ([M+H].sup.+), 559 ([M+H-Boc].sup.+).
(269) Synthesis of the Protected Macrolactam Ex.115
(270) According to procedure F.1.1, amino acid 101 (3.44 g, 5.2 mmol) in dry CH.sub.2Cl.sub.2 (50 mL) was treated with T3P (50% in EtOAc, 6.2 mL, 10 mmol) and i-Pr.sub.2NEt (3.6 mL, 21 mmol) in dry CH.sub.2Cl.sub.2 (470 mL) to give after FC(CH.sub.2Cl.sub.2/MeOH 95:5) macrolactam Ex.115 (2.95 g, 90%).
(271) Data of Ex.115: C.sub.33H.sub.41FN.sub.4O.sub.8 (640.7). HPLC (20% CH.sub.3CN): R.sub.t=4.05 (93). LC-MS (method 9c): R.sub.t=2.08; 641 ([M+H].sup.+). .sup.1H-NMR (DMSO-d.sub.6): complex spectrum, mixture of isomers, 7.38 (s, 5 H), 7.35-6.95 (several m, 2 H), 6.81-6.72 (several m, 0.4 H), 6.64 (dd, J=3.1, 8.2, 0.25 H), 6.39 (dd, J=3.2, 7.7, 0.25 H), 6.30 (dd, J=3.3, 8.2, 0.1 H), 5.37-4.99 (m, 3 H), 4.60-3.60 (several m, 9 H), 3.20-2.60 (several m and s, 8 H), 2.40-1.70 (several m, 4 H), 1.45, 1.43, 1.42, 1.38 (4 s, Boc).
(272) Synthesis of the Acid Ex.116:
(273) According to procedure H, the ester Ex.115 (1.2 g, 1.9 mmol) was hydrogenated in MeOH (120 mL) in the presence of the catalyst (0.6 g) for 2 h to afford the acid Ex.116 (1.02 g, 99%).
(274) Data of Ex.116: C.sub.26H.sub.35FN.sub.4O.sub.8 (550.6). HPLC (10% CH.sub.3CN): R.sub.t=3.47 (20), 3.55 (75). LC-MS: (method 9c): R.sub.t=1.53, 1.58; 551 ([M+H].sup.+).
Core 08: Synthesis of Ex.132 and Ex.133 (Scheme 12)
(275) Synthesis of the Mitsunobu Product 102
(276) Following procedure E.1.2, the reaction of phenol 56 (2.0 g, 4.7 mmol), alcohol 84 (2.08 g, 5.6 mmol), and CMBP (2.25 g, 9.3 mmol) in dry toluene (80 mL) afforded after 3 h and after FC (hexane/EtOAc 1:1 to 1:2) the protected amino acid 102 (2.06 g, 56%).
(277) Synthesis of the Amino Acid 103
(278) Following procedure E.2, the reaction of 102 (2.05 g, 2.6 mmol), 1,3-dimethylbarbituric acid (1.0 g, 6.3 mmol) and Pd(PPh.sub.3).sub.4 (0.15 g) in EtOAc/CH.sub.2Cl.sub.2 (55:45; 45 mL) yielded after 2 h and after FC (EtOAc, then CH.sub.2Cl.sub.2/MeOH 95:5 to 70:30) amino acid 103 (1.45 g, 85%).
(279) Data of 103: C.sub.33H.sub.43FN.sub.4O.sub.9 (658.7). HPLC (5% CH.sub.3CN): R.sub.t=4.04 (97). LC-MS (method 9c): R.sub.t=1.87, 659 ([M+H].sup.+).
(280) Synthesis of the Protected Macrolactam Ex.132
(281) According to procedure F.1.1, the amino acid 103 (1.44 g, 2.19 mmol) in dry CH.sub.2Cl.sub.2 (40 mL) was treated with T3P (50% in EtOAc, 2.6 mL, 4.37 mmol) and i-Pr.sub.2NEt (1.5 mL, 8.74 mmol) in dry CH.sub.2Cl.sub.2 (170 mL) to give after FC(CH.sub.2Cl.sub.2/MeOH 95:5) the macrolactam Ex.132 (1.36 g, 96%).
(282) Data of Ex.132: C.sub.33H.sub.41FN.sub.4O.sub.8 (640.7). LC-MS (method 2): R.sub.t=1.93 (100), 641 ([M+H].sup.+); LC-MS (method 9c): R.sub.t=2.12, 641 ([M+H].sup.+).
(283) .sup.1H-NMR (DMSO-d.sub.6): complex spectrum, mixture of isomers, 7.38 (s, 5 H), 7.35-6.99 (several m, 2 H), 6.85-6.73 (several m, 0.4 H), 6.65 (dd, J=3.1, 8.2, 0.25 H), 6.39 (dd, J=3.1, 7.9, 0.25 H), 6.30 (dd, J=3.3, 8.1, 0.1 H), 5.37-4.99 (m, 3 H), 4.6-3.6 (several m, 9 H), 3.2-2.6 (several m and s, 8 H), 2.4-1.7 (several m, 4 H), 1.45, 1.43, 1.41, 1.38 (4 s, Boc).
(284) Synthesis of the Acid Ex.133:
(285) According to procedure H, ester Ex.132 (1.13 g, 1.7 mmol) was hydrogenated in MeOH (110 mL) in the presence of the catalyst (0.56 g) for 4 h to afford acid Ex.133 (0.92 g, 94%).
(286) Data of Ex.133: C.sub.26H.sub.35FN.sub.4O.sub.8 (550.6). HPLC (5% CH.sub.3CN): R.sub.t=3.65 (27), 3.72 (71). LC-MS: (method 9c): R.sub.t=1.53, 551 ([M+H].sup.+); 1.57, 551 ([M+H].sup.+).
Core 09: Synthesis of Ex.142 and Ex.143 (Scheme 13)
(287) Synthesis of the Mitsunobu Product 104
(288) Following procedure E.1.2, the reaction of the phenol 54 (3.1 g, 7.2 mmol), alcohol 86 (3.34 g, 8.7 mmol), and CMBP (3.49 g, 14.4 mmol) in dry toluene (123 mL) afforded after 3 h and after FC (hexane/EtOAc 1:1 to 1:2) the protected amino acid 104 (4.11 g, 71%).
(289) Synthesis of the Amino Acid 105
(290) Following procedure E.2, the reaction of 104 (4.07 g, 5.1 mmol), 1,3-dimethylbarbituric acid (1.9 g, 12 mmol) and Pd(PPh.sub.3).sub.4 (0.3 g) in EtOAc/CH.sub.2Cl.sub.2 (45:55, 90 mL) yielded after 2 h and after FC (EtOAc, then CH.sub.2Cl.sub.2/MeOH 95:5 to 70:30) the amino acid 105 (3.19 g, 93%).
(291) Data of 105: C.sub.34H.sub.45FN.sub.4O.sub.9 (672.7). HPLC (5% CH.sub.3CN): R.sub.t=3.96 (88). LC-MS (method 9c): R.sub.t=1.83, 673 ([M+H].sup.+).
(292) Synthesis of the Protected Macrolactam Ex.142
(293) According to procedure F.1.1, amino acid 105 (2.4 g, 3.6 mmol) in dry CH.sub.2Cl.sub.2 (40 mL) was treated with T3P (50% in EtOAc, 4.2 mL, 7.1 mmol) and i-Pr.sub.2NEt (2.4 mL, 14.2 mmol) in dry CH.sub.2Cl.sub.2 (300 mL) to give after FC(CH.sub.2Cl.sub.2/MeOH 95:5) the macrolactam Ex.142 (1.92 g, 82%).
(294) Data of Ex.142: C.sub.34H.sub.43FN.sub.4O.sub.8 (654.7). HPLC (30% CH.sub.3CN): R.sub.t=3.50 (89). LC-MS (method 9b): R.sub.t=2.01; 655 ([M+H].sup.+), 599 ([M+H-tBu].sup.+), 555 ([M+H-Boc].sup.+). .sup.1H-NMR (DMSO-d.sub.6): complex spectrum, mixture of isomers, 7.41-7.38 (m, 5 H), 7.37-7.14 (m, 3 H), 6.80-6.67 (m, 1 H), 5.45-5.13 (m, 3 H), 4.60-3.30 (several m, 8 H), 3.10-2.50 (several m and s, 8 H), 2.50-1.80 (several m, 6 H), 1.39, 1.38, 1.36 (3 s, Boc).
(295) Synthesis of the Acid Ex.143:
(296) According to procedure H, ester Ex.142 (1.07 g, 1.6 mmol) was hydrogenated in MeOH (100 mL) in the presence of the catalyst (0.53 g) for 4 h to afford acid Ex.143 (0.92 g, 99%).
(297) Data of Ex.143: C.sub.27H.sub.37FN.sub.4O.sub.8 (564.6). LC-MS: (method 2): R.sub.t=1.54 (91), 565 ([M+H].sup.+).
Core 10: Synthesis of Ex.164 and Ex.165 (Scheme 14)
(298) Synthesis of the Mitsunobu Product 106
(299) Following procedure B.1.2, the reaction of phenol 63 (4.2 g, 9.8 mmol), alcohol 16 (4.4 g, 13 mmol), and CMBP (4.8 g, 20 mmol) in dry toluene (120 mL) afforded after 4 h and FC (hexane/EtOAc 50:50) the protected amino acid 106 (6.37 g, 86%).
(300) Synthesis of the Amino Acid 107
(301) Following procedure B.2, the reaction of 106 (1.18 g, 1.6 mmol), 1,3-dimethylbarbituric acid (0.6 g, 3.8 mmol) and Pd(PPh.sub.3).sub.4 (90 mg) in EtOAc/CH.sub.2Cl.sub.2 (60:40, 15 mL) yielded after 3 h and after FC(CH.sub.2Cl.sub.2/EtOH 100:0 to 80:20) the amino acid 107 (0.86 g, 87%).
(302) Data of 107: C.sub.31H.sub.44N.sub.4O.sub.8Si (628.8). LC-MS: (method 6): R.sub.t=1.08 (88), 629 ([M+H].sup.+).
(303) Synthesis of the Protected Macrolactam Ex.164
(304) According to procedure F.1.2, amino acid 107 (310 mg, 0.49 mmol) in dry DMF (5 mL) was treated with FDPP (379 mg, 0.99 mmol) in dry DMF (500 mL) to afford after FC (hexane/EtOAc/MeOH 50:50:0 to 0:95:5) the macrolactam Ex.164 (131 mg, 43%).
(305) Data of Ex.164: C.sub.31H.sub.42N.sub.4O.sub.7Si (610.8). LC-MS: (method 7): R.sub.t=1.34 (98), 611 ([M+H].sup.+). .sup.1H-NMR (DMSO-d.sub.6): 7.42-7.27 (m, 8 H), 6.98 (dd, J=1.4, 8.2, 1 H), 6.91 (d, J=7.5, 1 H), 6.84 (s, 1 H), 4.98 (s, 2 H), 4.50 (d, J=11.9, 1 H), 4.35-4.15 (m, 3 H), 4.06-3.96 (m, 4 H), 3.21 (m, 1 H), 3.10-2.95 (m, 2 H), 2.87 (s, 3 H), 2.30-1.80 (m, 4 H), 0.91 (t, J=8.3, 2 H), 0.00 (s, 9 H).
(306) Synthesis of the Amine Ex.165
(307) At 0 C., a solution of TBAF in THF (1 M, 3.9 mL, 3.9 mmol) was added to a solution of Ex.164 (1.2 g, 1.96 mmol) in THF (42 mL). The solution was allowed to stir at 0 C. to room temperature for 15 h, followed by the addition of more TBAF in THF (1 M, 1.18 mL, 1.18 mmol). Stirring was continued for 2 h. The solution was distributed between CH.sub.2Cl.sub.2 and H.sub.2O. The aqueous phase was repeatedly extracted with CH.sub.2Cl.sub.2. The combined organic phase was dried (Na.sub.2SO.sub.4), filtered and concentrated. FC (CH.sub.2Cl.sub.2/MeOH 100:0 to 90:10) afforded Ex.165 (0.76 g, 83%).
(308) Data of Ex.165:C.sub.25H.sub.30N.sub.4O.sub.5 (466.52). LC-MS: (method 4a): R.sub.t=1.49 (99), 467 ([M+H].sup.+).
Core 11: Synthesis of Ex.181 and Ex.182 (Scheme 15)
(309) Synthesis of the Mitsunobu Product 108
(310) Following procedure B.1.2, the reaction of phenol 65 (10.7 g, 24 mmol), alcohol 16 (10.0 g, 29 mmol), and CMBP (12.0 g, 49 mmol) in dry toluene (362 mL) afforded after FC (hexane/EtOAc 50:50 to 70:30) the protected amino acid 108 (14.55 g, 78%).
(311) Synthesis of the Amino Acid 109
(312) Following procedure B.2, the reaction of 108 (14.50 g, 19 mmol), 1,3-dimethylbarbituric acid (7.0 g, 47.0 mmol) and Pd(PPh.sub.3).sub.4 (0.1 g) in EtOAc/CH.sub.2Cl.sub.2 (55:45, 203 mL) yielded after 3 h and after FC(CH.sub.2Cl.sub.2/MeOH 99:1 to 90:10) the amino acid 109 (11.26 g, 92%).
(313) Data of 109: C.sub.32H.sub.46N.sub.4O.sub.8Si (642.8). LC-MS: (method 6): R.sub.t=1.13 (94), 643 ([M+H].sup.+).
(314) Synthesis of the Protected Macrolactam Ex.181
(315) According to procedure F.1.1, the amino acid 109 (4.0 g, 6.2 mmol) in dry CH.sub.2Cl.sub.2 (100 mL) was treated with T3P (50% in EtOAc, 7.4 mL, 12.4 mmol) and i-Pr.sub.2NEt (4.3 mL, 24.8 mmol) in dry CH.sub.2Cl.sub.2 (560 mL). Prior to aqueous workup, the CH.sub.2Cl.sub.2 was replaced by EtOAc. FC (hexane/EtOAc 50:50 to 0:100) afforded the macrolactam Ex.181 (2.11 g, 54%).
(316) Data of Ex.181: C.sub.32H.sub.44N.sub.4O.sub.7Si (624.8). LC-MS (method 7): R.sub.t=1.37 (99), 625 ([M+H].sup.+). .sup.1H-NMR (DMSO-d.sub.6): 7.46 (d, J=8.0, 1 H), 7.42 (d, J=7.2, 1 H), 7.34-7.23 (m, 6 H), 7.06 (d, J=8.2, 1 H), 6.82 (d, J=7.4, 1 H), 6.78 (s, 1 H), 5.02-4.86 (m, 3 H), 4.13 (t, J=8.5, 1 H), 4.06-3.67 (m, 7 H), 3.05 (br. m, 1 H), 2.88 (br. m, 1 H), 2.88 (s, 3 H), 2.15 (m, 2 H), 1.51 (br. m, 2 H), 1.33 (br. m, 1 H), 1.12 (br. m, 1 H), 0.91 (t-like m, J ca. 8.4, 2 H), 0.00 (s, 9 H).
(317) Synthesis of the Amine Ex.182
(318) According to procedure I.2, carbamate Ex.181 (844 mg, 1.3 mmol) in THF (34 mL) was treated with TBAF solution (4.1 mL) to afford after FC(CH.sub.2Cl.sub.2/MeOH 90:10) the amine Ex.182 (620 mg, 95%)
(319) Data of Ex.182: C.sub.26H.sub.32N.sub.4O.sub.5 (480.5). LC-MS: (method 2): R.sub.t=1.35 (99), 481 ([M+H].sup.+).
Core 12: Linear Synthesis of Ex.196 and Ex.197 (Scheme 16)
(320) Synthesis of the Mitsunobu Product 110
(321) Following procedure B.1.1, the reaction of phenol 59 (5.22 g, 12.6 mmol), alcohol 16 (5.2 g, 15.2 mmol), PPh.sub.3 (5.0 g, 19 mmol) in dry benzene (124 mL) and DEAD (40% in toluene, 7.0 mL, 15.2 mmol) in dry benzene (36 mL) afforded after FC (hexane/EtOAc 60:40 to 40:60) the protected amino acid 110 (8.3 g, 88%, contaminated with some triphenylphosphine oxide; acceptable for the use in the next stop without further purification).
(322) Synthesis of the Amino Acid 111
(323) Following procedure B.2, the reaction of 110 (4.15 g, 5.62 mmol), 1,3-dimethylbarbituric acid (2.19 g, 14.0 mmol) and Pd(PPh.sub.3).sub.4 (0.71 g) in EtOAc/CH.sub.2Cl.sub.2 1:1 (60 mL) yielded after 1 h and after FC(CH.sub.2Cl.sub.2/EtOH 95:5 to 90:10 then CH.sub.2Cl.sub.2/MeOH 90:10 to 70:30) amino acid 111 (2.75 g, 80%).
(324) Data of 111: C.sub.30H.sub.42N.sub.4O.sub.8Si (614.8). HPLC (10% CH.sub.3CN): R.sub.t=3.82 (99). LC-MS (method 9a): R.sub.t=1.81; 615 ([M+H].sup.+).
(325) Synthesis of the Alloc Protected Amino Acid 112
(326) Following procedure C.1, the reaction of the amino acid 111 (1.5 g, 2.4 mmol), allyl choroformate (0.29 mL, 2.68 mmol) and Na.sub.2CO.sub.3 (0.72 g, 6.83 mmol) in dioxane (40 mL) and H.sub.2O (40 mL) gave acid 112 (1.7 g, 100%).
(327) Synthesis of the Protected Amino Acid 113
(328) Following procedure C.2, the acid 112 (1.7 g, 2.4 mmol) was reacted with sarcosine allylester p-toluenesulfonate (46.p-TsOH, 0.88 g, 2.9 mmol), HOAt (0.5 g, 3.6 mmol), HATU (1.4 g, 3.6 mmol) and i-Pr.sub.2NEt (2.1 mL, 12 mmol) in DMF (25 mL) to afford the protected amino acid 113 (1.51 g, 75%).
(329) Data of 113: C.sub.40H.sub.55N.sub.5O.sub.22Si (809.9). HPLC (40% CH.sub.3CN): R.sub.t=4.43 (91). LC-MS (method 9c): R.sub.t=2.51, 810 ([M+H].sup.+).
(330) Deprotection to Amino Acid 114
(331) Following procedure C.3, the reaction of the protected amino acid 113 (1.5 g, 1.85 mmol), 1,3-dimethylbarbituric acid (0.72 g, 4.6 mmol) and Pd(PPh.sub.3).sub.4 (0.23 g) in EtOAc/CH.sub.2Cl.sub.2 (1:1, 25 mL) yielded amino acid 114 (1.05 g, 83%).
(332) Data of 114: C.sub.33H.sub.47N.sub.5O.sub.9Si (685.8). HPLC (10% CH.sub.3CN): R.sub.t=3.85 (95). LC-MS (method 9c): R.sub.t=1.78, 686 ([M+H].sup.+).
(333) Synthesis of the Protected Macrolactam Ex.196
(334) According to procedure F.1.2, amino acid 114 (1.0 g, 1.46 mmol) in dry DMF (20 mL) was treated with FDPP (1.12 g, 2.92 mmol) in dry DMF (130 mL) to yield after FC (EtOAc) the macrolactam Ex.196 (0.61 g, 63%).
(335) Data of Ex.196: C.sub.33H.sub.45N.sub.5O.sub.8Si (667.8). LC-MS (method 1a): R.sub.t=2.66 (100), 668 ([M+H].sup.+). LC-MS (method 9c): R.sub.t=2.12, 668 ([M+H].sup.+), 640. .sup.1H-NMR (CDCl.sub.3): 7.34-7.26 (m, 6 H), 7.17 (d, J=7.6, 1 H), 7.02 (s, 1 H), 6.91 (d, J=9.5, 1 H), 5.49 (d, J=9.5, 2 H), 5.10 (m, 1 H), 5.06 (s, 2 H), 4.39-4.13 (m, 5 H), 4.00-3.95 (m, 2 H), 3.65 (m, 1 H), 3.36 (br. s, 2 H), 3.14 (m, 2 H), 3.09 (s, 3 H), 2.74 (s, 3 H), 2.45 (m, 1 H), 2.08 (m, 1 H), 0.98 (m, 2 H), 0.00 (s, 9 H). .sup.1H-NMR (DMSO-d.sub.6): 7.98 (d, J=9.9, 1 H), 7.52 (d, J=7.9, 1 H), 7.36-7.27 (m, 6 H), 7.18 (s, 1 H), 7.06 (dd, J=1.8, 8.1, 1 H), 6.83 (d, J=7.5, 1 H), 5.12 (d, J=12.5, 1 H), 5.04 (d, J=12.5, 1 H), 4.87 (d, J=8.8, 1 H), 4.25-3.89 (m, 8 H), 3.71-3.66 (m, 2 H), 3.20 (m, 1 H), 3.02 (m, 1 H), 2.97 (s, 3 H), 2.65 (s, 3 H), 2.20 (m, 1 H), 2.09 (m, 1 H), 0.92 (t, J=8.2, 2 H), 0.00 (s, 9 H).
(336) Synthesis of the Amine Ex.197
(337) According to procedure I.1, carbamate Ex.196 (120 mg, 0.18 mmol) in dioxane (3 mL) was treated with 4 M HCl-dioxane (3 mL) to afford Ex.197.HCl (59 mg, 58%).
(338) Data of Ex.197.HCl: C.sub.27R.sub.33N.sub.5O.sub.6.HCl (523.5, free base). HPLC (5% CH.sub.3CN): R.sub.t=3.05 (83). LC-MS (method 9c): R.sub.t=1.12, 524 ([M+H].sup.+). .sup.1H-NMR (DMSO-d.sub.6): 8.53 (br. s, NH.sub.3), 8.03 (d, =9.9, 1 H), 7.41-7.31 (m, 7 H), 7.15 (m, 1 H), 6.85 (d, J=7.5, 1 H), 5.14 (d, J=12.5, 1 H), 5.04 (d, J=12.5, 1 H), 4.86 (dd, J ca. 2.2, 11.0, 1 H), 4.42-4.13 (m, 2 H), 4.05 (t, J=8.5, 1 H), 3.96 (d, J=17.8, 1 H), 3.85-3.75 (m, 2 H), 3.65 (br. m, 1 H), ca. 3.3-3.1 (m, 3 H, partially superimposed by the H.sub.2O signal), 2.97 (s, 3 H), 2.67 (s, 3 H), 2.42 (m, 1 H), 2.18 (br. q, J ca. 11.1, 1 H).
Core 12: Convergent Synthesis of Ex.197 and Ex.198 (Scheme 17)
(339) Synthesis of the Mitsunobu Product 115
(340) Following procedure E.1.1, phenol 59 (4.6 g, 11 mmol) was treated for 40 h with alcohol 81 (5.0 g, 13 mmol), DEAD (40% in toluene, 6.1 mL, 13 mmol) and PPh.sub.3 (4.4 g, 17 mmol) in dry benzene (150 mL). After 2 h and after 18 h, more PPh.sub.3 (1.82 g, 6.9 mmol), alcohol 81 (2.04 g, 5.5 mmol) in benzene (50 mL), and DEAD (40% in toluene, 2.55 mL, 5.6 mmol) in benzene (13 mL) were added. FC (hexane/EtOAc 50:50 to 90:10) afforded the protected amino acid 115.1 (2.5 g, 29%).
(341) Following procedure E.1.2, the reaction of phenol 59 (2.9 g, 7.0 mmol), alcohol 81, (5.7 g, 15 mmol) and CMBP (5.1 g, 21 mmol) in dry toluene (121 mL) afforded after FC (hexane/EtOAc 20:80 to 90:10) the protected amino acid 115.2 (2.92 g, 54%).
(342) Synthesis of the Amino Acid 116
(343) Following procedure E.2, the reaction of 115.1 (3.17 g, 4.14 mmol), 1,3-dimethylbarbituric acid (1.62 g, 10.3 mmol) and Pd(PPh.sub.3).sub.4 (0.53 g) in EtOAc/CH.sub.2Cl.sub.2 (1:1, 46 mL) yielded after 1 h and after FC(CH.sub.2Cl.sub.2/MeOH 90:10 to 70:30) the amino acid 116.1 (1.86 g, 70%).
(344) Data of 116.1: C.sub.32H.sub.43N.sub.5O.sub.9 (641.7). HPLC (5% CH.sub.3CN): R.sub.t=3.65 (100). LC-MS (method 9c): R.sub.t=1.60, 642 ([M+H].sup.+).
(345) Following procedure E.2, the reaction of 115.2 (2.9 g, 3.8 mmol), 1,3-dimethylbarbituric acid (1.5 g, 9.5 mmol) and Pd(PPh.sub.3).sub.4 (0.48 g) in EtOAc/CH.sub.2Cl.sub.2 (1:1, 46 mL) yielded after 1 h and after FC(CH.sub.2Cl.sub.2/MeOH 90:10 to 70:30) the amino acid 116.2 (2.0 g, 83%).
(346) Data of 116.2: C.sub.32H.sub.43N.sub.5O.sub.9 (641.7). HPLC (5% CH.sub.3CN): R.sub.t=3.73 (98). LC-MS (method 9c): R.sub.t=1.61, 642 ([M+H].sup.+).
(347) Synthesis of the Protected Macrolactam Ex.198
(348) According to procedure F.1.1, the amino acid 116.1 (1.0 g, 1.6 mmol) in dry CH.sub.2Cl.sub.2 (200 mL) was treated with T3P (50% in EtOAc, 1.8 mL, 3.1 mmol) and i-Pr.sub.2NEt (1.1 mL, 6.2 mmol) in dry CH.sub.2Cl.sub.2 (1400 mL) to afford after FC (EtOAc/MeOH 95:5 to 80:20) the macrolactam Ex.198 (containing 15% of the epimer Ex.231; 0.38 g, 39%).
(349) Data of Ex.198: C.sub.32H.sub.41N.sub.5O.sub.8 (623.7). LC-MS: (method 2): R.sub.t=1.78 (84), 624 ([M+H].sup.+); 1.82 (15). LC-MS (method 9c): R.sub.t=1.87, 624 ([M+H].sup.+).
(350) .sup.1H-NMR (CDCl.sub.3): 7.42-7.25 (m, 7 H), 7.07 (s, 1 H), 7.00 (d, J=8.2, 1 H), 5.59 (d, J=9.5, 1 H), 5.38 (br. d, J ca 7.9, 1 H), 5.18 (dd, J=2.5, 12.2, 1 H), 5.13 (s, 2 H), 4.43-4.01 (m, 5 H), 3.73 (m, 1 H), 3.47 (d, J=17.7, 1 H), 3.33 (d, J=17.7, 1 H), 3.20-3.11 (m, 2 H), 3.17 (s, 3 H), 2.81 (s, 3 H), 2.50 (m, 1 H), 2.15 (m, 1 H), 1.51 (s, Boc, major isomer), 1.45 (s, Boc, minor isomer); .sup.1H-NMR (DMSO-d.sub.6): 7.97 (d, J=10.3, 1 H), 7.41-7.30 (m, 7 H), 7.18 (s, 1 H), 7.09 (d, J=8.2, 1 H), 6.85 (J=7.6, 1 H), 5.12 (d, J=12.5, 1 H), 5.05 (d, J=12.6, 1 H), 4.89 (J=9.6, 1 H), 4.30-3.55 (m, 6 H), 3.40 (2 H, superimposed by H.sub.2O signal), 3.25-3.00 (m, 2 H), 2.99 (s, 3 H), 2.65 (s, 3 H), 2.22 (m, 1 H), 2.05 (br. q, 1 H), 1.41, (s, 9 H).
(351) According to procedure F.1.1, amino acid 116.2 (0.85 g, 1.3 mmol) in dry CH.sub.2Cl.sub.2 (170 mL) was treated with T3P (50% in EtOAc, 1.56 mL, 2.6 mmol) and i-Pr.sub.2NEt (0.91 mL, 5.3 mmol) in dry CH.sub.2Cl.sub.2 (1190 mL) to afford after FC (EtOAc/MeOH 95:5 to 80:20) the macrolactam Ex.198 and its epimer Ex.231 (ca 1:1 mixture; 0.61 g, 73%).
(352) Data of the mixture Ex.198/Ex.231: C.sub.32H.sub.41N.sub.5O.sub.8 (623.7). LC-MS: (method 2): R.sub.t=1.78 (44), 624 ([M+H].sup.+); 1.82 (56), 624 ([M+H].sup.+). .sup.1H-NMR (CDCl.sub.3): complex spectrum, mixture of epimers, 7.41-7.20 (m, 6 H), 7.07-6.92 (m, 3 H) 5.8-4.8 (several m, 5 H), 4.3-3.0 (several m, 10 H), 3.16 (s, NCH.sub.3), 2.81 (s, NCH.sub.3), 2.58-2.45 (m, 1 H), 2.19-2.03 (m, 1 H), 1.51, 1.41 (2 s, 9 H)
(353) Synthesis of the Amine Ex.197
(354) According to procedure J, carbamate Ex.198/Ex.231 (ca. 85:15, 749 mg, 1.2 mmol) in dioxane (7.5 mL) was treated with 4 M HCl-dioxane (15 mL) to afford Ex.197.HCl/Ex.232.HCl (607 mg, 90%). Data of Ex.197.HCl/Ex.232.HCl: C.sub.27H.sub.33N.sub.5O.sub.6.HCl (523.5, free base). LC-MS (method 2): R.sub.t=1.26 (75), 1.33 (14); 524 ([M+H].sup.+).
(355) .sup.1H-NMR (DMSO-d.sub.6), major component Ex.197HCl: spectrum identical with the one described above for compound Ex.197HCl (cf. Scheme 16).
(356) According to procedure J, carbamate Ex.198/Ex.231 (ca. 1:1, 1.32 g, 2.12 mmol) in dioxane (13 mL) was treated with 4 M HC1-dioxane (26 mL) to afford after separation of the isomers by preparative RP-HPLC (method 1) Ex.197 TFA (460 mg, 34%) and Ex.232 TFA (470 mg, 35%).
(357) Data of Ex.197 TFA: C.sub.27H.sub.33N.sub.5O.sub.6.C.sub.2HF.sub.3O.sub.2 (523.5, free base). LC-MS (method 2): R.sub.t=1.25 (99), 524 ([M+H].sup.+). LC-MS (method 7): R.sub.t=0.74 (97), 524 ([M+H].sup.+). .sup.1H-NMR (DMSO-d.sub.6): 8.34 (br. s, NH.sub.3.sup.+), 8.07 (d, J=9.9, 1 H), 7.43-7.33 (m, 6 H), 7.20 (s, 1 H), 7.10 (dd, J=1.5, 8.2, 1 H), 6.87 (d, J=7.4, 1 H), 5.17 (d, J=12.5, 1 H), 5.05 (d, J=12.5, 1 H), 4.87 (br. dd, 1 H), 4.27-4.16 (m, 2 H), 4.06 (t, J=8.6, 1 H), 4.01-3.91 (m, 2 H), 3.82 (t-like dd, J ca. 8.1, 1 H), 3.70 (br. m, 1 H), 3.35-3.20 (m, 3 H), 2.98 (s, 3 H), 2.70 (s, 3 H), 2.49 (m, 1 H), 2.18 (br. q, J ca 11.0, 1 H).
(358) Data of Ex.232 TFA: See below; Core 14.
Core 13: Synthesis of Ex.215 and Ex.216 (Scheme 18)
(359) Synthesis of the Mitsunobu Product 117
(360) Following procedure B.1.1, the reaction of phenol 59 (2.1 g, 5.1 mmol), alcohol 20 (2.1 g, 6.1 mmol), PPh.sub.3 (2.0 g, 7.6 mmol) in dry benzene (50 mL) and DEAD (40% in toluene, 2.8 mL, 6.1 mmol) in dry benzene (14 mL) afforded, after further addition of PPh.sub.3 (0.84 g, 3.2 mmol), alcohol 20 (0.88 g, 2.6 mmol) in benzene (21 mL) and DEAD (40% in toluene, 1.2 mL, 2.6 mmol) in benzene (6 mL) and after FC (hexane/EtOAc 50:50) the protected amino acid 117 (3.8 g, 100%).
(361) Synthesis of the Amino Acid 118
(362) Following procedure B.2, the reaction of 117 (7.63 g, 10.3 mmol), 1,3-dimethylbarbituric acid (4.03 g, 25.8 mmol) and Pd(PPh.sub.3).sub.4 (1.31 g) in EtOAc/CH.sub.2Cl.sub.2 (1:1, 110 mL) yielded after 1 h and after FC(CH.sub.2Cl.sub.2/MeOH 95:5 to 70:30) the amino acid 118 (3.48 g, 60%).
(363) Data of 118: C.sub.30H.sub.42N.sub.4O.sub.8Si (614.8). HPLC (10% CH.sub.3CN): R.sub.t=3.88 (100). LC-MS (method 9a): R.sub.t=1.80, 615 ([M+H].sup.+).
(364) Synthesis of the Alloc Protected Amino Acid 119
(365) Following procedure C.1, the reaction of the amino acid 118 (3.36 g, 5.5 mmol), allyl choroformate (0.64 mL, 6.0 mmol) and Na.sub.2CO.sub.3 (0.87 g, 8.2 mmol) in dioxane (51 mL) and H.sub.2O (51 mL) gave the acid 119 (3.51 g, 92%).
(366) Synthesis of the Protected Amino Acid 120
(367) Following procedure C.2, acid 119 (3.47 g, 5.0 mmol) was reacted with sarcosine allylester p-toluenesulfonate (46.p-TsOH, 1.8 g, 6.0 mmol), HOAt (1.0 g, 7.4 mmol), HATU (2.8 g, 7.4 mmol) and i-Pr.sub.2NEt (4.2 mL, 25 mmol) in DMF (108 mL) to afford the protected amino acid 120 (3.52 g, 88%).
(368) Data of 120: C.sub.40H.sub.55N.sub.5O.sub.22Si (809.9). LC-MS: (method 4b): R.sub.t=2.51 (95), 810 ([M+H].sup.+)
(369) Deprotection to Amino Acid 121
(370) Following procedure C.3, the reaction of the protected amino acid 120 (3.49 g, 4.31 mmol), 1,3-dimethylbarbituric acid (1.68 g, 10.8 mmol) and Pd(PPh.sub.3).sub.4 (0.55 g) in EtOAc/CH.sub.2Cl.sub.2 (1:1; 50 mL) yielded the amino acid 121 (2.72 g, 92%).
(371) Data of 121: C.sub.33H.sub.47N.sub.5O.sub.9Si (685.8). LC-MS: (method 4b): R.sub.t=1.84 (94), 686 ([M+H].sup.+)
(372) Synthesis of the Protected Macrolactam Ex.215
(373) According to procedure F.1.2, amino acid 121 (1.33 g, 1.94 mmol) in dry DMF (27 mL) was treated with FDPP (1.49 g, 3.88 mmol) in dry DMF (164 mL) to yield after FC (EtOAc/MeOH 95:5) macrolactam Ex.215 (0.89 g, 68%).
(374) Data of Ex.215: C.sub.33H.sub.45N.sub.5O.sub.8Si (667.8). LC-MS: (method 1b): R.sub.t=2.60 (99), 668 ([M+H].sup.+). LC-MS: (method 9c): R.sub.t=2.14, 668 ([M+H].sup.+). .sup.1H-NMR (DMSO-d.sub.6): 7.94 (d, J=9.8, 1 H), 7.39-7.27 (m, 7 H), 7.11 (s, 1 H), 6.97 (dd, J=1.5, 8.2, 1 H), 6.82 (d, J=7.5, 1 H), 5.05 (s, 2 H), 4.83 (br. d, 1 H), 4.25 (br. m, 1 H), 4.17-3.96 (m, 5 H), 3.73 (br. q, J ca. 16.8, 2 H), 3.47 (m, 1 H), 3.33 (m, 1 H), 3.19 (m, 2 H), 2.96 (s, 3 H), 2.67 (s, 3 H), 2.20 (m, 1 H), 2.00 (m, 1 H), 0.91 (t, J=8.4, 2 H), 0.00 (s, 9 H).
(375) Synthesis of the Amine Ex.216
(376) According to procedure I.1, carbamate Ex.215 (881 mg, 1.3 mmol) in dioxane (16 mL) was treated with 4 M HCl-dioxane (16 mL) to afford Ex.216.HCl (666 mg, 90%).
(377) Data of Ex.216.HCl: C.sub.27H.sub.33N.sub.5O.sub.6.HCl (523.5, free base). HPLC (5% CH.sub.3CN): R.sub.t=3.11 (91). LC-MS (method 9c): R.sub.t=1.19, 524 ([M+H].sup.+).
Core 14: Synthesis of Ex.231 and Ex.232 (Scheme 19)
(378) Synthesis of the Mitsunobu Product 122
(379) A mixture of phenol 61 (4.6 g, 11.2 mmol) and PPh.sub.3 (5.27 g, 20.1 mmol) was dissolved in benzene. The solution was concentrated and the residue was dried i.v. for 20 min. A solution of the alcohol 81, (7.46 g, 20.1 mmol) in dry, degassed benzene (120 mL) was added. The resulting mixture was cooled to 0 C. DEAD (40% in toluene, 11.5 mL, 25.1 mmol) in benzene (10 mL) was slowly added. The solution was stirred at room temperature for 16 h. More PPh.sub.3 (1.46 g, 5.6 mmol), alcohol 81 (1.04 g, 2.8 mmol) and at 0 C., a solution of DEAD (40% in toluene, 2.6 mL, 5.7 mmol) in benzene (2 mL) were added and stirring at room temperature was continued for 7 h. More PPh.sub.3 (1.46 g, 5.6 mmol), alcohol 81 (1.04 g, 2.8 mmol), and at 0 C., a solution of DEAD (40% in toluene, 2.6 mL, 5.7 mmol) in benzene (2 mL) were added. Stirring at room temperature was continued for 16 h. The mixture was concentrated. FC (hexane/EtOAc 30:70 to 0:100) afforded 122 (12.8 g, contaminated with ca 40% triphenylphosphinoxide, yield ca 90%). The material was used for the next step without further purification)
(380) Synthesis of the Amino Acid 123
(381) Following procedure E.2, the reaction of the protected amino acid 122 (contaminated with ca 40% of triphenylphosphine oxide, 12.8 g, ca 10 mmol), 1,3-dimethylbarbituric acid (3.91 g, 25.1 mmol) and Pd(PPh.sub.3).sub.4 (1.27 g) in EtOAc/CH.sub.2Cl.sub.2 (1:1, 120 mL) yielded after 1 h and after FC(CH.sub.2Cl.sub.2/MeOH 100:0 to 70:30 then CHCl.sub.3/MeOH 70:30) the amino acid 123 (2.80 g, 44%).
(382) Data of 123: C.sub.32H.sub.43N.sub.5O.sub.9 (641.7). LC-MS: (method 2): R.sub.t=1.56 (94), 642 ([M+H].sup.+).
(383) Synthesis of the Protected Macrolactam Ex.231
(384) According to procedure F.1.2, amino acid 123 (3.29 g, 5.13 mmol) in dry DMF (150 mL) was added within 4 h at 60 C. to FDPP (3.94 g, 10.3 mmol) in dry DMF (4980 mL) to afford after 16 h at 60 C. and after FC (EtOAc/MeOH 100:0 to 95:5) the macrolactam Ex.231 (contained ca 15% of its epimer Ex.198; 2.5 g, 78%).
(385) Data of Ex.231: C.sub.32H.sub.41N.sub.5O.sub.8 (623.7). LC-MS: (method 2): R.sub.t=1.78 (12), 1.82 (83), 624 ([M+H].sup.+). LC-MS: (method 7): R.sub.t=1.16 (18), 624 ([M+H].sup.+); 1.18 (80), 624 ([M+H].sup.+). .sup.1H-NMR (CDCl.sub.3): complex spectrum, two epimers; 7.38-7.22 (m, 6H), 7.06-6.90 (m, 3 H), 5.80-4.80 (several m, 4 H), 5.08, 5.12 (2s, 2 H), 4.43-2.80 (several br. m, 15 H), 2.51 (m, 1 H), 2.19-2.03 (m, 1 H), 1.50, 1.42 (2 s, 9 H).
(386) Synthesis of the Amine Ex.232
(387) According to procedure J, carbamate Ex.231 (containing 15% of the epimer Ex.198; 1.42 g, 2.3 mmol) in dioxane (30 mL) was treated with 4 M HCl-dioxane (45 mL) to afford after preparative RP-HPLC (method 1) Ex.232 TFA (1.10 g, 71%) and Ex.197 TFA (0.27 g, 17%).
(388) Data of Ex.232 TFA: C.sub.27H.sub.33N.sub.5O.sub.6.C.sub.2HF.sub.3O.sub.2 (523.5, free base). LC-MS (method 2): R.sub.t=1.32 (99), 524 ([M+H].sup.+). .sup.1H-NMR (DMSO-d.sub.6): complex spectrum, mixture of isomers; 8.40 (br. s), 8.20 (br. s), 7.84 (d, J=7.1), 7.50-6.80 (several m), 5.25-3.40 (several m, partially superimposed by the H.sub.2O signal), 3.30-2.80 (m), 3.04 (s, NCH.sub.3), 2.98 (s, NCH.sub.3), 2.67 (s, NCH.sub.3), 2.64 (s, NCH.sub.3), 2.6-1.9 (several m).
(389) Data of Ex.197 TFA: See above; Core 12.
Core 15 and Core 16
Synthesis of Ex.238 and Ex.239 (Scheme 20)
(390) Synthesis of the Mitsunobu Product 124
(391) Following procedure E.1.1, phenol 77 (1.63 g, 8.5 mmol), alcohol 85 (5.72 g, 12.8 mmol) and PPh.sub.3 (4.02 g, 15.3 mmol) in dry benzene (80 mL) were treated with DEAD (40% in toluene, 8.79 mL, 19.2 mmol) for 20 h. Purification by FC (hexane/EtOAc 20:80 to 100:0) then (hexane/EtOAc 50:50 to 20:80) afforded the protected amino acid 124 (1.96 g, 37%).
(392) Synthesis of the Macrocycle Ex.238
(393) Dichloro-[1,3-bis(mesityl)-2-imidazoldinylidene]-(3-phenyl-1H-inden-1-ylidene) (tricyclohexylphosphine)ruthenium (II) (Umicore M2 catalyst; 88 mg) was added to a solution of 124 (1160 mg, 1.29 mmol) in dry, degassed CH.sub.2Cl.sub.2 (170 mL). The solution was stirred in a sealed tube at 40 C. for 68 h, followed by 45 h at room temperature. During this period further equal portions of catalyst (in total 350 mg) were added after 20 h, 28 h, 44 h, and 52 h. The solution was concentrated. FC (hexane/EtOAc 70:30 to 0:100) gave Ex.238 (350 mg, 46%, mixture of two isomers, ratio>9:1, acceptable for the use in the next step). An analytical sample (69 mg) was further purified by preparative RP-HPLC (method 2) to afford pure Ex.238 (major isomer; 45 mg).
(394) Data of Ex.238 (major isomer): C.sub.32H.sub.40N.sub.4O.sub.7 (592.6). LC-MS: (method 4a): R.sub.t=2.23 (92), 593 ([M+H].sup.+). .sup.1H-NMR (CDCl.sub.3): 7.62-7.31 (m, 6 H), 7.07 (d, J=7.6, 1 H), 6.99 (dd, J=2.0, 7.9, 1 H), 6.85 (s, 1 H), 5.69-5.61 (m, 2 H), 5.48 (d, J=8.2, 1 H), 5.21 (m, 1 H), 5.10 (s, 2 H), 4.76 (d, J=10.1, 1 H), 4.54 (dt, J=3.5, 7.9, 1 H), 4.41-4.25 (m, 2 H), 4.13 (d, J=10.7, 1 H), 3.97 (m, 1 H), 3.62 (m, 2 H), 3.48 (m, 1 H), 3.10 (s, 3 H), 2.73 (m, 1 H), 2.60-2.45 (m, 2 H), 2.02 (m, 1 H), 1.46 (s, 9 H).
(395) Synthesis of Amine Ex.239
(396) A solution of Ex.238 (430 mg, 0.73 mmol) in MeOH/THF 1:3, 36 mL) was hydrogenated for 3.5 h at room temperature and at normal pressure in the presence of palladium hydroxide on activated charcoal (moistened with 50% H.sub.2O; 215 mg). The mixture was filtered through a pad of celite. The filtrate was concentrated to give Ex.239 (355 mg, quantitative; used in the next step without further purification).
(397) An analytical sample (68 mg) was purified by preparative RP-HPLC (method 2) to afford pure Ex.239 (37 mg).
(398) Data of Ex.239: C.sub.24H.sub.36N.sub.4O.sub.5 (460.6): LC-MS (method 7): R.sub.t=0.88 (97), 461 ([M+H].sup.+). .sup.1H-NMR (DMSO-d.sub.6): 7.36 (t, J=7.8, 1 H), 7.25 (d, J=6.1, 1 H), 7.03 (dd, J=1.6, 8.2, 1 H), 6.88-6.65 (m, 2 H), 4.51 (d, J=8.3, 1 H), 4.18 (t, J=10.3, 2 H), 4.09 (br. s, 1 H), 3.96 (br. m, 2 H), 3.19-2.72 (m, 3 H), 2.92 (s, 3 H), 2.34 (m, 2 H), 2.05 (br. q, 1 H), 1.82 (m, 1 H), 1.60-0.85 (m, 5 H), 1.40 (s, 9 H), 0.82 (m, 1 H).
Core 17: Synthesis of Ex.248 and Ex.249 (Scheme 21)
(399) Synthesis of the Mitsunobu Product 125
(400) Following procedure E.1.1, phenol 68 (6.0 g, 14.6 mmol), alcohol 82 (9.75 g, 26.2 mmol), and PPh.sub.3 (6.88 g, 26.2 mmol) were treated in dry benzene (160 mL) with DEAD (40% in toluene, 15 mL, 32.8 mmol) for 40 h. After 18 h and after 25 h, more PPh.sub.3 (1.27 g, 4.8 mmol) and DEAD (40% in toluene, 2.23 mL, 4.9 mmol) in benzene (2 mL) were added. FC (hexane/EtOAc 30:70 to 20:80) afforded the protected amino acid 125 (16.85 g, contaminated with ca 40% triphenylphosphinoxide, yield ca 85%). The material was used for the next step without further purification)
(401) Synthesis of the Amino Acid 126
(402) Following procedure E.2, the reaction of 125 (16.8 g, contaminated with ca 40% of triphenylphosphine oxide, ca. 12 mmol), 1,3-dimethylbarbituric acid (4.80 g, 30.8 mmol) and Pd(PPh.sub.3).sub.4 (1.56 g) in EtOAc/CH.sub.2Cl.sub.2 (1:1, 170 mL) yielded after 1 h and after FC(CH.sub.2Cl.sub.2/MeOH 0:100 to 70:30, then CHCl.sub.3/MeOH 70:30) amino acid 126 (4.15 g, ca. 52%).
(403) Data of 126: C.sub.33H.sub.44N.sub.4O.sub.9 (640.7). HPLC (10% CH.sub.3CN): R.sub.t=3.67 (69). LC-MS (method 9c): R.sub.t=1.75, 641 ([M+H].sup.+).
(404) Synthesis of the Protected Macrolactam Ex.248
(405) According to procedure F.1.1, amino acid 126 (4.55 g, 7.1 mmol) in dry CH.sub.2Cl.sub.2 (120 mL) was added within 3 h to T3P (50% in EtOAc, 8.37 ml, 14.2 mmol) and i-Pr.sub.2NEt (4.83 ml, 28.4 mmol) in dry CH.sub.2Cl.sub.2 (6660 mL). Prior to aqueous workup, CH.sub.2Cl.sub.2 was replaced with EtOAc. FC(CH.sub.2Cl.sub.2/MeOH 100:0 to 95:5) yielded the macrolactam Ex.248 (2.38 g, 54%).
(406) Data of Ex.248: C.sub.33H.sub.42N.sub.4O.sub.8 (622.7). LC-MS: (method 2): R.sub.t=1.83 (100), 623 ([M+H].sup.+). LC-MS: (method 9c): R.sub.t=1.97, 623 ([M+H].sup.+). .sup.1H-NMR (DMSO-d.sub.6): 7.45-7.34 (m, 5 H), 7.15-6.78 (m, 5 H), 5.25 (s, 2 H), 5.08 (d, J=12.8, 1 H), 4.62 (d, J=13.5, 2 H), 4.29 (m, 1 H), 4.09 (d, J=7.3, 1 H), 3.89 (d, J=12.4, 1 H), 3.54 (br. t, 1 H), 3.27 (m, 1 H), 3.07 (s, 3 H), 2.80 (m, 1 H), 2.71 (s, 3 H), 2.28-2.06 (m, 4 H), 1.94 (m, 1 H), 1.71 (m, 1 H), 1.39 (s, 9 H).
(407) Synthesis of the Acid Ex.249:
(408) According to procedure H, the ester Ex.248 (2.16 g, 3.5 mmol) was hydrogenated in MeOH (130 mL)/THF (40 mL) in the presence of the catalyst (1.09 g) for 2.5 h to afford the acid Ex.249 (1.83 g, 99%).
(409) Data of Ex.249: C.sub.26H.sub.36N.sub.4O.sub.8 (532.6). LC-MS: (method 2): R.sub.t=1.42 (95), 533 ([M+H].sup.+).
Core 18: Synthesis of Ex.272, Ex.273, and Ex.274 (Scheme 22)
(410) Synthesis of the Mitsunobu Product 127
(411) Following procedure E.1.1, the reaction of phenol 71 (6.47 g, 15.7 mmol), the alcohol 81 (10.5 g, 28.2 mmol), DEAD (40% in toluene, 26 mL, 56.3 mmol), and PPh.sub.3 (14.8 g, 56.3 mmol) in dry benzene (380 mL) afforded after 2 h at room temperature and after aqueous workup (EtOAc, sat. aq. Na.sub.2CO.sub.3 soln, sat. aq. NaCl soln), drying (Na.sub.2SO.sub.4), concentration of the organic layer and FC (hexane/EtOAc 30:70, 0:100, then CH.sub.2Cl.sub.2/MeOH 90:10) the protected amino acid 127 (12.0 g, 99%).
(412) Synthesis of the Amino Acid 128
(413) Following procedure E.2, the reaction of 127 (12.0 g, 16 mmol), 1,3-dimethylbarbituric acid (5.9 g, 38.0 mmol) and Pd(PPh.sub.3).sub.4 (0.9 g) in EtOAc/CH.sub.2Cl.sub.2 (55:45, 275 mL) yielded after 2 h and after FC (EtOAc, then CH.sub.2Cl.sub.2/MeOH 90:10 to 60:40) the amino acid 128 (9.05 g, 90%).
(414) Data of 128: C.sub.31H.sub.42N.sub.6O.sub.9 (642.7). LC-MS: (method 7): R.sub.t=0.90 (94), 643 ([M+H].sup.+).
(415) Synthesis of the Protected Macrolactam Ex.272
(416) According to procedure F.1.1, the amino acid 128 (5.04 g, 7.8 mmol) in dry CH.sub.2Cl.sub.2 (100 mL) was treated with T3P (50% in EtOAc, 9.2 mL, 16 mmol) and i-Pr.sub.2NEt (5.4 mL, 31 mmol) in dry CH.sub.2Cl.sub.2 (700 mL) to afford after FC(CH.sub.2Cl.sub.2/MeOH 39:1 to 19:1) the epimeric macrolactams Ex.272 (1.90 g, 38%).
(417) Data of Ex.272: C.sub.34H.sub.40N.sub.6O.sub.8 (624.7). LC-MS: (method 2): R.sub.t=1.61 (99), 625 ([M+H].sup.+). LC-MS: (method 7): R.sub.t=1.01 (99), 625 ([M+H].sup.+). .sup.1H-NMR (DMSO-d.sub.6): 8.47 (d, J=2.6, 1 H), 8.12 (s, 1 H), 7.95 (d, J=9.6, 1 H), 7.61 (s, 1 H), 7.40-7.29 (m, 6 H), 5.10 (d, J=12.6, 1 H), 5.04 (d, J=12.6, 1 H), 4.98 (br. d, J=10.7, 1 H), 4.16 (br. d, J=11.8, 1 H), 4.10-3.90 (m, 4 H), 3.71 (br. t, J ca. 8.4, 1 H), 3.65-3.40 (m, 2 H), 3.23 (br. dd, J=11.1, 15.2, 1 H), 3.04 (s, 3 H), 2.92 (t, J=9.6, 1 H), 2.66 (s, 3 H), 2.12 (m, 1 H), 2.09 (br. q, 1 H), 1.42 (s, 9 H).
(418) Synthesis of the Amine Ex.273
(419) According to procedure J, carbamate Ex.272 (3.12 g, 5 mmol) in dioxane (31 mL) was treated with 4 M HCl-dioxane (62 mL) to afford Ex.273.2HCl (2.9 g, 97%)
(420) Data of Ex.273.2HCl: C.sub.26H.sub.32N.sub.6O.sub.6.2HCl (524.5, free base). LC-MS (method 2): R.sub.t=1.31 (92), 525 ([M+H].sup.+).
(421) Synthesis of the Amine Ex.274
(422) According to procedure K, carbamate Ex.272 (200 mg, 0.32 mmol) was hydrogenated in MeOH (20 mL) in the presence of the catalyst (100 mg) to afford Ex.274 (154 mg, 97%).
(423) Data of Ex.274: C.sub.23H.sub.34N.sub.6O.sub.6. (490.5). LC-MS (method 2): R.sub.t=1.26 (98), (491 ([M+H].sup.+).
Core 19: Synthesis of Ex.297 and Ex.298 (Scheme 23)
(424) Synthesis of the Mitsunobu Product 129
(425) Following procedure E.1.2, the reaction of phenol 75 (4.58 g, 9.9 mmol), alcohol 81 (5.5 g, 15 mmol), and CMBP (4.8 g, 20 mmol) in dry toluene (24 mL) afforded after FC (hexane/EtOAc 1:3) the protected amino acid 129 (5.54 g, 68%).
(426) Synthesis of the Amino Acid 130
(427) Following procedure E.2, the reaction of 129 (5.53 g, 6.8 mmol), 1,3-dimethylbarbituric acid (2.5 g, 16 mmol) and Pd(PPh.sub.3).sub.4 (0.39 g) in EtOAc/CH.sub.2Cl.sub.2 55:45 (118 mL) yielded after 2 h and after FC(CH.sub.2Cl.sub.2/MeOH 95:5 to 70:30) the amino acid 130 (1.45 g, 85%).
(428) Data of 130: C.sub.36H.sub.45N.sub.5O.sub.9 (691.7). LC-MS (method 7): R.sub.t=1.09 (96), 692 ([M+H].sup.+).
(429) Synthesis of the Protected Macrolactam Ex.297
(430) According to procedure F.1.1, amino acid 130 (2.57 g, 3.7 mmol) in dry CH.sub.2Cl.sub.2 (40 mL) was treated with T3P (50% in EtOAc, 4.4 mL, 7.4 mmol) and i-Pr.sub.2NEt (2.5 mL, 14.9 mmol) in dry CH.sub.2Cl.sub.2 (330 mL) to give after FC(CH.sub.2Cl.sub.2/MeOH 99:1 to 90:10) the macrolactam Ex.297 (2.5 g, contaminated with ca 20% i-Pr.sub.2NEt; yield 80%).
(431) Data of Ex.297: C.sub.36H.sub.43N.sub.5O.sub.8 (673.7). LC-MS: (method 7): R.sub.t=1.18 (93), 674 ([M+H].sup.+).
(432) Aqueous workup (EtOAc, 1 M aq. NaH PO.sub.4 soln) of ananalytical sample (100 mg) afforded pure Ex.297 (81 mg).
(433) LC-MS: (method 2): R.sub.t=2.20 (93), 674 ([M+H].sup.+). .sup.1H-NMR (DMSO-d.sub.6):complex spectrum, several isomers, 8.51 (d, J=8.5, 0.2; H), 8.47 (d, J=8.7, 0.1 H), 8.40 (d, J=8.5, 0.55 H), 8.32 (d, J=8.5, 0.15 H), 7.68-7.10 (several m, 10 H), 5.96 (br. s, 0.3 H), 5.90 (br. s, 0. 3 H), 5.4-5.0 (m, 2.4 H), 4.8-3.8 (several m, 8 H), 3.3-2.5 (several m and s, 8 H), 2.5-1.6 (several m, 4 H), 1.42, 1.41, 1.36, 1.26 (4 s, Boc).
(434) Synthesis of the Acid Ex.298:
(435) According to procedure H, the ester Ex.297 (2.0 g, contaminated with ca 20% i-Pr.sub.2NEt 2.4 mmol) was hydrogenated in MeOH (200 mL) in the presence of the catalyst (1 g) for 3 h.
(436) The crude product was suspended in diethyl ether (20 mL) stirred for 20 min, filtered, washed (diethyl ether) and dried to afford Ex.298 (1.63 g, contaminated with 15% i-Pr.sub.2NEt, quantitative yield).
(437) Aqueous workup (CH.sub.2Cl.sub.2, 1 M aq. NaH PO.sub.4 soln) of an analytical sample (200 mg) afforded pure Ex.298 (135 mg).
(438) Data of Ex.298: C.sub.29H.sub.37N.sub.5O.sub.8 (583.6). LC-MS: (method 4a): R.sub.t=1.78 (86), 584 ([M+H].sup.+).
Core 20: Synthesis of Ex.311 (Scheme 24)
(439) Synthesis of the Mitsunobu Product 131
(440) A solution of phenol 72 (200 mg, 0.34 mmol), alcohol 16 (178 mg, 0.52 mmol) and PPh.sub.3 (180 mg, 0.69 mmol) in benzene (5 mL) was degassed. At 0 C., DEAD (40% in toluene, 0.32 mL, 0.69 mmol) was added. The mixture was stirred at room temperature for 15 h. More of alcohol 16 (178 mg, 0.52 mmol) and PPh.sub.3 (180 mg, 0.69 mmol) were added. DEAD (40% in toluene, 0.32 mL, 0.69 mmol) was added at 0 C. The mixture was stirred for 20 h and concentrated. FC(CH.sub.2Cl.sub.2/EtOAc 100:0 to 80:20) afforded 131 (containing ca. 20% of diethyl hydrazine-1,2-dicarboxylate; used without any further purification).
(441) Synthesis of the Amino Acid 132
(442) Following procedure B.2, the reaction of 131 (250 mg, ca. 80%, 0.22 mmol), 1.3-dimethylbarbituric acid (107 mg, 0.69 mmol) and Pd(PPh.sub.3).sub.4 (16 mg) in EtOAc/CH.sub.2Cl.sub.2 (55:45, 4.8 mL) yielded after 3 h and after FC (EtOAc/MeOH 100:0 to 90:10, then CH.sub.2Cl.sub.2/MeOH 90:10 to 80:20) 132 (177 mg, yield over the two steps: 73%).
(443) Data of 132: C.sub.39H.sub.57N.sub.5O.sub.10Si (784.0): LC-MS: (method 7): R.sub.t=1.31, 784.2 ([M+H].sup.+).
(444) Synthesis of the Alloc Protected Amino Acid 133
(445) Following procedure C.1, the reaction of 132 (150 mg, 0.19 mmol), allyl chloroformate (23 L, 0.21 mmol) and Na.sub.2CO.sub.3 (61 mg, 0.57 mmol) in dioxane (1.5 mL) and H.sub.2O (1.5 mL) gave, after 2 h at 0 C., acid 133 (154 mg, 92%).
(446) Synthesis of the Protected Amino Acid 134
(447) Following procedure C.2, acid 133 (140 mg, 0.16 mmol) was reacted with sarcosine allylester p-toluenesulfonate (46 pTsOH, 58 mg, 0.194 mmol), HOAt (33 mg, 0.24 mmol), HATU (92 mg, 0.24 mmol) and i-Pr.sub.2NEt (0.138 mL, 0.81 mmol) in DMF (2.4 mL) to afford the protected amino acid 134 (106 mg, 67%).
(448) Data of 134: C.sub.49H.sub.70N.sub.6O.sub.13Si (979.2). LC-MS: (method 7): R.sub.t=1.68, 979.3 ([M+H].sup.+).
(449) Synthesis of Amino acid 135
(450) Following procedure C.3, the reaction of the protected amino acid 134 (100 mg, 0.10 mmol), 1.3-dimethylbarbituric acid (38 mg, 0.25 mmol) and Pd(PPh.sub.3).sub.4 (6 mg) in EtOAc/CH.sub.2Cl.sub.2 (45:55, 1.9 mL) yielded after 16 h and after FC (EtOAc, then CH.sub.2Cl.sub.2/MeOH 90:10) 135 (70 mg, 80%).
(451) Data of 135: C.sub.42H.sub.62N.sub.6O.sub.11Si (855.1). LC-MS: (method 7): R.sub.t=1.30, 855.5 ([M+H].sup.+).
(452) Synthesis of the Protected Macrolactam Ex.311
(453) According to procedure F.1.1, a solution of the amino acid 135 (60 mg, 0.07 mmol) in dry CH.sub.2Cl.sub.2 (2 mL), was added within 2 h to T3P (50% in EtOAc; 84 L, 0.14 mmol) and i-Pr.sub.2NEt (48 L, 0.28 mmol) in CH.sub.2Cl.sub.2 (5 mL). Then sat. aq. NaHCO.sub.3 solution was added and the mixture was extracted with CH.sub.2Cl.sub.2. The organic phase was dried (Na.sub.2SO.sub.4), filtered and concentrated. FC (EtOAc) afforded Ex.311 (26 mg, 44%).
(454) Data of Ex.311: (C.sub.42H.sub.60N.sub.6O.sub.10Si (837.0). LC-MS: (method 7): R.sub.t=1.51 (90), 837.4 ([M+H].sup.+). .sup.1H-NMR (CDCl.sub.3): 7.26 (s, 5 H), 7.09 (t, J=8.4, 1 H), 6.78 (d-like m, 1 H), 6.61 (d, J=7.4, 1 H), 5.50-4.90 (several br. m, 5 H), 4.90-3.80 (several br. m, 8 H), 3.69 (br. t, J ca. 8.5, 1 H), 3.6-2.3 (several br. m, 14 H), 2.12 (m, 1 H), 1.61 (m, 1 H), 1.38 (s, 9 H), 1.24 (s, 2 H), 0.93 (br. t, J ca. 8.0, 2 H), 0.00, 0.03 (2 s, 9 H).
Core 21: Synthesis of Ex.312 and Ex.313 (Scheme 25)
(455) Synthesis of the Mitsunobu Product 136
(456) Alcohol 82 (217 mg, 0.58 mmol) and CMBP (212 mg, 0.88 mmol) were dissolved in dry degassed toluene (7 mL) and heated at 100 C. for 30 min. A solution of 80 (250 mg, 0.58 mmol) in toluene (2 mL) was added dropwise. Stirring at 100 C. was continued for 1 h. The volatiles were evaporated. FC (hexane/EtOAc 2:1 to 1:1) yielded 136 (290 mg, 63%).
(457) Synthesis of Amino Acid 137
(458) Following procedure E.2 the reaction of 136 (250 mg, 0.32 mmol), 1,3-dimethylbarbituric acid (120 mg, 0.77 mmol) and Pd(PPh.sub.3).sub.4 (18 mg) in EtOAc/CH.sub.2Cl.sub.2 (45:55, 5.5 mL) yielded after 0.5 h and after FC(CH.sub.2Cl.sub.2/MeOH 95:5 to 70:30) the aminoacid 137 (164 mg, 78%).
(459) Data of 137: C.sub.33H.sub.44N.sub.4O.sub.8S (656.8). LC-MS (method 7): R.sub.t=1.15 (95), 657 ([M+H].sup.+).
(460) Synthesis of the Protected Macrolactam Ex.312
(461) According to procedure F.1.1, a solution of the amino acid 137 (100 mg, 0.15 mmol) in dry CH.sub.2Cl.sub.2 (2 mL) was added over 2 h to T3P (50% in EtOAc, 0.18 mL, 0.31 mmol) and i-Pr.sub.2NEt (0.1 mL, 0.61 mmol) in dry CH.sub.2Cl.sub.2 (13 mL). Stirring at room temperature was continued for 1 h, followed by aqueous workup (EtOAc, sat. aq. NaHCO.sub.3 soln, Na.sub.2SO.sub.4) and FC (EtOAc) to afford Ex.312 (56 mg, 57%).
(462) Data of Ex.312: C.sub.33H.sub.42N.sub.4O.sub.7S (638.7). LC-MS (method 7): R.sub.t=1.33 (95), 639 ([M+H].sup.+). .sup.1H-NMR (CDCl.sub.3): 7.37-7.23 (m, 8 H), 6.92 (br. s, 1 H), 5.25 (m, 2 H), 5.17 (s, 1 H), 4.88 (d, J=16.2, 1 H), 4.62 (br. m, 1 H), 4.46 (br. t-like m, 1 H), 4.31 (br. m, 1 H), 4.17 (dd, J=4.1, 14.2, 1 H), 3.72 (dd, J=4.8, 10.7, 1 H), 3.50 (m, 1 H), 3.30-2.80 (several m, 2 H), 3.14 (s, 3 H), 3.01 (s, 3 H), 2.60-1.90 (several m, 6 H), 1.46 (s, 9 H).
(463) Synthesis of Sulfon Ex.313
(464) m-CPBA (70% w/w; 10 mg, 41 mol) was added at 0 C. to a solution of Ex.312 (20 mg, 31 mol) in CH.sub.2Cl.sub.2 (0.5 mL). The mixture was stirred for 15 min followed by the addition of m-CPBA (9 mg, 37 mol). The mixture was allowed to warm to room temperature over 1 h, diluted with CH.sub.2Cl.sub.2 and washed with aq. Na.sub.2S.sub.2O.sub.3 soln and with aq. NaHCO.sub.3 soln. The organic phase was dried (Na.sub.2SO.sub.4), filtered and concentrated. FC (EtOAc/MeOH 100:0 to 90:10) afforded Ex.313 (8 mg, 38%).
(465) Data of Ex.313: C.sub.33H.sub.42N.sub.4O.sub.9S (670.7). LC-MS (method 6): R.sub.t=1.24 (95), 671 ([M+H].sup.+). .sup.1H-NMR (CDCl.sub.3): 7.89 (td, J=1.7, 7.3, 1 H), 7.71 (s, 1 H), 7.43-7.28 (m, 7 H), 5.17 (d, J=12.0, 1 H), 5.10 (d, J=12.0, 1 H), 5.01 (dd, J=5.9, 9.1, 1 H), 4.96-4.85 (m, 2 H), 4.71 (d, J=15.4, 1 H), 4.57 (br. m, 1 H), 4.33 (br. m, 2 H), 3.85 (dd, J=7.8, 12.3, 1 H), 3.25 (s, 3 H), 3.20 (m, 1 H), 3.10 (m, 1 H), 2.97 (s, 3 H), 2.73-2.54 (m, 2 H), 2.45-2.23 (m, 2 H), 2.17 (m, 1 H), 1.99 (m, 1 H), 1.46 (s, 9 H).
(466) Synthesis of Final Products
(467) Advanced macrocyclic intermediates and final products depicted in Tables 21a-36a (Scheme 26) and were prepared starting from the suitable precursor macrocyclic acid or macrocyclic amine applying the general procedures (HN) described above. Deviations from general procedures are indicated in Tables 21a-36a.
(468) Analytical data of these intermediates and final products are depicted in Tables 21b-36b.
(469) IUPAC names of all examples are listed in Tables 20, 21c-36c, and 37.
Detailed Description of Selected Examples
Core 03
Synthesis of Selected Advanced Intermediates and Final Products (Scheme 27)
(470) Synthesis of Amide Ex.27
(471) A mixture of Ex.4 (432 mg, 0.79 mmol), HATU (597 mg, 1.57 mmol) and HOAt (214 mg, 1.57 mmol) was dissolved in DMF (6 mL). N,N-dimethylethylenediamine (173 L, 1.57 mmol) and i-Pr.sub.2NEt (537 L, 3.14 mmol) were added. The solution was stirred at room temperature for 15 h and concentrated. The residue was dissolved in CHCl.sub.3 and washed with sat. aq. NaHCO.sub.3 solution and with H.sub.2O. The organic phase was dried (Na.sub.2SO.sub.4), filtered and concentrated. FC(CH.sub.2Cl.sub.2/MeOH/conc. aq. NH.sub.3 soln 100:0:0 to 90:10:0.5) afforded Ex.27 (405 mg, 83%).
(472) Data of Ex.27: Cf. Table 21b
(473) Synthesis of Amine Ex.28
(474) A solution of Ex.27 (400 mg, 0.64 mmol) in dioxane (4 mL) was treated at room temperature with 4 M HCl-dioxane (8 mL) for 2 h. The volatiles were evaporated. The residue was dissolved in CH.sub.2Cl.sub.2/MeOH, concentrated and dried i.v. to afford Ex.28.HCl (343 mg, 90%).
(475) Data of Ex.28: Cf. Table 21b
(476) Synthesis of Amide Ex.11
(477) A mixture of Ex.28.HCl (75 mg, 0.126 mmol), .sup.1H-indole-3-acetic acid (44 mg, 0.253 mmol), HATU (96 mg, 0.253 mmol) and HOAt (34 mg, 0.253 mmol) was dissolved in DMF (2 mL). i-Pr.sub.2NEt (87 L, 0.505 mmol) was added. The solution was stirred at room temperature for 15 h and concentrated. The residue was dissolved in CHCl.sub.3 and washed with sat. aq. NaHCO.sub.3 solution and with H.sub.2O. The organic phase was dried (Na.sub.2SO.sub.4), filtered and concentrated. FC(CH.sub.2Cl.sub.2/MeOH/conc. aq. NH.sub.3 soln 100:0:0 to 90:10:1) afforded Ex.11 (50 mg, 58%).
(478) Data of Ex.11: Cf. Table 21b
(479) .sup.1H-NMR (DMSO-d.sub.6): 10.81 (s, 1 H), 8.26 (d, J=7.4, 1 H), 7.62 (t, J=5.5, 1 H), 7.46 (d, J=7.9, 1 H), 7.37-7.15 (m, 4 H), 7.09 (d, J=2.2, 1 H), 7.04 (t, J=7.5, 1 H), 6.92 (t, J ca. 7.4, 1 H), 5.08 (d, J ca. 12.5, 1 H), 4.74 (d, J=8.9, 1 H), 4.37 (d, J=11.0, 1 H), 4.25 (d, J=17.7, 1 H), 4.22-4.13 (m, 2 H), 3.97 (d, J=17.6, 1 H), 3.78 (t, J=8.3, 1 H), 3.41 (s, 2 H), 3.24 (m, 1 H), 3.15 (m, 1 H), 2.98 (t, J=9.2, 1 H), 2.88 (s, 3 H), 2.53 (s, 3 H), 2.41-2.27 (m, 4 H), 2.17 (s, 6 H), 2.04 (m, 1 H), 1.83 (t-like m, 2 H), 1.69 (q-like m, 1 H).
(480) Synthesis of Amide Ex.49
(481) A mixture of Ex.28.HCl (60 mg, 0.101 mmol), 1-naphthylacetic acid (23 mg, 0.121 mmol), and HOBt.H.sub.2O (19 mg, 0.121 mmol) was dissolved in CH.sub.2Cl.sub.2 (1 mL). N-Cyclohexyl-carbodiimide-N-methylpolystyrene (1.9 mmol/g; 80 mg, 0.152 mmol) and i-Pr.sub.2NEt (52 L, 0.303 mmol) were added. The mixture was stirred for 15 h at room temperature. (Polystyrylmethyl)-trimethylammonium bicarbonate (3.5 mmol/g; 87 mg, 0.303 mmol) was added and stirring was continued for 1 h. The mixture was diluted with CH.sub.2Cl.sub.2/MeOH 9:1 (2 mL) and filtered. The polymer was washed with twice with CH.sub.2Cl.sub.2/MeOH 8:2 (5 mL). The combined filtrate and washings were concentrated. Purification of the crude product by FC(CH.sub.2Cl.sub.2/MeOH/conc. aq. NH.sub.3 soln. 100:0:0 to 90:10:1) afforded Ex.49 (58 mg, 83%).
(482) Data of Ex.49: Cf. Table 21b
(483) .sup.1H-NMR (DMSO-d.sub.6): 8.45 (d, J=7.3, 1 H), 8.00-7.87 (m, 2 H), 7.79 (d, J=8.0, 1 H), 7.62 (t, J=5.5, 1 H), 7.53-7.25 (m, 6 H), 7.19 (dd, J=3.0, 8.4, 1 H), 5.10 (d, J=12.3, 1 H), 4.75 (d, J=8.9, 1 H), 4.39 (d, J=10.8, 1 H), 4.27 (d, J=17.8, 1 H), 4.28-4.08 (m, 2 H), 3.95 (d, J=17.9, 1 H), 3.83 (m, 1 H), 3.81 (s, 2 H), 3.24 (m, 1 H), 3.16 (m, 1 H), 3.03 (t, J=9.2, 1 H), 2.87 (s, 3 H), 2.54 (s, 3 H), 2.42-2.27 (m, 4 H), 2.16 (s, 6 H), 2.02 (m, 1 H), 1.84 (t-like m, 2 H), 1.71 (q, J ca. 9.4, 1 H).
(484) Synthesis of Amide Ex.30
(485) A mixture of Ex.4 (400 mg, 0.73 mmol), HATU (552 mg, 1.45 mmol), HOAt (198 mg, 1.45 mmol) and tryptamine (233 mg, 1.45 mmol) was dissolved in DMF (6 mL). i-Pr.sub.2NEt (497 L, 2.91 mmol) was added. The solution was stirred at room temperature for 15 h followed by aqueous workup (CHCl.sub.3, sat. aq. NaHCO.sub.3 soln, H.sub.2O). The organic phase was dried (Na.sub.2SO.sub.4), filtered and concentrated. FC(CH.sub.2Cl.sub.2/MeOH 100:0 to 95:5) afforded Ex.30 (410 mg, 81%).
(486) Data of Ex.30: Cf. Table 21b
(487) .sup.1H-NMR (DMSO-d.sub.6): 10.80 (s, 1 H), 7.91 (t, J=5.6, 1 H), 7.56 (d, J=7.7, 1 H), 7.32 (d, J=8.0, 1 H), 7.27-7.12 (m, 5 H), 7.06 (t, J=7.5, 1 H), 6.97 (t, J=7.4, 1 H), 5.08 (d, J=12.4, 1 H), 4.75 (d, J=9.3, 1 H), 4.34 (d, J=10.9, 1 H), 4.24 (d, J=17.8, 1 H), 4.10 (t-like m, 1 H), 3.97 (d, J=17.7, 1 H), 3.86 (m, 1 H), 3.77 (m, 1 H), 3.42-3.30 (m, 2 H), 2.96-2.83 (m, 3 H), 2.89 (s, 3 H), 2.50 (s, 3 H, superimposed by DMSO-d signal), 2.27 (m, 2 H), 2.08 (m, 1 H), 1.84 (t-like m, 2 H), 1.65 (q, J=10.8, 1 H), 1.34 (s, 9 H).
(488) Synthesis of Amine Ex.55
(489) A solution of Ex.30 (380 mg, 0.55 mmol) in dioxane (4 mL) was treated at room temperature with 4 M HCl-dioxane (8 mL) for 4 h. The volatiles were evaporated. The residue was dissolved in dioxane (4 mL) and treated again for 2 h with 4 M HCl-dioxane (8 mL). The volatiles were evaporated. The residue was washed with diethyl ether and purified by FC(CH.sub.2Cl.sub.2/MeOH/conc. aq. NH.sub.3 soln 90:10:0 to 90:10:1) to afford Ex.55 (136 mg, 42%).
(490) Data of Ex.55: Cf. Table 21b
(491) Synthesis of Amide Ex.12
(492) A mixture of Ex.55 (68 mg, 0.092 mmol), 1H-indole-3-acetic acid (32 mg, 0.184 mmol), HATU (70 mg, 0.184 mmol) and HOAt (25 mg, 0.184 mmol) was dissolved in DMF (2 mL). i-Pr.sub.2NEt (63 L, 0.367 mmol) was added. The solution was stirred at room temperature for 15 h and concentrated. The residue was dissolved in CHCl.sub.3 and washed with sat. aq. NaHCO.sub.3 solution and with H.sub.2O. The organic phase was dried (Na.sub.2SO.sub.4), filtered and concentrated. Purification by prep. HPLC, method 1, afforded Ex.12 (38 mg, 55%).
(493) Data of Ex.12: Cf. Table 21b
(494) .sup.1H-NMR (DMSO-d.sub.6): 10.81 (s, 2 H), 8.26 (d, J=7.2, 1 H), 7.93 (t, J=5.7, 1 H), 7.57 (d, J=7.8, 1 H), 7.46 (d, J=7.7, 1 H), 7.38-6.90 (m, 11 H); 5.10 (d, J=12.1, 1 H), 4.76 (d, J=9.3, 1 H), 4.38 (d, J=10.8, 1 H), 4.26 (d, J=17.8, 1 H), 4.23-4.11 (m, 2 H), 3.96 (d, J=18.0, 1 H), 3.78 (t, J=8.3, 1 H), 3.7-3.25 (m, 3 H), 3.60 (s, 2 H), 3.01-2.81 (m, 2 H), 2.88 (s, 3 H), ca. 2.5 (s, 3 H, superimposed by DMSO-d signal), 2.33 (m, 2 H), 2.06 (m, 1 H), 1.85 (t-like m, 2 H), 1.63 (q, J ca. 10.7, 1 H).
(495) Synthesis of Amide Ex.16
(496) A mixture of Ex.55 (68 mg, 0.092 mmol), N,N-dimethyl glycine (19 mg, 0.184 mmol), HATU (70 mg, 0.184 mmol) and HOAt (25 mg, 0.184 mmol) was dissolved in DMF (2 mL). i-Pr.sub.2NEt (63 L, 0.367 mmol) was added. The solution was stirred at room temperature for 15 h and concentrated. The residue was dissolved in CHCl.sub.3 and washed with sat. aq. NaHCO.sub.3 solution and with H.sub.2O. The organic phase was dried (Na.sub.2SO.sub.4), filtered and concentrated. Purification by prep. HPLC, method 1, afforded Ex.16 TFA (40 mg, 55%).
(497) Data of Ex.16 TFA: Cf. Table 21b
(498) .sup.1H-NMR (DMSO-d.sub.6): 10.81 (s, 1 H), 9.66 (br. s, NH.sup.+), 8.75 (d, J=6.9, 1 H), 7.90 (t, J=5.6, 1 H), 7.56 (d, J=7.8, 1 H), 7.34-7.14 (m, 5 H), 7.06 (t, J ca. 7.5, 1 H), 6.97 (t, J=7.4, 1 H), 5.08 (d, J=12.3, 1 H), 4.78 (d, J=9.2, 1 H), 4.39 (d, J=10.7, 1 H), 4.24 (d, J=17.8, 1 H), 4.24-4.14 (m, 2 H), 4.00 (d, J=17.8, 1 H), 3.96-3.75 (m, 3 H), 3.45-3.35 (m, 2 H), 3.0-2.67 (m, 3 H), 2.90 (s, 3 H), 2.75 (s, 6 H), 2.50 (s, 3 H, superimposed by DMSO-d signal), 2.5-2.27 (m, 2 H), 2.08 (m, 1 H), 1.85 (t-like m, 2 H), 1.64 (q, J=10.8, 1 H).
(499) Synthesis of Amide Ex.53
(500) Pyridine (2 mL) and acetic anhydride (0.14 mL, 1.48 mmol) were added to a solution of Ex.5.HCl (95 mg, 0.15 mmol) in dry CH.sub.2Cl.sub.2 (2 mL). The solution was stirred at room temperature for 20 h. The solution was diluted with EtOAc and washed with 1 M aq. HCl soln, sat. aq. NaCl soln, sat. aq. NaHCO.sub.3 soln, and sat. aq. NaCl soln. The organic phase was dried (Na.sub.2SO.sub.4), filtered and concentrated. FC of the crude product afforded Ex.53 (60 mg, 70%).
(501) Data of Ex.53: Cf. Table 21b
(502) Synthesis of Acid Ex.54
(503) A solution of Ex.53 (58 mg, 0.01 mmol) in MeOH (5 mL) was hydrogenated at room temperature and normal pressure for 2 h in the presence of palladium hydroxide on activated charcoal (moistened with 50% H.sub.2O; 50 mg). The mixture was filtered through a pad of celite. The residue was washed (MeOH). The combined filtrate and washings were concentrated and dried i.v. to yield Ex.54 (45 mg, 92%).
(504) Data of Ex.54: Cf. Table 21b
(505) Synthesis of Amide Ex.9
(506) A mixture of Ex.54 (45 mg, 0.091 mmol), HATU (52 mg, 0.137 mmol) HOAt (19 mg, 0.137 mmol) and tryptamine (22 mg, 0.137 mmol) was dissolved in DMF (1 mL). i-Pr.sub.2NEt (47 L, 0.274 mmol) was added. The solution was stirred at room temperature for 20 h followed by aqueous workup (CHCl.sub.3, sat. aq. NaHCO.sub.3 soln, H.sub.2O). The organic phase was dried (Na.sub.2SO.sub.4), filtered and concentrated. FC(CH.sub.2Cl.sub.2/MeOH 100:0 to 86:14) afforded Ex.9 (36 mg, 62%).
(507) Data of Ex.9: Cf. Table 21b
(508) .sup.1H-NMR (DMSO-d.sub.6): 10.81 (s, 1 H), 8.06 (d, J=7.0, 1 H), 7.93 (t, J=5.6, 1 H), 7.56 (d, J=7.8, 1 H), 7.34-7.14 (m, 5 H), 7.05 (t, J ca. 7.5, 1 H), 6.97 (t, J ca. 7.4, 1 H), 5.09 (d, J=12.4, 1 H), 4.75 (d, J=9.1, 1 H), 4.38 (d, J=10.8, 1 H), 4.26 (d, J=17.7, 1 H), 4.19-4.10 (m, 2 H), 3.97 (d, J=17.9, 1 H), 3.78 (t, J=8.3, 1 H), 3.43-3.30 (m, 2 H), 2.96-2.83 (m, 3 H), 2.89 (s, 3 H), 2.50 (s, 3 H, superimposed by DMSO-d signal), 2.40-2.27 (m, 2 H), 2.08 (m, 1 H), 1.85 (m, 2 H), 1.71 (s, 3 H), 1.62 (q, J ca. 10.6, 1 H).
Core 11 and Core 12
Synthesis of Selected Advanced Intermediates and Final Products (Scheme 28)
(509) Synthesis of Amide Ex.184
(510) A mixture of Ex 182 (500 mg, 1.04 mmol), 2-naphthylacetic acid (232 mg, 1.25 mmol), HATU (791 mg, 2.08 mmol) and HOAt (283 mg, 2.08 mmol) was dissolved in DMF (15 mL). i-Pr.sub.2NEt (712 L, 4.16 mmol) was added. The solution was stirred at room temperature for 20 h and concentrated. The residue was dissolved in CHCl.sub.3 and washed with sat. aq. NaHCO.sub.3 solution and with H.sub.2O. The organic phase was dried (Na.sub.2SO.sub.4), filtered and concentrated. FC (EtOAc, then CH.sub.2Cl.sub.2/MeOH 95:5) afforded Ex.184 (637 mg, 94%).
(511) Data of Ex.184: Cf. Table 29b
(512) .sup.1H-NMR (DMSO-d.sub.6): 8.41 (d, J=7.0, 1 H), 7.90-7.83 (m, 3 H), 7.77 (s, 1 H), 7.53-7.44 (m, 4 H), 7.32-7.22 (m, 6 H), 7.04 (d, J=8.4, 1 H), 6.86 (d, J=7.4, 1 H), 6.81 (s, 1 H), 5.02-4.90 (m, 3 H), 4.19 (t, J ca. 8.6, 1 H), 4.14-3.96 (m, 2 H), 3.83 (t-like m, 2 H), 3.63 (s, 2 H), ca. 3.3 (m, 1 H, superimposed by H.sub.2O signal), 3.05 (m, 1 H), 2.95 (m, 1 H), 2.91 (s, 3 H), 2.27 (m, 1 H), 2.16 (br. q, J ca. 11.3, 1 H), 1.54 (m, 2 H), 1.31 (m, 1 H), 1.15 (m, 1 H).
(513) Synthesis of Amide Ex.200
(514) A mixture of Ex.197 TFA (60 mg, 0.094 mmol), 1H-indole-3-acetic acid (25 mg, 0.14 mmol), HATU (54 mg, 0.14 mmol) and HOAt (19 mg, 0.14 mmol) was dissolved in DMF (1.5 mL). i-Pr.sub.2NEt (81 L, 0.471 mmol) was added. The solution was stirred for 18 h at room temperature and concentrated. The residue was dissolved in CHCl.sub.3 and washed (sat. aq. NaHCO.sub.3 soln, H.sub.2O). The organic phase was dried (Na.sub.2SO.sub.4), filtered and concentrated, followed by FC (EtOAc, then CH.sub.2Cl.sub.2/MeOH 95:5) to afford Ex.200 (50 mg, 78%).
(515) Data of Ex.200: Cf. Table 30b
(516) .sup.1H-NMR (DMSO-d.sub.6): 10.86 (s, 1 H), 8.42 (d, J=7.8, 1 H), 8.01 (d, J=10.0, 1 H), 7.58 (d, J=7.8, 1 H), 7.36-7.19 (m, 9 H), 7.07-7.02 (m, 2 H), 6.97 (t, J=7.1, 1 H), 6.86 (d, J=7.6, 1 H), 5.08 (s, 2 H), 4.88 (d, J=8.7, 1 H), 4.30-4.10 (m, 2 H), 4.13 (d, J=10.9, 1 H), 4.01 (t-like m, 1 H), 3.95 (d, J=18.0, 1 H), 3.75-3.70 (m, 2 H), 3.56 (s, 2 H), 3.4-3.2 (m, 2 H, partially superimposed by H.sub.2O signal), 3.04 (t, J=9.9, 1 H), 2.98 (s, 3 H), 2.65 (s, 3 H), 2.27 (m, 1 H), 2.09 (q, J=11.7, 1 H).
(517) Synthesis of Amine Ex.202
(518) A solution of Ex.200 (320 mg, 0.47 mmol) in MeOH (28 mL) was hydrogenated at normal pressure and at room temperature for 4 h in the presence of palladium hydroxide on activated charcoal (moistened with 50% H.sub.2O; 158 mg). The mixture was filtered through a pad of celite. The residue was washed (MeOH). The combined filtrate and washings were concentrated and dried i.v. to yield Ex.202 (250 mg, 97%).
(519) Data of Ex.202: Cf. Table 30b
(520) Synthesis of Amide Ex.213
(521) A solution of Ex.202 (60 mg, 0.11 mmol) in dry CH.sub.2Cl.sub.2 (1 mL) was treated with pyridine (89 L, 1.1 mmol). Decanoyl chloride (46 L, 0.22 mmol) was slowly added at 0 C. The mixture was stirred at 0 C. to room temperature for 18 h followed by the addition of MeOH (0.1 mL). Stirring was continued for 10 min. The volatiles were evaporated. The residue was three times treated with toluene and evaporated. Purification by prep. HPLC, method 1 and subsequent FC (EtOAc/MeOH 90:10 to 80:20) afforded Ex.213 (27 mg, 35%).
(522) Data of Ex.213: Cf. Table 30b
(523) .sup.1H-NMR (DMSO-d.sub.6): 10.86 (s, 1 H), 8.53 (d, J=9.8, 1 H), 8.44 (d, J=7.7, 1 H), 7.57 (d, J=7.7, 1 H), 7.35-7.30 (m, 3 H), 7.27 (s, 1 H), 7.19-6.95 (m, 3 H), 6.84 (d, J=7.5, 1 H), 4.86 (dd, J=2.4, 11.2, 1 H), 4.60 (q, J=8.4, 1 H), 4.25 (q-like m, 1 H), 4.14 (d, J=10.7, 1 H), 4.04-3.82 (m, 3 H), 3.73 (t, J ca. 8.5, 1 H), 3.55 (s, 2 H), 3.24 (d, J=7.8, 2 H), 3.09 (t, J=9.5, 1 H), 2.99 (s, 3 H), 2.67 (s, 3 H), 2.26 (m, 1 H), 2.15 (t, J=7.2, 2 H), 2.09 (m, 1 H), 1.51 (t-like m, 2 H), 1.24 (s, 12 H), 0.85 (t, J=6.6, 3 H).
Core 11
Synthesis of Ex.186 on Solid Support (Scheme 29)
(524) Synthesis of Amine 139
(525) A solution of Ex.181 (2.0 g, 3.2 mmol) in MeOH (200 mL) was hydrogenated for 3 h at room temperature and at normal pressure in the presence of palladium hydroxide on activated charcoal (15-20% Pd, moistened with 50% H.sub.2O; 400 mg). The mixture was filtered through a pad of celite. The residue was washed (MeOH). The combined filtrate and washings were concentrated and dried i.v. to give the corresponding amine (1.57 g), which was dissolved in CH.sub.2Cl.sub.2 (8 mL) and treated with sat. aqueous NaHCO.sub.3 solution (2.9 mL) and allyl chloroformate (0.36 mL, 3.43 mmol). The mixture was stirred at room temperature for 2 h. The organic phase was separated and concentrated. Purification of the residue by FC (EtOAc) afforded the allyl carbamate 138 (1.65 g, 92%).
(526) TBAF solution (1 M in THF, 7 mL, 7 mmol) was added at 0 C. to a solution of 138 (1.29 g, 2.24 mmol) in THF (53 mL). The solution was stirred at 0 C. to room temperature for 3 h and concentrated. The residue was distributed between CH.sub.2Cl.sub.2 and sat. aq. NaHCO.sub.3 solution. The aqueous phase was separated and extracted with CH.sub.2Cl.sub.2. The combined organic phase was dried (Na.sub.2SO.sub.4), filtered and concentrated. The residue was dissolved in CH.sub.2Cl.sub.2 (10 mL) and treated for 20 min with 25% aq. HCl solution (0.29 mL). The volatiles were evaporated and the residue was dried i.v. to afford 139.HCl (1.14 g; contaminated with ca 15% tetrabutylammonium salt and used without further purification; yield ca 90%)
(527) Data of 139.HCl: C.sub.22H.sub.30N.sub.4O.sub.5.HCl (430.5, free base). LC-MS (method 4a): R.sub.t=1.22 (92), 431.3 [M+H].sup.+.
(528) Synthesis of the Resin 140
(529) DFPE polystyrene (1% DVB, 100-200 mesh, loading 0.89 mmol/g; 200 mg, 0.178 mmol) was swollen in DCE (2 mL) for 1 h. The resin was filtered. A solution of amine hydrochloride 139.HCl (ca 85% w/w, 166 mg, 0.303 mmol) in DCE (1.33 mL) and trimethyl orthoformate (0.66 mL, 6.02 mmol) were added. The resin was shaken for 1 h at room temperature, followed by the addition of sodium triacetoxyborohydride (75 mg, 0.356 mmol). The mixture was shaken for 15 h and the resin was filtered. The resin was successively washed three times each with DMF, 10% i-Pr.sub.2NEt in DMF, DMF, CH.sub.2Cl.sub.2 and dried i.v. to afford resin 140 (293 mg).
(530) Synthesis of the Resin 141
(531) 1st Acid coupling step: The resin 140 (loading 0.77 mmol/g; 50 mg, 0.038 mmol) was swollen in DMF (1 mL) for 30 min and filtered. CH.sub.2Cl.sub.2 (0.5 mL), DMF (0.5 mL), 2-naphthylacetic acid (65 mg, 0.35 mmol), i-Pr.sub.2NEt (0.13 mL, 0.76 mmol) and HATU (144 mg, 0.38 mmol) were successively added. The resin was shaken for 1 h, filtered and washed with DMF. CH.sub.2Cl.sub.2 (0.5 mL), DMF (0.5 mL) 2-naphthylacetic acid (65 mg, 0.35 mmol), i-Pr.sub.2NEt (0.13 mL, 0.76 mmol) and then HATU (144 mg, 0.38 mmol) were added to the resin. The mixture was shaken for 1 h and filtered. The resin was washed three times with DMF and two times with CH.sub.2Cl.sub.2.
(532) Cleavage of the Alloc group: CH.sub.2Cl.sub.2 (1 mL), phenylsilane (41 mg, 0.375 mmol) and Pd(PPh.sub.3).sub.4 (9 mg) were added to the resin. The mixture was shaken for 15 min and filtered. The resin was washed with CH.sub.2Cl.sub.2 and treated again for 15 min with CH.sub.2Cl.sub.2 (1 mL), phenylsilane (41 mg, 0.375 mmol) and Pd(PPh.sub.3).sub.4 (9 mg). The resin was filtered, washed three times each with CH.sub.2Cl.sub.2, DMF and twice with MeOH and CH.sub.2Cl.sub.2.
(533) 2nd Acid coupling step: DMF (0.5 mL), CH.sub.2Cl.sub.2 (1 mL), 2-naphthylacetic acid (70 mg, 0.375 mmol), i-Pr.sub.2NEt (0.13 mL, 0.75 mmol) and PyBOP (195 mg, 0.375 mmol) were added to the resin. The mixture was shaken for 1 h and filtered. The resin was washed three times each with DMF and CH.sub.2Cl.sub.2 to afford resin 141, which was immediately used in the next step.
(534) Release of the Amide Ex.186
(535) The resin 141 was treated with 20% TFA in CH.sub.2Cl.sub.2 (1 mL) for 10 min, filtered and washed with CH.sub.2Cl.sub.2. The resin was treated again for 10 min with 20% TFA in CH.sub.2Cl.sub.2 (1 mL), filtered and washed three times with CH.sub.2Cl.sub.2. The combined filtrates and washings were concentrated. The residue was treated with CH.sub.3CN, evaporated and dried i.v. Purification of the crude product by prep. HPLC, method 3, afforded Ex.186 (11 mg, yield: overall 32% based on 139).
(536) Data of Ex.186: C.sub.42H.sub.42N.sub.4O.sub.5 (682.8). LC-MS (method 4a): R.sub.t=2.26 (98). .sup.1H-NMR (DMSO-d.sub.6): 8.38 (d, J=7.0, 2 H), 7.91-7.69 (m, 8 H), 7.54-7.27 (m, 7 H), 7.03 (dd, J=1.5, 8.2, 1 H), 6.86-6.82 (m, 2 H), 4.94 (d, J=12.7, 1 H), 4.19 (t, J=8.6, 1 H), 4.11-3.94 (m, 3 H), 3.71 (dd, J=9.2, 16.5, 1 H), 3.62 (s, 2 H), 3.58 (s, 2 H), 3.08 (m, 1 H), 2.89 (m, 1 H), 2.89 (s, 3 H), 2.5 (m, 1 H, superimposed by DMSO-d signal), 2.30 (m, 1 H), 2.14 (q-like m, 1 H), 1.64-1.49 (m, 2 H), 1.34 (m, 1 H), 1.14 (m, 1 H).
(537) The .sup.1H-NMR spectrum is identical with the spectrum of the sample prepared in solution, cf. Table 29
(538) TABLE-US-00030 TABLE 20 Examples of Core 01 and Core 02 (Ex. 1-Ex. 2) No IUPAC name Core 01 R2 R50 Ex. 1
(539) TABLE-US-00031 TABLE 21a Examples of Core 03 (Ex. 3-Ex. 55,) General Yield, Starting Pro- Purification (isolated No R50 R2 material cedure Reagent Method salt) Ex. 3-Ex. 5: cf. experimental description Ex. 6 OH
(540) TABLE-US-00032 TABLE 21b Examples of Core 03 (Ex. 3-Ex. 55,) Monoisotopic Rt (purity [M + H].sup.+ LC-MS- No R50 R2 Formula Mass at 220 nm) found Method Ex. 3-Ex. 5: cf. experimental description Ex. 6 OH
(541) TABLE-US-00033 TABLE 21c Examples of Core 03 (Ex. 3-Ex.55,) No R50 R2 IUPAC name Ex. 3 OCH.sub.2Ph
(542) TABLE-US-00034 TABLE 22a Examples of Core 04 (Ex. 56-Ex. 84,) Yield, (iso- Starting General Purification lated No R50 R2 material Procedure Reagent Method salt) Ex. 56-Ex 57: cf. experimental description Ex. 58
(543) TABLE-US-00035 TABLE 22b Examples of Core 04 (Ex. 56-Ex. 84,) Monoisotopic Rt (purity [M + H].sup.+ LC-MS- No R50 R2 Formula Mass at 220 nm) found Method Ex. 56-Ex 57: cf. experimental description Ex. 58
(544) TABLE-US-00036 TABLE 22c Examples of Core 04 (Ex. 56-Ex. 84,) No R50 R2 IUPAC name Ex. 56 OCH.sub.2Ph
(545) TABLE-US-00037 TABLE 23a Examples of Core 05 (Ex. 85-Ex. 103,) Yield, (iso- Starting General Purification lated No R2 R50 material Procedure Reagent Method salt) Ex. 85-Ex 86: cf. experimental description Ex. 87
(546) TABLE-US-00038 TABLE 23b Examples of Core 05 (Ex. 85-Ex. 103,) Monoiso- Rt [M + topic (purity at H].sup.+ LC-MS- No R2 R50 Formula Mass 220 nm) found Method Ex. 85-Ex 86: cf. experimental description Ex. 87
(547) TABLE-US-00039 TABLE 23c Examples of Core 05 (Ex. 85-Ex. 103,) No R2 R50 IUPAC name Ex. 85
(548) TABLE-US-00040 TABLE 24a Examples of Core 06 (Ex. 104-Ex. 114,) Yield, Starting General Purification (isolated No R2 R50 material Procedure Reagent Method salt) Ex. 104-Ex. 105: cf. experimental description Ex. 106
(549) TABLE-US-00041 TABLE 24b Examples of Core 06 (Ex. 104-Ex. 114,) Rt Monoisotopic (purity at [M + H].sup.+ LC-MS- No R2 R50 Formula Mass 220 nm) found Method Ex. 104-Ex. 105: cf. experimental description Ex. 106
(550) TABLE-US-00042 TABLE 24c Examples of Core 06 (Ex. 104-Ex. 114,) No R2 R50 IUPAC name Ex. 104
(551) TABLE-US-00043 TABLE 25a Examples of Core 07 (Ex. 115-Ex. 131,) Yield, Starting General Purification (isolated No R11 R50 material Procedure Reagent Method salt) Ex. 115-Ex. 116: cf. experimental description Ex. 117
(552) TABLE-US-00044 TABLE 25b Examples of Core 07 (Ex. 115-Ex. 131,) Monoisotopic Rt (purity at [M + H].sup.+ LC-MS- No R11 R50 Formula Mass 220 nm) found Method Ex. 115-Ex. 116: cf. experimental description Ex. 117
(553) TABLE-US-00045 TABLE 25c Examples of Core 07 (Ex. 115-Ex. 131,) No R11 R50 IUPAC name Ex. 115
(554) TABLE-US-00046 TABLE 26a Examples of Core 08 (Ex. 132-Ex. 141,) Yield, Starting General Purification (isolated No R11 R50 material Procedure Reagent Method salt) Ex. 132-Ex. 133: cf. experimental description Ex. 134
(555) TABLE-US-00047 TABLE 26b Examples of Core 08 (Ex. 132-Ex. 141,) Rt Monoisotopic (purity at [M + H].sup.+ LC-MS- No R11 R50 Formula Mass 220 nm) found Method Ex. 132-Ex. 133: cf. experimental description Ex. 134
(556) TABLE-US-00048 TABLE 26c Examples of Core 08 (Ex. 132-Ex. 141,) No R11 R50 IUPAC name Ex. 132
(557) TABLE-US-00049 TABLE 27a Examples of Core 09 (Ex. 142-Ex. 163,) Yield, Starting General Purification (isolated No R2 R50 material Procedure Reagent Method salt) Ex. 142-Ex. 143: cf. experimental description Ex. 144
(558) TABLE-US-00050 TABLE 27b Examples of Core 09 (Ex. 142-Ex. 163,) Rt Monoisotopic (purity at [M + H].sup.+ LC-MS- No R2 R50 Formula Mass 220 nm) found Method Ex. 142-Ex. 143: cf. experimental description Ex. 144
(559) TABLE-US-00051 TABLE 27c Examples of Core 09 (Ex. 142-Ex. 163,) No R2 R50 IUPAC name Ex. 142
(560) TABLE-US-00052 TABLE 28a Examples of Core 10 (Ex. 164-Ex. 180,) Yield, Starting General Purification (isolated No R2 R5 material Procedure Reagent Method salt) Ex. 164-Ex. 165: cf. experimental description Ex. 166
(561) TABLE-US-00053 TABLE 28b Examples of Core 10 (Ex. 164-Ex. 180,) Rt Monoisotopic (purity at [M + H].sup.+ LC-MS- No R2 R5 Formula Mass 220 nm) found Method Ex. 164-Ex. 165: cf. experimental description Ex. 166
(562) TABLE-US-00054 TABLE 28c Examples of Core 10 (Ex. 164-Ex. 180,) No R2 R5 IUPAC name Ex. 164
(563) TABLE-US-00055 TABLE 29a Examples of Core 11 (Ex. 181-Ex. 195) Yield, Starting General Purification (isolated No R2 R5 material Procedure Reagent Method salt) Ex. 181-Ex. 182: cf. experimental description Ex. 183
(564) TABLE-US-00056 TABLE 29b Examples of Core 11 (Ex. 181-Ex. 195, continued on the following pages) Mono- Rt isotopic (purity at [M + H].sup.+ LC-MS- No R2 R5 Formula Mass 220 nm) found Method Ex. 181-Ex. 182: of. experimental description Ex. 183
(565) TABLE-US-00057 TABLE 29c Examples of Core 11 (Ex. 181-Ex. 195, continued on the following pages) No R2 R5 IUPAC name Ex. 181
(566) TABLE-US-00058 TABLE 30a Examples of Core 12 (Ex. 196-Ex. 214 continued on the following pages) General Yield, Starting Pro- Purification (isolated No R2 R5 material cedure Reagent Method salt) Ex. 196-Ex. 198: cf. experimental description Ex. 199
(567) TABLE-US-00059 TABLE 30b Examples of Core 12 (Ex. 196-Ex. 214, continued on the following pages) Monoisotopic Rt (purity at [M + H].sup.+ LC-MS- No R2 R5 Formula Mass 220 nm) found Method Ex. 196-Ex. cf.198: experimental description Ex. 199
(568) TABLE-US-00060 TABLE 30c Examples of Core 12 (Ex. 196-Ex. 214, continued on the following pages) No R2 R5 IUPAC name Ex. 196
(569) TABLE-US-00061 TABLE 31a Examples of Core 13 (Ex. 215-Ex. 230, continued on the following pages) Yield, Starting General Purification (isolated No R2 R5 material Procedure Reagent Method salt) Ex. 215-Ex. 216: cf experimental description Ex. 217
(570) TABLE-US-00062 TABLE 31b Examples of Core 13 (Ex. 215-Ex. 230, continued on the following pages) Monoisotopic Rt (purity at [M + H].sup.+ LC-MS- No R2 R5 Formula Mass 220 nm) found Method Ex. 215-Ex. 216: cf. experimental description Ex. 217
(571) TABLE-US-00063 TABLE 31c Examples of Core 13 (Ex. 215-Ex. 230) No R2 R5 IUPAC name Ex. 215
(572) TABLE-US-00064 TABLE 32a Examples of Core 14 (Ex. 231-Ex. 237) Yield, Starting General Purification (isolated No R2 R5 material Procedure Reagent Method salt) Ex. 231-Ex. 232: cf. experimental description Ex. 233
(573) TABLE-US-00065 TABLE 32b Examples of Core 14 (Ex. 231-Ex. 237) Monoisotopic Rt (purity at [M + H].sup.+ LC-MS- No R2 R5 Formula Mass 220 nm) found Method Ex. 231-Ex. 232: cf. experimental description Ex. 233
(574) TABLE-US-00066 TABLE 32c Examples of Core 14 (Ex. 231-Ex. 237) No R2 R5 IUPAC name Ex. 231
(575) TABLE-US-00067 Table 33a Examples of Core 15 and Core 16 (Ex. 238-Ex. 247) Yield, Starting General Purification (isolated No R2 R5 material Procedure Reagent Method salt) Ex. 238-Ex. 239: cf. experimental description Ex. 240
(576) TABLE-US-00068 TABLE 33b Examples of Core 15 and Core 16 (Ex. 238-Ex. 247) Monoisotopic Rt (purity at [M + H].sup.+ LC-MS- No R2 R5 Formula Mass 220 nm) found Method Ex. 238-Ex. 239: cf. experimental description Ex. 240
(577) TABLE-US-00069 TABLE 33c Examples of Core 15 and Core 16 (Ex. 238-Ex. 247) No R2 R5 IUPAC name Core 15 Ex. 238
(578) TABLE-US-00070 TABLE 34a Examples of Core 17 (Ex. 248-Ex. 271) Yield, Starting General Purification (isolated No R2 R50 material Procedure Reagent Method salt) Ex. 248-Ex. 249: cf. experimental description Ex. 250
(579) TABLE-US-00071 TABLE 34b Examples of Core 17 (Ex. 248-Ex. 271) Monoisotopic Rt (purity [M + H].sup.+ LC-MS- No R2 R50 Formula Mass at 220 nm) found Method Ex. 248-Ex. 249: cf. experimental description Ex. 250
(580) TABLE-US-00072 TABLE 34c Examples of Core 17 (Ex. 248-Ex. 271) No R2 R50 IUPAC name Ex. 248
(581) TABLE-US-00073 TABLE 35a Examples of Core 18 (Ex. 272-Ex. 296) Yield, Starting General Purification (isolated No R2 R5 material Procedure Reagent Method salt) Ex. 272-Ex. 274: cf. experimental description Ex. 275
(582) TABLE-US-00074 TABLE 35b Examples of Core 18 (Ex. 272-Ex. 296) Monoisotopic Rt (purity at [M + H].sup.+ LC-MS- No R2 R5 Formula Mass 220 nm) found Method Ex. 272-Ex. 274: cf. experimental description Ex. 275
(583) TABLE-US-00075 TABLE 35c Examples of Core 18 (Ex. 272-Ex. 296) No R2 R5 IUPAC name Ex. 272
(584) TABLE-US-00076 TABLE 36a Examples of Core 19 (Ex. 297-Ex. 310) Yield, Starting General Purification (isolated No R2 R50 material Procedure Reagent Method salt) Ex. 297-Ex. 298: cf. experimental description Ex. 299
(585) TABLE-US-00077 TABLE 36b Examples of Core 19 (Ex. 297-Ex. 310) Rt Monoisotopic (purity at [M + H] .sup.+ LC-MS- No R2 R5 Formula Mass 220 nm) found Method Ex. 297-Ex. 298: cf. experimental description Ex. 299
(586) TABLE-US-00078 TABLE 36c Examples of Core 19 (Ex. 297-Ex. 310) No R2 R5 IUPAC name Ex. 297
(587) TABLE-US-00079 TABLE 37 Examples of Core 20 and Core 21 (Ex. 311-Ex. 313) No Core 20 R2 R5 R38 IUPAC name Ex. 311
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Biological Methods
1. Preparation of the Example Compounds
(617) Example compounds were weighed on a Microbalance (Mettler MX5) and dissolved in 100% DMSO to a final concentration of 2.5 mM for Ca.sup.2+ assays.
(618) Example compounds were dissolved in DMSO/H.sub.2O 90:10 to a final concentration of 10 mM for plasma stability determination and metabolic stability determination.
(619) 2. Ca.sup.2+ Assays: GPCR Assays for Motilin Receptor, Prostaglandin F (FP) Receptor and 5-Hydroxytryptamine 2B (5-HT.sub.2B) Receptor
(620) Assays were performed using a FLIPR Tetra (Molecular Devices); the data analysis and FLIPR Tetra Operating-Soft ware was ScreenWorks version 2 (Molecular Devices).
(621) Dose dependent agonist and antagonist activities were determined. Percentage activation and percentage inhibition values were determined.
(622) Percentage activation was determined upon initial addition of the sample compounds followed by 10 minutes incubation at 25 C. Following compound incubation, reference agonists were added at EC.sub.80 to determine percentage inhibition.
(623) Reference agonists were purchased from reputable commercial vendors and prepared according to specifications specific to each ligand. All handling of ligands were done to ensure proper control throughout the experiments.
(624) Test compounds were serially diluted with DMSO. Once the appropriate concentrations were attained, the compounds were diluted into assay buffer.
(625) GPCR Assay Buffer:
(626) Assay buffer was a supplemented HBSS (Hank's Balanced Salt Solution). HBSS was supplemented with 20 mM HEPES (4-(2-hydroxyethyl)-piperazin-1-ethansulfonic acid) and 2.5 mM Probenecid (Sigma P8761).
(627) Assay Plate Seeding:
(628) GPCR assays were performed using Ca.sup.2+ optimized hematopoietic cell lines (rat) with cultures never exceeding 90% confluency. Cells were harvested and seeded (from cultures at less than 90% confluency) at 50000 cells/well for a 96-well plate (12500 cells/well for 384). After seeding, the assay plates were incubated for forty-five (45) minutes at room temperature. After room temperature incubation, the assay plates were incubated at 37 C. 5% CO.sub.2 for 24 hours prior to assaying.
(629) Calcium Dye Loading:
(630) All GPCR assays were performed using Fluo-8 Ca.sup.2+ dye. Ca.sup.2+ dye was prepared at 1 dye concentration in GPCR assay buffer. After 24 hours of incubation, cells were washed with GPCR assay buffer, then Ca.sup.2+-dye (100 L/well) was added. The plates were incubated for 90 minutes at 30 C. 5% CO.sub.2 prior to FLIPR assay.
(631) Agonist Assay:
(632) Compound plates were prepared to add 50 L/well during the agonist assay mode. During the FLIPR assay, 50 L/well from the compound plate was diluted 3-fold into the existing 100 L/well from the dye loading step. Therefore all compounds were prepared as 3 the final concentration desired in the assay.
(633) After completion of the first single addition assay run, assay plate was removed from the FLIPR Tetra and placed at 25 C. for seven (7) minutes before antagonist assay.
(634) Antagonist Assay:
(635) Using the EC.sub.80 values determined during the agonist assay, all pre-incubated sample compound and reference antagonist (if applicable) wells were stimulated with EC.sub.80 of reference agonist (motilin; prostaglandin F2).
(636) After the addition of the reference agonist fluorescence was monitored for 180 sec using FLIPR Tetra.
(637) Data Analysis:
(638) From the FLIPR data, with negative control correction enabled, the maximum statistic for each well was exported and percentage activation relative to E.sub.max control was calculated.
(639) 3. Plasma Stability
(640) Human plasma (3-5 donors, Blutspendedienst SRK, Basel) and CD-1 mouse plasma (mixed gender pool>50 animals, Innovative Research, CA, USA) are both sodium citrate stabilized. The assay is performed in triplicates at 10 M compound concentration and 37 C. Samples are taken at 0, 15, 60, and 240 minutes and stopped by precipitation with 2 volumes of acetonitrile. The supernatant is collected, evaporated and reconstituted in a 5% acetonitrile solution to be analyzed by HPLC/MS/MS. The resulting peak area counts are expressed in percent of the 0 value and used to determine the endpoint stability in % and the half life T in minutes. In order to monitor assay integrity the degradation of propantheline is assayed with every experimental set
(641) 4. Metabolic Stability
(642) Microsomes from a human 50 donor mixed gender pool and 1:1 mixtures of microsomes from CD-1 mouse single-gender pools are purchased from Celsis (Belgium). The enzymatic reaction is performed in a buffer containing an NADPH regeneration system and microsomes with the following end concentrations: 100 mM potassium phosphate buffer (all from Sigma), 1 mg/mL glucose-6-phosphate, 1 mg/mL -nicotinamide adenine dinucleotide phosphate (NADP), 0.65 mg/mL magnesium chloride, 0.8 units/mL of glucose-6-phosphate dehydrogenase (prediluted with 5 mM citrate buffer), 10 M compound and 1 mg/ml microsomal protein. Compounds are incubated at 37 C. in duplicates and samples are taken after 0, 20 and 60 minutes. After acetonitrile precipitation (2 volumes) and HPLC/MS/MS analysis metabolic turnover is expressed in % of the initial 0 minutes value and half life T (min) is calculated. Verapamil for human and propranolol for mouse are used as reference and are assayed with every experimental set. F. P. Guengerich, Analysis and Characterization of Enzymes; in: Principles and Methods of Toxicology; A. W. Hayes (Ed.) Raven Press: New York, 1989, 777-813. R. Singh et al., In vitro metabolism of a potent HIV-protease inhibitor (141W94) using rat, monkey and human liver S9, Rapid Commun. Mass Spectrom. 1996, 10, 1019-1026.
5. Results
(643) The results of the experiments described under 1.-4. (above) are indicated in Table 38 and Table 39 herein below.
(644) TABLE-US-00080 TABLE 38 Biological Data Motilin receptor FP receptor 5-HT.sub.2B receptor antagonist activity Motilin receptor antagonist activity FP receptor agonist activity 5-HT.sub.2B receptor [% inhibition antagonist activity [% inhibition antagonist activity [% activation agonist activity No at 10 M] IC.sub.50 [M]] at 10 M] IC.sub.50 [M] at 12.5 M] EC.sub.50[M] Ex. 9 n.d. n.d. n.d. n.d. 48 12 Ex. 11 79 0.78 n.d. n.d. n.d. n.d. Ex. 12 95 2.7 44 n.d. 36 3.3 Ex. 16 n.d. n.d. n.d. n.d. 45 6.6 Ex. 30 n.d. n.d. n.d. n.d. 48 3.3 Ex. 49 98 0.16 n.d. n.d. n.d. n.d. Ex. 184 n.d. n.d. 74 0.52 n.d. n.d. Ex. 200 n.d. n.d. 38 28 n.d. n.d. Ex. 213 n.d. n.d. 78 1.7 n.d. n.d. n.d. not determined
(645) TABLE-US-00081 TABLE 39 Plasma Stability and Metabolic Stability Plasma Stability Metabolic Stability T [min] 240 min T min] 240 min T [min] 60 min T [min] 60 min No hum hum mouse mouse hum hum mouse mouse Ex. 9 240 99 240 93 32 20 60 80 Ex. 11 240 100 240 100 60 74 60 77 Ex. 12 240 99 240 100 17 7 35 33 Ex. 16 240 95 240 97 38 31 60 79 Ex. 30 240 85 240 100 22 2 60 55 Ex. 37 240 77 240 100 60 100 60 100 Ex. 43 240 82 240 89 60 100 60 98 Ex. 49 240 83 240 96 24 10 60 83 Ex. 78 240 65 240 100 60 78 60 100 Ex. 91 240 96 240 88 60 91 60 95 Ex. 93 240 100 240 100 24 1 29 15 Ex. 95 240 100 240 93 60 76 60 94 Ex. 98 240 100 240 78 60 97 60 100 Ex. 102 240 65 240 75 23 4 42 39 Ex. 103 240 97 240 75 35 22 36 25 Ex. 138 240 89 240 78 60 62 60 99 Ex. 184 240 66 240 58 15 0 22 0 Ex. 200 240 91 240 100 27 12 30 26 Ex. 208 240 98 240 90 60 84 60 100 Ex. 213 n.d. n.d. n.d. n.d. 16 0 17 4 Ex. 230 240 100 240 95 60 61 60 90 Ex. 260 240 100 240 100 37 19 60 69 Ex. 262 240 100 240 93 21 0 23 4 Ex. 264 240 94 240 88 n.d. n.d. 42 32 Ex. 266 240 92 240 74 60 100 60 98 Ex. 267 240 75 240 79 60 99 60 100 Ex. 272 240 95 240 92 60 65 60 81 n.d. not determined