WO2003054000A1 - Template-fixed peptidomimeticsas inhibitors of serine proteases - Google Patents

Template-fixed peptidomimeticsas inhibitors of serine proteases Download PDF

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Publication number
WO2003054000A1
WO2003054000A1 PCT/EP2001/014528 EP0114528W WO03054000A1 WO 2003054000 A1 WO2003054000 A1 WO 2003054000A1 EP 0114528 W EP0114528 W EP 0114528W WO 03054000 A1 WO03054000 A1 WO 03054000A1
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Prior art keywords
lower alkyl
chr
alkenyl
alkyl
lower alkenyl
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PCT/EP2001/014528
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French (fr)
Inventor
Daniel Obrecht
John Anthony Robinson
Anne Descours
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Polyphor Ltd.
Universität Zürich
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Priority to CNA2007101543999A priority Critical patent/CN101157924A/en
Priority to CNA018238807A priority patent/CN1582296A/en
Priority to BRPI0117190A priority patent/BRPI0117190B1/en
Priority to PCT/EP2001/014528 priority patent/WO2003054000A1/en
Priority to CN2007101537108A priority patent/CN101157720B/en
Priority to AU2002234564A priority patent/AU2002234564C1/en
Priority to DE60137654T priority patent/DE60137654D1/en
Priority to BR0117190-9A priority patent/BR0117190A/en
Application filed by Polyphor Ltd., Universität Zürich filed Critical Polyphor Ltd.
Priority to AT01985390T priority patent/ATE422501T1/en
Priority to CA2466591A priority patent/CA2466591C/en
Priority to JP2003554716A priority patent/JP4098244B2/en
Priority to EP01985390A priority patent/EP1458747B1/en
Priority to US10/498,468 priority patent/US7417024B2/en
Publication of WO2003054000A1 publication Critical patent/WO2003054000A1/en
Priority to HK08107766.9A priority patent/HK1117541A1/en

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/04—Linear peptides containing only normal peptide links
    • C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/04—Linear peptides containing only normal peptide links
    • C07K7/08—Linear peptides containing only normal peptide links having 12 to 20 amino acids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/64—Cyclic peptides containing only normal peptide links
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • the present invention provides template-fixed ⁇ -haiipin peptidomimetics incorporating a template-fixed chain of 7 or 11 ⁇ -amino acid residues which, depending on their position in the chain, are Gly, or Pro, or of certain types, as defined hereinbelow.
  • These template-fixed ⁇ -hairpin peptidomimetics are useful as inhibitors of protease enzymes. They are especially valuable as inhibitors of various serine proteases such as trypsin, human cathepsin G, and thrombin.
  • the present invention provides an efficient process by which these compounds can, if desired, be made in library-format. This library-approach constitutes an efficient novel tool to identify specific serine protease inhibitors.
  • Inhibitors of proteases are emerging with promising therapeutic uses in the treatment of diseases such as cancers (R. P. Beckett, A. Davidson, A. H. Drummond, M. Whittaker, Drug Disc. Today 1996, 1, 16-26; L. L. Johnson, R. Dyer, D. J. Hupe, Curr. Opin. Chem. Biol. 1998, 2, 466-71; D. Leung, G. Abbenante, and D. P. Fairlie, J. Med. Chem. 2000, 43, 305-341), parasitic, fungal, and viral infections [e.g. schistosomiasis (M. M. Becker, S. A. Harrop, J. P. Dalton, B. H. Kalinna, D. P.
  • Serine proteases constitute important therapeutic targets. Serine proteases are classified by their substrate specificity, particularly by the type of residue found at P 1 , as either trypsin-like (positively charged residues Lys/Arg preferred at PI), e-f ⁇ -st ⁇ - ⁇ e-like (small hydrophobic residues Ala/Val at PI), or chymotrypsin-like (large hydrophobic residues Phe/Tyr/Leu at PI).
  • Serine proteases for which protease-inhibitor X-ray crystal data is available on the PDB data base include trypsin, a-chymotrypsin, ⁇ - chymotrypsin, human neutrophil elastase, thrombin, subtilisin, human cytomegalovirus, proteinase A, achromobacter, human cathepsin G, glutamic acid-specific protease, carbopeptidase D, blood coagulation factorVIIa, porcine factor 1XA, mesentericopeptidase, HCV protease, and thermitase.
  • serine proteases which are of therapeutic interest include tryptase, complement convertase, hepatitis C-NS3 protease.
  • Inhibitors of thrombin e.g. J. L. Metha, L. Y. Chen, W. W. Nichols, C. Mattsson, D. Gustaffson, T. G. P. Saldeen, J. Cardiovasc. Pharmacol. 1998, 31, 345-51; C. Lila, P. Gloanec, L. Cadet, Y. Herve, J. Fournier, F. Leborgne, T. J. Verbeuren, G. DeNanteuil, Synth. Comm. 1998, 28, 4419-29) and/ ⁇ ctorZ ⁇ (e.g. J. J.
  • cathepsin G and elastase are intimately involved in the modulation of activities of cytokines and their receptors. Particularly at sites of inflammation, high concentration of cathepsin G, elastase and proteinase 3 are released from infiltrating polymorphonuclear cells in close temporal correlation to elevated levels of inflammatory cytokines, strongly indicating that these proteases are involved in the control of cytokine bioactivity and availability (U. Bank, S. Ansorge, J. Leukoc. Biol. 2001, 69, 177-90). Thus inhibitors of thrombin and cathepsin G constitute valuable targets for novel drug candidates.
  • sunflower trypsin inhibitor S. Luckett, R. Santiago Garcia, J. J. Barker, A. V. Konarev, P. R. Shewry, A. R. Clarke, R. L. Brady, /. Mol. Biol. 1999, 290, 525-533; Y.-Q. Long, S.-L. Lee, C.-Y. Lin, I. J. Enyedy, S. Wang, P. Li, R. B. Dickson, P. P. Roller, Biorg. & Med. Chem. Lett.
  • SFTI-1 sunflower trypsin inhibitor
  • SFTI-1 inhibited ⁇ -trypsin (Ki ⁇ 0.1nM), cathepsin G, elastase (Kj ⁇ 105 ⁇ M), chymottypsin (K; ⁇ 7.4 ⁇ M) and thrombin (Kj ⁇ 136mM).
  • ⁇ -hairpin peptidomimetics of the present invention are compounds of the general formula
  • R > 1 is- H; lower alkyl; or aryl-lower alkyl;
  • R 2 is H; alkyl; alkenyl; -(CH 2 ) m (CHR 6I ) s 0R 55 ; -(CH 2 ) m (CHR 61 ) s SR 56 ; -
  • R 3 is H; alkyl; alkenyl; -(CH 2 ) m (CHR 61 ) s OR 55 ; -(CH 2 ) m (CHR 61 ) s SR 56 ; -
  • R 4 is H; alkyl; alkenyl; -(CH 2 ) m (CHR 61 ) s OR 55 ; -(CH 2 ) m (CHR 61 ) s SR 56 ; -
  • R 5 is alkyl; alkenyl; -(CH 2 ) 0 (CHR 61 ) s OR 55 ; -(CH 2 ) 0 (CHR 61 ) S SR 56 ; -(CH 2 ) 0 (CHR 61 ) S NR 33 R 34 ; -
  • R 6 is H; alkyl; alkenyl; -(CH 2 ) 0 (CHR 61 ) s 0R 55 ; -(CH 2 ) 0 (CHR 61 ) S SR 56 ; -(CH 2 ) 0 (CHR 61 ) S NR 33 R 34 ; -(CH 2 ) o (CHR ⁇ l ) s COOR 57 ; -(CH 2 ) 0 (CHR 61 ) s CONR 58 R 59 ; -(CH 2 ) o (CHR 61 ) s PO(OR 60 ) 2 ;
  • R 7 is alkyl; alkenyl; -(CH 2 ) q (CHR 6I ) s OR 55 ; -(CH 2 ) q (CHR 61 ) s NR 33 R 34 ; -(CH 2 ) r (CHR 61 ) s COOR 57 ; -(CH 2 ) r (CHR ⁇ I ) s CONR 58 R 59 ; -(CH 2 ) r (CHR ⁇ l ) s S0 2 R 62 ; or R 8 is H; Cl; F; CF 3 ; N0 2 ; lower alkyl; lower alkenyl; aryl; aryl-lower alkyl; -
  • R 9 is alkyl; alkenyl; -(CH 2 ) 0 (CHR 61 ) S OR 55 ; -(CH 2 ) 0 (CHR ⁇ I ) s SR 5 ⁇ ; -(CH 2 ) 0 (CHR ⁇ l ) s NR 33 R 34 ;
  • R 10 is alkyl; alkenyl; -(CH 2 ) 0 (CHR ⁇ l ) s OR 55 ; -(CH 2 ) 0 (CHR 61 ) s SR 5 ⁇ ; -(CH 2 ) 0 (CHR ⁇ l ) s NR 33 R 34 ;
  • R n is H; alkyl; alkenyl; -(CH 2 ) m (CHR ⁇ l ) s OR 55 ; -(CH 2 ) m (CHR 61 ) s SR 5 ⁇ ; - (CH 2 ) m (CHR ⁇ I ) s NR 33 R 34 ; -(CH 2 ) m (CHR 6I ) s OCONR 33 R 78 ; -
  • R 12 is H; alkyl; alkenyl; -(CH 2 ) m (CHR ⁇ l ) s OR 55 ; -(CH 2 ) m (CHR ⁇ l ) s SR 5 ⁇ ; -
  • R 13 is alkyl; alkenyl; -(CH 2 ) q (CHR ⁇ l ) s 0R 55 ; -(CH 2 ) q (CHR ⁇ , ) s SR 5 ⁇ ; -(CH 2 ) q (CHR ⁇ l ) s NR 33 R 34 ;
  • R 14 is H; alkyl; alkenyl; -(CH 2 ) m (CHR 61 ) s OR 55 ; -(CH 2 ) m (CHR 61 ) s NR 33 R 34 ; - (CH 2 ) q (CHR 61 ) s COOR 57 ; -(CH 2 ) q (CHR ⁇ l ) s CONR 58 R 59 ; -(CH 2 ) q (CHR ⁇ l ) s PO(OR ⁇ 0 ) 2 ; -
  • R 15 is alkyl; alkenyl; -(CH 2 ) 0 (CHR ⁇ l ) s OR 55 ; -(CH 2 ) 0 (CHR ⁇ l ) s SR 56 ; -(CH 2 ) 0 (CHR ⁇ l ) s NR 33 R 34 ;
  • R 16 is alkyl; alkenyl; -(CH 2 ) 0 (CHR ⁇ I ) s 0R 55 ; -(CH 2 ) 0 (CHR 61 ) S SR 56 ; -(CH 2 ) 0 (CHR ⁇ l ) s NR 33 R 34 ;
  • R' 7 is alkyl; alkenyl; -(CH 2 ) m (CHR ⁇ I ) s OR 55 ; -(CH 2 ) m (CHR ⁇ I ) s SR 56 ; -(CH 2 ) m (CHR ⁇ l ) s NR 33 R 34 ;
  • R 18 is alkyl; alkenyl; -(CH 2 ) p (CHR 6I ) s OR 55 ; -(CH 2 ) p (CHR ⁇ I ) s SR 5 ⁇ ; -(CH 2 ) P (CHR 61 ) S NR 33 R 34 ;
  • R 19 is lower alkyl; -(CH 2 ) p (CHR ⁇ l ) s OR 55 ; -(CH 2 ) P (CHR 6I ) S SR 56 ; -(CH 2 ) p (CHR ⁇ l ) s NR 33 R 34 ;
  • -(CH 2 ) P (CHR 61 ) S S0 2 R 62 ; or -(CH 2 ) 0 (CHR 61 ) S C 6 H 4 R 8 ; or R 18 and R 19 taken together can form: -(CH 2 ) 2 . 6 -; -(CH 2 ) 2 0(CH 2 ) 2 -; -(CH 2 ) 2 S(CH 2 ) 2 -; or
  • R 20 is H; alkyl; alkenyl; or aryl-lower alkyl;
  • R 21 is H; alkyl; alkenyl; -(CH 2 ) 0 (CHR ⁇ I ) s OR 55 ; -(CH 2 ) 0 (CHR 6I ) S SR 56 ; -(CH 2 ) 0 (CHR 6I ) S NR 33 R 34 ;
  • R 22 is H; alkyl; alkenyl; -(CH 2 ) 0 (CHR ⁇ !
  • R 23 is alkyl; alkenyl; -(CH 2 ) 0 (CHR 61 ) s OR 55 ; -(CH 2 ) 0 (CHR 61 ) s SR 5 ⁇ ; -(CH 2 ) 0 (CHR 61 ) S NR 33 R 34 ;
  • R 24 is alkyl; alkenyl; -(CH 2 ) 0 (CHR 61 ) S 0R 55 ; -(CH 2 ) 0 (CHR ⁇ l ) s SR 56 ; -(CH 2 ) 0 (CHR ⁇ l ) s NR 33 R 34 ;
  • R 25 is H; alkyl; alkenyl; -(CH 2 ) m (CHR 61 ) s OR 55 ; -(CH 2 ) m (CHR 61 ) s SR 5 ⁇ ; -
  • R 2 ⁇ is H; alkyl; alkenyl; -(CH 2 ) m (CHR 61 ) s OR 55 ; -(CH 2 ) m (CHR ⁇ l ) s SR 5 ⁇ ; -
  • R 27 is H; alkyl; alkenyl; -(CH 2 ) 0 (CHR ⁇ l ) s OR 55 ; -(CH 2 ) 0 (CHR ⁇ l ) s SR 5 ⁇ ; -(CH 2 ) 0 (CHR ⁇ l ) s NR 33 R 34 ;
  • R 28 is alkyl; alkenyl; -(CH 2 ) 0 (CHR 6I ) S -OR 55 ; - ⁇ x ⁇ 2 0 ⁇ -dR 61 ) s SR 5 ⁇ ; -(CH 2 ) 0 (CHR 61 ) S NR 33 R 34 ;
  • R 29 is alkyl; alkenyl; -(CH 2 ) 0 (CHR 6I ) S OR 55 ; -(CH 2 ) 0 (CHR ⁇ l ) s SR 5 ⁇ ; -(CH 2 ) 0 (CHR ⁇ l ) s NR 33 R 34 ; -(CH 2 ) o (CHR ⁇ I ) s COOR 57 ; -(CH 2 ) 0 (CHR ⁇ l ) s CONR 58 R 59 ; -(CH 2 ) o (CHR 6I ) s PO(OR 60 ) 2 ;
  • R 30 is H; alkyl; alkenyl; or aryl-lower alkyl; R 31 is H; alkyl; alkenyl; -(CH 2 ) p (CHR ⁇ l ) s OR 55 ; -(CH 2 ) p (CHR ⁇ l ) s NR 33 R 34 ; -
  • R 32 is H; lower alkyl; or aryl-lower alkyl; R 33 is H; alkyl, alkenyl; -(CH 2 ) m (CHR 61 ) s 0R 55 ; -(CH 2 ) m (CHR ⁇ l ) s SR 5 ⁇ ; -
  • R 34 is H; lower alkyl; aryl, or aryl-lower alkyl;
  • R 35 is H; alkyl; alkenyl; -(CH 2 ) ra (CHR ⁇ l ) s 0R 55 ; -(CH 2 ) m (CHR 61 ) s NR 33 R 34 ; -
  • R 36 is H, alkyl; alkenyl; -(CH 2 ) 0 (CHR ⁇ l ) s OR 55 ; -(CH 2 ) p (CHR ⁇ l ) s NR 33 R 34 ; -
  • R 37 is H; F; Br; Cl; N0 2 ; CF 3 ; lower alkyl; -(CH 2 ) p (CHR 6I ) s OR 55 ; -(CH 2 ) p (CHR ⁇ l ) s NR 33 R 34 ;
  • R 38 is H; F; Br; Cl; N0 2 ; CF 3 ; alkyl; alkenyl; -(CH 2 ) p (CHR ⁇ I ) s OR 55 ; -(CH 2 ) p (CHR ⁇ l ) s NR 33 R 34 ;
  • R 39 is H; alkyl; alkenyl; or aryl-lower alkyl; R 40 is H; alkyl; alkenyl; or aryl-lower alkyl;
  • R 41 is H; F; Br; Cl; N0 2 ; CF 3 ; alkyl; alkenyl; -(CH 2 ) P (CHR ⁇ l ) s OR 55 ; -(CH 2 ) P (CHR 61 ) S NR 33 R 34 ;
  • R 42 is H; F; Br; Cl; N0 2 ; CF 3 ; alkyl; alkenyl; -(CH 2 ) p (CHR 61 ) s OR 55 ; -(CH 2 ) p (CHR ⁇ I ) s NR 33 R 34 ; -(CH 2 ) 0 (CHR ⁇ l ) s COOR 57 ; -(C_i 2 ) 0 (C ⁇ ss. -(CH 2 ) o (CHR ⁇ l ) s PO(OR ⁇ 0 ) 2 ;
  • R 43 is H; alkyl; alkenyl; -(CH 2 ) m (CHR ⁇ l ) s OR 55 ; -(CH 2 ) m (CHR ⁇ l ) s NR 33 R 34 ; -
  • R 44 is alkyl; alkenyl; -(CH 2 ) r (CHR ⁇ I ) s OR 55 ; -(CH 2 ) r (CHR ⁇ I ) s SR 5 ⁇ ; -(CH 2 ) r (CHR 6I ) s NR 33 R 34 ;
  • R 45 is H; alkyl; alkenyl; -(CH 2 ) 0 (CHR ⁇ l ) s OR 55 ; -(CH 2 ) 0 (CHR ⁇ l ) s SR 5 ⁇ ; -(CH 2 ) 0 (CHR ⁇ l ) s NR 33 R 34 ; -(CH 2 ) o (CHR ⁇ I ) s COOR 57 ; -(CH 2 ) s (CHR 6I ) s CONR 58 R 59 ; -(CH 2 ) s (CHR ⁇ , ) s PO(OR ⁇ 0 ) 2 ;
  • R 4 ⁇ is H; alkyl; alkenyl; or -(CH 2 ) 0 (CHR 61 ) P C 6 H 4 R 8 ;
  • R 47 is H; alkyl; alkenyl; or -(CH 2 ) 0 (CHR ⁇ I ) s 0R 55 ;
  • R 48 is H; lower alkyl; lower alkenyl; or aryl-lower alkyl;
  • R 49 is H; alkyl; alkenyl; -(CHR ⁇ l ) s C00R 57 ; (CHR ⁇ l ) s CONR 58 R 59 ; (CHR ⁇ l ) s PO(OR 60 ) 2 ;
  • R 50 is H; lower alkyl; or aryl-lower alkyl; R 51 is H; alkyl; alkenyl; -(CH 2 ) m (CHR ⁇ l ) s OR 55 ; -(CH 2 ) m (CHR ⁇ l ) s SR 56 ; -
  • R 52 is H; alkyl; alkenyl; -(CH 2 ) m (CHR ⁇ I ) s OR 55 ; -(CH 2 ) m (CHR ⁇ l ) s SR 5 ⁇ ; -
  • R 53 is H; alkyl; alkenyl; -(CH 2 ) m (CHR ⁇ l ) s OR 55 ; -(CH 2 ) m (CHR ⁇ l ) s SR 5 ⁇ ; -
  • R 54 is H; alkyl; alkenyl; -(CH 2 ) m (CHR 61 ) s OR 55 ; -(CH 2 ) ra (CHR ⁇ I ) s NR 33 R 34 ; - (CH 2 ) 0 (CHR ⁇ I )COOR 57 ; -(CH 2 ) 0 (CHR ⁇ , ) s CONR 58 R 59 ; or -(CH 2 ) 0 (CHR ⁇ l ) s C 6 H 4 R 8 ;
  • R 55 is H; lower alkyl; lower alkenyl; aryl-lower alkyl; -(CH 2 ) m (CHR ⁇ l ) s OR 57 ;
  • R 5 ⁇ is H; lower alkyl; lower alkenyl; aryl-lower alkyl; -(CH 2 ) m (CHR ⁇ l ) s OR 57 ; -(CH 2 ) m (CHR ⁇ l ) s NR 34 R 63 ; or -(CH 2 ) 0 (CHR ⁇ l ) s CONR 58 R 59 ;
  • R 57 is H; lower alkyl; lower alkenyl; aryl lower alkyl; or heteroaryl lower alkyl;
  • R 58 is H; lower alkyl; lower alkenyl; aryl; heteroaryl; aryl-lower alkyl; or heteroaryl-lower alkyl;
  • R 59 is H; lower alkyl; lower alkenyl; aryl; heteroaryl; aryl-lower alkyl; or heteroaryl-lower alkyl;
  • R 59 is H; lower alkyl; lower al
  • R 60 is H; lower alkyl; lower alkenyl; aryl; or aryl-lower alkyl; R ⁇ l is alkyl; alkenyl; aryl; heteroaryl; aryl-lower alkyl; heteroaryl-lower alkyl; -(CH 2 ) m OR 55 ;
  • R 62 is lower alkyl; lower alkenyl; aryl, heteroaryl; or aryl-lower alkyl; R 63 is H; lower alkyl; lower alkenyl; aryl, heteroaryl; aryl-lower alkyl; heteroaryl-lower alkyl;
  • R 64 is H; lower alkyl; lower alkenyl; aryl; heteroaryl; aryl-lower alkyl; heteroaryl-lower alkyl;
  • R 65 is H; lower alkyl; lower alkenyl; aryl, aryl-lower alkyl; heteroaryl-lower alkyl; -COR 57 ;
  • -COOR 57 or -CONR 58 R 59 ;
  • R ⁇ is H; lower alkyl; lower alkenyl; aryl; aryl-lower alkyl; heteroaryl-lower alkyl; or - CONR 58 R 59 ;
  • m is 2-4; o is 0-4; p is 1-4; q is 0-2; r is 1 or 2; s is 0 or 1;
  • Z is a chain of n ⁇ -amino acid residues, n being the integer 7 or 11, the positions of said amino acid residues in said chain being counted starting from the N-terminal amino acid, whereby these amino acid residues are, depending on their position in the chains, Gly, or Pro, or of formula -A- CO-, or of one of the types C: -NR 20 CH(R 72 )CO- D: -NR 20 CH(R 73 )CO- E: -NR 20 CH(R 74 )CO- F: -NR 20 CH(R 84 )CO-; and
  • R 71 is H; lower alkyl; lower alkenyl; , s R 75 ; -(CH 2 ) p (CHR ⁇ I ) s SR 75 ; -(CH 2 ) P NR 78 R 79 ;
  • R 72 is H; lower alkyl; lower alkenyl; -(CH 2 ) p (CHR ⁇ l ) s 0R 85 ; or -(CH 2 ) p (CHR ⁇ l ) s SR 85 ;
  • R 73 is -(CH 2 ) 0 R 77 ; -(CH2) r O(CH 2 ) 0 R 77 ; -(CH 2 ) r S(CH 2 ) 0 R 77 ; or -(CH 2 ) r NR 20 (CH 2 ) o R 77 ;
  • R 75 is lower alkyl; lower alkenyl; or aryl-lower alkyl; R 76 is H; lower alkyl; lower alkenyl; aryl-lower alkyl; -(CH 2 ) 0 OR 72 ; -(CH 2 ) 0 SR 72 ; -
  • R 78 is H; lower alkyl; aryl; or aryl-lower alkyl;
  • R 79 is H; lower alkyl; aryl; or aryl-lower alkyl; or
  • R 78 and R 79 taken together, can be -(CH 2 ) 2 . 7 -; -(CH 2 ) 2 0(CH 2 ) 2 -; or -(CH 2 ) 2 NR 33 (CH 2 ) 2 -;
  • R 80 is H; or lower alkyl
  • R 81 is H; lower alkyl; or aryl-lower alkyl;
  • R 82 is H; lower alkyl; aryl; heteroaryl; or aryl-lower alkyl;
  • R w is H; lower alkyl; aryl; or -NR 7'S 8 ⁇ R, 7 / 9 y ;.
  • R 84 is -(CH 2 ) m (CHR 61 ) s OH; -(CH 2 ) p C0NR 78 R 79 ; -(CH 2 ) P NR 80 CONR 78 R 79 ; -(CH 2 ) p C 6 H 4 CONR 78 R 79 ;
  • R 85 is lower alkyl; or lower alkenyl
  • amino acid residues in positions 1 to 7 are: - PI : of type C or of type F or of type D;
  • P2 of type E or of type C or of type D or of type F;
  • P3 of type F or of type C, or the residue is Gly or Pro;
  • P4 of type C or of type D or of type F, or the residue is Gly or Pro;
  • P5 of type F or of formula -A-CO-, or the residue is Gly or Pro
  • P6 of type C or of type E or o ⁇ -luiium- ⁇ -A-CO-, or the residue is Pro
  • P7 of type C or of type F or of type D
  • amino acid residues in positions 1 to 11 are: - PI: of type E or of type F or of type C;
  • P2 of type C or of type F or of type E;
  • P3 oftype C or oftype F;
  • P4 of type E or of type C or of type D or of type F, or the residue is Gly or Pro;
  • P5 of type F or of type C, or the residue is Gly or Pro
  • - P6 oftype C or of type D or of type F, or the residue is Gly or Pro
  • P7 of type F or of formula -A-CO-, or the residue is Gly or Pro;
  • P8 of type C or of type E or of formula -A-CO-, or the residue is Gly or Pro;
  • P9 of type C or of type F;
  • P 10 of type F or of type C
  • - Pl l of type D or of type E or of type F or of type C
  • P2 and P 10 taken together, can form a group of type H;
  • ⁇ -hairpin peptidomimetics can be prepared by a process which comprises
  • X is an N-protecting group or, if
  • Template is to be group (al) or (a2), above, alternatively
  • step (fa) coupling the product obtained in step (d) or (e) with an appropriately N-protected derivative of an amino acid of the general formula HOOC-B-H III or HOOC-A-H IV wherein B and A are as defined above, any functional group which may be present in said N-protected amino acid derivative being likewise appropriately protected;
  • alkyl designates saturated, straight-chain or branched hydrocarbon radicals having up to 24, preferably up to 12, carbon atoms.
  • alkenyl designates straight chain or branched hydrocarbon radicals having up to 24, preferably up to 12, carbon atoms and containing at least one or, depending on the chain length, up to four olefinic double bonds.
  • lower designates radicals and compounds having up to 6 carbon atoms.
  • lower alkyl designates saturated, straight-chain or branched hydrocarbon radicals having up to 6 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl and the like.
  • aryl designates aromatic carbocyclic hydrocarbon radicals containing one or two six-membered rings, such as phenyl or naphthyl, which may be substituted by up to three substituents such as Br, Cl, F, CF 3 , N0 2 , lower alkyl or lower alkenyl.
  • heteroaryl designates aromatic heterocyclic radicals containing one or two five- and/or six-membered rings, at least one of them containing up to three heteroatoms selected from the group consisting of O, S and N and said ring(s) being optionally substituted; representative examples of such optionally substituted heteroaryl radicals are indicated hereinabove in connection with the definition of R 77 .
  • the structural element -A-CO- designates amino acid building blocks which in combination with the structural element -B-CO- form templates (al) and (a2).
  • Templates (a) through (p) constitute building blocks which have an N-terminus and a oriented in space in such a way that the distance between those two groups may lie between 4.0-5.5A.
  • a peptide chain is linked to the C-terminus and the N-terminus of the templates (a) through (p) via the corresponding N- and C- termini so that the template and the chain form a cyclic structure such as that depicted in formula I.
  • template and peptide chain form a ⁇ -hairpin mimetic.
  • the ⁇ -hairpin conformation is highly relevant for the protease inhibitory activities of the ⁇ - hairpin mimetics of the present invention.
  • the ⁇ -hairpin stabilizing conformational properties of the templates (a) through (p) play a key role not only for protease inhibitory activity but also for the synthesis process defined hereinabove, as incorporation of the templates near the middle of the linear protected peptide precursors enhance significantly cyclization yields.
  • Building blocks A1-A69 belong to a class of amino acids wherein the N-terminus is a secondary amine forming part of a ring. Among the genetically encoded amino acids only proline falls into this class..
  • the configuration of building block Al through A69 is (D), and they are combined with a building block -B-CO- of (L)-configuration.
  • Preferred combinations for templates (al) are- D Al-CO- B-CO- to D A69-CO- L B-CO-.
  • D Pro- L Pro constitutes the prototype of templates (al).
  • templates (a2) are - Al-CO- D B-CO- to L A69-CO- D B-CO-.
  • L Pro- D Pro constitutes a less preferred prototype of template (a2).
  • building blocks -Al-CO- to -A69-CO- in which A has (D)- configuration are carrying a group R 1 at the ⁇ -position to the N-terminus.
  • the preferred values for R 1 are H and lower alkyl with the most preferred values for R 1 being H and methyl.
  • A1-A69 are shown in (D)-configuration which, for R 1 being H and methyl, corresponds to the (Reconfiguration.
  • this configuration may also have to be expressed as (S).
  • R 2 building blocks -Al-CO- to -A69-CO- can carry an additional substituent designated as R 2 to R 17 .
  • This additional substituent can be H, and if it is other than H, it is preferably a small to medium-sized aliphatic or aromatic group.
  • R 2 to R 17 are: R 2 : H; lower alkyl; lower alkenyl; (CH 2 , m --. s.
  • R 55 lower alkyl; or lower alkenyl
  • (CH 2 ) m SR 5 ⁇ where R 5 ⁇ : H; or lower alkyl; or lower alkenyl
  • (CH 2 ) m NR 33 R 34 where R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl)
  • (CH 2 ) m OCONR 33 R 78 where R 33 : lower alkyl; or lower alkenyl; R 78 : H; or lower alkyl
  • (CH 2 ) m NR 20 CONR 33 R 78 (where R 20 : H or lower alkyl; R 33 : lower alkyl; or lower alkenyl; R 78 : H; or lower alkyl); (CH 2 ) o N(R 20 )COR ⁇ 4 (where: R 20 : H; or lower alkyl; R 64 : lower alkyl; or lower alkenyl); (CH 2 )
  • R 3 H; lower alkyl; lower alkenyl; (CH 2 ) m OR 55 (where R 55 : lower alkyl; or lower alkenyl); (CH 2 ) m SR 56 (where R 5 ⁇ : H; or lower alkyl; or lower alkenyl); (CH 2 ) m NR 33 R 34 (where R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl) ; (CH 2 ) o N(R 20 )COR 64 (where: R 20 : H; or lower alkyl; R 64 : lower alkyl; or lower alkenyl); (CH 2 ) 0 COOR 57 (where R 57 : lower alkyl; or lower alkenyl); (CH 2 ) 0 CONR 58 R 59 (where R 58 : lower alkyl; or lower alkenyl; and R 59 : H; lower alkyl); (CH 2 ) o PO(OR ⁇
  • R 4 H; lower alkyl; lower alkenyl; (CH 2 ) m OR 55 (where R 55 : lower alkyl; or lower alkenyl); (CH 2 ) m SR 56 (where R 5 ⁇ : H; or lower alkyl; or lower alkenyl); (CH 2 ) m NR 33 R 34 ( where R 33 : lower alkyl; or lower alkenyl; R 34 : Ho r lower alkyl) ; (CH 2 ) ra N(R 20 )COR ⁇ (where: R 20 : H; or lower alkyl; R 64 : lower alkyl; or lower alkenyl); (CH 2 ) 0 COOR 57 (where R 57 : lower alkyl; or lower alkenyl); (CH 2 ) 0 CONR 58 R 59 (where R 58 : lower alkyl; or lower alkenyl; and R 59 : H; or lower alkyl); (CH 2 ) o PO(
  • R s lower alkyl; lower alkenyl; (CH 2 ) 0 OR 55 (where R 55 : lower alkyl; or lower alkenyl); (CH 2 ) 0 SR 5 ⁇ (where R 5 ⁇ : H; or lower alkyl; or lower alkenyl); (CH 2 ) 0 NR 33 R 34 ( where R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl) ; (CH 2 ) m OCONR 33 R 78 (where R 33 : lower alkyl; or lower alkenyl; R 78 : H; or lower alkyl); (CH 2 ) m NR 20 CONR 33 R 78 (where R 20 : H or lower alkyl; R 33 : lower alkyl; or lower alkenyl; R 78 : H; or lower alkyl); (CH 2 ) o N(R 20 )COR ⁇ 4 (where: R 20 : H; or lower alkyl
  • R 6 H; lower alkyl; lower alkenyl; (CH 2 ) 0 OR 55 (where R 55 : lower alkyl; or lower alkenyl); (CH 2 ) 0 SR 56 (where R 56 : H; or lower alkyl; or lower alkenyl); (CH 2 ) 0 NR 3 R 34 (where R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl) ; (CH 2 ) 0 N(R 20 )COR 64 (where: R 20 : H; or lower alkyl; R 64 : lower alkyl; or lower alkenyl); (CH 2 ) 0 COOR 57 (where R 57 : lower alkyl; or lower alkenyl); (CH 2 ) 0 CONR 58 R 59 (where R 58 : lower alkyl; or lower alkenyl; and R 59 : H; or lower alkyl); (CH 2 ) o PO(OR 60
  • R 7 lower alkyl; lower alkenyl; (CH 2 ) q OR 55 (where R 55 : lower alkyl; or lower alkenyl); (CH 2 ) q SR 56 (where R 56 : H or lower alkyl; or lower alkenyl); (CH 2 ) q NR 33 R 34 ( where R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl) ; (CH 2 ) q N(R 20 )COR 64 (where: R 20 : H; or lower alkyl; R 64 : lower alkyl; or lower alkenyl); (CH 2 ) r COOR 57 (where R 57 : lower alkyl; or lower alkenyl); (CH 2 ) q CONR 58 R 59 (where R 58 : lower alkyl; or lower alkenyl; and R 59 : H; or lower alkyl); (CH 2 ) r PO(OR 60 ) 2 (CH
  • R s H; F; Cl; CF 3 ; lower alkyl; lower alkenyl; (CH 2 ) 0 OR 55 (where R 55 : lower alkyl; or lower alkenyl); (CH 2 ) 0 SR 56 (where R 56 : H; or lower alkyl; or lower alkenyl); (CH 2 ) 0 NR 33 R 34 ( where R 33 : lower alkyl; or lower alkenyl; R 34 : H or lower alkyl) ; (CH 2 ) 0 OCONR 33 R 78 (where R 33 : lower alkyl; or lower alkenyl; R 78 : H; or lower alkyl); (CH 2 ) 0 NR 20 CONR 33 R 78 (where R 20 : H or lower alkyl; R 33 : lower alkyl; or lower alkenyl; R 78 : H; or lower alkyl); (CH 2 ) 0 N(R 20 )COR 64 (where: R 20 : H
  • R 9 lower alkyl; lower alkenyl; (CH 2 ) 0 OR 55 (where R 55 : lower alkyl; or lower alkenyl); (CH 2 ) 0 SR 5 ⁇ (where R 56 : H; or lower alkyl; or lower alkenyl); (CH 2 ) 0 NR 33 R 34 ( where R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl) ; (CH 2 ) 0 N(R 20 )COR 64 (where: R 20 : H; or lower alkyl; R 64 : lower alkyl; or lower alkenyl); (CH 2 ) 0 COOR 57 (where R 57 : lower alkyl; or lower alkenyl); (CH 2 ) 0 CONR 5S R 59 (where R 58 : lower alkyl; or lower alkenyl; and R 59 : H; or lower alkyl); (CH 2 ) o PO(OR 60 )
  • R ;o lower alkyl; lower alkenyl; (CH 2 ) 0 OR 55 (where R 55 : lower alkyl; or lower alkenyl); (CH 2 ) 0 SR 56 (where R 56 : H; or lower alkyl; or lower alkenyl); (CH 2 ) 0 NR 33 R 34 ( where R 33 : lower alkyl; or lower alkenyl; R 34 : lower alkenyl); (CH 2 ) 0 COOR 57 (where R 57 : lower alkyl; or lower alkenyl); (CH 2 ) 0 CONR 58 R 59 (where R 58 : lower alkyl; or lower alkenyl; and R 59 : H; lower alkyl); (CH 2 ) 0 PO(OR 60 ) 2 (where R 60 : lower alkyl; or lower alkenyl); (CH 2 ) 0 S0 2 R 62 (where R 62 : lower alkyl; or lower alkenyl);
  • R 13 lower alkyl; lower alkenyl; (CH 2 ) q OR 55 (where R 55 : lower alkyl; or lower alkenyl); (CH 2 ) q SR 56 (where R 56 : H; or lower alkyl; or lower alkenyl); (CH 2 ) q NR 33 R 34 ( where R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl) ; (CH 2 ) q N(R 20 )COR 64 (where: R 20 : H; or lower alkyl; R 64 : lower alkyl; or lower alkenyl); (CH 2 ) r C00 57 (where R 57 : lower alkyl; or lower alkenyl); (CH 2 ) q C0NR 58 R 59 (where R 58 : lower alkyl; or lower alkenyl; and R 59 : H; or lower alkyl); (CH 2 ) r PO(OR 60 )
  • R 14 H; lower alkyl; lower alkenyl; (CH 2 ) m OR 55 (where R 55 : lower alkyl; or lower alkenyl); (CH 2 ) m SR 56 (where R 56 : H; or lower alkyl; or lower alkenyl); (CH 2 ) m NR 33 R 34 ( where R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl) ; (CH 2 ) m N(R 20 )COR 64 (where: R 20 : H; lower alkyl; R 64 : lower alkyl; or lower alkenyl); (CH 2 ) 0 COOR 57 (where R 57 : lower alkyl; or lower alkenyl); (CH 2 ) 0 CONR 58 R 59 (where R 58 : lower alkyl; or lower alkenyl; and R 59 : H; or lower alkyl); (CH 2 ) o PO(OR 60 )
  • R 58 lower alkyl, or lower alkenyl; andR 59 : H; lower alkyl
  • (CH 2 ) o PO(OR 60 ) 2 where R 60 : lower alkyl; or lower alkenyl
  • (CH 2 ) 0 S0 2 R 62 where R 62 : lower alkyl; or lower alkenyl
  • (CH 2 ) q C 6 H 4 R 8 where R 8 : H; F; Cl; CF 3 ; lower alkyl; lower alkenyl; or lower alkoxy).
  • R 16 lower alkyl; lower alkenyl; (CH 2 ) 0 OR 55 (where R 55 : lower alkyl; or lower alkenyl); (CH 2 ) 0 SR 56 (where R 5 ⁇ : H; or lower alkyl; or lower alkenyl); (CH 2 ) 0 NR 33 R 34 (where R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl) ; (CH 2 ) o N(R 20 )COR 64 (where: R 20 : H; or lower alkyl; R 64 : lower alkyl; or lower alkenyl); (CH 2 ) 0 COOR 57 (where R 57 : lower alkyl; or lower alkenyl); (CH 2 ) 0 CONR 58 R 59 (where R 58 : lower alkyl; or lower alkenyl; and R 59 : H; or lower alkyl); (CH 2 ) 0 PO(OR 60 )
  • R 77 lower alkyl; lower alkenyl; (CH 2 ) q OR 55 (where R 55 : lower alkyl; or lower alkenyl); (CH 2 ) q SR 56 (where R 56 : H; or lower alkyl; or lower alkenyl); (CH 2 ) q NR 33 R 34 ( where R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl) ; (CH 2 ) q N(R 0 )COR 6 (where: R 20 : H; or lower alkyl; R 64 : lower alkyl; or lower alkenyl); (CH 2 ) r COOR 57 (where R 57 : lower alkyl; or lower alkenyl); (CH 2 ) q CONR 58 R 59 (where R 58 : lower alkyl; or lower alkenyl; and R 59 : H; lower alkyl); (CH 2 ) r PO(OR 60 )
  • R 20 is H or lower alkyl; and R 64 is alkyl; alkenyl; aryl; aryl-lower alkyl; or heteroaryl- lower alkyl; and R 75 is lower alkyl; lower alkenyl; or aryl-lower alkyl; especially those wherein R 75 is allyl (A8'-l) and R 64 is n-hexyl (A8"-l).
  • Building block A70 belongs to the class of open-chained ⁇ -substituted ⁇ -amino acids, building blocks A71 and A72 to the corresponding ⁇ -amino acid analogues and building blocks A73-A104 to the cyclic analogues of A70.
  • Such amino acid derivatives have been shown to constrain small peptides in well defined reverse turn or U-shaped conformations (C. M. Venkatachalam, Biopolymers, 1968, 6, 1425-1434; W. Kabsch, C Sander, Biopolymers 1983, 22, 2577).
  • Such building blocks or templates are ideally suited for the stabilization of ⁇ -hairpin conformations in peptide loops (D. Obrecht, M. Altorfer, J. A. Robinson, "Novel Peptide Mimetic Building Blocks and Strategies for Efficient Lead Finding", Adv. Med Chem. 1999, Vo , 1-68; P. Balaram, "Non- standard amino acids in peptide design and protein engineering", Curr. Opin. Struct. Biol.
  • templates (al) can also consist of -A70-CO- to A104-CO- where building block A70 to A104 is of either (D)- or (L)-configuration, in combination with a building block -B-CO- of (L)- configuration.
  • R 20 in A70 to A104 are H or lower alkyl with methyl being most preferred.
  • Preferred values for R 18 , R 19 and R 2I -R 29 in building blocks A70 to A104 are the following: R ,s : lower alkyl R 19 : lower alkyl; lower alkenyl; (CH 2 ) p OR 55 (where R 55 : lower alkyl; or lower alkenyl); (CH 2 ) P SR 56 (where R 56 : H; or lower alkyl; or lower alkenyl); (CH 2 ) P NR 33 R 34 ( where R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl) ; (CH 2 ) P N(R 20 )COR 64 (where: R 20 : H; or lower alkyl; R 64 : lower alkyl; or lower alkenyl); (CH 2 ) p COOR 57 (where R 57 : lower alkyl; or lower alkenyl
  • R 2 ' H ; lower alkyl; lower alkenyl; (CH 2 ) 0 OR 55 (where R 55 : lower alkyl; or lower alkenyl); (CH 2 ) 0 SR 56 (where R 56 : H; or lower alkyl; or lower alkenyl); (CH 2 ) 0 NR 33 R 34 ( where R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl) ; (CH 2 ) 0 N(R 20 )COR 64 (where: R 20 : H; or lower alkyl; R 64 : lower alkyl; or lower alkenyl); (CH 2 ) 0 COOR 57 (where R 57 : lower alkyl; or lower alkenyl); (CH 2 ) 0 CONR 58 R 59 (where R 58 : lower alkyl, or lower alkenyl; and R 59 : H; lower alkyl); (CH 2 ) 0 PO(OR 60
  • R 23 H; lower alkyl; lower alkenyl; (CH 2 ) 0 OR 55 (where R 55 : lower alkyl; or lower alkenyl); (CH 2 ) 0 SR 56 (where R 5 ⁇ : H; or lower alkyl; or lower alkenyl); (CH 2 ) 0 NR 33 R 34 (where R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl) ; (CH 2 ) o N(R 20 )COR 64 (where: R 20 : H; or lower alkyl; R 64 : lower alkyl; or lower alkenyl); particularly favoured are NR 20 COlower alkyl
  • R 20 H; or lower alkyl
  • (CH 2 ) 0 COOR 57 (where R 57 : lower alkyl; or lower alkenyl);
  • R 24 lower alkyl; lower alkenyl; (CH 2 ) 0 OR 55 (where R 55 : lower alkyl; or lower alkenyl);
  • R 58 lower alkyl, or lower alkenyl; and R 59 : H; lower alkyl
  • (CH 2 ) 0 PO(OR 60 ) 2 where R 60 : lower alkyl; or lower alkenyl
  • (CH 2 ) 0 S0 2 R 62 where R 62 : lower alkyl; or lower alkenyl
  • (CH 2 ) q C 6 H 4 R 8 where R 8 : H; F; Cl; CF 3 ; lower alkyl; lower alkenyl; or lower alkoxy).
  • R 25 H; lower alkyl; lower alkenyl; (CH 2 ) m OR 55 (where R 55 : lower alkyl; or lower alkenyl); (CH 2 ) m NR 33 R 34 (where R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl);
  • R 8 H; F; Cl; CF 3 ; lower alkyl; lower alkenyl; or lower alkoxy.
  • R 26 H; lower alkyl; lower alkenyl; (CH 2 ) m OR 55 (where R 55 : lower alkyl; or lower alkenyl); (CH 2 ) m NR 33 R 3 (where R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl) ; (CH 2 ) m N(R 20 )COR 64 (where: R zo : H; or lower alkyl; R 64 : lower alkyl; or lower alkenyl);
  • R 8 H; F; Cl; CF 3 ; lower alkyl; lower alkenyl; or lower alkoxy.
  • R 25 and R 26 taken togethc-. .an uc -(CH 2 ) 2 . 6 -; -(CH 2 ) 2 0(CH 2 ) 2 -; - (CH 2 ) 2 S(CH 2 ) 2 -; or -(CH 2 ) 2 NR 34 (CH 2 ) 2 -;
  • R 27 H; lower alkyl; lower alkenyl; (CH 2 ) 0 OR 55 (where R 55 : lower alkyl; or lower alkenyl); (CH 2 ) 0 SR 56 (where R 56 : H; or lower alkyl; or lower alkenyl); (CH 2 ) 0 NR 33 R 34 ( where R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl) ; (CH 2 ) o N(R 20 )COR 64 (where: R 20 : H; or lower alkyl; R 64 : lower alkyl; or lower alkenyl); (CH 2 ) 0 COOR 57 (where R 57 : lower alkyl; or lower alkenyl); (CH 2 ) 0 CONR 58 R 59 (where R 58 : lower alkyl, or lower alkenyl; and R 59 : H; lower alkyl); (CH 2 ) 0 PO(OR eo
  • R 28 lower alkyl; lower alkenyl; (CH 2 ) 0 OR 55 (where R 55 : lower alkyl; or lower alkenyl); (CH 2 ) 0 SR 56 (where R 56 : H; or lower alkyl; or lower alkenyl); (CH 2 ) 0 NR 33 R 34 ( here R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl) ; (CH 2 ) 0 N(R 20 )COR 64 (where: R 20 : H; or lower alkyl; R 64 : lower alkyl; or lower alkenyl); (CH 2 ) 0 COOR 57 (where R 57 : lower alkyl; or lower alkenyl); (CH 2 ) 0 CONR 58 R 59 (where R 58 : lower alkyl, or lower alkenyl; and R 59 : H; lower alkyl); (CH 2 ) o PO(OR 60 ) 2 (CH
  • R 20 H; or lower alkyl - R 30 : H, methyl
  • R 3 ' H; lower alkyl; lower alkenyl; (CH 2 ) p OR 55 (where R 55 : lower alkyl; or lower alkenyl); (CH 2 ) P NR 33 R 34 (where R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl) ; (CH 2 ) P N(R 20 )COR 64 (where: R 20 : H; or lower alkyl; R 64 : lower alkyl; or lower alkenyl); (CH 2 ) 0 COOR 57 (where R 57 : lower alkyl; or lower alkenyl); (CH 2 ) 0 CONR 58 R 59 (where R 58 : lower alkyl, or lower alkenyl; and R 59 : H; lower alkyl); (CH 2 ) o PO(OR 60 ) 2 (where R 60 : lower alkyl; or lower alkenyl); (CH 2 ) 0 S0 2 R 62
  • R 32 H, methyl - R 33 : lower alkyl; lower alkenyl; (CH 2 ) m OR 55 (where R 55 : lower alkyl; or lower alkenyl);
  • R 34 H; or lower alkyl.
  • R 35 H; lower alkyl; lower alkenyl; (CH 2 ) m OR 55 (where R 55 : lower alkyl; or lower alkenyl); (CH 2 ) m N(R 20 )COR 64 (where: R 20 : H; or lower alkyl; R 64 : lower alkyl; or lower alkenyl); (CH 2 ) 0 COOR 57 (where R 57 : lower alkyl; or lower alkenyl); (CH 2 ) 0 CONR 58 R 59 (where R 58 : lower alkyl; or lower alkenyl; and R 59 : H; lower alkyl).
  • R 36 lower alkyl; lower alkenyl; or aryl-lower alkyl .
  • R 37 H; lower alkyl; lower alkenyl; (CH 2 ) p OR 55 (where R 55 : lower alkyl; or lower alkenyl); (CH 2 ) P NR 33 R 34 (where R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl) ; (CH 2 ) p N(R 20 )COR 64 (where: R 20 : H; or lower alkyl; R fi4 : lower alkyl; or lower alkenyl);
  • R 3S H; lower alkyl; lower alkenyl; (G-.jy p .- ⁇ ⁇ where R 55 : lower alkyl; or lower alkenyl); (CH 2 ) P NR 33 R 34 ( where R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl) ;
  • R 8 H; F; Cl; CF 3 ; lower alkyl; lower alkenyl; or lower alkoxy.
  • R 39 H; lower alkyl; lower alkenyl; (CH 2 ) m OR 55 (where R 55 : lower alkyl; or lower alkenyl); (CH 2 ) m N(R 20 )COR 64 (where: R 20 : H; or lower alkyl; R 64 : lower alkyl; or lower alkenyl); (CH 2 ) 0 COOR 57 (where R 57 : lower alkyl; or lower alkenyl); (CH 2 ) 0 CONR 58 R 59 (where R 58 : lower alkyl; or lower alkenyl; and R 59 : H; lower alkyl).
  • R 40 lower alkyl; lower alkenyl; or aryl-lower alkyl.
  • R 4 ' H; lower alkyl; lower alkenyl; (CH 2 ) p OR 55 (where R 55 : lower alkyl; or lower alkenyl); (CH 2 ) P NR 33 R 34 (where R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl) ; (CH 2 ) p N(R 20 )COR 64 (where: R 20 : H; or lower alkyl; R 64 : lower alkyl; or lower alkenyl);
  • R 8 H; F; Cl; CF 3 ; lower alkyl; lower alkenyl; or lower alkoxy.
  • R 42 H; lower alkyl; lower alkenyl; (CH 2 ) p OR 55 (where R 55 : lower alkyl; or lower alkenyl); (CH 2 ) P NR 33 R 34 (where R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl) ;
  • R 8 H; F; Cl; CF 3 ; lower alkyl; lower alkenyl; or lower alkoxy.
  • R 43 H; lower alkyl; lower alkenyl; (CH 2 ) m OR 55 (where R 55 : lower alkyl; or lower alkenyl); (CH 2 ) m SR S ⁇ (where R 56 : lower alkyl; or lower alkenyl); (CH 2 ) m NR 33 R 34 ( where R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl) ; (CH 2 ) m N(R 20 )COR 64 (where: R 20 : H; or lower alkyl; R 64 : lower alkyl; or lower alkenyl); (CH 2 ) 0 COOR 57 (where R 57 : lower alkyl; or lower alkenyl); (CH 2 ) 0 CONR 58 R 59 (where R 58 : lower alkyl; or lower alkenyl; and R 59 : H; lower alkyl);
  • R 44 lower alkyl; lower alkenyl; doctrine e R 55 : lower alkyl; or lower alkenyl); (CH 2 ) P SR 56 (where R 56 : H; or lower alkyl; or lower alkenyl); (CH 2 ) P NR 33 R 34 (where R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl) ; (CH 2 ) P N(R 20 )COR 64 (where: R 20 : H; or lower alkyl; R 64 : lower alkyl; or lower alkenyl); (CH 2 ) p COOR 57 (where R 57 : lower alkyl; or lower alkenyl); (CH 2 ) p CONR 58 R 59 (where R 58 : lower alkyl; or lower alkenyl; and R 59 : H; lower alkyl); or (CH 2 ) 0 C 6 H 4 R 8 (where R 8 : H; F; Cl;
  • R 45 H; lower alkyl; lower alkenyl; (CH 2 ) s OR 55 (where R 55 : lower alkyl; or lower alkenyl); (CH 2 ) S SR 56 (where R 5 ⁇ : H; or lower alkyl; or lower alkenyl); (CH 2 ) 0 NR 33 R 34 ( where R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl) ; (CH 2 ) o N(R 20 )COR 64 (where: R 20 : H; or lower alkyl; R 64 : lower alkyl; or lower alkenyl); (CH 2 ) 0 COOR 57 (where R 57 : lower alkyl; or lower alkenyl); (CH 2 ) 0 CONR 58 R 59 (where R 58 : lower alkyl; or lower alkenyl; and R 59 : H; lower alkyl); or (CH 2 ) S C 6 H R 8 (
  • R 4S H; lower alkyl; lower alkenyl; (CH 2 ) s OR 55 (where R 55 : lower alkyl; or lower alkenyl); (CH 2 ) S SR 56 (where R 56 : H; or lower alkyl; or lower alkenyl); (CH 2 ) 0 NR 33 R 34 (where R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl) ; (CH 2 ) o N(R 20 )COR 64 (where: R 20 : H; or lower alkyl; R 64 : lower alkyl; or lower alkenyl); (CH 2 ) 0 COOR 57 (where R 57 : lower alkyl; or lower alkenyl); (CH 2 ) 0 CONR 58 R 59 (where R 58 : lower alkyl; or lower alkenyl; and R 59 : H; lower alkyl); or (CH 2 ) S C 6 H R 8 (CH
  • R 47 H; or OR 55 (where R 55 : lower alkyl; or lower alkenyl).
  • R 4S H; or lower alkyl.
  • R 49 H;lower alkyl; (CH 2 ) 0 COOR 57 (where R 57 : lower alkyl; or lower alkenyl); (CH 2 ) 0 CONR 58 R 59 (where R 58 : lower alkyl; or lower alkenyl; and R 59 : H; lower alkyl); or (CH 2 ) s C ⁇ H R 8 (where R 8 : H; F; Cl; CF 3 ; lower alkyl; lower alkenyl; or lower alkoxy).
  • R 50 H; methyl - R 5/ : H; lower alkyl; lower alkenyl; (CH 2 ) ra OR 55 (where R 55 : lower alkyl; or lower alkenyl); (CH 2 ) m NR 33 R 34 (where R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl) ; (CH 2 ) m N(R 20 )COR 64 (where: R 20 : H; or lower alkyl; R 64 : lower alkyl; or lower alkenyl); (CH 2 ) p COOR 57 (where R 57 : lower alkyl; or lower alkenyl); (CH 2 ) p CONR 58 R 59 (where R 58 : lower alkyl; or lower alkenyl; and R 59 : H; lower alkyl); or (CH 2 ) r C 6 H 4 R 8 (where R 8 : H; F; Cl; CF 3 ; lower
  • R 52 H; lower alkyl; lower alkenyl; (CH 2 ) m OR 55 (where R 55 : lower alkyl; or lower alkenyl); (CH 2 ) m NR 33 R 34 (where R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl) ; (CH 2 ) m N(R 20 )COR 64 (where: R 20 : H; or lower alkyl; R 64 : lower alkyl; or lower alkenyl); (CH 2 ) p COOR 57 (where R 57 : lower alkyl; or lower alkenyl); (CH 2 ) p CONR 58 R 59 (where R 58 : lower alkyl; or lower alkenyl; and R 59 : H; lower alky _,, w-. (where R 8 : H; F; Cl; CF 3 ; lower alkyl; lower alkenyl; or lower alkoxy).
  • R 53 H; lower alkyl; lower alkenyl; (CH 2 ) m OR 55 (where R 55 : lower alkyl; or lower alkenyl); (CH 2 ) m NR 33 R 34 (where R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl) ; (CH 2 ) m N(R 20 )COR 64 (where: R 20 : H; or lower alkyl; R 64 : lower alkyl; or lower alkenyl); (CH 2 ) p COOR 57 (where R 57 : lower alkyl; or lower alkenyl); (CH 2 ) p CONR 58 R 59 (where R 58 : lower alkyl; or lower alkenyl; and R 59 : H; lower alkyl); or (CH 2 ) r C 6 H 4 R 8 (where R 8 : H; F; Cl; CF 3 ; lower alkyl; lower alkenyl; or lower al
  • R 54 lower alkyl; lower alkenyl; or aryl-lower alkyl.
  • building blocks A70 to A104 the following are preferred: A74 with R 22 being H, A75, A76, A77 with R 22 being H, A78 and A79.
  • the building block -B-CO- within template (al) and (a2) designates an L-amino acid residue.
  • Preferred values for B are: -NR 20 CH(R 71 )- and enantiomers of groups A5 with R 2 being H, A8, A22, A25, A38 with R 2 being H, A42, A47, and A50. Most preferred are
  • R 20 is H or lower alkyl and R is alkyl; alkenyl; aryl; aryl-lower alkyl; or heteroaryl- lower alkyl; especially those wherein R 64 . is n-hexyl (A8"'-l)
  • the peptidic chains Z of the ⁇ -hairpin mimetioo uw ⁇ uucJ herein is generally defined in terms of amino acid residues belonging to one of the following groups:
  • amino acid residues in chain Z can also be of formula -A-CO- wherein A is as defined above.
  • Group C comprises amino acid residues with small to medium-sized hydrophobic side chain groups according to the general definition for substituent R 72 .
  • a hydrophobic residue refers to an amino acid side chain that is uncharged at physiological pH and that is repelled by aqueous solution .
  • these side chains generally do not contain hydrogen bond donor groups, such as (but not limited to) primary and secondary amides, primary and secondary amines and the corresponding protonated salts thereof, thiols, alcohols, phosphonates, phosphates, ureas or thioureas.
  • ethers such as ethers, thioethers, esters, tertiary amides, alkyl- or aryl phosphonates and phosphates or tertiary amines.
  • Genetically encoded small-to-medium-sized amino acids include alanine, isoleucine, leucine, methionine and valine.
  • Group D comprises amino acid residues with aromatic and heteroaromatic side chain groups according to the general definition for substituent R 73 .
  • An aromatic amino acid residue refers to a hydrophobic amino acid having a side chain containing at least one ring having a conjugated ⁇ - electron system (aromatic group).
  • hydrogen bond donor groups such as (but not limited to) primary and secondary amides, primary and secondary amines and the corresponding protonated salts thereof, thiols, alcohols, phosphonates, phosphates, ureas or thioureas, and hydrogen bond acceptor groups such as (but not limited to) ethers, thioethers, esters, tetriary amides, alkyl- or aryl phosphonates -and phosphates or tertiary amines.
  • Genetically encoded aromatic amino acids include phenylalanine and tyrosine.
  • a heteroaromatic amino acid residue refers to a hydrophobic amino acid having a side chain containing at least one ring having a conjugated ⁇ -system incorporating at least one heteroatom such as (but not limited to) O, S and N according to the general definition for substituent R 77 .
  • they may contain hydrogen bond donor groups such as (but not limited to) primary and secondary amides, primary and secondary amines and the corresponding protonated salts thereof, thiols, alcohols, phosphonates, phosphates, ureas or thioureas, and hydrogen bond acceptor groups such as (but not limited to) ethers, thioethers, esters, tetriary amides, alkyl- or aryl phosphonates -and phosphates or tertiary amines.
  • Hydro bond donor groups such as (but not limited to) primary and secondary amides, primary and secondary amines and the corresponding protonated salts thereof, thiols, alcohols, phosphonates, phosphates, ureas or thioureas, and hydrogen bond acceptor groups such as (but not limited to) ethers, thioethers, esters, tetriary amides, alkyl- or aryl phosphonates -
  • Group E comprises amino acids containing side chains with polar-cationic and urea-derived residues according to the general definition for substituen R 74 .
  • Polar-cationic refers to a basic side chain which is protonated at physiological pH.
  • Genetically encoded polar-cationic amino acids include arginine, lysine and histidine.
  • Citrulline is an example for an amino acid containing a urea-derived residue.
  • Group F comprises amino acids containing side chains with polar-non-charged or anionic residues according to the general definition for substituent R 84 .
  • a polar-non-charged or anionic residue refers to a hydrophilic side chain that is uncharged and, respectively, anionic at physiological pH (carboxylic acids are included), but that is not repelled by aqueous solutions.
  • Such side chains typically contain hydrogen bond donor groups such as (but not limited to) primary and secondary amides, carboxylic acids and esters, primary and secondary amines, thiols, alcohols, phosphonates, phosphates, ureas or thioureas. These groups can form hydrogen bond networks with water molecules.
  • polar-non-charged and anionic amino acids include asparagine, cysteine, glutamine, serine and threonine but also aspartic acid and glutamic acid.
  • Group H comprises side chains of preferably (L)-amino acids at opposite positions of the ⁇ - strand region that can form an interstrand linkage.
  • the most widely known linkage is the disulfide bridge formed by cysteines and homo-cysteines positioned at opposite positions of the ⁇ -strand.
  • Various methods are known to form disulfide linkages including those described by: J. P. Tarn et al. Synthesis 1979, 955-957; Stewart et al. , Soaa rnase eptide Synthesis, 2d Ed., Pierce Chemical Company, III., 1984; Ahmed et al. J. Biol. Chem.
  • disulfide linkages can be prepared as described hereinafter in the pertinent Examples (procedure 3), using acetamidomethyl (Acm)- protective groups for cysteine.
  • a well established interstrand linkage consists in linking ornithines and lysines, respectively, with glutamic and aspartic acid residues located at opposite ⁇ -strand positions by means of an amide bond formation.
  • Preferred protective groups for the side chain amino-groups of ornithine and lysine are allyloxycarbonyl (Alloc) and allylesters for aspartic and glutamic acid.
  • interstrand linkages can also be established by linking the amino groups of lysine and ornithine located at opposite ⁇ -strand positions with reagents such as N,N-carbonylimidazole to form cyclic ureas.
  • positions for interstrand linkages are the following:
  • Such interstrand linkages are known to stabilize the ⁇ -hairpin conformations and thus constitute an important structural element for the design of ⁇ -hairpin mimetics.
  • amino acid residues in chain Z are those derived from natural ⁇ -amino acids.
  • amino acids which, or the residues of which, are suitable for the purposes of the present invention, the abbreviation corresponding to generally adopted usual practice:
  • residues for group C are:
  • Particularlily preferred residues for group D are:
  • the positions P 1 to P n of each amino acid residue in the chain Z are unequivocally defined as follows: P 1 represents the first amino acid in the chain Z that is coupled with its N-terminus to the C-terminus of the templates (b)-(p) or of group -B-CO- in template (al), or of Group -A-CO- in template (A2) and P n represents the last amino acid in the chain Z that is coupled with its C-terminus to the N-terminus of the templates (b)-(p) or of group -A-CO- in template (al) or of group -B-CO- in template (A2).
  • Each of the positions P 1 to P n will preferably contain an amino acid residue belonging to one or two or three of above types C to F, or being Pro, as follows: if n is 7, the amino acid residues in position 1 - 7 are preferably: PI: oftype C or oftype F; P2: of type E or of type D --,-. ⁇ J . ⁇ C C;
  • P3 oftype F or of type C;
  • P4 of type C or type F or of type D;
  • P6 of type C or of type E, or the residue is Pro;
  • P7 of type C or of type F; if n is 11, the amino acid residues in position 1 - 11 are preferably:
  • P2 of type C or of type F;
  • P4 of type E or of type D or of type C;
  • P5 oftype F or of type C;
  • P6 of type C, or of type D;
  • P7 of type F, or the residue is Pro
  • P8 of type C or of type E, or the residue is Pro;
  • P9 of type C or of type F;
  • P10 oftype F or oftype C;
  • PI 1 of type D or of type E; or P2 and P10, taken together can form a group of type H;
  • ⁇ -peptidomimetics of the invention include those described in Examples 1, 4, 7, 8 and 15.
  • the process of the invention can advantageously be carried out as parallel array synthesis to yield libraries of template-fixed ⁇ -hairpin peptidomimetics of the above general formula I.
  • Such parallel synthesis allows one to obtain arrays of numerous (normally 24 to 192, typically 96) compounds of general formula I in high yields and defined purities, minimizing the formation of dimeric and polymeric by-products.
  • the proper choice of the functionalized solid-support (i.e. solid support plus linker molecule), templates and site of cyclization play thereby key roles.
  • the functionalized solid support is conveniently derived from polystyrene crosslinked with, preferably 1-5%, divinylbenzene; polystyrene coated with polyethyleneglycol spacers (Tentagel R ); and polyacrylamide resins (see also Obrecht, D.; Villalgordo, J.-M, "Solid- Supported Combinatorial and Parallel Synthet, -- - ⁇ .-. ⁇ -.-.-Molecular-Weight Compound
  • the solid support is functionalized by means of a linker, i.e. a bifunctional spacer molecule which contains on one end an anchoring group for attachment to the solid support and on the other end a selectively cleavable functional group used for the subsequent chemical transformations and cleavage procedures.
  • a linker i.e. a bifunctional spacer molecule which contains on one end an anchoring group for attachment to the solid support and on the other end a selectively cleavable functional group used for the subsequent chemical transformations and cleavage procedures.
  • the linker must be designed to eventually release the carboxyl group under mild acidic conditions which do not affect protecting groups present on any functional group in the side-chains of the various amino acids.
  • Linkers which are suitable for the purposes of the present invention form acid-labile esters with the carboxyl group of the amino acids, usually acid-labile benzyl, benzhydryl and trityl esters; examples of linker structures of this kind include 2-methoxy-4-hydroxymethylphenoxy (Sasrin R linker), 4-(2,4-dimethoxyphenyl-hydroxymethyl)-phenoxy (Rink linker), 4-(4-hydroxymethyl-3- methoxyphenoxy)butyric acid (HMPB linker), trityl and 2-chlorotrityl.
  • Sasrin R linker 2-methoxy-4-hydroxymethylphenoxy
  • Rink linker 4-(2,4-dimethoxyphenyl-hydroxymethyl)-phenoxy
  • HMPB linker 4-(4-hydroxymethyl-3- methoxyphenoxy)butyric acid
  • trityl 2-chlorotrityl
  • the support is derived from polystyrene crosslinked with, most preferably 1-5%, divinylbenzene and functionalized by means of the 2-chlorotrityl linker.
  • reaction vessels normally 24 to 192, typically 96
  • 25 to 1000 mg preferably 100 mg
  • of the appropriate functionalized solid support preferably 1 to 3% cross linked polystyrene or tentagel resin.
  • the solvent to be used must be capable of swelling the resin and includes, but is not limited to, dichloromethane (DCM), dimethylformamide (DMF), N-methylpyrrolidone (NMP), dioxane, toluene, tetrahydrofuran (THF), ethanol (EtOH), trifluoroethanol (TFE), isopropylalcohol and the like.
  • Solvent mixtures containing as at least one component a polar solvent e. g. 20% TFE/DCM, 35% THF/NMP
  • Tetrahedron Lett. 1988, 294005-4008) is cleavable with diluted trifluoroacetic acid (0.5-1% TFA in DCM) and is stable to Fmoc deprotection conditions during the peptide synthesis, Boc/tBu- based additional protecting groups being compatible with this protection scheme.
  • Other linkers which are suitable for the process of the invention include the super acid labile 4-(2,4- dimethoxyphenyl-hydroxymethyl)-phenoxy linker (Rink linker, Rink, H. Tetrahedron Lett.
  • Suitable protecting groups for amino acids and, respectively, for their residues are, for example,
  • amino group (as is present e. g. also in the side-chain of lysine)
  • Ts tosyl i. e. p-toluenesulfonyl
  • the 9-fluorenylmethoxycarbonyl- (Fmoc)-protected amino acid derivatives are preferably used as the building blocks for the construction of the template-fixed ⁇ -hai ⁇ in loop mimetics of formula I.
  • For the deprotection i. e. cleaving off of the Fmoc group, 20%> piperidine in DMF or 2%> DBU/2% piperidine in DMF can be used.
  • the quantity of the reactant i. e.
  • reaction tube in combination with the holder block and the manifold, are reinserted into the reservoir block and the apparatus is fastened together. Gas flow through the manifold is initiated to provide a controlled environment, for example, nitrogen, argon, air and the like. The gas flow may also be heated or chilled prior to flow through the manifold.
  • Heating or cooling of the reaction wells is achieved by heating the reaction block or cooling externally with isopropanol/dry ice and the like to bring about the desired synthetic reactions. Agitation is achieved by shaking or magnetic stirring (within the reaction tube).
  • the preferred workstations are Labsource's Combi-chem station and MultiSyn Tech's-Syro synthesizer.
  • Amide bond formation requires the activation of the ⁇ -carboxyl group for the acylation step.
  • this activation is being carried out by means of the commonly used carbodiimides such as dicyclohexylcarbodiimide (DCC, Sheehan & Hess, J. Am. Chem. Soc. 1955, 77, 1067-1068) or diisopropylcarbodiimide (DIC, Sarantakis et al Biochem. Biophys. Res. Commun.1916, 73, 336- 342)
  • DCC dicyclohexylcarbodiimide
  • DIC Diisopropylcarbodiimide
  • 1-hydroxybenzotriazole (HOBt, K ⁇ nig & Geiger, Chem. Ber 1970, 103, 788-798) is included as an additive to the coupling mixture.
  • HOBt prevents dehydration, suppresses racemization of the activated amino acids and acts as a catalyst to improve the sluggish coupling reactions.
  • Certain phosphonium reagents have been used as direct coupling reagents, such as benzotriazol-1-yl-oxy-tris- (dimethylamino)-phosphonium hexafluorophosphate (BOP) (Castro et al., Tetrahedron Lett.
  • these phosphonium reagents are also suitable for in situ formation of HOBt esters with the protected amino acid derivatives. More recently diphenoxyphosphoryl azide (DPP A) or 0-(7-aza- benzotriazol-l-yl)-N,N,N',N'-tetramethyluroniu tetrafluoroborate (TATU) or 0-(7-aza- benzotriazol-l-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU)/7-aza-l- hydroxy benzotriazole (HO At, Ca ⁇ ino et al., leiranearon Lett. 1994, 35, 2279-2281) have also been used as coupling reagents.
  • DPP A diphenoxyphosphoryl azide
  • TATU 0-(7-aza- benzotriazol-l-yl)-N,N,N',N'-tetramethyluroniu te
  • reaction wells are filled with solvent (preferably 5 ml), the reaction tubes, in combination with the holder block and manifold, are immersed and agitated for 5 to 300 minutes, preferably 15 minutes, and drained by gravity followed by gas pressure applied through the manifold inlet (while closing the outlet) to expel the solvent;
  • solvent preferably 5 ml
  • Both of the above washing procedures are repeated up to about 50 times (preferably about 10 times), monitoring the efficiency of reagent, solvent, and byproduct removal by methods such as TLC, GC, or inspection of the washings.
  • (allyloxycarbonyl) is an example for such a protecting group for amino which can be selectively removed, e.g. by means of Pd° and phenylsilane in CH 2 C1 2 , without affecting the remaining protecting groups, such as Fmoc, present in the molecule.
  • the reactive group thus liberated can then be treated with an agent suitable for introducing the desired substituent.
  • an amino group can be acylated by means of an acylating agent corresponding to the acyl substituent to be introduced.
  • Detachment of the fully protected linear peptide from the solid support is achieved by immersion of the reaction tubes, in combination with the holder block and manifold, in reaction wells containing a solution of the cleavage reagent (preferably 3 to 5 ml). Gas flow, temperature control, agitation, and reaction monitoring are implemented as described above and as desired to effect the detachment reaction.
  • the reaction tubes, in combination with the holder block and manifold are disassembled from the reservoir block and raised above the solution level but below the upper lip of the reaction wells, and gas pressure is applied through the manifold inlet (while closing the outlet) to efficiently expel the final product solution into the reservoir wells.
  • the resin remaining in the reaction tubes is then washed 2 to 5 times as above with 3 to 5 ml of an appropriate solvent to extract (wash out) as much of the detached product as possible.
  • the product solutions thus obtained are combined, taking care to avoid cross-mixing.
  • the individual solutions/extracts are then manipulated as needed to isolate the final compounds. Typical manipulations include, but are not limited to, evaporation, concentration, liquid/liquid extraction, acidification, basification, neutralization or additional reactions in solution.
  • interstrand linkages between side-chains of appropriate amino acid residues at opposite positions of the ⁇ -strand region.
  • Interstrand linkages and their formation have been discussed above, in connection with the explanations made regarding groups of the type H which can, for example, be disulfide bridges formed by cysteines and homocysteines at opposite positions of the ⁇ -strand, or glutamic and aspartic acid residues linking ornithines and, respectively, lysines located at opposite ⁇ -strand positions by amide bond formation.
  • groups of the type H which can, for example, be disulfide bridges formed by cysteines and homocysteines at opposite positions of the ⁇ -strand, or glutamic and aspartic acid residues linking ornithines and, respectively, lysines located at opposite ⁇ -strand positions by amide bond formation.
  • the formation of such interstrand linkages can be effected by methods well known in the art.
  • the fully protected peptide derivative of type I is treated with 95% TFA, 2.5% H 2 0, 2.5% TIS or another combination of scavengers for effecting the cleavage of the protecting groups.
  • the cleavage reaction time is commonly 30 minutes to 12 hours, preferably about 2 hours.
  • most of the TFA is evaporated and the product is precipitated with ether/hexane (1:1) or other solvents which are suitable therefor.
  • the cyclic peptide derivative obtained as end-product can be isolated.
  • this peptide derivative can be used directly for biological assays, or it has to be further purified, for example by preparative HPLC.
  • Hydrolytic enzymes involve hydrolysis of amides and nitriles by aminopeptidases or nitrilases, cleavage of N-acyl groups by acylases, and ester hydrolysis by lipases or proteases. It is well documented that certain enzymes will lead specifically to pure (L)-enantiomers whereas others yield the corresponding (D)-enantiomers (e.g. : R. Duthaler, Tetrahedron Report 1994, 349, 1540-1650; R. M. Williams, "Synthesis of optically active ⁇ -amino acids", Tetrahedron Organic Chemistry Series, Vol.7, J. E. Baldwin, P. D. Magnus (Eds.), Pergamon Press., Oxford 1989).
  • R 6 OH
  • A14 Amino acids of type A14 can be made according to Scheme 1.
  • i NaH, BrCH(R )COOMe, DMF; ii: LiOHxl H 2 0, MeOH, H 2 0; iii: polyphosphoric acid(PPA); iv: NaH, CICOOMe, THF; v: enzymatic resolution (e.g.lipase); vi: NaOH, MeOH, H 2 0, heat; vii: FmocOSu, Na 2 C0 3 aq court dioxane
  • A23 See Beyerman, Boekee, Reel. Trav. Ch . u ⁇ _-_,us 1959, 78, 648-653; D. R. Adams, P. D . Bailey, I. D. Collier, J. D. Heffernan, S. Stokes J. Chem. Soc. Chem. Commun. 1996, 349-350; J. E. Baldwin, R. M. Adlington, C.
  • A30 and A31 can be prepared according to Schemes 4 and 5.
  • i NaH, tert.-butyl N-benzoyl glycinate, DMF; ii: NaH, Pd(0), toluene; iii: TFA, CH 2 CI 2 ; iv: polypho- sphoric acid; v: NaOHaq.,MeOH, 75°; then HCIaq.; vi: DBU, Mel, DMF; vii: lithium hexamethyl- disilazide,THF, chloro trimethylsilane, -78°; then R 1 -X; viii: enzymatic resolution(e.g. lipase); then isolation as methylester: DBU, Mel, DMF; ix: NaOHaq., MeOH, heat; x: FmocOSu,
  • i Boc 2 0, Na 2 C0 3 aq., dioxane; ii: DBU, Mel, DMF; iii: lithium hexamethyldisilazide, THF, chlorotrimethylsilane, -78°; then R 2 -X; iv: LiOHx1H 2 0, MeOH, H 2 0; v:TFA, CH 2 CI 2 ; vi: FmocOSu,
  • A32 can be prepared according to P. W. Schiller, G. Weltrowska, T. M.-D. Nguyen, C. Lemieux, N. Nga, /. Med. Chem. 1991, 34, 3125-3132; V. S. Goodfellow, M. V. Marathe, K. G. Kuhlman, T. D. Fitzpatrick, D. Cuadrato, J. Med. Chem. 1996, 39, 1472-1484; G. Caliendo, F. Fiorino, P. Grieco, E. Perissutti, S. DeLuca, A. Guiliano, G. Santelli, D. Califano, B. Severino, V.
  • 26 27 28 i: BocNHNH 2 , NaCNBH 3 , MeOH, AcOH; ii: CbzCI, Et 3 N, CH 2 CI 2 ; iii: TFA, CH 2 CI 2 ; then pyridine,
  • DMAP heat; iv: resolution (e.g. lipase); v: DBU, Mel, DMF; vi: La esson reagent, toluene, 75°; vii: DBU, Mel, DMF; viii: NaBH 4 or NaCNBH 3 , MeOH; ix: R 3 introduced by reductive amination, alkylation or acylation; x: LiOHx1H 2 0, MeOH, H 2 0; xi: Pd/C, H 2 , EtOH; xii: FmocOSu, Na C0 3 aq., dioxane
  • a 2 to A46 Compounds of this typj? can be prepared according to Scheme, 8 to 12.
  • Key intermediate 34" and ⁇ -amino acid synthesis involving this building block include: R. M. Williams, M.-N. Im, Tetrahedron Lett. 1988, 29, 6079-6082; R. M. Williams, M.-N. Im, J. Am. Chem. Soc.1991, 113, 9276-9286; j. F. Dellaria, B. D. Santarsiero, Tetrahedron Lett. 1988, 29, 6079-6082; j. F. Dellaria, B. D. Santarsiero, J. Org. Chem. 1989, 54, 3916-3926; j. E. Baldwin, V. Lee, C. J. Schofield, Synlett 1992, 249-251; j. E. Baldwin, V. Lee, C. J. Schofield, Heterocycles 1992, 34, 903-906.
  • i HBr; ii: DBU, Mel, DMF; iii: DIBAL-H, THF; iv: EtOH, pyridinium p-toluenesulfonate, mol. sieves 4A; v: lithium hexamethyldisilazide, THF, -78°, 43; vi: Pd/C, H 2 , EtOH; then DBU,
  • i HBr; ii: DBU, Mel, DMF; iii: DIBAL-H, THF; iv: EtOH, pyridinium p-toluenesulfonate, mol. sieves 4A; v: lithium hexamethyldisilazide, THF, -78°, 47; vi: Pd/C, H 2 , EtOH; then DBU.Mel,
  • i HBr; ii: DBU, Mel, DMF; iii: DIBAL-H, THF; iv: EtOH, pyridinium p-toluenesulfonate, mol. sieves 4A; v: lithium hexamethyldisilazide, THF, -78°, 51; vi: Pd/C, H 2 , EtOH; then DBU,
  • i HBr; ii: DBU, Mel, DMF; iii: DIBAL-H, THF; iv: EtOH, pyridinium p-toluenesulfonate, mol. sieves 4A; v: lithium hexamethyldisilazide, THF, -78°, 59; vi: Pd/C, H 2 , EtOH; then DBU,
  • i HBr; ii: DBU, Mel, DMF; iii: DIBAH, THF; iv: EtOH, pyridinium p-toluenesuifonate, mol. sieves 4A; v: lithium hexamethyldisilazide, THF, -78°, 63 vi: Pd/C, H 2 , EtOH; then DBU,
  • i iBuMgCI.THF; ii: NaH, THF; iii: lithium hexamethyldisilazide, THF, chlorotrimetylsilane, -78°; then R 6 -X; iv: NaOHaq., MeOH, 75°; then HCIaq.; v: DBU, Mel, DMF; vi: lithium hexamethyldisilazide , THF, chlorotrimetylsilane, -78°; then R 1 -X; vii: resolution (e.g.
  • i NaOMe, MeOH; ii: NaH, THF; iii: NaOHaq., MeOH, 75°; then HCIaq.; iv: DBU, Mel, DMF; v: lithium hexamethyldisilazide, THF, chlorotrimethylsilane, -78°; then R 1 -X; vi: resolution (e.g. lipase); then isolation of methylester: DBU, Mel, DMF; vii: LiOHx1H 2 O, MeOH, H 2 O; viii.TFA,
  • A59 can be prepared according to Scheme 21.
  • i NaOMe, MeOH; ii: NaH, THF; iii: NaOHaq., MeOH, 75°; then HCIaq.; iv: DBU, Mel, DMF; v: lithium hexamethyldisilazide, THF, chlorotrimethylsilane, -78°; then R 1 -X; vi: resolution (e.g. lipase); then isolation of methylester: DBU, Mel, DMF; vii: LiOHx1 H 2 O, MeOH, H 2 O; viii.TFA,
  • R 1 -X Raney-Ni, H 2 , EtOH; vi: CbzCI, Et 3 N, CH 2 CI 2 ; vii: NaH, Br(CH 2 ) 2 Br, THF; viii: resolution
  • i resolution (e.g. lipase); then DBU, Mel, DMF; ii: lithium hexamethyldisilazide, THF, chlorotrimethylsilane, -78°; then R 6 -X; iii: LiOHxl H 2 O, MeOH, H 2 O; iv: TFA, CH 2 CI 2 ; v: FmocOSu,
  • DMF lithium diisopropylamide, THF, chlorotrimethylsilane, -78°; then R 1 -X; ix: resolution (e.g. lipase); then isolation of methylester: DBU, Mel, DMF; x: LiOHxl H 2 0, MeOH, H 2 0; xi: TFA,
  • building blocks A70 belong to the class of open-chain ⁇ -substituted ⁇ -amino acids, A71 and A72 to the class of the the corresponding ⁇ -amino acid analogues and A73-A104 to the class of the cyclic analogues of A70.
  • FmocCI The method depicted in Scheme 27 consists in treatment of the appropriate ketones 126 with KCN, (NH 4 ) 2 CO 3 in a mixture of ethanol/water (E. Ware, J. Chem. Res. 1950, 46, 403; L. H. Goodson, I. L. Honigberg, J. j. Lehmann, W. H. Burton, J. Org. Chem. 1960, 25, 1920; S. N. Rastogi, j. S. Bindra, N. Anand, Ind. J. Chem. 1971, 1175) to yield the corresponding hydantoins 127, which were hydrolyzed with Ba(OH) 2 in water at 120- 140° (R. Sarges, R. C.
  • azlactones 129 could also be prepared starting from amino acids 130 and 131, Schotten-Baumann acylation and cyclization with N,N'- dicyclohexyl carbodiimide to azlactones 132 and 133 and alkylation to yield 129 (D. Obrecht, U. Bohdal, C. Broger, D. Bur, C. Lehmann, R. Ruffieux, P. Schonholzer, C.
  • A73-A104 can be prepared starting from the corresponding ketones 138, hydantoin formation (139) (E. Ware, J. Chem. Res. 1950, 46, 403; L. H. Goodson, I. L. Honigberg, J. J. Lehmann, W. H. Burton, J. Org. Chem. 1960, 25, 1920; S. N. Rastogi, J. S. Bindra, N. Anand, Ind. J. Chem. 1971, 1175; D. Obrecht, U. Bohdal, C. Broger, D. Bur, C. Lehmann, R. Ruffieux, P. Schonholzer, C. Spiegler, Helv. Chim.
  • Amino acid building blocks of this type can be conveniently prepared from the corresponding disubstituted succinates 146 by Cwrttw-s-rearrangement as shown in Scheme 29.
  • Amino acid building blocks of type A72 can be conveniently prepared by Arndt-Eistert Cl- homologation of compounds of type A70 according to Scheme 30.
  • A75 and A76 Compounds of this type can be prepared using the following methods: P. Hughes,
  • A79 Compounds of this type can be prepared according to general method described in Scheme 28 starting from the corresponding pyrrolidine-3-ones.
  • A80-A82 Compounds of this type can be prepared according to D. M. Walker, E. W. Logusch, Tetrahedron Lett. 1989, 30, 1181-1184; Y. Morimoto, K. Achiwa, Chem. Pharm. Bull. 1989, 35, 3845-3849; J. Yoshimura, S. Kondo, M. Diara, H. Hashimoto, Carbohydrate Res. 1982, 99, 129- 142.
  • A87 and A88 Compounds of this type can be prepared according to L. Munday, J. Chem. Soc. 1961, 4372; J. Ansell, D. Morgan, H. C. Price, Tetrahedron Lett. 1978, 47, 4615-4616.
  • A90 Compounds of this type can be prepared according to general method described in Scheme 28 starting from the corresponding tetrahydrothiapyran-3-ones.
  • A91 Compounds of this type can be prepared according to general method described in Scheme 28 starting from the corresponding tetrahydropyran-3-ones.
  • A101 Compounds of this type can be prepared according to general method described in Scheme 28 starting from the corresponding tetrahydroquinoline-2,4-diones.
  • A102 Compounds of this type can be prepared according to K. Ishizumi, N. Ohashi, N. Tanno, J. Org. Chem. 1987, 52, 4477-4485; D. Obrecht, U. Bohdal, C. Broger, D. Bur, C. Lehmann, R. Ruffieux, P. Schonholzer, C. Spiegler, Helv. Chim. Acta 1995, 78, 563-580; D. Obrecht, C. Spiegler, P. Schonholzer, K. M ⁇ ller, H. Heimgartner, F.
  • Templates of type (bl) can be prepared according to Schemes 31 and 32.
  • Boc e.g. using di-tert. -butyl dicarbonate and triethylamine in a suitable solvent such as dichloromethane; any other suitable N-protecting group (not shown in
  • Reaction Scheme 31 can be introduced in an analogous manner.
  • Reaction of formed product with phthalimide, diethyl diazodicarboxylate and triphenylphoshine under standard Mitsunobu conditions (Mitsunobu, O.; Wada, M.; Sano,
  • v The resulting product is hydrogenated using H 2 and a suitable catalyst such as palladium on charcoal in a solvent such as ethyl acetate, DMF or ethanol; subsequently separation of diastereomers takes place and yields 156.
  • a suitable catalyst such as palladium on charcoal in a solvent such as ethyl acetate, DMF or ethanol; subsequently separation of diastereomers takes place and yields 156.
  • Cyclization conveniently with DBU in DMF to yield 157.
  • viii The phthalimide group is cleaved off from resulting product, conveniently by hydrazinolysis, e.g. treatment with methylhydrazine in a suitable solvent such as DMF.
  • ix The formed product is conveniently protected with reagents such as 9- fluorenylmethoxcarbonyl chloride or 9-fluorenylmethoxcarbonyl succinimide using a base such as sodium carbonate or triethylamine in a suitable solvent or mixture of solvents such as dioxane and water, or dichloromethane to yield 158.
  • x Standard removal of an allyl ester group using e.g. palladium(0) as catalyst gives 159.
  • Templates of type (b2) can be prepared according to Scheme 33.
  • i 160 (obtainable from Vitamin C as described by Hubhelen, C. (Synthesis 1986, 962) is treated with phthalimide, diethyl diazodicarboxylate and triphenylphoshine under standard Mitsunobu conditions (Mitsunobu, O.; Wada, M.; Sano, T. J. J. Am. Chem. Soc. 1972, 94, 672).
  • ii The phthalimide group is cleaved off from the product, conveniently by hydrazinolysis, e.g. by treatment with methylhydrazine in a suitable solvent such as DMF.
  • iii The amino group is protected by treatment with a benzoylating reagent such as benzoic acid anhydride or benzoylchloride and a base such as triethylamine or 4-dimethylaminopyridine in a suitable solvent such as dichloromethane or DMF.
  • a benzoylating reagent such as benzoic acid anhydride or benzoylchloride and a base such as triethylamine or 4-dimethylaminopyridine in a suitable solvent such as dichloromethane or DMF.
  • iv Removal of the 2,4-dimethoxybenzyl group, e.g. with K 2 S 2 O 8 and Na 2 HP0 4 in aqueous acetonitrile at an elevated temperature, e.g. at about 80° C.
  • v Introduction of a ter -butoxycarbonyl group using e.g.
  • xi Cleavage of the tert.-butyl ester and tert-butyloxycarbonyl groups, conveniently using trifluoracetic acid in dichloromethane or 4N hydrochloric acid in dioxane.
  • xii The intermediate free amino acid formed is conveniently protected with reagents such as
  • Templates of type (cl) can be prepared according to Schemes 34 to 37.
  • 166 can be synthesized from 165 according to P. Waldmeier, "Solid-supported synthesis of highly substituted xanthene-derived templates for the synthesis of ⁇ -turn stabilized cyclic peptide libraries", PhD-thesis, University of Zurich, 1996.
  • a suitable solvent such as ethanol
  • ii The intermediate aminonitrile is saponified, conveniently under basic conditions, e.g. with aqueous sodium hydroxide in a suitable solvent such as ethanol at an elevated temperature, conveniently under reflux, to yield 167.
  • the intermediate free amino acid formed is conveniently protected with reagents such as 9-fluorenylmethoxcarbonyl chloride or 9-fluorenylmethoxcarbonyl succinimide using a base such as sodium carbonate or triethylamine in a suitable solvent or mixture of solvents such as dioxane and water, or dichloromethane to yield 168 as described by P. Waldmeier, "Solid-supported synthesis of highly substituted xanthene-derived templates for the synthesis of ⁇ -turn stabilized cyclic peptide libraries", PhD-thesis, University of Zurich, 1996.
  • x The intermediate free amino acid formed is conveniently protected with reagents such as 9-fluorenylmethoxcarbonyl chloride or 9-fluorenylmethoxcarbonyl succinimide using a base such as sodium carbonate or triethylamine in a suitable solvent or mixture of solvents such as dioxane and water, or dichloromethane to yield 170.
  • reagents such as 9-fluorenylmethoxcarbonyl chloride or 9-fluorenylmethoxcarbonyl succinimide using a base such as sodium carbonate or triethylamine in a suitable solvent or mixture of solvents such as dioxane and water, or dichloromethane to yield 170.
  • Double ortho- bromination of 171 is performed preferably with excess bromine in acetic acid and dichloromethane.
  • ii The amino group is protected, conveniently Cbz-protected, with reagents such as benzyloxycarbonyl chloride or succinimide in a suitable solvent such as dioxane in the presence of a base such as aqueous sodium hydroxide.
  • iii The carboxylic acid group is esterif ⁇ ed, preferably with DBU and methyl iodide in DMF to yield 172.
  • R 38 e.g. by palladium(O)- catalyzed Stille- (Stille, J.K. Angew. Chem.1986, 68, 504) and
  • aqueous hydrochloric acid in a suitable solvent such as dioxane at an elevated temperature, conveniently at about 100° C.
  • reagents such as 9-fluorenylmethoxcarbonyl chloride or 9-fluorenylmethoxcarbonyl succinimide using a base such as sodium carbonate or triethylamine in a suitable solvent or mixture of solvents such as dioxane and water, or dichloromethane to yield 174.
  • i Cleavage of the methoxy groups of 166, preferably by treatment with an excess of boron tribromide in a suitable solvent such as dichloromethane.
  • ii Hydrolysis of the cyano group under acidic conditions, preferably with 25% aqueous hydrochloric acid in a suitable solvent such as dioxane at an elevated temperature, conveniently at about 100° C.
  • iii The resulting acid is treated with a dehydrating agent such as thionyl chloride in a suitable solvent such as dioxane to yield 175.
  • Any other functionalization known for phenol groups can be employed for introduction of substituents R 35 .
  • Any other functionalization known for aryl bromides can be employed for introduction of substituents R 36 .
  • viii Hydrolysis of the ester group under acidic conditions, conveniently with 25% aqueous hydrochloric acid in a suitable solvent such as dioxane at an elevated temperature, e.g. at about 100° C.
  • ix Cleavage of the phthalimido group, conveniently by hydrazinolysis, e.g. with hydrazine hydrate in a suitable solvent such as ethanol.
  • x The intermediate free amino acid formed is conveniently protected with reagents such as 9-fluorenylmethoxcarbonyl chloride or 9-fluorenylmethoxcarbonyl succinimide using a base such as sodium carbonate or triethylamine in a suitable solvent or mixture of solvents such as dioxane and water, or dichloromethane to yield 179.
  • xi The ester group is hydrolyzed under acidic conditions, conveniently with 25% aqueous hydrochloric acid in a suitable solvent such as dioxane at an elevated temperature, e.g. at about 100° C.
  • xii The phthali ido group is cleaved, e.g. by hydrazinolysis, conveniently with hydrazine hydrate in a suitable solvent such as ethanol.
  • xiii The intermediate free amino acid formed is conveniently protected with reagents such as
  • 3,7-Dimethoxyphenothiazine 186 is prepared and converted into 187 according to M ⁇ ller, K.; Obrecht, D.; Knierzinger, A.; Spiegler, C; Bannwarth, W.; Trzeciak, A.; Englert, G; Labhardt, A.; Schonholzer, P. Perspectives in Medicinal Chemistry, Editor Testa, B.; Kyburz, E.; Fuhrer, W.; Giger, R., Weinheim, New York, Basel, Cambridge: Verlag Helvetica Chimica Acta, 1993, 513-531; Bannwarth, W.; Gerber, F.; Grieder, A.; Knierzinger, A.; M ⁇ ller, K.; Obrecht.
  • the benzyl group is cleaved off from 187 conveniently by hydrogenation, e.g. with H 2 and a catalyst such as palladium on charcoal in a suitable solvent such as ethanol, DMF or ethyl acetate.
  • a catalyst such as palladium on charcoal in a suitable solvent such as ethanol, DMF or ethyl acetate.
  • ii Introduction of lower alkyl (R 43 ) by alkylation using an appropriate alkylating agent (R 43 -X'; X - OTf, Br, I) and strong bases such as sodium amide in liquid ammonia or sodium hydride in tetrahydrofuran, dioxan or DMF in the presence of a phase transfer catalyst such as TDA-I.
  • substituted lower alkyl R 43
  • R 43 CH 2 COOR 55 and CH 2 CH 2 COOR 55 can be introduced by treatment with the appropriate 2-halo acetic and, respectively, 3 -halo propionic acid derivatives.
  • Any other functionalization known for diarylamines can be employed for introduction of substituents R 43 .
  • iii Cleavage of the methoxy groups of 188, conveniently by treatment with an excess of boron tribromide in a suitable solvent such as dichloromethane at temperatures ranging from about -20° C to about room temperature
  • a suitable solvent such as dichloromethane
  • phenol groups can be employed for introduction of substituents R 39 and R 40 .
  • v The cyano group of 188 and, respectively, 189 is hydrolyzed, conveniently under acidic conditions, e.g. with 25% aqueous hydrochloric acid in a suitable solvent such as dioxane at an elevated temperature, e.g. at about 100° C.
  • vi The phthalimide group of the intermediate is cleaved, conveniently by hydrazinolysis, e.g. with hydrazine hydrate in a suitable solvent such as ethanol.
  • reagents such as 9- fluorenylmethoxcarbonyl chloride or 9-fluorenylmethoxcarbonyl succinimide using a base such as sodium carbonate or triethylamine in a suitable solvent or mixture of solvents such as dioxane and water, or dichloromethane to yield 190 and, respectively, 191.
  • cyano group of 188 is hydrolyzed, conveniently under acidic conditions, e.g. with
  • ii The phthalimide group of the intermediate is cleaved, conveniently by hydrazinolysis, e.g. with hydrazine hydrate in a suitable solvent such as ethanol to yield 192.
  • iii Double ortho- bromination of 192 is performed preferably with excess bromine in acetic acid and dichloromethane.
  • substituents R 41 and R 42 Any other functionalization by electrophilic aromatic substitution known can be employed for introduction of substituents R 41 and R 42 .
  • the amino group is protected, conveniently Cbz-protected, with reagents such as benzyloxycarbonyl chloride or succinimide in a suitable solvent such as dioxane in the presence of a base such as aqueous sodium hydroxide.
  • reagents such as benzyloxycarbonyl chloride or succinimide in a suitable solvent such as dioxane in the presence of a base such as aqueous sodium hydroxide.
  • v The carboxylic acid group is esterified, preferably with DBU and methyl iodide in DMF to yield 193.
  • Regioselective bromination of 192 is performed preferably with bromine in acetic acid and dichloromethane.
  • Any other functionalization by electrophilic aromatic substitution known can be employed for introduction of substituents R 41 .
  • the amino group is conveniently Cbz-protected with reagents such as benzyloxycarbonyl chloride or succinimide in a suitable solvent such as dioxane in presence of a base such as aqueous sodium hydroxide.
  • viii The carboxylic acid group is esterified, preferably with DBU and methyl iodide in DMF to yield 194.
  • ix Introduction of lower alkyl, substituted lower alkyl and aryl substituents (R 41 )for
  • aqueous hydrochloric acid in a suitable solvent such as dioxane at an elevated temperature, preferably at about 100° C.
  • a suitable solvent such as dioxane
  • reagents such as 9-fluorenylmethoxcarbonyl chloride or 9-fluorenylmethoxcarbonyl succinimide using a base such as sodium carbonate or triethylamine in a suitable solvent or mixture of solvents such as dioxane and water, or dichloromethane to yield 195 and 196.

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Abstract

Template-fixed β-hairpin peptidomimetics of the general formulae (1), wherein Z is a template-fixed chain of 7 to 11 α-amino acid residues which, depending on their positions in the chain (counted starting formteh N-terminal amino acid) are Gly, or Pro, or of certain types which, as the remaining symbols in the aove formulae, are defined in the description and the claims, and salts thereof, have the property to inhibit proteases, in particular serine proteases. These β-hairpin peptidomimetics can be manufactured by a process which is based on a mixed solid- and solution phase synthetic strategy.

Description

TEMPLATE-FIXED PEPTIDOMIMETICS AS INHIBITORS OF SERINE PROTEASES
The present invention provides template-fixed β-haiipin peptidomimetics incorporating a template-fixed chain of 7 or 11 α-amino acid residues which, depending on their position in the chain, are Gly, or Pro, or of certain types, as defined hereinbelow. These template-fixed β-hairpin peptidomimetics are useful as inhibitors of protease enzymes. They are especially valuable as inhibitors of various serine proteases such as trypsin, human cathepsin G, and thrombin. In addition the present invention provides an efficient process by which these compounds can, if desired, be made in library-format. This library-approach constitutes an efficient novel tool to identify specific serine protease inhibitors.
Inhibitors of proteases are emerging with promising therapeutic uses in the treatment of diseases such as cancers (R. P. Beckett, A. Davidson, A. H. Drummond, M. Whittaker, Drug Disc. Today 1996, 1, 16-26; L. L. Johnson, R. Dyer, D. J. Hupe, Curr. Opin. Chem. Biol. 1998, 2, 466-71; D. Leung, G. Abbenante, and D. P. Fairlie, J. Med. Chem. 2000, 43, 305-341), parasitic, fungal, and viral infections [e.g. schistosomiasis (M. M. Becker, S. A. Harrop, J. P. Dalton, B. H. Kalinna, D. P. McManus, D. P. Brindley, J. Biol. Chem. 1995, 270, 24496-501); malaria (A. M. Silva, A. Y. Lee, S. V. Gulnik, P. Maier, J. Collins, T. N. Bhat, P. J. Collins, R. E. Cachau, K. E. Luker, I. Y. Gluzman, S. E. Francis, A. Oksman, D. E. Goldberg, J. W. Erikson, Proc. Natl. Acad. Sci. U.S.A 1996, 93, 10034-9), C. albicans (C. Abad-Zapetero, R. Goldman, S. W. Muchmore, C. Hutchins, K. Stewart, J. Navaza, C. D. Payne, T. L. Ray, Protein Sci. 1996, 5, 640-52), HIV (A. Wlodawer, J. W. Erickson, Annu. Rev. Biochem. 1993, 62, 543-85; P. L. Darke, J. R. Huff, Adv. Pharmacol. 1994, 5, 399-454), hepatitis (J. L. Kim, K. A. Morgenstern, , C. Lin, T. Fox, M. D. Dwyer, J. A. Landro, S. P. Chambers , W. Markland, C. A. Lepre, E. T. O'Malley, S. L. Harbeson, C. M. Rice, M. A. Murcko, P. R. Caron, J. A. Thomson, Cell, 1996, 87, 343-55; R. A. Love, H. E. Parge, J. A. ickersham, Z. Hostomsky, N. Habuka, E. W. Moomaw, T. Adachi, Z. Hostomska, Cell, 1996, 87, 331-342), herpes (W. Gibson, M. R. Hall, Drug. Des. Discov. 1997, 15, 39-47)], and inflammatory, immunological, respiratory (P. R. Bernstein, P. D. Edwards, J. C. Williams, Prog. Med. Chem. 1994, 31, 59-120; T. Ε. Hugh, Trends Biotechnol. 1996, 14, 409-12), cardiovascular (M. T. Stubbs, W. A. Bode, Thromb. Res. 1993, 69, 1-58), and neurodegenerative defects including Alzheimer's disease (R. Vassar, B. D. Bennett, S. Babu-Kahn, S. Kahn, Ε. A. Mendiaz, Science, 1999, 286, 735-41). As most proteases bind their substrates in extenαeα or p-srrand conformations, good inhibitors must thus be able to mimick such a conformation. β-Hairpin mimetics are thus ideally suited to lock peptide sequences in an extended conformation.
Among proteases, serine proteases constitute important therapeutic targets. Serine proteases are classified by their substrate specificity, particularly by the type of residue found at P 1 , as either trypsin-like (positively charged residues Lys/Arg preferred at PI), e-fø-stα-ϊe-like (small hydrophobic residues Ala/Val at PI), or chymotrypsin-like (large hydrophobic residues Phe/Tyr/Leu at PI). Serine proteases for which protease-inhibitor X-ray crystal data is available on the PDB data base (PDB: www.rcsb.org pdb) include trypsin, a-chymotrypsin, γ- chymotrypsin, human neutrophil elastase, thrombin, subtilisin, human cytomegalovirus, proteinase A, achromobacter, human cathepsin G, glutamic acid-specific protease, carbopeptidase D, blood coagulation factorVIIa, porcine factor 1XA, mesentericopeptidase, HCV protease, and thermitase. Other serine proteases which are of therapeutic interest include tryptase, complement convertase, hepatitis C-NS3 protease. Inhibitors of thrombin (e.g. J. L. Metha, L. Y. Chen, W. W. Nichols, C. Mattsson, D. Gustaffson, T. G. P. Saldeen, J. Cardiovasc. Pharmacol. 1998, 31, 345-51; C. Lila, P. Gloanec, L. Cadet, Y. Herve, J. Fournier, F. Leborgne, T. J. Verbeuren, G. DeNanteuil, Synth. Comm. 1998, 28, 4419-29) and/αctorZα (e.g. J. P. Vacca, Annu. Rep. Med. Chem. 1998, 33, 81-90) are in clinical evaluation as anti-thrombotics, inhibitors of elastase (J. R. Williams, R. C. Falcone, C. Knee, R. L. Stein, A. M. Strimpler, B. Reaves, R. E. Giles, R. D. Krell, Am. Rev. Respir. Dis. 1991, 144, 875-83) are in clinical trials for emphysema and other pulmonary diseases whereas tryptase inhibitors are currently in phase II clinical trials for asthma (C. Seife, Science 1997, 277, 1602-3). Finally, cathepsin G and elastase are intimately involved in the modulation of activities of cytokines and their receptors. Particularly at sites of inflammation, high concentration of cathepsin G, elastase and proteinase 3 are released from infiltrating polymorphonuclear cells in close temporal correlation to elevated levels of inflammatory cytokines, strongly indicating that these proteases are involved in the control of cytokine bioactivity and availability (U. Bank, S. Ansorge, J. Leukoc. Biol. 2001, 69, 177-90). Thus inhibitors of thrombin and cathepsin G constitute valuable targets for novel drug candidates. Of the many occurring proteinaceous serine protease inhibitors, one is a 14 amino acid cyclic peptide from sunflower seeds, termed sunflower trypsin inhibitor (SFTI-1) (S. Luckett, R. Santiago Garcia, J. J. Barker, A. V. Konarev, P. R. Shewry, A. R. Clarke, R. L. Brady, /. Mol. Biol. 1999, 290, 525-533; Y.-Q. Long, S.-L. Lee, C.-Y. Lin, I. J. Enyedy, S. Wang, P. Li, R. B. Dickson, P. P. Roller, Biorg. & Med. Chem. Lett. 2001, 11, 2515-2519), which shows both sequence and conformational similarity with the uyμ&m-ieactive loop of the Bowman-Birk family of serine protease inhibitors. The inhibitor adopts a β-haiipin conformation when bound to the active site of bovine β-trypsin. SFTI-1 inhibited β-trypsin (Ki<0.1nM), cathepsin G, elastase (Kj~105μM), chymottypsin (K;~7.4μM) and thrombin (Kj~136mM).
We illustrate here an approach to inhibitor design which involves transplanting the β-hairpin loop from the naturally occurring peptide onto a hairpin-inducing template. Based on the well defined 3D-structure of the β-hairpin mimetics libraries of compounds can be designed which ultimately can lead to novel inhibitors showing different specificity profiles towards several classes of proteases.
Template-bound hairpin mimetic peptides have been described in the literature (D, Obrecht, M. Altorfer, J. A. Robinson, Adv. Med. Chem. 1999, 4, 1-68; J. A. Robinson, Syn. Lett. 2000, 4, 429- 441), but such molecules have not previously been evaluated for development of peptides which inhibit proteases and constitute mimetics of extended peptide conformations. However, the ability to generate β-hairpin peptidomimetics using combinatorial and parallel synthesis methods has now been established (L. Jiang, K. Moehle, B. Dhanapal, D. Obrecht, J. A. Robinson, Helv. Chim. Acta. 2000, 83, 3097-3112). This technology allows to rapidly synthesise libraries of protease inhibitors and to explore key residues which determine the specificity for a given serine protease.
The β-hairpin peptidomimetics of the present invention are compounds of the general formula
Figure imgf000004_0001
(I)
wherein
Figure imgf000004_0002
is a group of one of the formulae
Figure imgf000005_0001
(a1) (a2)
Figure imgf000005_0002
(c2) (c3) (d)
Figure imgf000005_0003
Figure imgf000006_0001
(h) (11) 02)
Figure imgf000006_0002
03) 04) (j)
Figure imgf000006_0003
(n) (o) (P)
wherein
Figure imgf000006_0004
is the residue of an L-α-amino acid with B being a resiαue of formula -NR 0CH(R71)- or the enantiomer of one of the groups Al to A69 as defined hereinafter;
is a group of one of the formulae
Figure imgf000007_0001
A1 A2 A3 A4
Figure imgf000007_0002
A5 A6 A7 A8 A9
Figure imgf000007_0003
Figure imgf000007_0004
A15 A16 A17 A18 A19
Figure imgf000008_0001
Figure imgf000008_0002
A34 A35 A36 A37
Figure imgf000008_0003
A38 A39 A40 A41 A42
Figure imgf000009_0001
A53 A54 A55 A56 A57
Figure imgf000009_0002
A63 A64 A65 A66
Figure imgf000010_0001
A80 A81 A82 A83 A84
Figure imgf000010_0002
A90 A91 A92 A93 A94
Figure imgf000011_0001
R > 1 : is- H; lower alkyl; or aryl-lower alkyl;
R2 is H; alkyl; alkenyl; -(CH2)m(CHR6I)s0R55; -(CH2)m(CHR61)sSR56; -
(CH2)m(CHR61)sNR33R34; -(CH2)m(CHR61)sOCONR33R78; - (CH2)m(CHR6I)sNR20CONR33R78; -(CH2)0(CHR6I)sCOOR57; -(CH2)0(CHR61)sCONR58R59;
-(CH2)o(CHR6I)sPO(OR60)2; -(CH2)0(CHR6!)S S02R62; or -(CH2)0(CHR61)SC6H4R8; R3 is H; alkyl; alkenyl; -(CH2)m(CHR61)sOR55; -(CH2)m(CHR61)sSR56; -
(CH2)m(CHR61)sNR33R34;
-(CH2)0(CHR61)SCOOR57 ; -(CH2)0(CHR61)sCONR58R59 ; -(CH2)o(CHR61)sPO(OR60)2; -(CH2)o(CHR61)s S02R62; or -(CH2)0(CHRδl)sC6H4R8;
R4 is H; alkyl; alkenyl; -(CH2)m(CHR61)sOR55; -(CH2)m(CHR61)sSR56; -
(CH2)m(CHR61)sNR33R34;
-(CH2)p(CHR6I)sCOOR57; -(CH2)p(CHR61)sCONR58R59; -(CH2)p(CHR61)sPO(OR60)2;
-(CH2)p(CHR61)s S02R62; or -(CH2)0(CHR61)SC6H4R8; R5 is alkyl; alkenyl; -(CH2)0(CHR61)sOR55; -(CH2)0(CHR61)SSR56; -(CH2)0(CHR61)SNR33R34; -
(CH2)m(CHR61)sOCONR33R78; -(CH2)m(CHR61)sNR20CONR33R78; -
(CH2)0(CHR61)sCOOR57; -(CH2)0(CHR61)sCONR58R59; -(CH2)o(CHR61)sPO(OR60)2; -
(CH2)o(CHR61)s S02R62; or -(CH2)0(CHR6I)SC6H4R8; R6 is H; alkyl; alkenyl; -(CH2)0(CHR61)s0R55; -(CH2)0(CHR61)SSR56; -(CH2)0(CHR61)SNR33R34; -(CH2)o(CHRδl)sCOOR57; -(CH2)0(CHR61)sCONR58R59; -(CH2)o(CHR61)sPO(OR60)2;
-(CH2)0(CHR61)S SO2R62; or -(CH2)0(CHR6,)SC6H4R8; R7 is alkyl; alkenyl; -(CH2)q(CHR6I)sOR55; -(CH2)q(CHR61)sNR33R34; -(CH2)r(CHR61)sCOOR57; -(CH2)r(CHRδI)sCONR58R59;
Figure imgf000012_0001
-(CH2)r(CHRδl)sS02R62; or
Figure imgf000012_0002
R8 is H; Cl; F; CF3; N02; lower alkyl; lower alkenyl; aryl; aryl-lower alkyl; -
(CH2)o(CHRδ,)sOR55; -(CH2)0(CHRδl)sSR5δ; -(CH2)0(CHRδI)NR33R34 ; - (CH2)0(CHRδl)sOCONR33R78; -(CH2)0(CHRδl)sNR20CONR33R78; -
(CH2)o(CHRδI)sCOOR57;-(CH2)o(CHR61)sCONR58R59; -(CH2)o(CHR6I)sPO(OR60)2; -
(CH2)0(CHRδl)sSO2R62; or -(CH2)0(CHRδI)sCOR64; R9 is alkyl; alkenyl; -(CH2)0(CHR61)SOR55; -(CH2)0(CHRδI)sSR5δ; -(CH2)0(CHRδl)sNR33R34;
-(CH2)o(CHRδl)sCOOR57; -(CH2)o(CHRδl)sCONR58R59; -(CH2)o(CHRδI)sPO(ORδ0)2; -(CH2)0(CHRδI)s S02R62; or -(CH2)0(CHRδI)sC6H4R8;
R10 is alkyl; alkenyl; -(CH2)0(CHRδl)sOR55; -(CH2)0(CHR61)sSR5δ; -(CH2)0(CHRδl)sNR33R34;
-(CH2)o(CHRδl)sCOOR57; -(CH2)o(CHRδ,)sCONR58R59; -(CH2)o(CHRδl)sPO(ORδ0)2;
-(CH2)0(CHRδl)s S02R62; or -(CH2)0(CHRδl)sC6H4R8; Rn is H; alkyl; alkenyl; -(CH2)m(CHRδl)sOR55; -(CH2)m(CHR61)sSR5δ; - (CH2)m(CHRδI)sNR33R34; -(CH2)m(CHR6I)sOCONR33R78; -
Figure imgf000012_0003
-(CH2)o(CHRδ,)sPO(OR60)2; -(CH2)0(CHRδl)sS02R62; or -(CH2)0(CHR61)S H4R8; R12 is H; alkyl; alkenyl; -(CH2)m(CHRδl)sOR55; -(CH2)m(CHRδl)sSR5δ; -
(CH2)m(CHR61)sNR33R34; -(CH2)r(CHRδl)sCOOR57; -(CH2)r(CHRδl)sCONR58R59; - (CH2)r(CHR61)sPO(ORδ0)2; -(CH2)r(CHR61)s S02R62; or -(CH2)r(CHRδl)sC6H4R8;
R13 is alkyl; alkenyl; -(CH2)q(CHRδl)s0R55; -(CH2)q(CHRδ,)sSR5δ; -(CH2)q(CHRδl)sNR33R34;
-(CH2)q(CHRδl)sCOOR57; -(CH2)q(CHR6I)sCONR58R59; -(CH2)q(CHR61)sPO(ORδ0)2;
-(CH2)q(CHRδl)s S02R62; or -(CH2)q(CHR61)sC6H4R8; R14 is H; alkyl; alkenyl; -(CH2)m(CHR61)sOR55; -(CH2)m(CHR61)sNR33R34; - (CH2)q(CHR61)sCOOR57; -(CH2)q(CHRδl)sCONR58R59; -(CH2)q(CHRδl)sPO(ORδ0)2; -
(CH2)q(CHRδl)sSOR62; or -(CH2)q(CHRδl)s H-iR8; R15 is alkyl; alkenyl; -(CH2)0(CHRδl)sOR55; -(CH2)0(CHRδl)sSR56; -(CH2)0(CHRδl)sNR33R34;
-(CH2)o(CHRδl)sCOOR57; -(CH2)0(CHR61)sCONR58R59; -(CH2)0(CHR61)sPO(OR6Q)2;
-(CH2)0(CHRδl)s S02R62; or -(CH2)0(CHR6I)SC6H4R8; R16 is alkyl; alkenyl; -(CH2)0(CHRδI)s0R55; -(CH2)0(CHR61)SSR56; -(CH2)0(CHRδl)sNR33R34;
-(CH2)o(CHRδl)sCOOR57; -(CH2)o(CHR6I)sCONR58R59; -(CH2)o(CHR61)sPO(ORδ0)2;
-(CH2)o(CHRδl)s S02R62; or -(CH2)0(CHRδl)sCsH4R8; R'7 is alkyl; alkenyl; -(CH2)m(CHRδI)sOR55; -(CH2)m(CHRδI)sSR56; -(CH2)m(CHRδl)sNR33R34;
-(CH2)q(CIDlδl)sCOOR57; -(CH2)q(CHR61)sCONR58R59; -(CH2)q(CHRδI)sPO(OR60)2; -(CH2)q(CHRδl)s S02Rδ2; or -(CH2)q(C-,--v ,S^,4R8; R18 is alkyl; alkenyl; -(CH2)p(CHR6I)sOR55; -(CH2)p(CHRδI)sSR5δ; -(CH2)P(CHR61)SNR33R34;
-(CH2)p(CHR61)sCOOR57; -(CH2)p(CHRδl)sCONR58R59; -(CH2)p(CHR61)sPO(OR60)2;
-(CH2)p(CHRδl)s S02R62; or -(CH2)0(CHRδl)sC6H4R8; R19 is lower alkyl; -(CH2)p(CHRδl)sOR55; -(CH2)P(CHR6I)SSR56; -(CH2)p(CHRδl)sNR33R34;
-(CH2)p(CHRδI)sCOOR57; -(CH2)p(CHRδl)sCONR58R59; -(CH2)p(CHRδI)sPO(ORδ0)2;
-(CH2)P(CHR61)S S02R62; or -(CH2)0(CHR61)SC6H4R8; or R18 and R19 taken together can form: -(CH2)2.6-; -(CH2)20(CH2)2-; -(CH2)2S(CH2)2-; or
-(CH2)2NR34(CH2)2-; R20 is H; alkyl; alkenyl; or aryl-lower alkyl;
R21 is H; alkyl; alkenyl; -(CH2)0(CHRδI)sOR55; -(CH2)0(CHR6I)SSR56; -(CH2)0(CHR6I)SNR33R34;
-(CH2)o(CHRδl)sCOOR57; -(CH2)o(CHR61)sCONR58R59; -(CH2)o(CHR61)sPO(ORδ0)2;
-(CH2)0(CHR61)S S02R62; or -(CH2)0(CHRδl)sC6H4R8; R22 is H; alkyl; alkenyl; -(CH2)0(CHRδ!)sOR55; -(CH2)0(CHRδl)sSR5δ; -(CH2)0(CHRδl)sNR33R34; -(CH2)o(CHR61)sCOOR57; -(CH2)0(CHRδl)sCONR58R59; -(CH2)0(CHR6I)sPO(ORδo)2;
-(CH2)0(CHRδl)s S02R62; or -(CH2)0(CHRδI)sC6H4R8; R23 is alkyl; alkenyl; -(CH2)0(CHR61)sOR55; -(CH2)0(CHR61)sSR5δ; -(CH2)0(CHR61)SNR33R34;
-(CH2)o(CHR6I)sCOOR57; -(CH2)o(CHR61)sCONR58R59; -(CH2)o(CHR61)sPO(OR60)2;
-(CH2)o(CHRδl)s S02R62; or -(CH2)0(CHRδl)sC6H4R8; R24 is alkyl; alkenyl; -(CH2)0(CHR61)S0R55; -(CH2)0(CHRδl)sSR56; -(CH2)0(CHRδl)sNR33R34;
-(CH2)0(CHR6I)sCOOR57; -(CH2)0(CHR6I)sCONR58R59; -(CH2)0(CHRδl)sPO(ORδo)2;
-(CH2)0(CHRδ,)s S02R62; or -(CH2)0(CHR61)SC6H4R8; R25 is H; alkyl; alkenyl; -(CH2)m(CHR61)sOR55; -(CH2)m(CHR61)sSR5δ; -
(CH2)m(CHRδl)sNR33R34; -(CH2)0(CHR6I)sCOOR57; -(CH2)0(CHR6I)SCONR58R59; -(CH2)o(CHRδI)sPO(ORδ0)2;
-(CH2)o(CHRδl)sS02R62; or -(CH2)0(CHR61)SC6H4R8; R2δ is H; alkyl; alkenyl; -(CH2)m(CHR61)sOR55; -(CH2)m(CHRδl)sSR5δ; -
(CH2)m(CHR61)sNR33R34; -(CH2)0(CHRδl)sCOOR57; -(CH2)0(CHRδl)sCONR58R59; -
(CH2)o(CHR6I)sPO(OR60)2; -(CH2)0(CHRδl)s S02R62; or -(CH^CHR^ H-R8; or R25 and R26 taken together can form: -(CH2)2.6-; -(CH2)rO(CH2)r; -(CH2)rS(CH2)r-; or
-(CH2)rNR34(CH2)r-; R27 is H; alkyl; alkenyl; -(CH2)0(CHRδl)sOR55; -(CH2)0(CHRδl)sSR5δ; -(CH2)0(CHRδl)sNR33R34;
-(CH2)o(CHRδl)sCOOR57; -(CH2)o(CHR61)sCONR58R59; -(CH2)o(CHRδI)sPO(OR60)2;
-(CH2)o(CHR6I)s S02R62; or -(CH2)o(CHRδI)sC6H4R8; R28 is alkyl; alkenyl; -(CH2)0(CHR6I)S-OR55; -^xα2 0^-dR61)s SR5δ; -(CH2)0(CHR61)S NR33R34;
-(CH2)o(CHRδl)s COOR57; -(CH2)0(CHRδl)s CONR58R59; -(CH2)0(CHRδI)s PO(OR60)2;
-(CH2)o(CHRδI)s S02R62; or -(CH2)0(CHRδI)s C6H4R8; R29 is alkyl; alkenyl; -(CH2)0(CHR6I)SOR55; -(CH2)0(CHRδl)sSR5δ; -(CH2)0(CHRδl)sNR33R34; -(CH2)o(CHRδI)sCOOR57; -(CH2)0(CHRδl)sCONR58R59; -(CH2)o(CHR6I)sPO(OR60)2;
-(CH2)o(CHRδl)s S02R62; or -(CH2)0(CHRδl)sC6H4R8; R30 is H; alkyl; alkenyl; or aryl-lower alkyl; R31 is H; alkyl; alkenyl; -(CH2)p(CHRδl)sOR55; -(CH2)p(CHRδl)sNR33R34; -
(CH2)0(CHR61)sC00R57; -(CH2)0(CHRδl)sCONR58R59; -(CH2)o(CHRδl)sPO(ORδ0)2; - (CH2)o(CHRδI)sS02R62; or -(CH2)0(CHRδl)s Ht 8;
R32 is H; lower alkyl; or aryl-lower alkyl; R33 is H; alkyl, alkenyl; -(CH2)m(CHR61)s0R55; -(CH2)m(CHRδl)sSR5δ; -
(CH2)m(CHRδI)sNR34Rδ3; -(CH2)m(CHRδl)sOCONR34R78;-
(CH2)m(CHRδl)sNR20CONR34R78; -(CH2)0(CHRδl)sCOR64; -(CH2)0(CHR61)s-CONR58R59, -(CH2)o(CHRδl)sPO(ORδ0)2; -(CH2)0(CHRδl)s S02R62; or -(CH2)0(CHRδl)sC6H4R8;
R34 is H; lower alkyl; aryl, or aryl-lower alkyl; R35 is H; alkyl; alkenyl; -(CH2)ra(CHRδl)s0R55; -(CH2)m(CHR61)sNR33R34; -
(CH2)p(CHR61)sC00R57; -(CH2)p(CHR61)sCONR58R59; -(CH2)p(CHRδI)sPO(ORδ0)2; -
(CH2)p(CHRδl)sS02R62; or -(CH2)p(CHRδl)s C6H4R8; R36 is H, alkyl; alkenyl; -(CH2)0(CHRδl)sOR55; -(CH2)p(CHRδl)sNR33R34; -
(CH2)p(CHRδI)sCOOR57; -(CH2)p(CHR61)sCONR58R59; -(CH2)p(CHRδl)sPO(OR60)2; -
(CH2)p(CHR61)sS02R62; or -(CH2)0(CHRδl)s C6H4R8; R37 is H; F; Br; Cl; N02; CF3; lower alkyl; -(CH2)p(CHR6I)sOR55; -(CH2)p(CHRδl)sNR33R34;
-(CH2)o(CHRδl)sCOOR57; -(CH2)o(CHRδl)sCONR58R59; -(CH2)0(CHR6t)sPO(OR60)2; -(CH2)0(CHRδl)sS02R62; or -(CH2)0(CHRδI)s C6H4R8;
R38 is H; F; Br; Cl; N02; CF3; alkyl; alkenyl; -(CH2)p(CHRδI)sOR55; -(CH2)p(CHRδl)sNR33R34;
-(CH2)o(CHR61)sCOOR57; -(CH2)o(CHRδl)sCONR58R59; -(CH2)0(CHR61)sPO(ORδ0)2;
-(CH2)0(CHR61)sSO2R62; or -(CH2)0(CHRδl)sC6H4R8; R39 is H; alkyl; alkenyl; or aryl-lower alkyl; R40 is H; alkyl; alkenyl; or aryl-lower alkyl;
R41 is H; F; Br; Cl; N02; CF3; alkyl; alkenyl; -(CH2)P(CHRδl)sOR55; -(CH2)P(CHR61)SNR33R34;
-(CH2)o(CHR61)sCOOR57; -(CH2)o(CHRδl)sCONR58R59; -(CH2)o(CHRδl)sPO(OR60)2;
-(CH2)0(CHR61)sS02R62; or -(CH2)0(CHRδI)s C6H4R8; R42 is H; F; Br; Cl; N02; CF3; alkyl; alkenyl; -(CH2)p(CHR61)sOR55; -(CH2)p(CHRδI)sNR33R34; -(CH2)0(CHRδl)sCOOR57; -(C_i2)0(CΗss.
Figure imgf000015_0001
-(CH2)o(CHRδl)sPO(ORδ0)2;
-(CH2)0(CHRδl)sS02R62; or -(CH2)0(CHRδl)s C6H4R8; R43 is H; alkyl; alkenyl; -(CH2)m(CHRδl)sOR55; -(CH2)m(CHRδl)sNR33R34; -
(CH2)0(CHRδI)sCOOR57; -(CH2)0(CHR61)sCONR58R59; -(CH2)o(CHRδl)sPO(ORδ0)2; - (CH2)o(CHR61)sS02R62; or -(CH2)0(CHRδI)s C6H4R8;
R44 is alkyl; alkenyl; -(CH2)r(CHRδI)sOR55; -(CH2)r(CHRδI)sSR5δ; -(CH2)r(CHR6I)sNR33R34;
-(CH2)r(CHR61)sCOOR57; -(CH2)r(CHR61)sCONR58R59; -(CH2)r(CHRδI)sPO(OR60)2;
-(CH2)r(CHRδl)s S02R62; or -(CH2)r(CHR61)sC6H4R8; R45 is H; alkyl; alkenyl; -(CH2)0(CHRδl)sOR55; -(CH2)0(CHRδl)sSR5δ; -(CH2)0(CHRδl)sNR33R34; -(CH2)o(CHRδI)sCOOR57; -(CH2)s(CHR6I)sCONR58R59; -(CH2)s(CHRδ,)sPO(ORδ0)2;
-(CH2)s(CHRδl)s S02R62; or -(CH2)s(CHRδl)sC6H4R8; R4δ is H; alkyl; alkenyl; or -(CH2)0(CHR61)PC6H4R8; R47 is H; alkyl; alkenyl; or -(CH2)0(CHRδI)s0R55; R48 is H; lower alkyl; lower alkenyl; or aryl-lower alkyl; R49 is H; alkyl; alkenyl; -(CHRδl)sC00R57; (CHRδl)sCONR58R59; (CHRδl)sPO(OR60)2;
-(CHRδI)sSOR62; or -(CHRδl)sC6H4R8; R50 is H; lower alkyl; or aryl-lower alkyl; R51 is H; alkyl; alkenyl; -(CH2)m(CHRδl)sOR55; -(CH2)m(CHRδl)sSR56; -
(CH2)m(CHRδl)sNR33R34; -(CH2)0(CHRδl)sCOOR57; -(CH2)0(CHRδl)sCONR58R59; - (CH2)o(CHRδl)pPO(ORδ0)2; -(CH2)p(CHRδ,)s S02R62; or -(CH2)p(CHRδl)sC6H4R8;
R52 is H; alkyl; alkenyl; -(CH2)m(CHRδI)sOR55; -(CH2)m(CHRδl)sSR5δ; -
(CH2)m(CHRδl)sNR33R34;
-(CH2)o(Cm61)sCOOR57; -(CH2)o(C-HR6I)sCONR58R59; -(CH2)o(CHR61)pPO(OR60)2;
-(CH2)p(CHRδl)s S02R62; or -(CH2)p(CHRδl)sC6H4R8; R53 is H; alkyl; alkenyl; -(CH2)m(CHRδl)sOR55; -(CH2)m(CHRδl)sSR5δ; -
(CH2)m(CHRδl)sNR33R34;
-(CH2)o(CHR61)sCOOR57; -(CH2)o(CHR61)sCONR58R59; -(CH2)o(CHR61)pPO(OR60)2;
-(CH2)p(CHRδl)s S02R62; or -(CH2)p(CHRδl)sC6H4R8; R54 is H; alkyl; alkenyl; -(CH2)m(CHR61)sOR55; -(CH2)ra(CHRδI)sNR33R34; - (CH2)0(CHRδI)COOR57; -(CH2)0(CHRδ,)sCONR58R59; or -(CH2)0(CHRδl)s C6H4R8;
R55 is H; lower alkyl; lower alkenyl; aryl-lower alkyl; -(CH2)m(CHRδl)sOR57;
-(CH2)m(CHRδl)sNR34R63; -(CH2)0(CHRδl)s-COR64; -(CH2)0(CHRδl)COOR57; or
-(CH2)0(CHRδI)sCONR58R59; R5δ is H; lower alkyl; lower alkenyl; aryl-lower alkyl; -(CH2)m(CHRδl)sOR57; -(CH2)m(CHRδl)sNR34R63;
Figure imgf000016_0001
or -(CH2)0(CHRδl)sCONR58R59; R57 is H; lower alkyl; lower alkenyl; aryl lower alkyl; or heteroaryl lower alkyl; R58 is H; lower alkyl; lower alkenyl; aryl; heteroaryl; aryl-lower alkyl; or heteroaryl-lower alkyl; R59 is H; lower alkyl; lower alkenyl; aryl; heteroaryl; aryl-lower alkyl; or heteroaryl-lower alkyl; or R58 and R59 taken together can form: -(CH2)2.6-; -(CH2)2θ(CH2)2-; -(CH2)2S(CH2)2-; or
-(CH2)2NR34(CH2)2-; R60 is H; lower alkyl; lower alkenyl; aryl; or aryl-lower alkyl; Rδl is alkyl; alkenyl; aryl; heteroaryl; aryl-lower alkyl; heteroaryl-lower alkyl; -(CH2)mOR55;
-(CH2)mNR33R34; -(CH2)0COOR37; -(CH2)0NR58R59; or -(CH2)0PO(CORδ0)2; R62 is lower alkyl; lower alkenyl; aryl, heteroaryl; or aryl-lower alkyl; R63 is H; lower alkyl; lower alkenyl; aryl, heteroaryl; aryl-lower alkyl; heteroaryl-lower alkyl;
-COR64; -COOR57; -CONR58R59; -S02R62; or -PO(ORδ0)2; R64 is H; lower alkyl; lower alkenyl; aryl; heteroaryl; aryl-lower alkyl; heteroaryl-lower alkyl;
-(CH2)p(CHRδl)sOR65; -(CH2)P(CHR61)SSR66; or -(CH2)p(CHRδl)sNR34R63; R65 is H; lower alkyl; lower alkenyl; aryl, aryl-lower alkyl; heteroaryl-lower alkyl; -COR57;
-COOR57; or -CONR58R59; Rδδ is H; lower alkyl; lower alkenyl; aryl; aryl-lower alkyl; heteroaryl-lower alkyl; or - CONR58R59; m is 2-4; o is 0-4; p is 1-4; q is 0-2; r is 1 or 2; s is 0 or 1;
Z is a chain of n α-amino acid residues, n being the integer 7 or 11, the positions of said amino acid residues in said chain being counted starting from the N-terminal amino acid, whereby these amino acid residues are, depending on their position in the chains, Gly, or Pro, or of formula -A- CO-, or of one of the types C: -NR20CH(R72)CO- D: -NR20CH(R73)CO- E: -NR20CH(R74)CO- F: -NR20CH(R84)CO-; and
H: -NR20-CH(CO-)-(CH2) -7-CH(CO-)-NR20-;
-NR20-CH(CO-)-(CH2)PSS(CH2)p-CH(CO-)-NR20-;
-NR20-CH(CO-)-(-(CH2)PNR20CO(CH2)p-CH(CO-)-NR20-; and
-NR20-CH(CO-)-(-(CH2)pNR20CONR20(CH2)p-CH(CO-)-NR20-; R71 is H; lower alkyl; lower alkenyl;
Figure imgf000017_0001
,s R75; -(CH2)p(CHRδI)sSR75; -(CH2)PNR78R79 ;
-(CH2)o(CHRδl)sCOOR75; -(CH2)pCONR78R79; -(CH2)pPO(OR62)2; -(CH2)pS02R62; or
-(CH2)o-C6R67Rδ8R69R70R76; R72 is H; lower alkyl; lower alkenyl; -(CH2)p(CHRδl)s0R85; or -(CH2)p(CHRδl)sSR85; R73 is -(CH2)0R77; -(CH2)rO(CH2)0R77; -(CH2)rS(CH2)0R77; or -(CH2)rNR20(CH2)oR77; R74 is -(CH2)PNR78R79; -(CH2)PC(=NR80)NR78R79; -(CH2)PC(=NOR50)NR78R79;
Figure imgf000017_0002
-(CH2)PC6H4C(=NNR78R79)NR78R79; -(CH2)PC6H4NR80C(=NR80)NR78R79; -(CH2)rO(CH2)mNR78R79; -(CH2)rO(CH2)pC(=NR80)NR78R79; -
(CH2)rO(CH2)pC(=NOR50)NR78R79;
-(CH2)rO(CH2)pC(=NNR78R79)NR78R79; -(CH2)rO(CH2)mNR80C(=NR80)NR78R79;
-(CH2)rO(CH2)pC6H4CNR78R79; -(CH2)rO(CH2)pC6H4C(=NR80)NR78R79;
-(CH2)rO(CH2)pC6H4C(=NOR50)NR78R79;- (CH2)rO(CH2)pC6H4C(=NNR78R79)NR78R79; -(CH2)rO(CH2)pC6H4NR80C(=NR80)NR78R79; -(CH2)rS(CH2)mNR78R79;
-(CH2)rS(CH2)pC(=NR80)NR78R79; -(CH2)rS(CH2)pC(=NOR50)NR78R79;
-(CH2)rS(CH2)pC(=NNR78R79)NR78R79; -(CH2)rS(CH2)mNR80C(=NR80)NR78R79;
-(CH2)rS(CH2)pC6H4CNR78R79; -(CH2)rS(CH2)pC6H4C(=NR80)NR78R79;
-(CH2)rS(CH2)pC6H4C(=NOR50)NR78R79; -(CH2)rS(CH2)pC6H4C(=NNR78R79)NR78R79; -(CH2)rS(CH2)pC6H4NR80C-(=NR80)NR78R79; -(CH2)PNR80CONR78R79; or
-(CH2)PC6H4NR80CONR78R79; R75 is lower alkyl; lower alkenyl; or aryl-lower alkyl; R76 is H; lower alkyl; lower alkenyl; aryl-lower alkyl; -(CH2)0OR72; -(CH2)0SR72; -
(CH2)0NR33R34; -(CH2)QCOOR75; -(CH2)0CONR58R59; -(CH2)oPO(ORδ0)2; -(CH2)pS02R62; or -
(CH2)0COR64; R77 is -C.5R67R68R69R70R76; or a heteroaryl group of one of the formulae
Figure imgf000017_0003
Figure imgf000018_0001
H6 H7 H8 H9 H10
Figure imgf000018_0002
H11 H12 H13 H14 H15
Figure imgf000018_0003
H16 H17 H18 H19 H20
Figure imgf000018_0004
H26 H27 H28 H29
Figure imgf000018_0005
H30 H31 H32 H33
Figure imgf000018_0006
H38 H39 H40 H41
Figure imgf000019_0001
H42 H43 H44 H45
Figure imgf000019_0002
H46 H47 H48 H49
Figure imgf000019_0003
H50 H51
R78 is H; lower alkyl; aryl; or aryl-lower alkyl;
R79 is H; lower alkyl; aryl; or aryl-lower alkyl; or
R78 and R79, taken together, can be -(CH2)2.7-; -(CH2)20(CH2)2-; or -(CH2)2NR33(CH2)2-;
R80 is H; or lower alkyl;
R81 is H; lower alkyl; or aryl-lower alkyl;
R82 is H; lower alkyl; aryl; heteroaryl; or aryl-lower alkyl;
Rw is H; lower alkyl; aryl; or -NR 7'S8τR, 7/9y;.
R84 is -(CH2)m(CHR61)sOH; -(CH2)pC0NR78R79; -(CH2)PNR80CONR78R79; -(CH2)pC6H4CONR78R79;
-(CH2)pCOOR80 or -(CH2)PC6H4NR80CONR78R79; R85 is lower alkyl; or lower alkenyl;
with the proviso that in said chain of n α-amino acid residues Z if n is 7, the amino acid residues in positions 1 to 7 are: - PI : of type C or of type F or of type D;
P2: of type E or of type C or of type D or of type F;
P3 : of type F or of type C, or the residue is Gly or Pro;
P4: of type C or of type D or of type F, or the residue is Gly or Pro;
P5 : of type F or of formula -A-CO-, or the residue is Gly or Pro; P6: of type C or of type E or o± -luiium-α -A-CO-, or the residue is Pro; P7: of type C or of type F or of type D;
if n is 11, the amino acid residues in positions 1 to 11 are: - PI: of type E or of type F or of type C;
P2: of type C or of type F or of type E;
P3: oftype C or oftype F;
P4: of type E or of type C or of type D or of type F, or the residue is Gly or Pro;
P5: of type F or of type C, or the residue is Gly or Pro; - P6: oftype C or of type D or of type F, or the residue is Gly or Pro;
P7: of type F or of formula -A-CO-, or the residue is Gly or Pro;
P8: of type C or of type E or of formula -A-CO-, or the residue is Gly or Pro;
P9: of type C or of type F;
P 10: of type F or of type C; - Pl l: of type D or of type E or of type F or of type C; or
P2 and P 10, taken together, can form a group of type H;
and pharmaceutically acceptable salts thereof.
In accordance with the present invention these β-hairpin peptidomimetics can be prepared by a process which comprises
(a) coupling an appropriately functionalized solid support with an appropriately N-protected derivative of that amino acid which in the desired end-product is in position 72+V2 or 72- 2, any functional group which may be present in said N-protected amino acid derivative being likewise appropriately protected;
(b) removing the N-protecting group from the product thus obtained;
(c) coupling the product thus obtained with an appropriately N-protected derivative of that amino acid which in the desired end-product is one position nearer the N-terminal amino acid residue, any functional group which may be present in said N-protected amino acid derivative being likewise appropriately protected;
(d) removing the N-protecting group from the product thus obtained;
(e) repeating, if necessary, steps (c) and (d) until the N-terminal amino acid residue has been introduced;
(f) coupling the product thus obtained to a compound of the general formula
Figure imgf000021_0001
wherein
Figure imgf000021_0002
is as defined above and X is an N-protecting group or, if
°
Template is to be group (al) or (a2), above, alternatively
(fa) coupling the product obtained in step (d) or (e) with an appropriately N-protected derivative of an amino acid of the general formula HOOC-B-H III or HOOC-A-H IV wherein B and A are as defined above, any functional group which may be present in said N-protected amino acid derivative being likewise appropriately protected;
(fb) removing the N-protecting group from the product thus obtained; and
(fc) coupling the product thus obtained with an appropriately N-protected derivative of an amino acid of the above general formula IV and, respectively, HI, any functional group which may be present in said N-protected amino acid derivative being likewise appropriately protected;
(g) removing the N-protecting group from the product obtained in step (f) or (fc);
(h) coupling the product thus obtained to an appropriately N-protected derivative of that amino acid which in the desired end-product is in position n, any functional group which may be present in said N-protected amino acid derivative being likewise appropriately protected;
(i) removing the N-protecting group from the product thus obtained;
0) coupling the product thus obtained to an appropriately N-protected derivative of that amino acid which in the desired end-product is one position farther away from position n, any functional group which may be present in said N-protected amino acid derivative being likewise appropriately protected; (k) removing the N-protecting group from w.- pj-uuuU thus obtained;
(1) repeating, if necessary, steps (j) and (k) until all amino acid residues have been introduced;
(m) if desired, selectively deprotecting one or several protected functional group(s) present in the molecule and appropriately substituting the reactive group(s) thus liberated;
(o) detaching the product thus obtained from the solid support;
(p) cyclizing the product cleaved from the solid support;
(q) if desired, forming an interstrand linkage between side-chains of appropriate amino acid residues at opposite positions of the β-strand region; (r) removing any protecting groups present on functional groups of any members of the chain of amino acid residues and, if desired, any protecting group(s) which may in addition be present in the molecule; and
(r) if desired, converting the product thus obtained into a pharmaceutically acceptable salt or converting a pharmaceutically acceptable, or unacceptable, salt thus obtained into the corresponding free compound of formula I or into a different, pharmaceutically acceptable, salt.
As used in this description, the term "alkyl", taken alone or in combinations, designates saturated, straight-chain or branched hydrocarbon radicals having up to 24, preferably up to 12, carbon atoms. Similarly, the term "alkenyl" designates straight chain or branched hydrocarbon radicals having up to 24, preferably up to 12, carbon atoms and containing at least one or, depending on the chain length, up to four olefinic double bonds. The term "lower" designates radicals and compounds having up to 6 carbon atoms. Thus, for example, the term "lower alkyl" designates saturated, straight-chain or branched hydrocarbon radicals having up to 6 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl and the like. The term "aryl" designates aromatic carbocyclic hydrocarbon radicals containing one or two six-membered rings, such as phenyl or naphthyl, which may be substituted by up to three substituents such as Br, Cl, F, CF3, N02, lower alkyl or lower alkenyl. The term "heteroaryl" designates aromatic heterocyclic radicals containing one or two five- and/or six-membered rings, at least one of them containing up to three heteroatoms selected from the group consisting of O, S and N and said ring(s) being optionally substituted; representative examples of such optionally substituted heteroaryl radicals are indicated hereinabove in connection with the definition of R77.
The structural element -A-CO- designates amino acid building blocks which in combination with the structural element -B-CO- form templates (al) and (a2). Templates (a) through (p) constitute building blocks which have an N-terminus and a oriented in space in such a way that the distance between those two groups may lie between 4.0-5.5A. A peptide chain is linked to the C-terminus and the N-terminus of the templates (a) through (p) via the corresponding N- and C- termini so that the template and the chain form a cyclic structure such as that depicted in formula I. In a case as here where the distance between the N- and C- termini of the template lies between 4.0-5.5A the template will induce the H-bond network necessary for the formation of a β-hairpin conformation in the peptide chain Z. Thus template and peptide chain form a β-hairpin mimetic. The β-hairpin conformation is highly relevant for the protease inhibitory activities of the β- hairpin mimetics of the present invention. The β-hairpin stabilizing conformational properties of the templates (a) through (p) play a key role not only for protease inhibitory activity but also for the synthesis process defined hereinabove, as incorporation of the templates near the middle of the linear protected peptide precursors enhance significantly cyclization yields.
Building blocks A1-A69 belong to a class of amino acids wherein the N-terminus is a secondary amine forming part of a ring. Among the genetically encoded amino acids only proline falls into this class.. The configuration of building block Al through A69 is (D), and they are combined with a building block -B-CO- of (L)-configuration. Preferred combinations for templates (al) are- DAl-CO- B-CO- to DA69-CO-LB-CO-. Thus, for example, DPro-LPro constitutes the prototype of templates (al). Less preferred, but possible are combinations where templates (a2) are - Al-CO- DB-CO- to LA69-CO-DB-CO-. Thus, for example, LPro-DPro constitutes a less preferred prototype of template (a2).
It will be appreciated that building blocks -Al-CO- to -A69-CO- in which A has (D)- configuration, are carrying a group R1 at the α-position to the N-terminus. The preferred values for R1 are H and lower alkyl with the most preferred values for R1 being H and methyl. It will be recognized by those skilled in the art, that A1-A69 are shown in (D)-configuration which, for R1 being H and methyl, corresponds to the (Reconfiguration. Depending on the priority of other values for R1 according to the Cahn, Ingold and Prelog-rules, this configuration may also have to be expressed as (S).
In addition to R1 building blocks -Al-CO- to -A69-CO- can carry an additional substituent designated as R2 to R17. This additional substituent can be H, and if it is other than H, it is preferably a small to medium-sized aliphatic or aromatic group. Examples of preferred values for R2 to R17 are: R2: H; lower alkyl; lower alkenyl; (CH2,m --. s. eτe R55: lower alkyl; or lower alkenyl); (CH2)mSR5δ (where R5δ: H; or lower alkyl; or lower alkenyl); (CH2)mNR33R34 (where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl) ; (CH2)mOCONR33R78 (where R33: lower alkyl; or lower alkenyl; R78: H; or lower alkyl); (CH2)mNR20CONR33R78 (where R20: H or lower alkyl; R33: lower alkyl; or lower alkenyl; R78: H; or lower alkyl); (CH2)oN(R20)CORδ4(where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); (CH2)0COOR57 (where R57: lower; or lower alkenyl); (CH2)0CONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; or lower alkyl); (CH2)oPO(ORδ0)2 (where Rδ0 : lower alkyl; or lower alkenyl); (CH2)0S02R62 (where R62: lower alkyl; or lower alkenyl); or (CH2)qC6H4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
R3: H; lower alkyl; lower alkenyl; (CH2)mOR55 (where R55: lower alkyl; or lower alkenyl); (CH2)mSR56 (where R5δ: H; or lower alkyl; or lower alkenyl); (CH2)mNR33R34( where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl) ; (CH2)oN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); (CH2)0COOR57 (where R57: lower alkyl; or lower alkenyl); (CH2)0CONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; lower alkyl); (CH2)oPO(ORδ0)2 (where Rδ0: lower alkyl; or lower alkenyl); (CH2)0SO2R62 (where R62: lower alkyl; or lower alkenyl); or (CH2)qC6H R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
R4: H; lower alkyl; lower alkenyl; (CH2)mOR55 (where R55: lower alkyl; or lower alkenyl); (CH2)mSR56 (where R5δ: H; or lower alkyl; or lower alkenyl); (CH2)mNR33R34( where R33: lower alkyl; or lower alkenyl; R34: Ho r lower alkyl) ; (CH2)raN(R20)CORδ (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); (CH2)0COOR57 (where R57: lower alkyl; or lower alkenyl); (CH2)0CONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; or lower alkyl); (CH2)oPO(ORδ0)2 (where Rδ0: lower alkyl; or lower alkenyl); (CH2)0SO2R62 (where R62: lower alkyl; or lower alkenyl); or (CH2)qC6H4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl;or lower alkoxy).
Rs: lower alkyl; lower alkenyl; (CH2)0OR55 (where R55: lower alkyl; or lower alkenyl); (CH2)0SR5δ (where R5δ: H; or lower alkyl; or lower alkenyl); (CH2)0NR33R34 ( where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl) ; (CH2)mOCONR33R78 (where R33: lower alkyl; or lower alkenyl; R78: H; or lower alkyl); (CH2)mNR20CONR33R78 (where R20: H or lower alkyl; R33: lower alkyl; or lower alkenyl; R78: H; or lower alkyl); (CH2)oN(R20)CORδ4(where: R20: H; or lower alkyl; R64: alkyl; alkenyl; aryl; and aryl-lower alkyl; heteroaryl-lower alkyl); (CH2)0COOR57 (where R57: lower alkyl; or lower alkenyl); (CH2)0CONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; or lower alkyl); (CH2)oPO(ORδ0)2 (where Rδ0: lower alkyl; or lower alkenyl); (CH2)0S02R62 (where R62: lowc-. ui lower alkenyl); or (CH2)qC6H4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
R6: H; lower alkyl; lower alkenyl; (CH2)0OR55 (where R55: lower alkyl; or lower alkenyl); (CH2)0SR56 (where R56 : H; or lower alkyl; or lower alkenyl); (CH2)0NR 3R34 ( where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl) ; (CH2)0N(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); (CH2)0COOR57 (where R57: lower alkyl; or lower alkenyl); (CH2)0CONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; or lower alkyl); (CH2)oPO(OR60)2 (where R60 : lower alkyl; or lower alkenyl); (CH2)0S02R62 (where R62: lower alkyl; or lower alkenyl); or (CH2)qC6H4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
R7: lower alkyl; lower alkenyl; (CH2)qOR55 (where R55 : lower alkyl; or lower alkenyl); (CH2)qSR56 (where R56 : H or lower alkyl; or lower alkenyl); (CH2)qNR33R34( where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl) ; (CH2)qN(R20)COR64(where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); (CH2)rCOOR57 (where R57: lower alkyl; or lower alkenyl); (CH2)qCONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; or lower alkyl); (CH2)rPO(OR60)2 (where R60 : lower alkyl; or lower alkenyl); (CH2)rS02R62 (where R62: lower alkyl; or lower alkenyl); or (CH2)qC6H R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl;or lower alkoxy).
Rs: H; F; Cl; CF3; lower alkyl; lower alkenyl; (CH2)0OR55 (where R55: lower alkyl; or lower alkenyl); (CH2)0SR56 (where R56: H; or lower alkyl; or lower alkenyl); (CH2)0NR33R34( where R33: lower alkyl; or lower alkenyl; R34: H or lower alkyl) ; (CH2)0OCONR33R78 (where R33: lower alkyl; or lower alkenyl; R78: H; or lower alkyl); (CH2)0NR20CONR33R78 (where R20: H or lower alkyl; R33: lower alkyl; or lower alkenyl; R78: H; or lower alkyl); (CH2)0N(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); (CH2)0COOR57 (where R57 : lower alkyl; or lower alkenyl); (CH2)0CONR58R59 (where R58 : lower alkyl; or lower alkenyl; and R59: H; or lower alkyl); (CH2)oPO(OR60)2 (where R60: lower alkyl; or lower alkenyl); (CH2)0S02R62 (where R62: lower alkyl; or lower alkenyl); or (CH2)qC6H4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
R9: lower alkyl; lower alkenyl; (CH2)0OR55 (where R55: lower alkyl; or lower alkenyl); (CH2)0SR5δ (where R56: H; or lower alkyl; or lower alkenyl); (CH2)0NR33R34( where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl) ; (CH2)0N(R20)COR64(where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); (CH2)0COOR57 (where R57: lower alkyl; or lower alkenyl); (CH2)0CONR5SR59 (where R58 : lower alkyl; or lower alkenyl; and R59: H; or lower alkyl); (CH2)oPO(OR60)2 (where Rso: lower alkyl; or lower alkenyl); (CH2)0SO2R62 (where R62: lower alkyl; or lower alkenyl); or (CH2)qC H4k w-.j-t-.«. i- 8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
R;o: lower alkyl; lower alkenyl; (CH2)0OR55 (where R55: lower alkyl; or lower alkenyl); (CH2)0SR56 (where R56: H; or lower alkyl; or lower alkenyl); (CH2)0NR33R34 ( where R33: lower alkyl; or lower alkenyl; R34: lower alkenyl); (CH2)0COOR57 (where R57: lower alkyl; or lower alkenyl); (CH2)0CONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; lower alkyl); (CH2)0PO(OR60)2 (where R60: lower alkyl; or lower alkenyl); (CH2)0S02R62 (where R62: lower alkyl; or lower alkenyl); or (CH2)qC6H4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy). - R": H; lower alkyl; lower alkenyl; (CH2)mOR55 (where R55: lower alkyl; or lower alkenyl); (CH2)mSR56 (where R56: H; or lower alkyl; or lower alkenyl); (CH2)mNR33R34( where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl) ; (CH2)mOCONR33R78 ( where R33: lower alkyl; or lower alkenyl; R78: H; or lower alkyl) ; (CH2)mNR20CONR33R78 ( where R20: H; or lower alkyl; R33: lower alkyl; or lower alkenyl; R78: H; or lower alkyl) ; (CH2)mNR20COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); (CH2)0COOR57 (where R57: lower alkyl; or lower alkenyl); (CH2)0CONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; lower alkyl); (CH2)oPO(OR60)2 (where R60: lower alkyl; or lower alkenyl); (CH2)0S02R62 (where R62: lower alkyl; or lower alkenyl); or (CH2)qC6H4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy). - R'2: H; lower alkyl; lower alkenyl; (CH2)mOR55 (where R55: lower alkyl; or lower alkenyl); (CH2)mSR56 (where R56: H; or lower alkyl; or lower alkenyl); (CH2)mNR33R34 ( where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl) ; (CH2)mN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); (CH2)rCOOR57 (where R57: lower alkyl; or lower alkenyl); (CH2)rCONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; or lower alkyl); (CH2)rPO(OR60)2 (where R60: lower alkyl; or lower alkenyl); (CH2)0SO2R62 (where R62: lower alkyl; or lower alkenyl); or (CH2)qCβH4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
R13: lower alkyl; lower alkenyl; (CH2)qOR55 (where R55: lower alkyl; or lower alkenyl); (CH2)qSR56 (where R56: H; or lower alkyl; or lower alkenyl); (CH2)qNR33R34 ( where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl) ; (CH2)qN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); (CH2)rC0057 (where R57: lower alkyl; or lower alkenyl); (CH2)qC0NR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; or lower alkyl); (CH2)rPO(OR60)2 (where R60: lower alkyl; or lower alkenyl); (CH2)rS02R62 (where R62: lower alkyl; or lower alkenyl); or (CH2)qCsH4k v w-ucic ^8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
R14: H; lower alkyl; lower alkenyl; (CH2)mOR55 (where R55: lower alkyl; or lower alkenyl); (CH2)mSR56 (where R56: H; or lower alkyl; or lower alkenyl); (CH2)mNR33R34 ( where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl) ; (CH2)mN(R20)COR64 (where: R20: H; lower alkyl; R64: lower alkyl; or lower alkenyl); (CH2)0COOR57 (where R57: lower alkyl; or lower alkenyl); (CH2)0CONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; or lower alkyl); (CH2)oPO(OR60)2 (where R60: lower alkyl; or lower alkenyl); (CH2)0S02R62 (where R62: lower alkyl; or lower alkenyl); (CH2)qC6H4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
R!S: lower alkyl; lower alkenyl; (CH2)0OR55 (where R55: lower alkyl; or lower alkenyl); (CH2)0SR56 (where R56: H; or lower alkyl; or lower alkenyl); (CH2)0NR33R34 ( where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl) ; (CH2)oN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); particularly favoured are NR20COlower alkyl (R20=H; or lower alkyl); (CH2)0COOR57 (where R57: lower alkyl; or lower alkenyl); (CH2)0CONR58R59
(where R58: lower alkyl, or lower alkenyl; andR59: H; lower alkyl); (CH2)oPO(OR60)2 (where R60 : lower alkyl; or lower alkenyl); (CH2)0S02R62 (where R62: lower alkyl; or lower alkenyl); or (CH2)qC6H4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
R16: lower alkyl; lower alkenyl; (CH2)0OR55 (where R55: lower alkyl; or lower alkenyl); (CH2)0SR56 (where R5δ: H; or lower alkyl; or lower alkenyl); (CH2)0NR33R34( where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl) ; (CH2)oN(R20)COR64 (where: R20 : H; or lower alkyl; R64: lower alkyl; or lower alkenyl); (CH2)0COOR57 (where R57: lower alkyl; or lower alkenyl); (CH2)0CONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; or lower alkyl); (CH2)0PO(OR60)2 (where R60: lower alkyl; or lower alkenyl); (CH2)0S02R62 (where R62: lower alkyl; or lower alkenyl); or (CH2)qC6H4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
R77: lower alkyl; lower alkenyl; (CH2)qOR55 (where R55: lower alkyl; or lower alkenyl); (CH2)qSR56 (where R56: H; or lower alkyl; or lower alkenyl); (CH2)qNR33R34( where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl) ; (CH2)qN(R 0)COR6 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); (CH2)rCOOR57 (where R57: lower alkyl; or lower alkenyl); (CH2)qCONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; lower alkyl); (CH2)rPO(OR60)2 (where R60: lower alkyl; or lower alkenyl); (CH2)rS02R62 (where R62: lower alkyl; or lower alkenyl); or (CH2)qC6H4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy). Among the building blocks Al to A69 the following are preferred: A5 with R2 being H, A8, A22, A25, A38 with R2 being H, A42, A47, and A50. Most preferred are building blocks of type A8' and A8":
Figure imgf000028_0001
A8" A8"
wherein R20 is H or lower alkyl; and R64 is alkyl; alkenyl; aryl; aryl-lower alkyl; or heteroaryl- lower alkyl; and R75 is lower alkyl; lower alkenyl; or aryl-lower alkyl; especially those wherein R75 is allyl (A8'-l) and R64 is n-hexyl (A8"-l). Building block A70 belongs to the class of open-chained α-substituted α-amino acids, building blocks A71 and A72 to the corresponding β-amino acid analogues and building blocks A73-A104 to the cyclic analogues of A70. Such amino acid derivatives have been shown to constrain small peptides in well defined reverse turn or U-shaped conformations (C. M. Venkatachalam, Biopolymers, 1968, 6, 1425-1434; W. Kabsch, C Sander, Biopolymers 1983, 22, 2577). Such building blocks or templates are ideally suited for the stabilization of β-hairpin conformations in peptide loops (D. Obrecht, M. Altorfer, J. A. Robinson, "Novel Peptide Mimetic Building Blocks and Strategies for Efficient Lead Finding", Adv. Med Chem. 1999, Vo , 1-68; P. Balaram, "Non- standard amino acids in peptide design and protein engineering", Curr. Opin. Struct. Biol. 1992, 2, 845-851; M. Crisma, G. Valle, C. Toniolo, S. Prasad, R. B. Rao, P. Balaram, "β-turn conformations in crystal structures of model peptides containing α,α- disubstituted amino acids", Biopolymers 1995, 35, 1-9; V. J. Hruby, F. Al-Obeidi, W. Kazmierski, Biochem. J. 1990, 268, 249-262).
It has been shown that both enantiomers of building blocks -A70-CO- to A104-CO- in combination with a building block -B-CO- of L-configuration can efficiently stabilize and induce β-hairpin conformations (D. Obrecht, M. Altorfer, J. A. Robinson, "Novel Peptide Mimetic
Building Blocks and Strategies for Efficient Lead Finding", Adv. Med Chem. 1999, Vol.4, 1-68; D. Obrecht, C. Spiegler, P. Schonholzer, K. Mϋller, H. Heimgartner, F. Stierli, Helv. Chim. Ada 1992, 75, 1666-1696; D. Obrecht, U. Bohdal, J. Daly, C. Lehmann, P. Schonholzer, K. Mϋller, Tetrahedron 1995, 51, 10883-10900; D. Obre -.-, ^. juw-.nai-n, C. Ruffieux, P. Schonholzer, K. Muller, Helv. Chim. Acta 1995, 78, 1567-1587; D. Obrecht, U. Bohdal, C. Broger, D. Bur, C. Lehmann, R. Ruffieux, P. Schonholzer, C. Spiegler, Helv. Chim. Acta 1995, 78, 563-580; D. Obrecht, H. Karajiannis, C. Lehmann, P. Schonholzer, C. Spiegler, Helv. Chim. Acta 1995, 78, 703-714).
Thus, for the purposes of the present invention templates (al) can also consist of -A70-CO- to A104-CO- where building block A70 to A104 is of either (D)- or (L)-configuration, in combination with a building block -B-CO- of (L)- configuration.
Preferred values for R20 in A70 to A104 are H or lower alkyl with methyl being most preferred. Preferred values for R18, R19 and R2I-R29 in building blocks A70 to A104 are the following: R,s: lower alkyl R19: lower alkyl; lower alkenyl; (CH2)pOR55 (where R55: lower alkyl; or lower alkenyl); (CH2)PSR56 (where R56: H; or lower alkyl; or lower alkenyl); (CH2)PNR33R34 ( where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl) ; (CH2)PN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); (CH2)pCOOR57 (where R57: lower alkyl; or lower alkenyl); (CH2)pCONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; or lower alkyl); (CH2)0PO(OR60)2 (where R60: lower alkyl; or lower alkenyl); (CH2)pS02R62 (where R62: lower alkyl; or lower alkenyl); or (CH2)0C6H4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
R2': H ; lower alkyl; lower alkenyl; (CH2)0OR55 (where R55: lower alkyl; or lower alkenyl); (CH2)0SR56 (where R56: H; or lower alkyl; or lower alkenyl); (CH2)0NR33R34 ( where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl) ; (CH2)0N(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); (CH2)0COOR57 (where R57: lower alkyl; or lower alkenyl); (CH2)0CONR58R59 (where R58: lower alkyl, or lower alkenyl; and R59: H; lower alkyl); (CH2)0PO(OR60)2 (where R60: lower alkyl; or lower alkenyl); (CH2)0S02R62 (where R62: lower alkyl; or lower alkenyl); or (CH2)qC6H4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy). - R22: lower alkyl; lower alkenyl; (CH2)0OR55 (where R55: lower alkyl; or lower alkenyl);
(CH2)0SR56 (where R56: H; or lower alkyl; or lower alkenyl); (CH2)0NR33R34 ( where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl) ; (CH2)oN(R20)COR64(where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); (CH2)0COOR57 (where R57: lower alkyl; or lower alkenyl); (CH2)0CONR58R59 (where R58 : lower alkyl, or lower alkenyl; and R59: H; lower alkyl); (CH2)oPO(OR60)2 (where R60: lower alkyl; or lυ cr aικeπyl); (CH2)0SO2R62 (where R62: lower alkyl; or lower alkenyl); or (CH2)qC6H4R8 (where R8: H; F; Cl; CF; lower alkyl; lower alkenyl; or lower alkoxy).
R23: H; lower alkyl; lower alkenyl; (CH2)0OR55 (where R55: lower alkyl; or lower alkenyl); (CH2)0SR56 (where R5δ: H; or lower alkyl; or lower alkenyl); (CH2)0NR33R34 (where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl) ; (CH2)oN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); particularly favoured are NR20COlower alkyl
(R20=H; or lower alkyl); (CH2)0COOR57 (where R57: lower alkyl; or lower alkenyl);
(CH2)0CONR5sR59 (where R58: lower alkyl, or lower alkenyl; and R59: H; lower alkyl); (CH2)oPO(OR60)2 (where R60: lower alkyl; or lower alkenyl); (CH2)0S02R62 (where R62: lower alkyl; or lower alkenyl); or (CH2)qC6H4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
R24: lower alkyl; lower alkenyl; (CH2)0OR55 (where R55: lower alkyl; or lower alkenyl);
(CH2)0SR56 (where R56: H; or lower alkyl; or lower alkenyl); (CH2)0NR33R34 ( where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl) ; (CH2)oN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); particularly favoured are NR20COlower alkyl (R20=H ; or lower alkyl); (CH2)0COOR57 (where R57: lower alkyl; or lower alkenyl); (CH2)0CONR58R59
(where R58: lower alkyl, or lower alkenyl; and R59: H; lower alkyl); (CH2)0PO(OR60)2 (where R60: lower alkyl; or lower alkenyl); (CH2)0S02R62 (where R62: lower alkyl; or lower alkenyl); or (CH2)qC6H4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
R25: H; lower alkyl; lower alkenyl; (CH2)mOR55 (where R55: lower alkyl; or lower alkenyl); (CH2)mNR33R34( where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl);
(CH2)raN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl);
(CH2)0COOR57 (where R57: lower alkyl; or lower alkenyl); (CH2)0CONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; lower alkyl); (CH2)oPO(OR60)2 (where R60: lower alkyl; or lower alkenyl); (CH2)0S02R62 (where R62: lower alkyl; or lower alkenyl); or (CH2)qC6H4R8
(where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
R26: H; lower alkyl; lower alkenyl; (CH2)mOR55 (where R55 : lower alkyl; or lower alkenyl); (CH2)mNR33R3 ( where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl) ; (CH2)mN(R20)COR64(where: Rzo: H; or lower alkyl; R64: lower alkyl; or lower alkenyl);
(CH2)0COOR57 (where R57: lower alkyl; or lower alkenyl); (CH2)0CONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; lower alkyl); (CH2)0PO(OR60)2 (where R60: lower alkyl; or lower alkenyl); (CH2)0S02R62 (where R62: lower alkyl; or lower alkenyl); or (CH2)qC6H4R8
(where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy). Alternatively, R25 and R26 taken togethc-. .an uc -(CH2)2.6-; -(CH2)20(CH2)2-; - (CH2)2S(CH2)2-; or -(CH2)2NR34(CH2)2-;
R27: H; lower alkyl; lower alkenyl; (CH2)0OR55 (where R55: lower alkyl; or lower alkenyl); (CH2)0SR56 (where R56: H; or lower alkyl; or lower alkenyl); (CH2)0NR33R34 ( where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl) ; (CH2)oN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); (CH2)0COOR57 (where R57: lower alkyl; or lower alkenyl); (CH2)0CONR58R59 (where R58: lower alkyl, or lower alkenyl; and R59: H; lower alkyl); (CH2)0PO(OReo)2 (where R60: lower alkyl; or lower alkenyl); (CH2)0S02R62 (where R62: lower alkyl; or lower alkenyl); or (CH2)qCsH4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
R28 : lower alkyl; lower alkenyl; (CH2)0OR55 (where R55: lower alkyl; or lower alkenyl); (CH2)0SR56 (where R56: H; or lower alkyl; or lower alkenyl); (CH2)0NR33R34 ( here R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl) ; (CH2)0N(R20)COR64(where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); (CH2)0COOR57 (where R57 : lower alkyl; or lower alkenyl); (CH2)0CONR58R59 (where R58: lower alkyl, or lower alkenyl; and R59: H; lower alkyl); (CH2)oPO(OR60)2 (where Rδ0: lower alkyl; or lower alkenyl); (CH2)0S02R62 (where R62: lower alkyl; or lower alkenyl); or (CH2)qC„H4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
R29: lower alkyl; lower alkenyl; (CH2)0OR55 (where R55: lower alkyl; or lower alkenyl); (CH2)0SR56 (where R56: H; or lower alkyl; or lower alkenyl); (CH2)0NR33R34 ( where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl) ; (CH2)oN(R20)COR64(where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); particularly favoured are NR20COlower-alkyl (R20=H; or lower alkyl); (CH2)0COOR57 (where R57: lower alkyl; or lower alkenyl); (CH2)0CONR58R59 (where R58: lower alkyl, or lower alkenyl; and R59: H; lower alkyl); (CH2)oPO(OR60)2 (where R60: lower alkyl; or lower alkenyl); (CH2)0S02R62 (where R62: lower alkyl; or lower alkenyl); or (CH qCβHμ 8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
For templates (b) to (p), such as (bl) and (cl), the preferred values for the various symbols are the following: - R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; (CH2)0OR55 (where R55: lower alkyl; or lower alkenyl); (CH2)0SR56 (where R56: H; or lower alkyl; or lower alkenyl); (CH2)0NR33R34 ( where R33: lower alkyl; or lower alkenyl; R34: H or lower alkyl) ; (CH2)0N(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); (CH2)0COOR57 (where R57: lower alkyl; or lower alkenyl); (CH2)0CONR5sR59 (where R58: lower alkyl; or lower alkenyl; and R59: H; or lower alkyl); (CH2)oPO(OR60)2 (where -u . -.uwci αlkyl; or lower alkenyl); (CH2)0S02R62 (where R62: lower alkyl; or lower alkenyl); or (CH2)qC6H4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
R20 : H; or lower alkyl - R30: H, methyl
R3': H; lower alkyl; lower alkenyl; (CH2)pOR55 (where R55: lower alkyl; or lower alkenyl); (CH2)PNR33R34 ( where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl) ; (CH2)PN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); (CH2)0COOR57 (where R57: lower alkyl; or lower alkenyl); (CH2)0CONR58R59 (where R58: lower alkyl, or lower alkenyl; and R59: H; lower alkyl); (CH2)oPO(OR60)2 (where R60: lower alkyl; or lower alkenyl); (CH2)0S02R62 (where R62: lower alkyl; or lower alkenyl); or (CH2)rC6H4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy); most preferred are CH2CONR58R59 (R58: H; or lower alkyl; R59: lower alkyl; or lower alkenyl).
R32: H, methyl - R33: lower alkyl; lower alkenyl; (CH2)mOR55 (where R55: lower alkyl; or lower alkenyl);
(CH2)mSR56 (where R56: H; or lower alkyl; or lower alkenyl); (CH2)mNR33R34 (where R33 : lower alkyl; or lower alkenyl; R34: H; or lower alkyl) ; (CH2)mOCONR33R78 (where R33: lower alkyl; or lower alkenyl; R78: H; or lower alkyl); (CH2)nvNR20CONR33R78 (where R20: H or lower alkyl; R33: lower alkyl; or lower alkenyl; R78: H; or lower alkyl); (CH2)raN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); (CH2)0COOR57 (where R57: lower alkyl; or lower alkenyl); (CH2)0CONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; lower alkyl).
R34: H; or lower alkyl.
R35: H; lower alkyl; lower alkenyl; (CH2)mOR55 (where R55: lower alkyl; or lower alkenyl); (CH2)mN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); (CH2)0COOR57 (where R57: lower alkyl; or lower alkenyl); (CH2)0CONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; lower alkyl).
R36: lower alkyl; lower alkenyl; or aryl-lower alkyl .
R37: H; lower alkyl; lower alkenyl; (CH2)pOR55 (where R55: lower alkyl; or lower alkenyl); (CH2)PNR33R34 ( where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl) ; (CH2)pN(R20)COR64 (where: R20: H; or lower alkyl; Rfi4: lower alkyl; or lower alkenyl);
(CH2)0COOR57 (where R57 : lower alkyl; or lower alkenyl); (CH2)0CONR58R59 (where R58: lower alkyl, or lower alkenyl; and R59: H; lower alkyl); (CH2)oPO(OR60)2 (where R60: lower alkyl; or lower alkenyl); (CH2)0S02R62 (where R62: lower alky; or lower alkenyl); or (CH2)qC6H4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy). R3S: H; lower alkyl; lower alkenyl; (G-.jyp .- ι^ where R55: lower alkyl; or lower alkenyl); (CH2)PNR33R34 ( where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl) ;
(CH2)PN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl);
(CH2)0COOR57 (where R57: lower alkyl; or lower alkenyl); (CH2)0CONR58R59 (where R58: lower alkyl, or lower alkenyl; and R59: H; lower alkyl); (CH2)0PO(OR60)2 (where R60: lower alkyl; or lower alkenyl); (CH2)0S02R62 (where R62: lower alkyl; or lower alkenyl); or (CH2)qC6H4R8
(where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
R39: H; lower alkyl; lower alkenyl; (CH2)mOR55 (where R55: lower alkyl; or lower alkenyl); (CH2)mN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); (CH2)0COOR57 (where R57: lower alkyl; or lower alkenyl); (CH2)0CONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; lower alkyl).
R40: lower alkyl; lower alkenyl; or aryl-lower alkyl.
R4': H; lower alkyl; lower alkenyl; (CH2)pOR55 (where R55: lower alkyl; or lower alkenyl); (CH2)PNR33R34 ( where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl) ; (CH2)pN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl);
(CH2)0COOR57 (where R57: lower alkyl; or lower alkenyl); (CH2)0CONR58R59 (where R58: lower alkyl, or lower alkenyl; and R59: H; lower alkyl); (CH2)0PO(OR60)2 (where R60: lower alkyl; or lower alkenyl); (CH2)0SO2R62 (where R62: lower alkyl; or lower alkenyl); or (CH2)qC6H4R8
(where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy). - R42: H; lower alkyl; lower alkenyl; (CH2)pOR55 (where R55: lower alkyl; or lower alkenyl); (CH2)PNR33R34 ( where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl) ;
(CH2)PN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl);
(CH2)0COOR57 (where R57: lower alkyl; or lower alkenyl); (CH2)0CONR58R59 (where R58: lower alkyl, or lower alkenyl; and R59: H; lower alkyl); (CH2)0PO(OR60)2 (where R60: lower alkyl; or lower alkenyl); (CH2)0S02R62 (where R62: lower alkyl; or lower alkenyl); or (CH^qC^R8
(where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
R43: H; lower alkyl; lower alkenyl; (CH2)mOR55 (where R55: lower alkyl; or lower alkenyl); (CH2)mSRSδ (where R56 : lower alkyl; or lower alkenyl); (CH2)mNR33R34 ( where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl) ; (CH2)mN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); (CH2)0COOR57 (where R57 : lower alkyl; or lower alkenyl); (CH2)0CONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; lower alkyl);
(CH2)0PO(OR60)2 (where R60: lower alkyl; or lower alkenyl); (CH2)0S02R62 (where R62: lower alkyl; or lower alkenyl); or (CH^qCδHtR8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy). R44: lower alkyl; lower alkenyl; „ e R55: lower alkyl; or lower alkenyl); (CH2)PSR56 (where R56: H; or lower alkyl; or lower alkenyl); (CH2)PNR33R34( where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl) ; (CH2)PN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); (CH2)pCOOR57 (where R57: lower alkyl; or lower alkenyl); (CH2)pCONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; lower alkyl); or (CH2)0C6H4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
R45: H; lower alkyl; lower alkenyl; (CH2)sOR55 (where R55: lower alkyl; or lower alkenyl); (CH2)SSR56 (where R5δ: H; or lower alkyl; or lower alkenyl); (CH2)0NR33R34 ( where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl) ; (CH2)oN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); (CH2)0COOR57 (where R57: lower alkyl; or lower alkenyl); (CH2)0CONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; lower alkyl); or (CH2)SC6H R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
R4S: H; lower alkyl; lower alkenyl; (CH2)sOR55 (where R55: lower alkyl; or lower alkenyl); (CH2)SSR56 (where R56: H; or lower alkyl; or lower alkenyl); (CH2)0NR33R34 (where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl) ; (CH2)oN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); (CH2)0COOR57 (where R57: lower alkyl; or lower alkenyl); (CH2)0CONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; lower alkyl); or (CH2)SC6H R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
R47: H; or OR55 (where R55: lower alkyl; or lower alkenyl). - R4S: H; or lower alkyl.
R49: H;lower alkyl; (CH2)0COOR57 (where R57 : lower alkyl; or lower alkenyl); (CH2)0CONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; lower alkyl); or (CH2)sCβH R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
R50: H; methyl - R5/: H; lower alkyl; lower alkenyl; (CH2)raOR55 (where R55: lower alkyl; or lower alkenyl); (CH2)mNR33R34 ( where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl) ; (CH2)mN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); (CH2)pCOOR57 (where R57: lower alkyl; or lower alkenyl); (CH2)pCONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59 : H; lower alkyl); or (CH2)rC6H4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
R52: H; lower alkyl; lower alkenyl; (CH2)mOR55 (where R55: lower alkyl; or lower alkenyl); (CH2)mNR33R34 (where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl) ; (CH2)mN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); (CH2)pCOOR57 (where R57: lower alkyl; or lower alkenyl); (CH2)pCONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; lower alky _,, w-.
Figure imgf000035_0001
(where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
R53: H; lower alkyl; lower alkenyl; (CH2)mOR55 (where R55: lower alkyl; or lower alkenyl); (CH2)mNR33R34( where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl) ; (CH2)mN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); (CH2)pCOOR57 (where R57: lower alkyl; or lower alkenyl); (CH2)pCONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; lower alkyl); or (CH2)rC6H4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
R54: lower alkyl; lower alkenyl; or aryl-lower alkyl.
Among the building blocks A70 to A104 the following are preferred: A74 with R22 being H, A75, A76, A77 with R22 being H, A78 and A79.
The building block -B-CO- within template (al) and (a2) designates an L-amino acid residue. Preferred values for B are: -NR20CH(R71)- and enantiomers of groups A5 with R2 being H, A8, A22, A25, A38 with R2 being H, A42, A47, and A50. Most preferred are
Ala L-Alanine
Arg L-Arginine
Asn L-Asparagine Cys L-Cysteine
Gin L-Glutamine
Gly Glycine
His L-Histidine
He L-Isoleucine Leu L-Leucine
Lys L-Lysine
Met L-Methionine
Phe L-Phenylalanine
Pro L-Proline Ser L-Serine
Thr L-Threonine
Trp L-Tryptophan
Tyr L-Tyrosine
Val L-Valine Cit
Orn L-Ornithine tBuA L-t-Butylalanine
Sar Sarcosine t-BuG L-tert.-Butylglycine
4AmPhe L-para-Aminophenylalanine
3AmPhe L-meta-Aminophenylalanine
2AmPhe L-ortho-Aminophenylalanine
Phe(mC(NH2)=NH) L-meta-Amidinophenylalanine
Phe(pC(NH2)=NH) L-para-Amidinophenylalanine
Phe(mNHC (NH2)=NH) L-meta-Guanidinophenylalanine
Phe(pNHC (NH2)=NH) L-para-Guanidinophenylalanine
Phg L-Phenylglycine
Cha L-Cyclohexylalanine
C4al L-3-Cyclobutylalanine
C5al L-3 -Cy clopentylalanine
Nle L-Norleucine
2-Nal L-2-Naphthylalanine
1-Nal L- 1 -Naphthylalanine
4C1-Phe L-4-Chlorophenylalanine
3C1-Phe L-3 -Chlorophenylalanine
2C1-Phe L-2-Chlorophenylalanine
3,4Cl2.Phe L-3 ,4-Dichlorophenylalanine
4F-Phe L-4-Fluorophenylalanine
3F-Phe L-3 -Fluorophenylalanine
2F-Phe L-2-Fluorophenylalanine
Tic L- 1 ,2,3 ,4-Tetrahydroisoquinoline-3 ■ -carboxylic acid
Thi L-β-2-Thienylalanine
Tza L-2-Thiazolylalanine
Mso L-Methionine sulfoxide
AcLys L-N-Acetyllysine
Dpr L-2,3-Diaminopropionic acid
A2Bu L-2,4-Diaminobutyric acid
Dbu (S)-2,3-Diaminobutyric acid Abu γ-AmmoDuιyπc acid (GABA)
Aha ε-Aminohexanoic acid
Aib α-Aminoisobutyric acid
Y(Bzl) L-O-Benzyltyrosine
Bip L-Biphenylalanine
S(Bzl) L-O-Benzylserine
T(Bzl) L-O-Benzylthreonine hCha L-Homo-cyclohexylalanine hCys L-Homo-cysteine hSer L-Homo-serine hArg L-Homo-arginine hPhe L-Homo-phenylalanine
Bpa L-4-Benzoylphenylalanine
Pip L-Pipecolic acid
OctG L-Octylglycine
MePhe L-N-Methylphenylalanine
MeNle L-N-Methylnorleucine
MeAla L-N-Methylalanine
Melle L-N-Methylisoleucine
MeVal L-N-Methvaline
MeLeu L-N-Methylleucine
In addition, the most preferred values for B also include groups of type A8"' of (L)- configuration:
Figure imgf000037_0001
A8' wherein R20 is H or lower alkyl and R is alkyl; alkenyl; aryl; aryl-lower alkyl; or heteroaryl- lower alkyl; especially those wherein R 64 . is n-hexyl (A8"'-l) The peptidic chains Z of the β-hairpin mimetioo uwυuucJ herein is generally defined in terms of amino acid residues belonging to one of the following groups:
Group C -NR20CH(R72)CO-; «hydrophobic: small to medium-sized»
Group D -NR20CH(R73)CO-; «hydrophobic: large aromatic or heteroaromatic» - Group E -NR20CH(R74)CO-; "polar-cationic" and "urea-derived"
Group F -NR20CH(R84)CO-; "polar-non-charged" and "anionic"
Group H -NR20-CH(CO-)-(CH2)4.7-CH(CO-)-NR20-;
-NR20-CH(CO-)-(CH2)pSS(CH2)p-CH(CO-)-NR20-; -NR20-CH(CO-)-(-(CH2)pNR20CO(CH2)p-CH(CO-)-NR20-; and -NR20-CH(CO-)-(-(CH2)pNR20CONR20(CH2)p-CH(CO-)-NR20-;
"interstrand linkage" Furthermore, the amino acid residues in chain Z can also be of formula -A-CO- wherein A is as defined above.
Group C comprises amino acid residues with small to medium-sized hydrophobic side chain groups according to the general definition for substituent R72. A hydrophobic residue refers to an amino acid side chain that is uncharged at physiological pH and that is repelled by aqueous solution . Furthermore these side chains generally do not contain hydrogen bond donor groups, such as (but not limited to) primary and secondary amides, primary and secondary amines and the corresponding protonated salts thereof, thiols, alcohols, phosphonates, phosphates, ureas or thioureas. However, they may contain hydrogen bond acceptor groups such as ethers, thioethers, esters, tertiary amides, alkyl- or aryl phosphonates and phosphates or tertiary amines. Genetically encoded small-to-medium-sized amino acids include alanine, isoleucine, leucine, methionine and valine.
Group D comprises amino acid residues with aromatic and heteroaromatic side chain groups according to the general definition for substituent R73. An aromatic amino acid residue refers to a hydrophobic amino acid having a side chain containing at least one ring having a conjugated π- electron system (aromatic group). In addition they may contain hydrogen bond donor groups such as (but not limited to) primary and secondary amides, primary and secondary amines and the corresponding protonated salts thereof, thiols, alcohols, phosphonates, phosphates, ureas or thioureas, and hydrogen bond acceptor groups such as (but not limited to) ethers, thioethers, esters, tetriary amides, alkyl- or aryl phosphonates -and phosphates or tertiary amines. Genetically encoded aromatic amino acids include phenylalanine and tyrosine. A heteroaromatic amino acid residue refers to a hydrophobic amino acid having a side chain containing at least one ring having a conjugated π-system incorporating at least one heteroatom such as (but not limited to) O, S and N according to the general definition for substituent R77. In addition they may contain hydrogen bond donor groups such as (but not limited to) primary and secondary amides, primary and secondary amines and the corresponding protonated salts thereof, thiols, alcohols, phosphonates, phosphates, ureas or thioureas, and hydrogen bond acceptor groups such as (but not limited to) ethers, thioethers, esters, tetriary amides, alkyl- or aryl phosphonates -and phosphates or tertiary amines. Genetically encoded heteroaromatic amino acids include tryptophan and histidine.
Group E comprises amino acids containing side chains with polar-cationic and urea-derived residues according to the general definition for substituen R74. Polar-cationic refers to a basic side chain which is protonated at physiological pH. Genetically encoded polar-cationic amino acids include arginine, lysine and histidine. Citrulline is an example for an amino acid containing a urea-derived residue.
Group F comprises amino acids containing side chains with polar-non-charged or anionic residues according to the general definition for substituent R84. A polar-non-charged or anionic residue refers to a hydrophilic side chain that is uncharged and, respectively, anionic at physiological pH (carboxylic acids are included), but that is not repelled by aqueous solutions. Such side chains typically contain hydrogen bond donor groups such as (but not limited to) primary and secondary amides, carboxylic acids and esters, primary and secondary amines, thiols, alcohols, phosphonates, phosphates, ureas or thioureas. These groups can form hydrogen bond networks with water molecules. In addition they may also contain hydrogen bond acceptor groups such as (but not limited to) ethers, thioethers, esters, tetriary amides, carboxylic acids and carboxylates, alkyl- or aryl phosphonates -and phosphates or tertiary amines. Genetically encoded polar-non-charged and anionic amino acids include asparagine, cysteine, glutamine, serine and threonine but also aspartic acid and glutamic acid.
Group H comprises side chains of preferably (L)-amino acids at opposite positions of the β- strand region that can form an interstrand linkage. The most widely known linkage is the disulfide bridge formed by cysteines and homo-cysteines positioned at opposite positions of the β-strand. Various methods are known to form disulfide linkages including those described by: J. P. Tarn et al. Synthesis 1979, 955-957; Stewart et al. , Soaa rnase eptide Synthesis, 2d Ed., Pierce Chemical Company, III., 1984; Ahmed et al. J. Biol. Chem. 1975, 250, 8477-8482 ; and Pennington et al., Peptides, pages 164-166, Giralt and Andreu, Eds., ESCOM Leiden, The Netherlands, 1990. Most advantageously, for the scope of the present invention, disulfide linkages can be prepared as described hereinafter in the pertinent Examples (procedure 3), using acetamidomethyl (Acm)- protective groups for cysteine. A well established interstrand linkage consists in linking ornithines and lysines, respectively, with glutamic and aspartic acid residues located at opposite β-strand positions by means of an amide bond formation. Preferred protective groups for the side chain amino-groups of ornithine and lysine are allyloxycarbonyl (Alloc) and allylesters for aspartic and glutamic acid. Finally, interstrand linkages can also be established by linking the amino groups of lysine and ornithine located at opposite β-strand positions with reagents such as N,N-carbonylimidazole to form cyclic ureas.
As mentioned earlier, positions for interstrand linkages are the following:
if n=ll Positions P2 and P10 taken together.
Such interstrand linkages are known to stabilize the β-hairpin conformations and thus constitute an important structural element for the design of β-hairpin mimetics.
Most preferred amino acid residues in chain Z are those derived from natural α-amino acids. Hereinafter follows a list of amino acids which, or the residues of which, are suitable for the purposes of the present invention, the abbreviation corresponding to generally adopted usual practice:
three letter code one let ter code
Ala L-Alanine A Arg L-Arginine R Asn L-Asparagine N Asp L-Aspartic acid D Cys L-Cysteine C Glu L-Glutamic acid E Gin L-Glutamine Q Gly Glycή-.v, G
His L-Histidine H lie L-Isoleucine I
Leu L-Leucine L
Lys L-Lysine K
Met L-Methionine M
Phe L-Phenylalanine F
Pro L-Proline P
DPro D-Proline DP
Ser L-Serine S
Thr L-Threonine T
Trp L-Tryptophan W
Tyr L-Tyrosine Y
Val L-Valine V
Other α-amino acids which, or the residues of which, are suitable for the purposes of the present invention include:
Cit L-Citrulline
Orn L-Ornithine tBuA L-t-Butylalanine
Sar Sarcosine
Pen L-Penicillamine t-BuG L-tert.-Butylglycine
4AmPhe L-para-Aminophenylalanine
3AmPhe L-meta-Aminophenylalanine
2AmPhe L-ortho-Aminophenylalanine
Phe(mC(NH2)=NH) L-meta-Amidinophenylalanine
Phe(pC(NH2)=NH) L-para-Amidinophenylalanine
Phe(mNHC (NH2)=NH) L-meta-Guanidinophenylalanine
Phe(pNHC (NH2)=NH) L-para-Guanidinophenylalanine
Phg L-Phenylglycine
Cha L-Cyclohexylalanine
C4al L-3 -Cyclobutylalanine
C5al L-3 -Cyclopentylalanine Nle L-Noπeucme
2-Nal L-2-Naphthylalanine
1-Nal L- 1 -Naphthylalanine
4C1-Phe L-4-Chlorophenylalanine
3C1-Phe L-3 -Chlorophenylalanine
2C1-Phe L-2-Chlorophenylalanine
3,4C12-Phe L-3 ,4-Dichlorophenylalanine
4F-Phe L-4-Fluorophenylalanine
3F-Phe L-3 -Fluorophenylalanine
2F-Phe L-2-Fluorophenylalanine
Tic 1,2,3 ,4-Tetrahydroisoquinoline-3 -carboxylic acid
Thi L-β-2-Thienylalanine
Tza L-2-Thiazolylalanine
Mso L-Methionine sulfoxide
AcLys N-Acetyllysine
Dpr 2,3-Diaminopropionic acid
A2Bu 2,4-Diaminobutyric acid
Dbu (S)-2,3-Diaminobutyric acid
Abu γ-Aminobutyric acid (GABA)
Aha ε-Aminohexanoic acid
Aib α-Aminoisobutyric acid
Y(Bzl) L-O-Benzyltyrosine
Bip L-(4-phenyl)phenylalanine
S(Bzl) L-O-Benzylserine
T(Bzl) L-O-Benzylthreonine hCha L-Homo-cyclohexylalanine hCys L-Homo-cysteine hSer L-Homo-serine hArg L-Homo-arginine hPhe L-Homo-phenylalanine
Bpa L-4-Benzoylphenylalanine
4-AmPyrrl (2S,4S)-4-Amino-pyrτolidine-L-carboxylic acid
4-AmPyrr2 (2S,4R)-4-Amino-pyrrolidine-L-carboxylic acid
4-PhePyrrl (2S,5R)-4-Phenyl-pyrrolidine-L-carboxylic acid 42
4-PhePyrr2 (2S,5„, . J. -._.v.-. l-pyrrolidine-L-carboxylic acid
5-PhePyrrl (2S,5R)-5-Phenyl-ρyrrolidine-L-carboxylic acid
5-PhePyrr2 (2S,5S)-5-Phenyl-pyrrolidine-L-carboxylic acid
Pro(4-OH)l (4S)-L-Hydroxyproline
•" Prό(4-OH)2 (4R)-L-Hydroxyproline
Pip L-Pipecolic acid
DPip D-Pipecolic acid
OctG L-Octylglycine
MePhe L-N-Methylphenylalanine
MeNle L-N-Methylnorleucine
MeAla L-N-Methylalanine
MeDe L-N-Methylisoleucine
MeVal L-N-Methylvaline
MeLeu L-N-Methylleucine 5 -
Particularly preferred residues for group C are:
Ala L-Alanine lie L-Isoleucine
Leu L-Leucine 0 Met L-Methionine
Val L-Valine tBuA L-t-Butylalanine t-BuG L-tert. -Butylglycine
Cha L-Cyclohexylalanine 5 C4al L-3 -Cyclobutylalanine
C5al L-3 -Cyclopentylalanine
Nle L-Norleucine hCha L-Homo-cyclohexylalanine
OctG L-Octylglycine 0 MePhe L-N-Methylphenylalanine
MeNle L-N-Methylnorleucine
MeAla L-N-Methylalanine
Melle L-N-Methylisoleucine
MeVal L-N-Methylvaline MeLeu L-N- -. _. -. -ι-ιe
Particularlily preferred residues for group D are:
His L-Histidine Phe L-Phenylalanine
Trp L-Tryptophan
Tyr L-Tyrosine
Phg L-Phenylglycine
2-Nal L-2-Naphthylalanine 1-Nal L-1-Naphthylalanine
4C1-Phe L-4-Chlorophenylalanine
3C1-Phe L-3-Chlorophenylalanine
2C1-Phe L-2-Chlorophenylalanine
3 ,4C12-Phe L-3 ,4-Dichlorophenylalanine 4F-Phe L-4-Fluorophenylalanine
3F-Phe L-3-Fluorophenylalanine
2F-Phe L-2-Fluorophenylalanine
Thi L-β-2-Thienylalanine
Tza L-2-Thiazolylalanine Y(Bzl) L-O-Benzyltyrosine
Bip L-Biphenylalanine
S(Bzl) L-O-Benzylserine
T(Bzl) L-O-Benzylthreonine hPhe L-Homo-phenylalanine Bpa L-4-Benzoylphenylalanine
Particularly preferred residues for group E are
Arg L-Arginine
Lys L-Lysine Orn L-Ornithine
Dpr L-2,3-Diaminopropionic acid
A2Bu L-2,4-Diaminobutyric acid Dbu (S)-2,3-Diaminobutyric acid
Phe(pNH2) L-para-Aminophenylalanine Phe(m-NH2) L-me ,.-,,--- ---.-.i-.-.j-up-.-.enylalanine
Phe(oNH2) L-ortho-Aminophenylalanine hArg L-Homo-arginine
Phe(mC(NH2)=NH) L-meta-Amidinophenylalanine Phe(pC(NH2)=NH) L-para-Amidinophenylalanine
Phe(mNHC (NH2)==NH) L-meta-Guanidinophenylalanine
Phe(pNHC (NH2)=NH) L-para-Guanidinophenylalanine
Cit L-Citrulline
Particularly preferred residues for group F are
Asp L-Aspartic acid
Asn L-Asparagine
Cys L-Cysteine
Glu L-Glutamic acid Gin L-Glutamine
Ser L-Serine
Thr L-Threonine
Cit L-Citrulline
Pen L-Penicillamine AcLys L-Nε-Acetyllysine hCys L-Homo-cysteine hSer L-Homo-serine
Generally, the peptidic chain Z within the β-hairpin mimetics of the invention comprises 7 or 11 amino acid residues (n = 7 or 11). The positions P1 to Pn of each amino acid residue in the chain Z are unequivocally defined as follows: P1 represents the first amino acid in the chain Z that is coupled with its N-terminus to the C-terminus of the templates (b)-(p) or of group -B-CO- in template (al), or of Group -A-CO- in template (A2) and Pn represents the last amino acid in the chain Z that is coupled with its C-terminus to the N-terminus of the templates (b)-(p) or of group -A-CO- in template (al) or of group -B-CO- in template (A2). Each of the positions P1 to Pn will preferably contain an amino acid residue belonging to one or two or three of above types C to F, or being Pro, as follows: if n is 7, the amino acid residues in position 1 - 7 are preferably: PI: oftype C or oftype F; P2: of type E or of type D --,-. υJ. ι C C;
P3: oftype F or of type C;
P4: of type C or type F or of type D;
P5 : of type F, or the residue is Pro;
P6: of type C or of type E, or the residue is Pro;
P7: of type C or of type F; if n is 11, the amino acid residues in position 1 - 11 are preferably:
PI: of type E or of type F;
P2: of type C or of type F;
P3: of type C;
P4: of type E or of type D or of type C;
P5: oftype F or of type C;
P6: of type C, or of type D;
P7: of type F, or the residue is Pro;
P8: of type C or of type E, or the residue is Pro;
P9: of type C or of type F;
P10: oftype F or oftype C;
PI 1 : of type D or of type E; or P2 and P10, taken together can form a group of type H;
Particularly preferred β-peptidomimetics of the invention include those described in Examples 1, 4, 7, 8 and 15.
The process of the invention can advantageously be carried out as parallel array synthesis to yield libraries of template-fixed β-hairpin peptidomimetics of the above general formula I. Such parallel synthesis allows one to obtain arrays of numerous (normally 24 to 192, typically 96) compounds of general formula I in high yields and defined purities, minimizing the formation of dimeric and polymeric by-products. The proper choice of the functionalized solid-support (i.e. solid support plus linker molecule), templates and site of cyclization play thereby key roles.
The functionalized solid support is conveniently derived from polystyrene crosslinked with, preferably 1-5%, divinylbenzene; polystyrene coated with polyethyleneglycol spacers (TentagelR); and polyacrylamide resins (see also Obrecht, D.; Villalgordo, J.-M, "Solid- Supported Combinatorial and Parallel Synthet, -- -^.-.^-.-.-Molecular-Weight Compound
Libraries", Tetrahedron Organic Chemistry Serie , Vol. 17, Pergamon, Elsevier Science, 1998).
The solid support is functionalized by means of a linker, i.e. a bifunctional spacer molecule which contains on one end an anchoring group for attachment to the solid support and on the other end a selectively cleavable functional group used for the subsequent chemical transformations and cleavage procedures. For the purposes of the present invention the linker must be designed to eventually release the carboxyl group under mild acidic conditions which do not affect protecting groups present on any functional group in the side-chains of the various amino acids. Linkers which are suitable for the purposes of the present invention form acid-labile esters with the carboxyl group of the amino acids, usually acid-labile benzyl, benzhydryl and trityl esters; examples of linker structures of this kind include 2-methoxy-4-hydroxymethylphenoxy (SasrinR linker), 4-(2,4-dimethoxyphenyl-hydroxymethyl)-phenoxy (Rink linker), 4-(4-hydroxymethyl-3- methoxyphenoxy)butyric acid (HMPB linker), trityl and 2-chlorotrityl.
Preferably, the support is derived from polystyrene crosslinked with, most preferably 1-5%, divinylbenzene and functionalized by means of the 2-chlorotrityl linker.
When carried out as a parallel array synthesis the process of the invention can be advantageously carried out as described hereinbelow but it will be immediately apparent to those skilled in the art how this procedure will have to be modified in case it is desired to synthesize one single compound of the above formula I.
A number of reaction vessels (normally 24 to 192, typically 96) equal to the total number of compounds to be synthesized by the parallel method are loaded with 25 to 1000 mg, preferably 100 mg, of the appropriate functionalized solid support, preferably 1 to 3% cross linked polystyrene or tentagel resin.
The solvent to be used must be capable of swelling the resin and includes, but is not limited to, dichloromethane (DCM), dimethylformamide (DMF), N-methylpyrrolidone (NMP), dioxane, toluene, tetrahydrofuran (THF), ethanol (EtOH), trifluoroethanol (TFE), isopropylalcohol and the like. Solvent mixtures containing as at least one component a polar solvent (e. g. 20% TFE/DCM, 35% THF/NMP) are beneficial for ensuring high reactivity and solvation of the resin-bound peptide chains ( Fields, G. B., Fields, C. G., J. Am. Chem. Soc. 1991, 113, 4202-4207). With the development of various linkers that release the C-terminal carboxylic acid group under mild acidic conditions, not affecting acid-labile groups protecting functional groups in the side chain(s), considerable progresses have been made in the synthesis of protected peptide fragments. The 2-methoxy-4-hydroxybenzylalcohol-derived linker (SasrinR linker, Mergler et al.,
Tetrahedron Lett. 1988, 294005-4008) is cleavable with diluted trifluoroacetic acid (0.5-1% TFA in DCM) and is stable to Fmoc deprotection conditions during the peptide synthesis, Boc/tBu- based additional protecting groups being compatible with this protection scheme. Other linkers which are suitable for the process of the invention include the super acid labile 4-(2,4- dimethoxyphenyl-hydroxymethyl)-phenoxy linker (Rink linker, Rink, H. Tetrahedron Lett. 1987, 28, 3787-3790), where the removal of the peptide requires 10% acetic acid in DCM or 0.2% trifluoroacetic acid in DCM; the 4-(4-hydroxymethyl-3-methoxyphenoxy)butyric acid-derived linker (HMPB-linker, Flδrsheimer & Riniker, Peptides 1991,1990 131) which is also cleaved with 1%TFA/DCM in order to yield a peptide fragment containing all acid labile side- chain protective groups; and, in addition, the 2-chlorotritylchloride linker (Barlos et al., Tetrahedron Lett. 1989, 30, 3943-3946), which allows the peptide detachment using a mixture of glacial acetic acid/trifluoroethanol/DCM (1:2:7) for 30 min.
Suitable protecting groups for amino acids and, respectively, for their residues are, for example,
for the amino group (as is present e. g. also in the side-chain of lysine)
Cbz benzyloxycarbonyl
Boc tert.-butyloxycarbonyl Fmoc 9-fluorenylmethoxycarbonyl
Alloc allyloxycarbonyl
Teoc trimethylsilylethoxycarbonyl
Tec trichloroethoxycarbonyl
Nps o-nitrophenylsulfonyl; Trt triphenymethyl or trityl
for the carboxyl group (as is present e. g. also in the side-chain of aspartic and glutamic acid) by conversion into esters with the alcohol components tBu tert. -butyl
Bn benzyl
Me methyl
Ph phenyl
Pac Phenacyl
Allyl
Tse trimethylsilylethyl
Tee trichloroethyl;
- for the guanidino group (as is present e. g. in the side-chain of arginine)
Pmc 2,2,5,7,8-pentamethylchroman-6-sulfonyl
Ts tosyl (i. e. p-toluenesulfonyl)
Cbz benzyloxycarbonyl Pbf pentamethyldihydrobenzofuran-5-sulfonyl
for the hydroxy group (as is present e. g. in the side-chain of threonine and serine)
tBu tert.-butyl
Bn benzyl
Trt trityl
and for the mercapto group (as is present e. g. in the side-chain of cysteine) Acm acetamidomethyl tBu tert.-butyl
Bn benzyl
Trt trityl
Mtr 4-methoxytrityl.
The 9-fluorenylmethoxycarbonyl- (Fmoc)-protected amino acid derivatives are preferably used as the building blocks for the construction of the template-fixed β-haiφin loop mimetics of formula I. For the deprotection, i. e. cleaving off of the Fmoc group, 20%> piperidine in DMF or 2%> DBU/2% piperidine in DMF can be used. The quantity of the reactant, i. e. of the amino a.ιu uenvative, is usually 1 to 20 equivalents based on the milliequivalents per gram (meq/g) loading of the functionalized solid support (typically 0.1 to 2.85 meq/g for polystyrene resins) originally weighed into the reaction tube. Additional equivalents of reactants can be used if required to drive the reaction to completion in a reasonable time. The reaction tubes, in combination with the holder block and the manifold, are reinserted into the reservoir block and the apparatus is fastened together. Gas flow through the manifold is initiated to provide a controlled environment, for example, nitrogen, argon, air and the like. The gas flow may also be heated or chilled prior to flow through the manifold. Heating or cooling of the reaction wells is achieved by heating the reaction block or cooling externally with isopropanol/dry ice and the like to bring about the desired synthetic reactions. Agitation is achieved by shaking or magnetic stirring (within the reaction tube). The preferred workstations (without, however, being limited thereto) are Labsource's Combi-chem station and MultiSyn Tech's-Syro synthesizer.
Amide bond formation requires the activation of the α-carboxyl group for the acylation step. When this activation is being carried out by means of the commonly used carbodiimides such as dicyclohexylcarbodiimide (DCC, Sheehan & Hess, J. Am. Chem. Soc. 1955, 77, 1067-1068) or diisopropylcarbodiimide (DIC, Sarantakis et al Biochem. Biophys. Res. Commun.1916, 73, 336- 342), the resulting dicyclohexylurea is insoluble and, respectively, diisopropylurea is soluble in the solvents generally used. In a variation of the carbodiimide method 1-hydroxybenzotriazole (HOBt, Kδnig & Geiger, Chem. Ber 1970, 103, 788-798) is included as an additive to the coupling mixture. HOBt prevents dehydration, suppresses racemization of the activated amino acids and acts as a catalyst to improve the sluggish coupling reactions. Certain phosphonium reagents have been used as direct coupling reagents, such as benzotriazol-1-yl-oxy-tris- (dimethylamino)-phosphonium hexafluorophosphate (BOP) (Castro et al., Tetrahedron Lett. 1975, 14, 1219-1222; Synthesis, 1976, 751-752), or benzotriazol-1-yl-oxy-tris-pyrrolidino- phosphonium hexaflurophoshate (Py-BOP, Coste et al., Tetrahedron Lett. 1990, 31, 205-208), or 2-(lH-benzotriazol-l-yl-)l,l,3,3-tetramethyluronium terafluoroborate (TBTU), or hexafluorophosphate (HBTU, Knorr et al., Tetrahedron Lett. 1989, 30, 1927-1930); these phosphonium reagents are also suitable for in situ formation of HOBt esters with the protected amino acid derivatives. More recently diphenoxyphosphoryl azide (DPP A) or 0-(7-aza- benzotriazol-l-yl)-N,N,N',N'-tetramethyluroniu tetrafluoroborate (TATU) or 0-(7-aza- benzotriazol-l-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU)/7-aza-l- hydroxy benzotriazole (HO At, Caφino et al., leiranearon Lett. 1994, 35, 2279-2281) have also been used as coupling reagents.
Due to the fact that near-quantitative coupling reactions are essential it is desirable to have experimental evidence for completion of the reactions. The ninhydrin test (Kaiser et al, Anal. Biochemistry 1970, 34, 595), where a positive colorimetric response to an aliquot of resin-bound peptide indicates qualitatively the presence of the primary amine, can easily and quickly be performed after each coupling step. Fmoc chemistry allows the spectrophotometric detection of the Fmoc chromophore when it is released with the base (Meienhofer et al., Int. J. Peptide Protein Res. 1979, 13, 35-42).
The resin-bound intermediate within each reaction tube is washed free of excess of retained reagents, of solvents, and of by-products by repetitive exposure to pure solvent(s) by one of the two following methods:
1) The reaction wells are filled with solvent (preferably 5 ml), the reaction tubes, in combination with the holder block and manifold, are immersed and agitated for 5 to 300 minutes, preferably 15 minutes, and drained by gravity followed by gas pressure applied through the manifold inlet (while closing the outlet) to expel the solvent;
2) The manifold is removed from the holder block, aliquots of solvent (preferably 5 ml) are dispensed through the top of the reaction tubes and drained by gravity through a filter into a receiving vessel such as a test tube or vial.
Both of the above washing procedures are repeated up to about 50 times (preferably about 10 times), monitoring the efficiency of reagent, solvent, and byproduct removal by methods such as TLC, GC, or inspection of the washings.
The above described procedure of reacting the resin-bound compound with reagents within the reaction wells followed by removal of excess reagents, by-products, and solvents is repeated with each successive transformation until the final resin-bound fully protected linear peptide is prepared. Before this fully protected linear peptide is deιαv,ucu iiυ-ui the solid support, it is possible, if desired, to selectively deprotect one or several protected functional group(s) present in the molecule and to appropriately substitute the reactive group(s) thus liberated. To this effect, the functional group(s) in question must initially be protected by a protecting group which can be selectively removed without affecting the remaining protecting groups present. Alloc
(allyloxycarbonyl) is an example for such a protecting group for amino which can be selectively removed, e.g. by means of Pd° and phenylsilane in CH2C12, without affecting the remaining protecting groups, such as Fmoc, present in the molecule. The reactive group thus liberated can then be treated with an agent suitable for introducing the desired substituent. Thus, for example, an amino group can be acylated by means of an acylating agent corresponding to the acyl substituent to be introduced.
Detachment of the fully protected linear peptide from the solid support is achieved by immersion of the reaction tubes, in combination with the holder block and manifold, in reaction wells containing a solution of the cleavage reagent (preferably 3 to 5 ml). Gas flow, temperature control, agitation, and reaction monitoring are implemented as described above and as desired to effect the detachment reaction. The reaction tubes, in combination with the holder block and manifold, are disassembled from the reservoir block and raised above the solution level but below the upper lip of the reaction wells, and gas pressure is applied through the manifold inlet (while closing the outlet) to efficiently expel the final product solution into the reservoir wells. The resin remaining in the reaction tubes is then washed 2 to 5 times as above with 3 to 5 ml of an appropriate solvent to extract (wash out) as much of the detached product as possible. The product solutions thus obtained are combined, taking care to avoid cross-mixing. The individual solutions/extracts are then manipulated as needed to isolate the final compounds. Typical manipulations include, but are not limited to, evaporation, concentration, liquid/liquid extraction, acidification, basification, neutralization or additional reactions in solution.
The solutions containing fully protected linear peptide derivatives which have been cleaved off from the solid support and neutralized with a base, are evaporated. Cyclization is then effected in solution using solvents such as DCM, DMF, dioxane, THF and the like. Various coupling reagents which were mentioned earlier can be used for the cyclization. The duration of the cyclization is about 6-48 hours, preferably about 24 hours. The progress of the reaction is followed, e. g. by RP-HPLC (Reverse Phase High Performance Liquid Chromatography). Then the solvent is removed by evaporation, the fully protected cyclic peptide derivative is dissolved in a solvent which is not miscible with water, su.-*-. __ _,s__v_, and the solution is extracted with water or a mixture of water-miscible solvents, in order to remove any excess of the coupling reagent.
Before removing the protecting groups from the fully protected cyclic peptide, it is possible, if desired, to form an interstrand linkage between side-chains of appropriate amino acid residues at opposite positions of the β-strand region. Interstrand linkages and their formation have been discussed above, in connection with the explanations made regarding groups of the type H which can, for example, be disulfide bridges formed by cysteines and homocysteines at opposite positions of the β-strand, or glutamic and aspartic acid residues linking ornithines and, respectively, lysines located at opposite β-strand positions by amide bond formation. The formation of such interstrand linkages can be effected by methods well known in the art.
Finally, the fully protected peptide derivative of type I is treated with 95% TFA, 2.5% H20, 2.5% TIS or another combination of scavengers for effecting the cleavage of the protecting groups. The cleavage reaction time is commonly 30 minutes to 12 hours, preferably about 2 hours. Thereafter most of the TFA is evaporated and the product is precipitated with ether/hexane (1:1) or other solvents which are suitable therefor. After careful removal of the solvent, the cyclic peptide derivative obtained as end-product can be isolated. Depending on ist purity, this peptide derivative can be used directly for biological assays, or it has to be further purified, for example by preparative HPLC.
As mentioned earlier, it is thereafter possible, if desired, to convert a fully deprotected product thus obtained into a pharmaceutically acceptable salt or to convert a pharmaceutically acceptable, or unacceptable, salt thus obtained into the corresponding free compound of formula I or into a different, pharmaceutically acceptable, salt. Any of these operations can be carried out by methods well known in the art.
The starting materials used in the process of the invention, pre-starting materials therefor, and the preparation of these starting and pre-starting materials will now be discussed in detail.
Building blocks of type A can be synthesized according to the literature methods described below. The corresponding amino acids have been described either as unprotected or as Boc- or Fmoc-protected racemates, (D)- or (L)-isomers. It will be appreciated that unprotected amino acid building blocks can be easily transformed into the corresponding Fmoc-protected amino acid building blocks required for the present inven..-.v-.-.-. _j oiαudard protecting group manipulations. Reviews describing general methods for the synthesis of α-amino acids include: R. Duthaler, Tetrahedron (Report) 1994, 349, 1540-1650; R. M. Williams, "Synthesis of optically active α- amino acids", Tetrahedron Organic Chemistry Series, Vol.7, J. E. Baldwin, P. D. Magnus (Eds.), Pergamon Press., Oxford 1989. An especially useful method for the synthesis of optically active α-amino acids relevant for this invention includes kinetic resolution using hydrolytic enzymes (M. A. Verhovskaya, I. A. Yamskov, Russian Chem. Rev. 1991, 60, 1163-1179; R. M. Williams, "Synthesis of optically active α-amino acids", Tetrahedron Organic Chemistry Series, Vol.7, J. E. Baldwin, P. D. Magnus (Eds.), Pergamon Press., Oxford 1989, Chapter 7, p.257-279). Hydrolytic enzymes involve hydrolysis of amides and nitriles by aminopeptidases or nitrilases, cleavage of N-acyl groups by acylases, and ester hydrolysis by lipases or proteases. It is well documented that certain enzymes will lead specifically to pure (L)-enantiomers whereas others yield the corresponding (D)-enantiomers (e.g. : R. Duthaler, Tetrahedron Report 1994, 349, 1540-1650; R. M. Williams, "Synthesis of optically active α-amino acids", Tetrahedron Organic Chemistry Series, Vol.7, J. E. Baldwin, P. D. Magnus (Eds.), Pergamon Press., Oxford 1989).
Al: See D. Ben-Ishai, Tetrahedron 1977, 33, 881-883; K. Sato, A. P. Kozikowski, Tetrahedron Lett. 1989, 30, 4073-4076; J. E. Baldwin, C. N. Farthing, A. T. Russell, C. J. Schofield, A. C. Spirey, Tetrahedron Lett. 1996, 37, 3761-3767; J. E. Baldwin, R. M. Adlington, N. G. Robinson, J. Chem. Soc. Chem. Commun. 1987, 153-157; P. Wipf, Y. Uto, Tetrahedron Lett. 1999, 40, 5165-5170; J. E. Baldwin, R. M. Adlington, A. O'Neil, A. C. Spirey, J. B. Sweeney, J. Chem. Soc. Chem. Commun. 1989, 1852-1854 (for R'= H, R2= H); T. Hiyama, Bull. Chem. Soc. Jpn. 1974, 47, 2909-2910; T. Wakamiya, K. Shimbo, T. Shiba, K. Nakajima, M. Neya, K. Okawa, Bull. Chem. Soc. Jpn. 1982, 55, 3878-3881; I. Shi a, N. Shimazaki, K. Imai, K. Hemmi, M. Hashimoto, Chem. Pharm. Bull. 1990, 38, 564-566; H. Han, J. Yoon, K. D. Janda, J. Org. Chem. 1998, 63, 2045-2048 (R'= H, R2= Me); J. Legters, G. H. Willems, L. Thijs, B. Zwannenburg, Reel. Trav. Chim. Pays-Bos 1992, 111, 59-68 (R]= H, R2= hexyl); J. Legters, L. Thijs, B. Zwannenburg, Reel. Trav. Chim. Pays-Bas 1992, 111, 16-21; G. A. Molander, P. J. Stengel, J. Org. Chem. 1995, 21, 6660-6661 (RI= H, R2= Ph); I. Funaki, L. Thijs, B. Zwannenburg, Tetrahedron 1996, 52, 9909-9924 (R'= H, R2= Bn); A. S. Pepito, D. C. Ditt er, J. Org. Chem. 1997, 62, 7920-7925 ; (R'= H, R2= CH2OH); M. Egli, A. S. Dreiding, Helv. Chim. Acta 1986, 69, 1442-1460 (R2= CH(OH)CH2OH); M. Carduccu, S. Fioravanti, M. A. Loreto, L. Pellacani, P. A. Tardella, Tetrahedron Lett. 1996, 37, 3777-3778; F. J. Lakner, L. P. Hager, Tetrahedron: Asymmetry 1997, 21, 3547-3550 (R'= Me, R2= H, Me); G. A. Molander, P. J. Stengel, Tetrahedron 1997, 26, 8887-8912; M. A. OTOIU, _■. - υmpei, P. A. Tardella, D. Tofani, Tetrahedron 1997, 53, 15853-15858 (R'= Me, R2= CH2SiMe3); H. Shao, J. K. Rueter, M. Goodman, J. Org. Chem. 1998, 63, 5240-5244 (R'= Me, R2= Me).
A2: See A. Rao, M. K.Gurjar, V. Vivarr, Tetrahedron: Asymmetry 1992, 3, 859-862; R. L.
Johnson, G. Rayakumar, K.-L. Yu, R. K. Misra, J. Med. Chem. 1986, 29, 2104-2107 (R'= H, R2=
H); J. E. Baldwin, R. M. Adlington, R. H. Jones, C. J. Schofield, C. Zarcostas, J. Chem. Soc.
Chem. Commun. 1985, 194-196; J. E. Baldwin, R. M. Adlington, R. H. Jones, C. J. Schofield, C.
Zarcostas, Tetrahedron 1986, 42, 4879-4888 (R'= H, R2= CH2OH, CH2CHO, CH2CH2COOH, CH2CH2OH); A. P. Kozikowski, W. Tueckmantel, I. J. Reynolds, J. T. Wroblewski, /. Med.
Chem. 1990, 33, 1561-1571; A. P. Kozikowski, W. Tueckmantel, Y. Liao, H. Manev, S.
Ikonomovic J. T. Wroblenski, J. Med. Chem. 1993, 36, 2706-2708 (R'= H, R2= CH2OH,
CHCONH2, CONHCH2COOH, COOtBu); D. Seebach, T. Vettiger, H.-M. Mϋller, D. Plattner, W.
Petter, Liebigs Ann. Chem. 1990, 687-695 (Rl= ArylCH(OH), R2=H); D. Seebach, E. Dziadulewicz, L. Behrendt, S. Cantoreggi, R. Fitzi, Liebigs Ann. Chem. 1989, 1215-1232 (R1=
Me, Et, R2=H).
A3: See A. P. Kozikowski, Y. Liao, W. Tueckmantel, S. Wang, S. Pshsenichkin, Bioorg. Med. Chem. Lett. 1996, 6, 2559-2564 (R!= H; R = CHCHO, CH2OH, CH2CH2OH, CH2COOH, COOH); Isono, J. Am. Chem. Soc, 1969, 91, 7490 (R*= H; R2= Et); P. J. Blythin, M. J. Green, M. J. Mary, H. Shue, J. Org. Chem. 1994, 59, 6098-6100; S. Hanessian, N. Bernstein, R.-Y. Yang, R. Maquire, Bioorg. Chem. Lett. 1994, 9, 1437-1442 (R!= H; R2= Ph).
A4: See G. Emmer, Tetrahedron 1992, 48, 7165-7172; M. P. Meyer, P. L. Feldman, H. Rapoport, J. Org. Chem. 1985, 50, 5223-5230 (R'= H; R2= H); A. J. Bose, M. S. Manhas, J. E. Vincent, I. F. Fernandez, J. Org. Chem. 1982, 47, 4075-4081 (R'= H; R2= NHCOCH2OPh); D. L. Boger, J. B. Meyers, J. Org. Chem. 1991, 56, 5385-5390 (R'= H; R2= NHCOCH2Ph); K.-D. Kampe, Tetrahedron Lett. 1969, 117-120 (R = CH2OH; R2= Ph); M. D. Andrews, M. G. Maloney, K. L. Owen, J. Chem. Soc. Perkin Trans.1, 1996, 227-228 (R'= CH2OH; R2= H).
A5: See C. Bisang, C. Weber, J. Inglis, C. A. Schiffer, W. F. van Gunsteren, J. A. Robinson J. Am. Chem. Soc. 1995, 117, 7904 (Rl= CH3; R2= H); S. Takano, M. Morija, Y. Iwabuki, K. Ogasawara, Tetrahedron Lett. 1989, 30, 3805-3806 (R1= H; R2= COOH); M. D. Bachi, R. Breiman, H. Meshulam, J. Org. Chem. 1983, 48, 1439-1444 (R'= H; R2= CH(Et)COOH); D. S. Kemp, T. P. Curran, Tetrahedron Lett. 1988, _,_•, -r.-,--- .-.jS' A; D. S. Kemp, T. P. Curran, W. M. Davies, J. Org. Chem. 1991, 56, 6672-6682 (R'= H; R2= CH2OH); F. Manfre, J.-M. Kern, J.-F. Biellmann, J. Org. Chem. 1992, 57, 2060-2065 (R'= H; R2= H, CH=CH2, CCH); B. W. Bycroft, S. R. Chabra, J. Chem. Soc. Chem. Commun. 1989, 423-425 (R'= H; R2= CH2COOtBu; Y. Xu, J. Choi, M. I. Calaza, S. Turner, H. Rapoport, J. Org. Chem. 1999, 64, 4069-4078 (R'= H; R2= 3- pyridyl); E. M. Khalil, W. J. Ojala, A. Pradham, V. D. Nair, W. B. Gleason, J. Med. Chem. 1999, 42, 628-637; E. M. Khalil, N. L. Subasinghe, R. L. Johnson, Tetrahedron Lett. 1996, 37, 3441- 3444 (R'= allyl; R2= H); A. DeNicola, J.-L. Luche, Tetrahedron Lett. 1992, 33, 6461-6464; S. Thaisrivongs, D. T. Pals, J. A. Lawson, S. Turner, D. W. Harris, J. Med. Chem. 1987, 30, 536- 541; E. M. Khalil, N. L. Subasinghe, R. L. Johnson, Tetrahedron Lett. 1996, 37, 3441-3444; A. Lewis, J. Wilkie, T. J. Rutherford, D. Gani, J. Chem. Soc. Perkin Trans.l, 1998, 3777-3794 (R'= Me; R2= H); A. Lewis, J. Wilkie, T. J. Rutherford, D. Gani, J. Chem. Soc. Perkin Trans.l, 1998, 3777-3794 (R*= CH2COOMe; R2= H); N. L. Subasinghe, E. M. Khalil,R. L. Johnson, Tetrahedron Lett. 1997, 38, 1317-1320 (R'= CH2CHO; R2= H); D. J. Witter, S. J. Famiglietti, J. C. Gambier, A. L. Castelhano, Bioorg. Med. Chem. Lett. 1998, 8, 3137-3142; E. H. Khalil, W. H. Ojada, A. Pradham, V. D. Nair, W. B. Gleason, J. Med. Chem. 1999, 42, 628-637 (R = CH2CH2CHO; R2= H).
A6: See DeNardo, Farmaco Ed. Sci. 1977, 32, 522-529 (Rλ= H; R3= H); P. J. T. Floris, N. Terhuis, H. Hiemstra, N. W. Speckamp, Tetrahedron, 1993, 49, 8605-8628; S. Kanemasa, N. Tomoshige, O. Tsuge, Bull. Chem. Soc. Jpn. 1989, 62, 3944-3949 (R1= H; R3= H); Sucrow, Chem. Ber. 1979, 112, 1719.
A7: See Fichter, J. Prat. Chem. 1906, 74, 310 (R:=Me; R4= Ph).
A8: See L. Lapantsanis, G. Milias, K. Froussios, M. Kolovos, Synthesis 1983, 641-673; H.
Nedev, H. Naharisoa, Tetrahedron Lett. 1993, 34, 4201-4204; D. Y. Jackson, C. Quan, D. R.
Artis, T. Rawson, B. Blackburn, J. Med. Chem. 1997, 40, 3359-3368; D. Konopinska, H. Bartosz-
Bechowski, G. Rosinski, W. Sobotka, Bull. Pol. Acad. Sci. Chem. 1993, 41, 27-40; J. Hondrelis, G. Lonergan, S. Voliotis, J. Matsukas, Tetrahedron 1990, 46, 565-576; T. Nakamura, H.
Matsuyama, H. Kanigata, M. Iyoda, J. Org. Chem. 1992, 57, 3783-3789; C. E. O'Connell, K.
Ackermann, C. A. Rowell, A. Garcia, M. D. Lewis, C. E. Schwartz, Bioorg. Med. Chem. Lett.
1999, 9, 2095-2100; G. Lowe, T. Vilaivan, J. Chem. Soc. Perkin Trans. 1997, 547-554; B.
Bellier, I. McCourt-Tranchepain, B. Ducos, S. Danascimenta, H. Mundal, J. Med. Chem. 1997, 40, 3947-3956; M. Peterson, R. Vince J. Med. Chem. 1991, 34, 2787-2797; E. M. Smith, G. F. Swiss, B. R. Neustadt, E. H. Gold, J. A. Sorr-i -..-., „. fflH, Chem. 1988, 31, 875-885; E. Rubini, C.
Gilon, Z. Selinger, M. Chorev, Tetrahedron 1986, 42, 6039-6045 (R'= H; R5= OH); C. R. Noe,
M. Knollmueller, H. Voellenkle, M. Noe-Letschnig, A. Weigand, J. Mϋhl, Pharmazie, 1996, 51,
800-804 (R'= CH3; R5= OH); J. Kitchin, R. C. Berthell. N. Cammack, S. Dolan, D. N. Evans, J. Med. Chem. 1994, 37, 3703-3716; D. Y. Jackson, C. Quan, D. R. Artis, T. Rawson, B. Blackburn,
J. Med. Chem. 1997, 40, 3359-3368 (Rl= H; R5= OBn); J. E. Baldwin, A. R. Field, C. C.
Lawrence, K. D. Merritt, C. J. Schofield, Tetrahedron Lett. 1993, 34, 7489-7492; K. Hashimoto,
Y. Shima, H. Shirahama, Heterocycles 1996, 42, 489-492 (R'= H; R5= OTS); T. R. Webb, C.
Eigenbrot, J. Org. Chem. 1991, 56, 3009-3016; D. C. Cafferty, C. A. Slate, B. M. Nakhle, H. D. Graham, T. L. Anstell, Tetrahedron 1995, 51, 9859-9872 (R'= H; R5= NH2); T. R. Webb, C.
Eigenbrot, J. Org. Chem. 1991, 56, 3009-3016 (R*= H; R5= CH2NH2); J. K. Thottathil. J. L.
Moniot, Tetrahedron Lett. 1986, 27, 151-154 (R'= H; R5= Ph); K. Plucinska, T. Kataoka, M.
Yodo, W. Cody, J. Med. Chem. 1993, 36, 1902-1913 (R'= H; R5= SBn); J. Krapcho, C. Turk, D.
W. Cushman, J. R. Powell, J. Med. Chem. 1988, 31, 1148-1160 (R'= H; R5= SPh); A. J. Verbiscar, B. Witkop, J. Org. Chem. 1970, 35, 1924-1927 (R1= H; R5= SCH2(4-OMe)C6H4); S. I.
Klein, J. M. Denner, B. F. Molino, C. Gardner, R. D'Alisa, Bioorg. Med. Chem. Lett. 1996, 6,
2225-2230 (RJ= H; R5= 0(CH2)3Ph); R. Zhang, F. Brownewell, J. S. Madalengoita, Tetrahedron
Lett. 1999, 40, 2707-2710 (R!= H; R5- CH2COOBn).
A9: See Blake, J. Am. Chem. Soc. 1964, 86, 5293-5297; J. Cooper, R. T. Gallagher, D. T. Knight,
/. Chem. Soc. Chem. Perkin Trans.l, 1993, 1313-1318; D. W. Knight, A. W. Sibley, J. Chem.
Soc. Perkin Trans.l, 1997, 2179, 2188 (R'= H; R6= H); Blake, J. Am. Chem. Soc. 1964, 86, 5293-
5297; Y. Yamada, T. Ishii, M. Kimura, K. Hosaka, Tetrahedron Lett. 1981, 1353-1354 (R*= H;
R6= OH); Y. Umio, Yakugaku Zasshi, 1958, 78, 727 (R*= H; R6= iPr); Miyamoto, Yakugaku Zasshi, 1957, 77, 580-584; Tanaka, Proc. Jpn. Acad. 1957, 33, 47-50 (R - H; R6=
CH(CH3)CH2N(CH3)2); L. E. Overman, B. N. Rodgers, J. E. Tellew, W. C. Trenkle, J. Am. Chem.
Soc. 1997, 119, 7159-7160 (R!= H; R6= allyl); Ohki, Chem. Pharm. Bull. 1976, 24, 1362-1369
(R'= CH3; R6= H).
A10: See J. Mulzer, A. Meier, J. Buschmann, P. Luger, Synthesis 1996, 123-132 (R1= H; R7= CH=CH2); J. Cooper, P. T. Gallagher, D. W. Knight, J. Chem. Soc. Chem. Commun. 1988, 509- 510; E. Gδtschi, C. Jenny, P. Reindl, F. Ricklin, Helv. Chim. Acta 1996, 79, 2219-2234 (R'= H; R7= OH); N. A. Sasaki, R. Pauli, C. Fontaine, A. Chiaroni, C. Riche, P. Potier, Tetrahedron Lett. 1994, 35, 241-244 (Rl= H; R7= COOH); R. Cotton, A. N. C. Johnstone, M. North, Tetrahedron 1995, 51, 8525-8544 (R'= H; R7= COOMe); J. S. Sabol, G. A. Flynn, D. Friedrich, E. W. Huber, Tetrahedron Lett. 1997, 38, 3687-3690 (R'= h; Λ = <-.υNH2); P. P. Waid, G. A. Flynn, E. W. Huber, J. S. Sabol, Tetrahedron Lett. 1996, 37, 4091-4094 (R'= H; R7= (4-BnO)C6H4); N. A. Sasaki, R. Pauli, P. Potier, Tetrahedron Lett. 1994, 35, 237-240 (R'= H; R7= S02Ph); R. J. Heffner, J. Jiang, M. Jouillie, J. Am. Chem. Soc. 1992, 114, 10181-10189; U. Schmidt, H. Griesser, A. Lieberknecht, J. Hausler, Angew. Chem. 1981, 93, 272-273 (R'= H; R7= OAryl); H. Mosberg, A. L. Lomize, C. Wang, H. Kroona, D. L. Heyl, J. Med. Chem. 1994, 37, 4371-4383 (R'= H; R7= 4-OHC6H4); S. A. Kolodziej, G. V. Nikiforovich, R. Sceean, M.-F. Lignon, J. Martinez, G. R. Marshall, J. Med. Chem. 1995, 38, 137-149 (R'= H; R7= SCH2(4-Me)C6H4).
All: See Kuhn, Osswald, Chem. Ber. 1956, 89, 1423-1434; Patchett, Witkop, J. Am. Chem. Soc. 1957, 79, 185-189; Benz, Helv. Chim. Acta 1974, 57, 2459-2475; P. Wessig, Synlett, 1999, 9, 1465-1467; E. M. Smit, G. F. Swiss, B. R. Neustadt, E. H. Gold, J. A. Sommer, J. Med. Chem. 1988, 31, 875-885; J. Krapcho, C. Turk, D. W. Cushman, J. R. Powell, J. M. DeForrest, J. Med. Chem. 1988, 31, 1148 (R'= H; R6= H); D. Benlshai, S. Hirsh, Tetrahedron 1988, 44, 5441-5450 (R'= H; R6= CH3); M. W. HoUaday, C. W. Lin, C. S. Garvey, D. G. Witte, J. Med. Chem. 1991, 34, 455-457 (R'= H; R6= allyl); P. Barralough, P. Hudhomme, C. A. Spray, D. W. Young, Tetrahedron 1995, 51, 4195-4212 (R*= H; Rδ= Et); J. E. Baldwin, M. Rudolf, Tetrahedron Lett. 1994, 35, 6163-6166; J. E. Baldwin, S. J. Bamford, A. M. Fryer, M. Rudolf, M. E. Wood, Tetrahedron 1997, 53, 5233-5254 (R'= H; R6= CH2COOfBu); P. Gill, W. D. Lubell, J. Org. Chem. 1995, 60, 2658-2659 (β}= H; R6= CH3; Bn; allyl; CH2COOMe); M. J. Blanco, F. J. Sardina, J. Org. Chem. 1998, 63, 3411-3466 (R'= H; R6= OCH2OMe).
A12: See Ahmed, Cheeseman, Tetrahedron 1977, 33, 2255-2257; J. S. New, J. P. Yevich, J. Heterocycl. Chem. 1984, 21, 1355-1360; R. Kikumoto, Y. Tamao, K. Ohkubo, T. Tezuka, S. Tonomura, J. Med. Chem. 1980, 23, 1293-1299; C. J. Blankley, J. S. Kaltenbronn, D. E. DeJohn, A. Werner, L. R. Bennett, J. Med. Chem. 1987, 30, 992-998; S. Klutcho, C. J. Blankley, R. W. Fleming, J. M. Hinkley, R. E. Werner, Med. Chem. 1986, 29, 1953-1961 (R!= H; R8= H); L. J. Beeley, C. J. M. Rockwell, Tetrahedron Lett. 1990, 31, 417-420 (R*= COOEt; R8= H).
A13: See G. Flouret, W. Brieher, T. Majewski, K. Mahan, J. Med. Chem. 1991, 43, 2089-2094;
G. Galiendo, P. Grieco, E. Perissuti, V. Santagada, Farmaco, 1996, 51, 197-202; D. F.
McComsey, M. J. Hawkins, P. Andrade-Gordon, M. F. Addo, B. E. Maryanoff, Bioorg. Med.
Chem. Lett. 1999, 9, 1423-1428; G. B. Jones, S. B. Heaton, B. J. Chapman, M. Guzel,
Tetrahedron: Asymmetry 1997, 8, 3625-3636; M. Asa i, H. Watanabe, K. Honda, S. Inoue, Tetrahedron: Asymmetry 1998, 9, 4165-4174; K. Gross, Y. M. Yun, P. Beak, J. Org. Chem. 1997, 62, 7679-7689 (R'= H; R6= H; R8= H); k. --_uu&6, _ . M. Yun, P. Beak, J. Org. Chem. 1997, 62, 7679-7689 (R'= H; R6= H; R8= 6-Cl); Ch. Noe, M. Knollmueller, C. Schoedl, M. L. Berger, Sci. Pharm. 1996, 64, 577-590; E. Reiman, W. Erdle, H. Unger, Pharmazie, 1994, 54, 418-421 (R'= H; R6= CH2COOH; R8= H); V. Collot, M. Schmitt. A. K. Marwah, B. Norberg, J.-J. Bourgignon, Tetrahedron Lett. 1997, 38, 8033-8036 (R!= H; R6= Ph; R8= H); L. V. Dunkerton, H. Chen, B. P. McKillican, Tetrahedron Lett. 1988, 29, 2539-2542 (R'= C(CH3)2CH=CH2; R6= H; R8= H); E. J. Corey, J. Am. Chem. Soc. 1970, 92, 2476-2488; Neunhoeffer, Lehmann, Chem. Ber. 1961, 94, 2960-2963 (R'= CH3; R6= H; R8= H).
A14: Amino acids of type A14 can be made according to Scheme 1.
Scheme 1
Figure imgf000060_0001
i: NaH, BrCH(R )COOMe, DMF; ii: LiOHxl H20, MeOH, H20; iii: polyphosphoric acid(PPA); iv: NaH, CICOOMe, THF; v: enzymatic resolution (e.g.lipase); vi: NaOH, MeOH, H20, heat; vii: FmocOSu, Na2C03aq„ dioxane
A15: See D. S. Perlow, J. M. Erb, N. P. Gould, R. D. Tung, R. M. Freidinger, J. Org. Chem. 1992, 57, 4394-4400; D. Y. Jackson, C. Quan, D. R. Artis, T. Rawson, B. Blackburn, J. Med. Chem. 1997, 40, 3359-3368 (R'= H; R2= H); H. H. Wasserman, K. Rodrigues, K. Kucharczyk, Tetrahedron Lett. 1989, 30, 6077-6080 (R:= H; R2= COOH).
A16: See Beyerman, Boekee, Reel. Trav. Chim. Pays-Bas, 1959, 78, 648-653; M. E. Freed, A. R.
Day, J. Org. Chem. 1960, 25, 2105-2107; D. R. Adams, P. D. Bailey, I. D. Collier, J. D.
Heferman, S. Slokes, J. Chem. Soc. Chem. Commun. 1996, 349-350; J. E. Baldwin, R. M.
Adlington, C. R. A. Godfrey, D. W. Collins, J. D. Vaughan, J. Chem. Soc. Chem. Commun. 1993,
1434-1435; Y. Matsumura, Y. Takeshima, H. Ohita, Bull. Chem. Soc. Jpn. 1994, 67, 304-306 (R'= H; R6= H); C. Herdeis, W. Engel, Arch. Pharm. 1991, 324, 670 (R*= COOMe; R6= CH3).
A17, A18: See C. R. Davies, J. S. Davies, J. Chem. Soc. Perkin Trans 1, 1976, 2390-2394; K. Bevan, J. Chem. Soc. C, 1971, 514-522; K. Umezawa, K. Nakazawa, Y. Ikeda, H. Naganawa, S. Kondo, J. Org. Chem. 1999, 64, 3034-3038 (R'= R3= H); P. D. Williams, M. G. Bock, R. D. Tung, V. M. Garsky, D. S. Pariow, J. Med. Chem, 1992, 35, 3905-3918 ; K. Tamaki, K. Tanzawa, S. Kurihara, T. Oikawa, S. Monma, Chem. Pharm. Bull. 1995, 43, 1883-1893 (R'= R5= H ; R3= COOBn) ; K. J. Hale, J. Cai, V. Delisser, S. Manaviazar, S. A. Peak, Tetrahedron 1996, 52, 1047- 1068 ; M. H. Chen, O. P. Goel, J.-W. Hyun, J. Magano, J. R. Rubin, Bioorg. Med. Chem. Lett. 1999, 9, 1587-1592 (R = R5= H; R3= COOfBu,, ... ^^-.-..eli, I. Brun, P. Hall, R. Metternich, Tetrahedron Lett. 1999, 40, 2109-2112 (R*= R5= H; R3= COR); K. J. Hale, N. Jogiya, S. Manaviazar, Tetrahedron 1998, 39, 7163-7166 (R;= H; R3= COOBn; R5= OBn); T. Kamenecka, S. J. Danishewsky, v4ngew. Chem. Int. Ed. Engl. 1998, 37, 2995-2998(R1= H; R3= COO(CH2)2SiMe3; R5= OSiMe2tBu.
A19: See Beilstein, Registry Number 648833 (R'=R4=R8=H). Compounds of this type can be prepared according to Scheme 2.
Scheme 2.
Figure imgf000062_0001
i: NaH, CH2(COOMe)2, DMSO; ii: NaH, R1-X, DMSO; iii: NaOHaq., MeOH, 75°; iv: DBU, Mel, DMF; v: LDA, BocN=NBoc; vi: TFA, CH2CI2; vii: CbzCI, Na2C03aq., dioxane; viii: enzymatic resolution (e.g. lipase); then DBU, Mel, DMF; ix: NaH, R4-X, THF; x: Pd/C, H2, EtOH; xi: LiOHx1H20, MeOH,
H20; xii: FmocOSu,Na2Cθ3aq., dioxane
A20: See D. Hagiwara, H. Miyake, N. Igari, M. Karino, Y. Maeda, J. Med. Chem. 1994, 37, 2090-2099 (R'= H; R9= OH); Y. Arakawa, M. Yasuda, M. Ohnishi, S. Yoshifuji, Chem. Pharm. Bull. 1997, 45, 255-259 (R!= H; R9= COOH); P. j. Murray, I. D. Starkey, Tetrahedron Lett. 1996, 37, 1875-1878 (R!= H; R9= (CH2)2NHCOCH2Ph); K. Clinch, A. Vasella, R. Schauer, Tetrahedron Lett. 1987, 28, 6425-6428 (R'= H; R9= NHAC).
A21: See A. Golubev, N. Sewald, K. Burger, Tetrahedron Lett. 1995, 36, 2037-2040; F. Machetti, F. M. Cordero, F. DeSario, A. Guarna, A. Brandi, Tetrahedron Lett. 1996, 37, 4205- 4208; P. L. Ornstein, D. D. Schoepp, M. B. Arnold, j. D. Leander, D. Lodge, J. Med. Chem.
1991, 34, 90-97 ; R^R^H); P. D. Leeson, B. J. Williams, R. Baker, T. Ludduwahetty, K. W. Moore, M. Rowley, J. Chem. Soc. Chem. Commun. 1990, 1578-1580; D. I. C. Scopes, N. F. Hayes, D. E. Bays, D. Belton, J. Brain, J. Med. Chem. 1992, 35, 490-501; H. Kessler, M. Kuehn, T. Lδschner, Liebigs Ann. Chem. 1986, 1-20 (R^R^H); C. Herdeis, W. Engel, Arch. Pharm.
1992, 7, 419-424 (R'=R6=Bn); C. Herdeis, W. Engel, Arch. Pharm. 1992, 411-418 (R^COOMe; R6=H); C. Herdeis, W. Engel, Arch. Pharm. 1992, 419-424 (R!=COOMe; Rδ=Bn).
A22: See P. D. Leeson, B. J. Williams, R. Baker, T. Ladduwahetty, K. W. Moore, M. Rowley, J. Chem. Soc. Chem. Comm. 1990, 1578-1580 (R'= H; R10= NHOBn). A23: See Beyerman, Boekee, Reel. Trav. Ch . uγ_-_,us 1959, 78, 648-653; D. R. Adams, P. D . Bailey, I. D. Collier, J. D. Heffernan, S. Stokes J. Chem. Soc. Chem. Commun. 1996, 349-350; J. E. Baldwin, R. M. Adlington, C. Godfrey, D. W. Collins, J. G. Vaughan, J. Chem. Soc. Chem. Comm. 1993, 1434-1435 (R'=R6=H); C. Herdeis, W. Engel, Arch. Pharm. 1993, 297-302 (R'=COOMe; R6=H).
A24: See Plieninger, Leonhauser, Chem. Ber. 1959, 92, 1579-1584; D. W. Knight, N. Lewis, A. C. Share, D. Haigh, J. Chem. Soc. Perkin Trans.l 1998, 22, 3673-3684; J. Drummond, G. Johnson, D. G. Nickell, D. F. Ortwine, R. F. Bruns, B. Welbaum, J. Med. Chem. 1989, 32, 2116- 2128; M. P. Moyer, P. L. Feldman, H. Rapoport, J. Org. Chem. 1985, 50, 5223-5230 (R1=R6=H); McElvain, Laughton, J. Am. Chem. Soc. 1951, 73, 448-451 (R'=H; R6=Ph); McElvain, Laughton, /. Am. Chem. Soc. 1951, 73, 448-451 (R!=Ph; Rδ=H);
A25: See L.-Y. Hu, T. R. Ryder, S. S. Nikam, E. Millerman, B. G. Szoke, M. F. Rafferty, Bioorg. Med. Chem. Lett. 1999, 9, 1121-1126; W. C. Lumma, R. D. Hartman, W. S. Saari, E. L. Engelhardt, V. J. Lotti, C. A. Stone, J. Med. Chem. 1981, 24, 93-101; N. Hosten, M. J. O. Antenuis, Bull. Soc. Chim. Belg. 1988, 97, 48-50; C. F. Bigge, S. J. Hays, P. M. Novak, J. T. Drummond, G. Johnson, T. P. Bobovski, Tetrahedron Lett. 1989, 30, 5193-5191; B. Aebischer, P. Frey, H.-P. Haerter, P. L. Herrling, W. Mϋller, Helv. Chim. Acta 1989, 72, 1043-1051; W. J. Hoeckstra, B. E. Maryanoff, B. P. Damiano, P. Andrade-Gordon, J. H. Cohen, M. J. Constanzo, B. J. Haertlein, L. R. Hecker, B. L. Hulshizer, J. A. Kauffman, P. Keane, J. Med. Chem. 1999, 42, 5254-5265 (R'=H; RU=H) ; B. D. Dorsey, R. B. Levin, S. L. McDaniel, J. P. Vacca, J. P. Guare, . Med. Chem. 1994, 37, 3443-3451; M. Cheng, B. De, S. Pikul, N. G. Almstaed, M. G. Natchus, M. V. Anastasio, S. J. McPhail, C. J. Snider, Y. O. Taiwo, L. Chen, C. M. Dunaway, J. Med. Chem. 2000, 43, 369-380; R. Kuwano, Y. Ito, J. Org. Chem. 1999, 64, 1232-1237 (R'=H;
Ru=COOtBu); J. Kitchin, R. C. Bethell, N. Cammack, S. Dolan, D. N. Evans, J. Med. Chem. 1994, 37, 3707-3716 (R'=H; Rn=COOPh); C. F. Bigge, S. J. Hays, P. M. Novak, J. T. Drummond, G. Johnson, T. P. Bobovski, J. Med. Chem. 1990, 33, 2916-2924
Figure imgf000063_0001
(CH2)3COOEt; (CH2)3PO(Me)OH; CH2PO(OH)2; (CH2)2PO(OEt)2; (CH2)2PO(OH)2).
Compounds of type A25 can also be prepared according to Scheme 3: Scheme 3
Figure imgf000064_0001
12 13 14 i: Lawesson reagent, toluene, 80°; ii: DBU, Mel, DMF; iii: NaBH4 or aCNBH3, MeOH; iv: Boc20, THF; v: LiOHx1 H20, MeOH, H20; vi: Pd/C, H2, EtOH; vii: FmocOSu, Na2C03aq., dioxane
A26: See Koegel, J. Biol. Chem. 1953, 201, 547 (R -R'^H).
A27: See G. Makara, G. R. Marshall, Tetrahedron Lett. 1997, 38, 5069-5072; R. N. Patel, A. Banerjee, R. L. Hanson, D. B. Brzozowski, L. W. Parker, L. J. Szarka, Tetrahedron: Asymmetry 1999, 10, 31-36 (R'=H; R13=OH, OtBu); J. E. Johanson, B. D. Christie, H. Rapoport, J. Org. Chem. 1981, 46, 4914-4920; N. Moss, J.-S. Duceppe, J.-M- Feriand, J. Gauthier, J. Med. Chem.
1996, 39, 2178-2187 (R!= H ; R13= CONHMe); G. M. Makara, G. R. Marshall, Tetrahedron Lett.
1997, 38, 5069-5072 (R^H; R13= SCH2(4-MeO)C6H4).
A28: See A. Golubev, N. Sewald, K. Burger, Tetrahedron Lett. 1995, 36, 2037-2040; P. L. Omstein, D. D. Schoepp, M. B. Arnold, J. D. Leander, D. Lodge, J. Med. Chem. 1991, 34, 90-97
(R'=Rδ=H); P. D. Leeson, B. J. Williams, R. Baker, T. Ladduwahetty, K. W. Moore, M. Rowley,
J. Chem. Soc. Chem. Commun. 1990, 22, 1578-1580; C. Herdeis, W. Engel, Arch. Pharm. 1991,
324, 670 (R:=H ; R6=Me); C. Herdeis, W. Engel, Arch. Pharm. 1991, 324, 670 (R^COOMe;
R6=H, Me).
A29: See Kawase, Masami, Chem. Pharm. Bull. 1997, 45, 1248-1253; I. G. C. Courts, J. A.
Hadfield, P. R. Huddleston, J. Chem. Res. Minipήnt, 1987, 9, 2472-2500; I. G. C. Courts, J. A.
Hadfield, P. R. Huddleston, J. Chem. Res. Miniprint, 1987, 9, 2472-2500; V. J. Hrubi, W. L.
Cody, A. M. Castrucci, M. E. Hadley, Collect. Czech. Chem. Commun. 1988, 53, 2549-2573; R. T. Shuman, R. B. Rothenberger, C. S. Campbell, G. F. Smith, D. S. Gifford-Moore, P. D.
Gesellchen, J. Med. Chem. 1993, 36, 314-319; M. Kawase, Y. Okada, H. Miyamae, Heterocycles,
1998, 48, 285-294 (R'=R8=H); Kawase, Masami, Chem. Pharm. Bull. 1997, 45, 1248-1253 (R1=H; R8=6,7-(Me02); D. F. Ortwine, T. C. Malone, C. F. Bigge, J. T. Drummond, C. Humblet, J. Med. Chem. 1992, 55, 1345-1370 (R'=H; R8=7-CH2PO(OEt)2); E. J. Corey, D. Y. Gin, Tetrahedron Lett. 1996, 37, 7163-7166
Figure imgf000065_0001
P. Dostert, M. Varasi, A. DellaTorre, C. Monti, V. Rizzo, Eur. J. Med. Chim. Ther. 1992, 27, 57-59 (R'=Me; R8=6,7- (OH)2); Z. Czamocki, D. Suh, D. B. McLean, P. G. Hultin,W. A. Szarek, Can. J. Chem. 1992, 70, 1555-1561; B. Schonenberger, A. Brossi, Helv. Chim. Acta 1986, 69, 1486-1497 (R!=Me; R8=6- OH; 7-MeO); Hahn, Stiel, Chem. Ber. 1936, 69, 2627; M. Chrzanowska, B. Schonenberger, A. Brossi, J. L. Flippen-Anderson, Helv. Chim. Acta 1987, 70, 1721-1731; T. Hudlicky, J. Org. Chem. 1981, 46, 1738-1741 (R*=Bn; R8=6,7-(OH)2); A. I. Meyers, M. A. Gonzalez, V. Struzka, A. Akahane, J. Guiles, J. S. Warmus, Tetrahedron Lett. 1991, 32, 5501-5504 (R'=CH2(3,4- methylenedioxy)C6H3; R8=6,7-(OMe)2).
A30 and A31 can be prepared according to Schemes 4 and 5.
Scheme 4
Figure imgf000066_0001
Figure imgf000066_0002
i: NaH, tert.-butyl N-benzoyl glycinate, DMF; ii: NaH, Pd(0), toluene; iii: TFA, CH2CI2; iv: polypho- sphoric acid; v: NaOHaq.,MeOH, 75°; then HCIaq.; vi: DBU, Mel, DMF; vii: lithium hexamethyl- disilazide,THF, chloro trimethylsilane, -78°; then R1-X; viii: enzymatic resolution(e.g. lipase); then isolation as methylester: DBU, Mel, DMF; ix: NaOHaq., MeOH, heat; x: FmocOSu,
Na C03aq., dioxane
Scheme 5
Figure imgf000066_0003
20 (R°: H) 22 23
i: Boc20, Na2C03aq., dioxane; ii: DBU, Mel, DMF; iii: lithium hexamethyldisilazide, THF, chlorotrimethylsilane, -78°; then R2-X; iv: LiOHx1H20, MeOH, H20; v:TFA, CH2CI2; vi: FmocOSu,
Na2C03aq., dioxane
A32 can be prepared according to P. W. Schiller, G. Weltrowska, T. M.-D. Nguyen, C. Lemieux, N. Nga, /. Med. Chem. 1991, 34, 3125-3132; V. S. Goodfellow, M. V. Marathe, K. G. Kuhlman, T. D. Fitzpatrick, D. Cuadrato, J. Med. Chem. 1996, 39, 1472-1484; G. Caliendo, F. Fiorino, P. Grieco, E. Perissutti, S. DeLuca, A. Guiliano, G. Santelli, D. Califano, B. Severino, V. Santagada, Farmacao, 1999, 54, 785-790; V. S. Goodfellow, M. V. Marathe, K. G. Kuhlman, T. D. Fitzpatrick, D. Cuadro, J. Med. Chem. 1996, 39, 1472-1484 (R'= R8= H); D. Tourwe, E. Mannekens, N. T. Trang, P. Verheyden, H. Jaspers, J. Med. Chem. 1998, 41, 5167-5176; A.-K. Szardenings, M. Gordeev, D. V. Patel, Tetrahε^, .. --.--,-.,.. 1996, 37, 3635-3638; W. Wiczk, K. Stachowiak, P. Skurski, L. Lankiewicz, A. Michniewicz, A. Roy, J. Am. Chem. Soc. 1996, 118, 8300-8307; K. Verschuren, G. Toth, D. Tourwe, M. Lebl., G. van Binst, V. Hrubi, Synthesis 1992, 458-460 (R'= H; R8= 6-OH); P. L. Omstein, M. B. Arnold, N. K. Augenstein, J. W. Paschal, J. Org. Chem. 1991, 56, 4388-4392 (R'= H; R8= 6-MeO); D. Ma, Z. Ma, A. P.
Kozikowski, S. Pshenichkin, J. T. Wroblenski, Bioorg. Med. Lett. 1998, 8, 2447-2450 (R'= H; R8= 6-COOH); U. Schδllkopf, R. Hinrichs, R. Lonsky, Angew. Chem. 1987, 99, 137-138 (R'= Me; R8=H); B. O. Kammermeier, U. Lerch, C. Sommer, Synthesis 1992, 1157-1160 (R'= COOMe; R8=H); T. Gees, W. B. Schweizer, D. Seebach, Helv. Chim. Acta 1993, 76, 2640-2653 (R'= Me; R8=6,7-(Me02).
A33: See Hinton, Mann, J. Chem. Soc. 1959, 599-608.
A34: See G. P. Zecchini, M. P. Paradisi, J. Heterocycl. Chem. 1979, 16, 1589-1597; S. Cerrini, J. Chem. Soc. Perkin Trans.l, 1979, 1013-1019; P. L. Omstein, J. W. Paschal, P. D. Gesellchen, J.
Org. Chem. 1990, 55, 738-741; G. M. Ksander, A. M. Yan, C. G. Diefenbacher, J. L. Stanton, J.
Med. Chem. 1985, 28, 1606-1611; J. A. Robl, D. S. Karanewsky, M. M. Asaad, Tetrahedron Lett.
1995, 36, 1593-1596; S. Katayama, N. Ae, R. Nagata, Tetrahedron: Asymmetry 1998, 9, 4295-
4300 (R'=R8=H); K. Hino, Y. Nagai, H. Uno, Chem. Pharm. Bull. 1988, 36, 2386-2400 (R!=Me; R8=H).
A35: See Beilstein Registry Numbers: 530775, 883013 (R'=R8=H).
A36: See R. W. Carling, P. D. Leeson, A. M. Moseley, R. Baker, A. C. Foster, J. Med. Chem. 1992, 35, 1942-1953; S. Kano, T. Ebata, S. Shibuya, J. Chem. Soc. Perkin Trans.l, 1980, 2105- 2111 (R'=R8=H); R. W. Carling, P. D. Leeson, A. M. Moseley, R. Baker, A. C. Foster, J. Med. Chem. 1992, 35, 1942-1953 (R'=H; R8=5-C1; 7-C1).
A37: See Nagarajan, Indian J. Chem. 1973, 11, 112 (Rl=CH2COOMe; R -H).
A38: See R. Pauly, N. A. Sasaki, P. Potire, Tetrahedron Lett. 1994, 35, 237-240; J. Podlech, D. Seebach, Liebigs Ann. Org. Bioorg. Chem. 1995, 7, 1217-1228; K. C. Nicolaou, G.-Q. Shi, K. Namoto, F. Bemal, J. Chem. Soc. Chem. Commun. 1998, 1757-1758 (R - H; R2= H). A39: See Beilstein, Registry Number 782885.
A40: See F. P. J. C. Rutjes, N. M. Terhuis, H. Hiemstra, N. W. Speckamp, Tetrahedron 1993, 49, 8605-8628 (R1= H; R3= Bn); compounds of this type can be prepared according to Scheme 6.
Scheme 6
Figure imgf000069_0001
26 27 28 i: BocNHNH2, NaCNBH3, MeOH, AcOH; ii: CbzCI, Et3N, CH2CI2; iii: TFA, CH2CI2; then pyridine,
DMAP, heat; iv: resolution (e.g. lipase); v: DBU, Mel, DMF; vi: La esson reagent, toluene, 75°; vii: DBU, Mel, DMF; viii: NaBH4 or NaCNBH3, MeOH; ix: R3 introduced by reductive amination, alkylation or acylation; x: LiOHx1H20, MeOH, H20; xi: Pd/C, H2, EtOH; xii: FmocOSu, Na C03aq., dioxane
A41: Compounds of this type can be prepared according to Scheme 7.
Scheme 7
Hooς fmoc.
Figure imgf000069_0002
Figure imgf000069_0003
26 29 30 i: resolution (e.g. lipase); then isolation as methylester: DBU, Mel, DMF; ii: NaH, R4-X, THF; iii: LiOHx1 H20, MeOH, H20; iv: Pd/C, H2, EtOH; v: FmocOSu, Na2C03aq., dioxane
A 2 to A46: Compounds of this typj? can be prepared according to Scheme, 8 to 12. Key intermediate 34" and α-amino acid synthesis involving this building block include: R. M. Williams, M.-N. Im, Tetrahedron Lett. 1988, 29, 6079-6082; R. M. Williams, M.-N. Im, J. Am. Chem. Soc.1991, 113, 9276-9286; j. F. Dellaria, B. D. Santarsiero, Tetrahedron Lett. 1988, 29, 6079-6082; j. F. Dellaria, B. D. Santarsiero, J. Org. Chem. 1989, 54, 3916-3926; j. E. Baldwin, V. Lee, C. J. Schofield, Synlett 1992, 249-251; j. E. Baldwin, V. Lee, C. J. Schofield, Heterocycles 1992, 34, 903-906. Scheme 8
Figure imgf000070_0001
34 35 36 i: lithium hexamethyldisilazide, THF, chlorotrimethylsilane, -78°; then R5-X; ii: HBr; iii: DBU,
Mel, DMF; iv: DIBAL-H, THF; v: EtOH, pyridinium p-toluenesulfonate, mol.sieves 4A; vi: lithium hexamethyldisilazide, THF, -78°, 33; vii: Pd/C, H2, EtOH; then DBU, Mel, DMF; then TFA, CH2CI2; viii: HCIaq., THF; then Na(OAc)3BH, AcOH, dichloroethane; ix: LiOHx1 H20,
MeOH, H20; x: FmocOSu, Na2C03aq., dioxane
Scheme 9
Figure imgf000070_0002
34 40 41 i: lithium hexamethyldisilazide, THF, chlorotrimethylsilane, -78°; then R6-X; ii: HBr; iii: DBU, Mel, DMF; iv: DIBAL-H, THF; v: EtOH, pyridinium p-toluenesulfonate, mol.sieves 4A; vi: lithium hexamethyldisilazide, THF, -78°, 39; vii: Pd/C, H2, EtOH; then DBU, Mel, DMF; then TFA, CH2CI2; viii: HCIaq., THF; then Na(OAc)3BH, AcOH, dichloroethane; viii: Boc20, Et3N, CH2CI2; ix: Bu NFx10H2O, THF; ix: pyridinium chlorochromate; x: LiOHx1H20, MeOH, H20; xi: TFA, CH2CI2; xii: FmocOSu, Na2C03aq., dioxane Scheme 10
Figure imgf000071_0001
i: HBr; ii: DBU, Mel, DMF; iii: DIBAL-H, THF; iv: EtOH, pyridinium p-toluenesulfonate, mol. sieves 4A; v: lithium hexamethyldisilazide, THF, -78°, 43; vi: Pd/C, H2, EtOH; then DBU,
Mel, DMF; then TFA, CH2CI2; vii: HCIaq., THF; then Na(OAc)3BH, AcOH, dichloroethane; viii: LiOHx1 H2O, MeOH, H2O; ix: FmocOSu, Na2CO3aq., dioxane
Scheme 11
Figure imgf000071_0002
i: HBr; ii: DBU, Mel, DMF; iii: DIBAL-H, THF; iv: EtOH, pyridinium p-toluenesulfonate, mol. sieves 4A; v: lithium hexamethyldisilazide, THF, -78°, 47; vi: Pd/C, H2, EtOH; then DBU.Mel,
DMF; then TFA, CH2Cl2; vii: HCIaq., THF; then Na(OAc)3BH, AcOH, dichloroethane; viii Boc2O, Et3N, CH2CI2; ix: Bu4NFx10H2O, THF; x: pyridinium chlorochromate; xi: LiOHx1 H2O, MeOH, H2O; xii: TFA, CH2CI2; xiii: FmocOSu, Na2CO3aq., dioxane Scheme 12
Figure imgf000072_0001
i: HBr; ii: DBU, Mel, DMF; iii: DIBAL-H, THF; iv: EtOH, pyridinium p-toluenesulfonate, mol. sieves 4A; v: lithium hexamethyldisilazide, THF, -78°, 51; vi: Pd/C, H2, EtOH; then DBU,
Mel, DMF; then TFA, CH2CI2; vii: HCIaq., THF; then Na(OAc)3BH, AcOH, dichloroethane; viii:
Boc2O, Et3N, CH2CI2; ix: Bu4NFx10H2O, THF; x: pyridinium chlorochromate; xi: LiOHx1 H2O,
MeOH, H2O; xii: TFA, CH2CI2; xiϋ: FmocOSu, Na2CO3aq., dioxane
A47: See P. Barraclough, R. D. Farrant, D. Kettle, S. Smith, J. Chem. Res. Miniprint 1991, 11, 2876-2884 (R^R'^H, Bn, (CH2)2P0(0Et)2).
A48: See A. Nouvet, M. Binard, F. Lamaty, j. Martinez, R. Lazaro, Tetrahedron 1999, 55, 4685- 4698 (R'=RI2=H) .
A49: See M. Y. Kolleganov, I. G. Kolleganova, M. D. Mitrofanova, L. I. Martynenko, P. P. Nazarov, V. I. Spitsyn, Bull. Acad. Sci. USSR Div. Chem. Sci (Engl. Trans.) 1983, 32, 1293-1299; Izv. Akad. NaukSSSR Ser. Khim. 1983, 6, 1293-1299 ; V. P. Vasilev, T. D. Orlova, S. F. Ledenkov, J. Gen. Chem. USSR (Engl. Trans. 1989, 59, 1629-1634; Zh. Obshch. Khim. 1989, 59, 1828-1833 (R^H; Ri2= CH(COOH)CH2COOH). Compounds of type A49 can also be prepared according to Scheme 13. Scheme 13
Me B
Figure imgf000073_0001
i: NaH, CbzNH(CH2)2Br, THF; ii: Pd/C, H2, EtOH; iii: EDCI, CH2CI2, diisopropylethylamin; iv: NaH,
R -.1'2 -X, THF; v: LiOHx1H20, MeOH, H20; vi: TFA, CH2CI2; vii: FmocOSu, Na2C03aq., dioxane
A50 and A51: Compounds of these types can be prepared according to Schemes 14 and 15. Scheme 14
Figure imgf000073_0002
i: HBr; ii: DBU, Mel, DMF; iii: DIBAL-H, THF; iv: EtOH, pyridinium p-toluenesulfonate, mol. sieves 4A; v: lithium hexamethyldisilazide, THF, -78°, 59; vi: Pd/C, H2, EtOH; then DBU,
Mel, DMF; then TFA, CH2CI2; vii: HCIaq., THF; then Na(OAc)3BH, AcOH, dichloroethane; viii: LiOHx1 H2O, MeOH, H2O; ix: FmocOSu, Na2CO3aq., dioxane Scheme 15
Figure imgf000074_0001
i: HBr; ii: DBU, Mel, DMF; iii: DIBAH, THF; iv: EtOH, pyridinium p-toluenesuifonate, mol. sieves 4A; v: lithium hexamethyldisilazide, THF, -78°, 63 vi: Pd/C, H2, EtOH; then DBU,
Mel, DMF; then TFA, CH2CI2; vii: HCIaq., THF; then Na(OAc)3BH, AcOH, dichloroethane; viii: Boc2O, Et3N, CH2CI2; ix: Bu NFx10H2O, THF; x: pyridinium chlorochromate; xi: LiOHx1 H2O, MeOH, H2O; xii: TFA, CH2CI2; xiii: FmocOSu, Na2CO3aq., dioxane
A53: See P. Barraclough, R. D. Farrant, D. Kettle, S. Smith, J. Chem. Res. Miniprint 1991, 11, 2876-2884(RI=R1I=H ; R'=H ; Rπ= Bn, (CH2)3PO(OH)2); (CH2)3PO(Et)2); J. I. Levin, J. F. DiJoseph, L. M. Killar; A. Sung, T. Walter, Bioorg. Med. Chem. Lett. 1998, 8, 2657-2662 (R'=H;
Figure imgf000074_0002
A 52 and A54: Compounds of this type can be prepared according to Schemes 16 and 77.
Scheme 16
Figure imgf000075_0001
Figure imgf000075_0002
i: iBuMgCI.THF; ii: NaH, THF; iii: lithium hexamethyldisilazide, THF, chlorotrimetylsilane, -78°; then R6-X; iv: NaOHaq., MeOH, 75°; then HCIaq.; v: DBU, Mel, DMF; vi: lithium hexamethyldisilazide , THF, chlorotrimetylsilane, -78°; then R1-X; vii: resolution (e.g. lipase); then DBU, Mel, DMF; viii: LiOHx1 H2O, MeOH, H2O; ix: TFA, CH2CI2; x: FmocOSu, Na2CO3aq., dioxane
Scheme 17
Me
B
Figure imgf000076_0001
i:NaN3, DMSO; ii: NaH, THF, CH2=CHCOOBn; iii: Pd/C, H2, EtOH; iv: EDCI, CH2CI2, diisopropylethylamine; v: NaH, R12-X, THF; vi: LiOHx1 H2O, MeOH, H2O; vii: TFA,
CH2CI2; viii: FmocOSu, Na2CO3aq., dioxane
A55 and A56: Compounds of this type can be prepared according to Schemes 18 and 19.
Scheme 18
COOMe COOMe BocHN— -R1 BocN X— -R1
COOMe COOMe
79 CbzHN" 80
Figure imgf000076_0002
Figure imgf000076_0003
i:NaH, THF, CbzNH(CH2)3Br; ii: Pd/C, H2, EtOH; then toluene, heat; iii: resolution (e.g. lipase); iv: DBU, Mel, DMF; v: NaH, R12-X, THF; vi: LiOHx1H20, MeOH, H2O; vii: TFA, CH2CI2; viii:
FmocOSu, Na2C03aq., dioxane Scheme 19
Figure imgf000077_0001
85 86
Figure imgf000077_0002
34 87 88 i: HBr; ii: DBU, Mel, DMF; iii: DIBAL-H, THF; iv: EtOH, pyridinium p-toluenesulfonate, mol. sieves 4A; v: lithium hexamethyldisilazide, THF, -78°, 86; vi: Pd/C, H2, EtOH; then DBU,
Mel, DMF; then TFA, CH2CI2; vii: HCIaq., THF; then Na(OAc)3BH, AcOH, dichloroethane; viii: LiOHx1 H2O, MeOH, H2O; ix: FmocOSu, Na2CO3aq., dioxane
A57: Compounds of this type can be prepared according to Scheme 20.
Scheme 20
BocHN
Figure imgf000078_0001
Figure imgf000078_0002
i: NaOMe, MeOH; ii: NaH, THF; iii: NaOHaq., MeOH, 75°; then HCIaq.; iv: DBU, Mel, DMF; v: lithium hexamethyldisilazide, THF, chlorotrimethylsilane, -78°; then R1-X; vi: resolution (e.g. lipase); then isolation of methylester: DBU, Mel, DMF; vii: LiOHx1H2O, MeOH, H2O; viii.TFA,
CH2CI2; ix: FmocOSu, Na2CO3aq., dioxane
A58: See C.-H. Lee, H. Kohn, J. Org. Chem. 1990, 55, 6098-6104 (R'=R°=H).
A59: can be prepared according to Scheme 21.
Scheme 21
BocHN
Figure imgf000079_0001
Figure imgf000079_0002
i: NaOMe, MeOH; ii: NaH, THF; iii: NaOHaq., MeOH, 75°; then HCIaq.; iv: DBU, Mel, DMF; v: lithium hexamethyldisilazide, THF, chlorotrimethylsilane, -78°; then R1-X; vi: resolution (e.g. lipase); then isolation of methylester: DBU, Mel, DMF; vii: LiOHx1 H2O, MeOH, H2O; viii.TFA,
CH2CI2; ix: FmocOSu, Na2CO3aq., dioxane ^
0: Compounds of this type can be prepared according to Scheme 22.
Scheme 22
BocH
Figure imgf000080_0001
Figure imgf000080_0002
i: NaH, DMSO; ii: NaOHaq., MeOH, 75°; then HCIaq.; iii: DBU, Mel, DMF; iv: NaOMe (2.2equiv.),
R1-X; v: Raney-Ni, H2, EtOH; vi: CbzCI, Et3N, CH2CI2; vii: NaH, Br(CH2)2Br, THF; viii: resolution
(e.g. lipase); then DBU, Mel, DMF; ix: Pd/C, H2, EtOH; x: NaH, R14-X, THF; xi: LiOHx1H20, MeOH,
H20; xii: TFA, CH2CI2; xiii: FmocOSu, Na2C03aq„ dioxane
A61: See D. R. Armour, K. M. Morriss, M. S. Congreve, A. B. Hawcock, Bioorg. Med. Chem. Lett. 1997, 7, 2037-2042 (Rl=ΕLl2=Ε_).
A62: Compounds of this type can be prepared according to Scheme 23.
Scheme 23
Figure imgf000081_0001
i: resolution (e.g. lipase); then DBU, Mel, DMF; ii: lithium hexamethyldisilazide, THF, chlorotrimethylsilane, -78°; then R6-X; iii: LiOHxl H2O, MeOH, H2O; iv: TFA, CH2CI2; v: FmocOSu,
Na2CO3aq., dioxane A63: See S. E. Gibson, N. Guillo, R. J. Middleton, A. Thuilliez, M. J. Tozer, J. Chem. Soc.
Perkin Trans.l, 1997, 4, 447-456; S. E. Gibson, N. Guillo, S. B. Kalindjan, M. J. Tozer, Bioorg.
Med. Chem. Lett,. 1997, 7, 1289-1292 (R1==H; R - H); Beilstein Registry Number: 459155
(R'=H; R8= 4,5-Me02).
A64: Compounds of this type can be prepared according to Scheme 24. Scheme 24
BocHN
Figure imgf000081_0002
Figure imgf000081_0003
112 113 114 i: NaH, DMSO; ii: Pd/C, H2, EtOH; iii: iBuOCOCI, diisopropylethylamine, CH2CI2; then diazomethane; iv: HBr, CH2CI2; v: NaH, THF; vi: NaOHaq., MeOH, 75°; then HCIaq.; vii: DBU, Mel,
DMF; viii: lithium diisopropylamide, THF, chlorotrimethylsilane, -78°; then R1-X; ix: resolution (e.g. lipase); then isolation of methylester: DBU, Mel, DMF; x: LiOHxl H20, MeOH, H20; xi: TFA,
CH2CI2; xii: FmocOSu, Na2C03aq., dioxane A65 and A 67: Compounds of these types can be prepared according to Schemes 25 and 26. Scheme 25
Figure imgf000082_0001
115 116 117
Figure imgf000082_0002
118 119 i: NaH, DMSO, BrCH(R1)COOMe; ii: LiOHxl H20, MeOH, H2O; iii: polyphosphoric acid; iv: NaH,
ClCOOMe, THF; v: resolution (e.g. lipase); then isolation as methylester: DBU, Mel, DMF; vi:
LiOHxl H2O, MeOH, H20; vii: TFA, CH2CI2; viii: FmocOSu, Na2C03aq., dioxane
Scheme 26
COOMe
Figure imgf000083_0002
tBu BocHN— (
Figure imgf000083_0001
COOMe 120 121 122
III, IV 66
Figure imgf000083_0003
i: NaH, THF, CH2I2; ϋ: NaH, DMSO; iii: Bu4NFx10H2O, THF; iv: methanesulfonylchloride, Et3N,
CH2CI2; then NaH, THF; v: NaOHaq., MeOH, 75°; then HCIaq.; vi: DBU, Mel, DMF; vii: lithium hexamethyldisilazide, THF, chlorotrimethylsilane, -78°; then R1-X; viii: Pd/C, H2, EtOH; ix: NaH,
THF, R14-X; x: resolution (e.g. lipase); then isolation of methylester: DBU, Mel, DMF; xi:
LiOHxl H2O, MeOH, H20; xii: TFA, CH2CI2; xiii: FmocOSu, Na2C03aq., dioxane
A66: See G. L. Grunewald, L. H. Dahanukar, J. Heterocycl. Chem. 1994, 31, 1609-1618 (R1=H; R8=H, 8-N02 ; C(1)=0).
A68: See Griesbeck , H. Mauder, I. Mϋller, Chem. Ber. 1992, 11, 2467-2476; (R^R^H; C(1)=0). A69: R. Kreher, W. Gerhardt, Liebigs Ann. Chem. 1981, 240-247 (R^R^H).
As explained above, building blocks A70 belong to the class of open-chain α-substituted α-amino acids, A71 and A72 to the class of the the corresponding β-amino acid analogues and A73-A104 to the class of the cyclic analogues of A70.
Building blocks of types A70 and A73-A104 have been synthesized by several different general methods: by [2+2] cycloaddition of ketenes with imines (I. Ojima, H. J. C. Chen, X. Quin, Tetrahedron Lett. 1988, 44, 5307-5318); by asymmetric aldol reaction (Y. Ito, M. Sawamura, E. Shirakawa, K. Hayashikazi, T. Hayashi, Tetrahedron Lett. 1988, 29, 235-238; by the oxazolidinone method (J. S. Amato, L. M. Weinstock, S. Karady, US 4508921 A; M. Gander- Coquoz, D. Seebach, Helv. Chim. Acta 1988, 71, 224-236; A. K. Beck, D. Seebach, Chimia 1988, 42, 142-144; D. Seebach, J. D. Aebi, M. Gander-Coquoz, R. Naef, Helv. Chim. Acta 1987, 70, 1194-1216; D. Seebach, A. Fadel, Helv. Chim. Acta 1995, 68, 1243-1250; J. D. Aebi, D. Seebach, Helv. Chim. Acta 1985, 68, 1507-1518; A. Fadel, J. Salaun, Tetrahedron Lett. 1987, 28, 2243- 2246); by Schmidt- rearrangement of α,α-disubstituted α-ketoesters (G. I. Georg, X. Guan, J. Kant, Tetrahedron Lett. 1988, 29, 403-406); asymmetric synthesis via chiral Ni(II)- derived Schiff-bases (Y. N. Belokon, V. I. Bakhmutov, N. I. Chernoglazova, K. A. Kochetov, S. V. Vitt, N. S. Garbalinskaya, V. M. Belikov, J. Chem. Soc. Perkin Trans. 1, 1988, 305-312; M. Kolb, J. Barth, Liebigs Ann. Chem. 1983, 1668-1688); by the bis-lactim ether synthesis (U. Schollkopf, R. Hinrichs, R. Lonsky, Angew. Chem. 1987, 99, 137-138); by microbial resolution (K. Sakashita, I. Watanabe, JP 62/253397 A2) and by the hydantoin method combined with resolution of the racemic amino acids with chiral auxilliaries derived from L-phenylalanine amides (D. Obrecht, C. Spiegler, P. Schonholzer, K. Mϋller, H. Heimgartner, F. Stierli, Helv. Chim. Acta 1992, 75, 1666- 1696; D. Obrecht, U. Bohdal, J. Daly, C. Lehmann, P. Schonholzer, K. Mϋller, Tetrahedron
1995, 51, 10883-10900; D. Obrecht, C. Lehmann, C. Ruffieux, P. Schonholzer, K. Mϋller, Helv. Chim. Acta 1995, 78, 1567-1587; D. Obrecht, U. Bohdal, C. Broger, D. Bur, C. Lehmann, R. Ruffieux, P. Schonholzer, C. Spiegler, Helv. Chim. Acta 1995, 78, 563-580; D. Obrecht, H. Karajiannis, C. Lehmann, P. Schonholzer, C. Spiegler, Helv. Chim. Acta 1995, 78, 703-714; D. Obrecht, M. Altorfer, C. Lehmann, P. Schonholzer, K. Mϋller, J. Org. Chem. 1996, 61, 4080- 4086; D. Obrecht, C. Abrecht, M. Altorfer, U. Bohdal, A. Grieder, P. Pfyffer, K. Mϋller, Helv. Chim. Acta 1996, 79, 1315-1337). The latter method has been especially useful in preparing both enantiomers of building blocks of type A70 (see Scheme 27) and A73-A104 (see Scheme 28) in pure form.
Scheme 27
Figure imgf000085_0001
136a i: KCN, (NH )2CO3, EtOH/H2O; ii: Ba(OH)2, H2O; iii: aq.NaOH, PhCOCl, dioxane; then DCC,
CH2CI2; iv: NaH, DMF, R 8-X or R19-X; v: L-phenylalanine cyclohexylamide, N-methylpyrrolidone,
70°; vi: CH3SO3H, MeOH, 80°; vii: 6N HCIaq., dioxane, 100°; viii: Me3SiCI, DIEA, CH2CI2; then
FmocCI The method depicted in Scheme 27 consists in treatment of the appropriate ketones 126 with KCN, (NH4)2CO3 in a mixture of ethanol/water (E. Ware, J. Chem. Res. 1950, 46, 403; L. H. Goodson, I. L. Honigberg, J. j. Lehmann, W. H. Burton, J. Org. Chem. 1960, 25, 1920; S. N. Rastogi, j. S. Bindra, N. Anand, Ind. J. Chem. 1971, 1175) to yield the corresponding hydantoins 127, which were hydrolyzed with Ba(OH)2 in water at 120- 140° (R. Sarges, R. C. Schur, j. L. Belletire, M. J. Paterson, J Med. Chem. 1988, 31, 230) to give 128 in high yields. Schotten-Baumann acylation (Houben-Weyl, 'Methoden der Organischen Chemie', Volume XI72, Stickstoff-Verbindungen II und III', Georg Tieme Nerlag, Stuttgart, pp 339) followed by cyclization with Ν,Ν'-dicyclohexyl carbodiimide gave azlactones 129 (D. Obrecht, U. Bohdal, C. Broger, D. Bur, C. Lehmann, R. Ruffieux, P. Schonholzer, C. Spiegler, Helv. Chim. Acta 1995, 78, 563-580; D. Obrecht, C. Spiegler, P. Schonholzer, K. Mϋller, H. Heimgartner, F. Stierli, Helv. Chim. Acta 1992, 75, 1666-1696). Alternatively, azlactones 129 could also be prepared starting from amino acids 130 and 131, Schotten-Baumann acylation and cyclization with N,N'- dicyclohexyl carbodiimide to azlactones 132 and 133 and alkylation to yield 129 (D. Obrecht, U. Bohdal, C. Broger, D. Bur, C. Lehmann, R. Ruffieux, P. Schonholzer, C. Spiegler, Helv. Chim. Acta 1995, 78, 563-580; D. Obrecht, C. Spiegler, P. Schonholzer, K. Mϋller, H. Heimgartner, F. Stierli, Helv. Chim. Acta 1992, 75, 1666-1696)(see Scheme 1). Treatment of 129 with L-phenylalanine cyclohexylamide (D. Obrecht, U. Bohdal, C. Broger, D. Bur, C. Lehmann, R. Ruffieux, P. Schonholzer, C. Spiegler, Helv. Chim. Acta 1995, 78, 563-580) gave diastereomeric peptides 134 and 135, which could be conveniently separated by flash-chromatography or crystallisation. Treatment of 134 and 135 with methanesulphonic acid in methanol at 80° gave esters 136a and 136b which were converted into the corresponding Fmoc-protected final building blocks 137a and 137b.
Scheme 28
Figure imgf000087_0001
138 i: KCN, (NH4)2CO3, EtOH/H20; ii: Ba(OH)2, H2O; iii: aq.NaOH, PhCOCl, dioxane; then DCC,
CH2CI2; iv: L-phenylalanine cyclohexylamide, N-methylpyrrolidone, 70°; v: CH3SO3H, MeOH,
80°; vi: 6N HCIaq., dioxane, 100°; vii: Me3SiCI, DIEA, CH2CI2; the FmocCI According to the general method described in Scheme 28 (D. Obrecht, U. Bohdal, C. Broger, D. Bur, C. Lehmann, R. Ruffieux, P. Schonholzer, C. Spiegler, Helv. Chim. Acta 1995, 78, 563-580; D. Obrecht, C. Spiegler, P. Schonholzer, K. Mϋller, H. Heimgartner, F. Stierli, Helv. Chim. Acta 1992, 75, 1666-1696) A73-A104 can be prepared starting from the corresponding ketones 138, hydantoin formation (139) (E. Ware, J. Chem. Res. 1950, 46, 403; L. H. Goodson, I. L. Honigberg, J. J. Lehmann, W. H. Burton, J. Org. Chem. 1960, 25, 1920; S. N. Rastogi, J. S. Bindra, N. Anand, Ind. J. Chem. 1971, 1175; D. Obrecht, U. Bohdal, C. Broger, D. Bur, C. Lehmann, R. Ruffieux, P. Schonholzer, C. Spiegler, Helv. Chim. Acta 1995, 78, 563-580) and saponification (Ba(OH)2) to yield the racemic amino acids 140, which upon Schotten-Baumann- acylation and cyclization with N,N'-dicyclohexylcarbodiimide gave azlactones 141. Reaction with L-phenylalanine cyclohexylamide (D. Obrecht, U. Bohdal, C. Broger, D. Bur, C. Lehmann, R. Ruffieux, P. Schonholzer, C. Spiegler, Helv. Chim. Acta 1995, 78, 563-580) gave the diastereomeric peptides 142 and 143, which were separated by flash-chromatography or crystallization. Treatment of 142 and 143 with methanesulphonic acid in methanol at 80° gave esters 144a and 144b which were converted into the corresponding suitably protected amino acid precursors 145a and 145b, ready for peptide synthesis.
A71: Amino acid building blocks of this type (see formula 147) can be conveniently prepared from the corresponding disubstituted succinates 146 by Cwrttw-s-rearrangement as shown in Scheme 29.
Scheme 29
HOθC /COOMe " CbzHN" VC00Me
R18 R19 R1 R19
146 147 i: diphenylphosphoryl azide, toluene, 80°; then benzyl alcohol
A71: See D. Seebach, S. Abele. T. Sifferlen, M. Haenggi, S. Gruner, P. Seiler, Helv. Chim. Acta
1998, 81, 2218-2243 (R18 and R19 form: -(CH2)2-; -(CH2)3-; -(CH2)4-; -(CH2)5-; R20=H); L. Ducrie, S. Reinelt, P. Seiler, F. Diederich, D. R. Bolin, R. M. Campbell, G. L. Olson, Helv. Chim. Acta
1999, 82, 2432-2447; C. N. C. Drey, R. J. Ridge, J. Chem. Soc. Perkin Trans.l, 1981, 2468-2471; U. P. Dhokte, V. V. Khau, D. R. Hutchinson, M. J. Martinelli, Tetrahedron Lett. 1998, 39, 8771- 8774 (R18=RI9= Me; R20=H); D. L. Varie, D. A. Hay, S. L. Andis, T. H. Corbett, Bioorg. Med.
Chem. Lett. 1999, 9, 369-374 (R18=R19= Et); Testa, J. Org. Chem. 1959, 24, 1928-1936 (R18= Et;
R19= Ph); M. Haddad, C. Wakselman, J. Fluorine Chem. 1995, 73, 57-60 (R18= Me; R19= CF3;
R20=H); T. Shono, K. Tsubata, N. Okinaga, J. Org. Chem. 1984, 49, 1056-1059
(R,8=R19=R20=Me); K. Ikeda, Y. Terao, M. Sekiya, Chem. Pharm. Bull. 1981, 29, 1747-1749 (R18 and R19 form: -(CH2)S-; R20=Me).
Amino acid building blocks of type A72 can be conveniently prepared by Arndt-Eistert Cl- homologation of compounds of type A70 according to Scheme 30.
Scheme 30
Figure imgf000089_0001
148 149 i: iBuOCOCI, diisopropylethylamine, CH2CI2; then diazomethane, hv or Cu(l)
A72: See Y. V. Zeifman, J. Gen. Chem. USSR (Engl.Trans.) 1967, 37, 2355-2363 (R18=R19=CF3);
W. R. Schoen, J. M. Pisano, K. Pendergast, M. J. Wyvratt, M. H. Fisher, J. Med. Chem. 1994, 37, 897-906; S. Thaisrivongs, D. T. Pals, D. W. DuCharme, S. Turner, G. L. DeGraaf, J. Med. Chem.
1991, 34, 655-642; T. K. Hansen, H. Thoegersen, B. S. Hansen, Bioorg. Med. Chem. Lett. 1997, 7, 2951-2954; R. J. DeVita, R. Bochis, A. J. Frontier, A. Kotliar, M. H. Fisher, J. Med. Chem. 1998, 41, 1716-1728; D. Seebach, P. E. Ciceri, M. Overhand, B. Jaun, D. Rigo, Helv. Chim. Acta 1996, 79, 2043-2066; R. P. Nargund, K. H. Barakat, K. Cheng, W. Chan, B. R. Butler, A. A. Patchett, Bioorg. Med. Chem. Lett. 1996, 6, 1265-1270 (R18=R19=Me); E. Altmann, K. Nebel, M. Mutter, Helv. Chim. Acta 1991, 74, 800-806 (RI8=Me; R19=COOMe).
A73: Compounds of this type can be prepared according to C. Mapelli, G. Tarocy, F. Schwitzer, C. H. Stammer, J. Org. Chem. 1989, 54, 145-149 (R21= 4-OHC6H4); F. Elrod, E. M. Holt, C. Mapelli, C. H. Stammer, J. Chem. Soc. Chem. Commun. 1988, 252-253 (R21= CH2COOMe); R. E. Mitchell, M. C. Pirrung, G. M. McGeehan, Phytochemistry 1987, 26, 2695 (R21= CH2OH), J. Bland, A. Batolussi, C. H. Stammer, J. Org. Chem. 1988, 53, 992-995 (R21= CH2NH2). Additional derivatives of A73 have been described by T. Wakamiya, Y. Oda, H. Fujita, T. Shiba, Tetrahedron Lett. 1986, 27, 2143-2134; U. Schollkopf, B. Hupfeld, R. Gull, Angew. Chem. 1986, 98, 755-756; J. E. Baldwin, R. M. Adlington, B. J. Rawlings, Tetrahedron Lett. 1985, 26, 481- 484; D. Kalvin, K. Ramalinggam, R. Woodard, Synth. Comm. 1985, 15, 267-272 and L. M. Izquierdo, I. Arenal, M. Bernabe, E. Alvarez, Tetrahedron Lett. 1985, 41, 215-220.
A74: Compounds of this type can be prepared according to general method described in Scheme
28 starting from the corresponding cyclobutanones.
A75 and A76: Compounds of this type can be prepared using the following methods: P. Hughes,
J. Clardy, J. Org. Chem. 1988, 53, 4793-4796; E. A. Bell, M. Y. Qureshi, R. J. Pryce, D. H.
Janzen, P. Lemke, J. Clardy, J. Am. Chem. Soc. 1980, 102, 1409; Y. Gaoni, Tetrahedron Lett.
1988, 29, 1591-1594; R. D. Allan, J. R Haurahan, T. W. Hambley, G. A. R. Johnston, K. N. Mewett, A. D. Mitrovic, J. Med. Chem. 1990, 33, 2905-2915 (R23= COOH); G. W. Fleet, J. A.
Seijas, M. Vasquez Tato, Tetrahedron 1988, 44, 2077-2080 (R23= CH2OH).
A77: Compounds of this type can be prepared according to J. H. Burckhalter, G. Schmied, J. Pharm. Sci. 1966, 55, 443-445 (R23= aryl).
A78: Compounds of this type can be prepared according to J. C. Watkins, P. Kroosgard-Larsen, T. Honore, TIPS 1990, 11, 25-33; F. Trigalo, D. Brisson, R. Azerad, Tetrahedron Lett. 1988, 29, 6109 (R24= COOH). A79: Compounds of this type can be prepared according to general method described in Scheme 28 starting from the corresponding pyrrolidine-3-ones.
A80-A82: Compounds of this type can be prepared according to D. M. Walker, E. W. Logusch, Tetrahedron Lett. 1989, 30, 1181-1184; Y. Morimoto, K. Achiwa, Chem. Pharm. Bull. 1989, 35, 3845-3849; J. Yoshimura, S. Kondo, M. Diara, H. Hashimoto, Carbohydrate Res. 1982, 99, 129- 142.
A83: Compounds of this type can be prepared according to general method described in Scheme 28 starting from the corresponding pyrazoline-4-ones.
A84: Compounds of this type can be prepared according to R. M. Pinder, B. H. Butcher, D. H. Buxton, D. J. Howells, J. Med. Chem. 1971, 14, 892-893; D. Obrecht, U. Bohdal, C. Broger, D. Bur, C. Lehmann, R. Ruffieux, P. Schonholzer, C. Spiegler, Helv. Chim. Acta 1995, 78, 563-580.
A85: Compounds of this type can be prepared according to general method described in Scheme 28 starting from the corresponding indane-l,3-diones.
A86: Compounds of this type can be prepared according to general method described in Scheme 28 starting from the corresponding indane-2-ones.
A87: Compounds of this type and analogues thereof can be prepared according to C. Cativiela, M. D. Diaz de Villegas, A. Avenoza, J. M. Peregrina, Tetrahedron 1993, 47, 10987-10996; C. Cativiela, P. Lopez, J. A. Mayoral, Tetrahedron Assymmetry 1990, 1, 379; C. Cativiela, J. A. Mayoral, A. Avenoza, M. Gonzalez, M. A. Rey, Synthesis 1990, 1114.
A87 and A88: Compounds of this type can be prepared according to L. Munday, J. Chem. Soc. 1961, 4372; J. Ansell, D. Morgan, H. C. Price, Tetrahedron Lett. 1978, 47, 4615-4616.
A89: Compounds of this type can be prepared according to general method described in Scheme 28 starting from the corresponding piperidine-3-ones.
A90: Compounds of this type can be prepared according to general method described in Scheme 28 starting from the corresponding tetrahydrothiapyran-3-ones. A91 : Compounds of this type can be prepared according to general method described in Scheme 28 starting from the corresponding tetrahydropyran-3-ones.
A92: Compounds of this type can be prepared according to general method described in Scheme 28 starting from the corresponding piperidine-2,5-diones.
A93: Compounds of this type can be prepared according to general method described in Scheme 28 starting from the corresponding cyclohexanones.
A94: Compounds of this type can be prepared according to J. Org. Chem. 1990, 55, 4208.
A95: Compounds of this type can be prepared according to N. J. Lewis, R. L. Inloes, J. Hes, R. H. Matthews, G. Milo, J. Med. Chem. 1978, 21, 1070-1073.
A96: Compounds of this type can be prepared according to general method described in Scheme 28 starting from the corresponding tetrahydropyran-4-ones.
A97: Compounds of this type can be prepared according to general method described in Scheme 28 starting from the corresponding piperidine-2,4-diones.
A98: Compounds of this type can be prepared according to general method described in Scheme 28 starting from the corresponding 1-tetralones (D. Obrecht, C. Spiegler, P. Schonholzer, K. Mϋller, H. Heimgartner, F. Stierli, Helv. Chim. Acta 1992, 75, 1666-1696).
A99: Compounds of this type can be prepared according to general method described in Scheme 28 starting from the corresponding tetraline-l,4-dione mono-diethylacetals.
A100: Compounds of this type can be prepared according to general method described in Scheme 28 starting from the corresponding tetrahydroquinolin-4-ones.
A101: Compounds of this type can be prepared according to general method described in Scheme 28 starting from the corresponding tetrahydroquinoline-2,4-diones. A102: Compounds of this type can be prepared according to K. Ishizumi, N. Ohashi, N. Tanno, J. Org. Chem. 1987, 52, 4477-4485; D. Obrecht, U. Bohdal, C. Broger, D. Bur, C. Lehmann, R. Ruffieux, P. Schonholzer, C. Spiegler, Helv. Chim. Acta 1995, 78, 563-580; D. Obrecht, C. Spiegler, P. Schonholzer, K. Mϋller, H. Heimgartner, F. Stierli, Helv. Chim. Acta 1992, 75, 1666- 1696; D. R. Haines, R. W. Fuller, S. Ahmad, D. T. Vistica, V. E. Marquez, J. Med. Chem. 1987, 30, 542-547; T. Decks, P. A. Crooks, R. D. Waigh, J. Pharm. Sci 1984, 73, 457-460; I. A. Blair, L. N. Mander, «-ytr. J. Chem. 1979, 32, 1055-1065.
Overviews dealing with building blocks of types (b)-(p) are: S. Hanessian, G. McNaughton- Smith, H.-G. Lombart, W. D. Lubell, Tetrahedron 1997, 38, 12789-12854; D. Obrecht, M. Altorfer, J. A. Robinson, "Novel Peptide Mimetic Building Blocks and Strategies for Efficient Lead Finding", Adv. Med. Chem. 1999, Vo , 1-68
Templates of type (bl) can be prepared according to Schemes 31 and 32.
Scheme 31
Figure imgf000094_0001
153 154
i: Treatment of 150 with a dehydrating reagent such as thionylchloride in methanol at an elevated temperature, conveniently at reflux.
Introduction of Boc, e.g. using di-tert. -butyl dicarbonate and triethylamine in a suitable solvent such as dichloromethane; any other suitable N-protecting group (not shown in
Reaction Scheme 31) can be introduced in an analogous manner. in: Reaction of formed product with phthalimide, diethyl diazodicarboxylate and triphenylphoshine under standard Mitsunobu conditions (Mitsunobu, O.; Wada, M.; Sano,
T. J. J. Am. Chem. Soc. 1972, 94, 672) to conveniently yield 151. iv: Treatment of 151 with trifluoracetic acid in dichloromethane. v: 152 is coupled under standard peptide coupling conditions with Cbz-Asp(tBu)OH in
DMF with reagents such as HBTU and 1-hydroxybenztriazole (HOBt) with a base such as diisopropylethylamine to yield 153. vi : Removal of the Cbz-group, conveniently by hydrogenation using H2 and a catalyst such as Palladium on charcoal, in solvents such as ethanol, DMF and ethyl acetate. vu: The phthalimide group is cleaved off from the resulting product, conveniently by treatment with hydrazine in a suitable solvent such as ethanol at an elevated temperature, suitably at about 80° C and cleavage of the formed product with trifluoracetic acid in
CH2C12. viii: The formed amino acid is conveniently protected with reagents such as 9- fluorenylmethoxcarbonyl chloride or 9-fluorenylmethoxcarbonyl succinimide using a base such as sodium carbonate or triethylamine in a suitable solvent or mixture of solvents such as dioxane and water, or dichloromethane to yield 154 as described by Bisang, C; Weber, C; Robinson, J. A. Helv. Chim. Acta 1996, 79, 1825-1842.
Scheme 32
Figure imgf000096_0001
159
Treatment of 150 with a dehydrating reagent such as thionyl chloride in a suitable solvent such as methanol at an elevated temperature, conveniently at reflux. n: The resulting amino acid ester is N-protected under standard conditions for introducing the Cbz-group, e.g. using benzyloxycarbonyl chloride and triethylamine in a suitable solvent such as dichloromethane. in: The Cbz-protected amino acid methyl ester is treated with trimethylsilylchloride and a base such as triethylamine in a solvent such as tetrahydrofuran, cooled, conveniently to about -78° C, followed by reaction with a strong base such as lithium diisopropylamide or lithium hexamethyldisilylazide and tert.-butyl bromoacetate yielding 155 as a mixture of diastereomers as described by Bisang, C; Jiang, L.; Freund, E.; Emery, F.; Bauch, C; Matile, H,; Pluschke, G.; Robinson, J. A. J. Am. Chem. Soc. 1998, 120, 7439-7449; Emery, F.; Bisang, C; Favre, M.; Jiang, L.; Robinson, J. A. J. Chem. Soc. Chem. Commun. 1996, 2155-2156. iv: Reaction of 155 with phthalimide, diethyl diazodicarboxylate and triphenylphosphine under standard Mitsunobu conditions (Mitsunobu, O.; Wada, M.; Sano, T. J. J. Am.
Chem. Soc. 1972, 94, 672). v: The resulting product is hydrogenated using H2 and a suitable catalyst such as palladium on charcoal in a solvent such as ethyl acetate, DMF or ethanol; subsequently separation of diastereomers takes place and yields 156. vi: 156 is coupled with Fmoc-Asp(allyl)OH under standard peptide coupling conditions using reagents such as HATU, HO At and a base such as diisopropylethylamine in a suitable solvent such as DMF. vii: Cyclization, conveniently with DBU in DMF to yield 157. viii: The phthalimide group is cleaved off from resulting product, conveniently by hydrazinolysis, e.g. treatment with methylhydrazine in a suitable solvent such as DMF. ix: The formed product is conveniently protected with reagents such as 9- fluorenylmethoxcarbonyl chloride or 9-fluorenylmethoxcarbonyl succinimide using a base such as sodium carbonate or triethylamine in a suitable solvent or mixture of solvents such as dioxane and water, or dichloromethane to yield 158. x: Standard removal of an allyl ester group using e.g. palladium(0) as catalyst gives 159.
Templates of type (b2) can be prepared according to Scheme 33.
Scheme 33
Vitamin C
Figure imgf000098_0001
Figure imgf000098_0002
164
i : 160 (obtainable from Vitamin C as described by Hubschwerlen, C. (Synthesis 1986, 962) is treated with phthalimide, diethyl diazodicarboxylate and triphenylphoshine under standard Mitsunobu conditions (Mitsunobu, O.; Wada, M.; Sano, T. J. J. Am. Chem. Soc. 1972, 94, 672). ii: The phthalimide group is cleaved off from the product, conveniently by hydrazinolysis, e.g. by treatment with methylhydrazine in a suitable solvent such as DMF. iii: The amino group is protected by treatment with a benzoylating reagent such as benzoic acid anhydride or benzoylchloride and a base such as triethylamine or 4-dimethylaminopyridine in a suitable solvent such as dichloromethane or DMF. iv: Removal of the 2,4-dimethoxybenzyl group, e.g. with K2S2O8 and Na2HP04 in aqueous acetonitrile at an elevated temperature, e.g. at about 80° C. v: Introduction of a ter -butoxycarbonyl group using e.g. di-tert.-butyloxycarbonyl dicarbonate, triethylamine and a catalytic amount of 4-dimethylarninopyridine in a suitable solvent such as dichloromethane. vi: Reaction with aqueous sodium carbonate in tetrahydrofuran followed by acidification, vii: Esterification of the carboxylic acid group, conveniently with diazomethane in a suitable solvent such as diethylether yielding 161. viii Removal of the Cbz-group, conveniently by hydrogenation with H2 in the presence of a catalyst such as palladium on charcoal in a solvent such as DMF to yield 161 as described by Pfeifer, M.; Robinson, J. A. J. Chem. Soc. Chem. Commun. 1998, 1977. ix: 161 is coupled under standard peptide coupling conditions with Cbz-Asp(tBu)OH in
DMF with reagents such as HBTTJ and 1-hydroxybenztriazole with a base such as diisopropylethylamine to yield 162 as described by Pfeifer, M.; Robinson, J. A. J. Chem. Soc. Chem. Commun. 1998, 1977. x: Removal of the Cbz-group, e.g. by hydrogenation using H2 and a catalyst such as palladium on charcoal under standard conditions, yields 163 as described by Pfeifer, M.;
Robinson, J. A. J. Chem. Soc. Chem. Commun. 1998, 1977. xi: Cleavage of the tert.-butyl ester and tert-butyloxycarbonyl groups, conveniently using trifluoracetic acid in dichloromethane or 4N hydrochloric acid in dioxane. xii: The intermediate free amino acid formed is conveniently protected with reagents such as
9-fluorenylmethoxcarbonyl chloride or 9-fluorenylmethoxcarbonyl succinimide using a base such as sodium carbonate or triethylamine in a suitable solvent or mixture of solvents such as dioxane and water, or dichloromethane to yield 164 as described by Pfeifer, M.; Robinson, J. A. J. Chem. Soc. Chem. Commun. 1998, 1977.
Templates of type (cl) can be prepared according to Schemes 34 to 37.
Scheme 34
Figure imgf000100_0001
Vll-X
Figure imgf000100_0002
168 170
i: 166 can be synthesized from 165 according to P. Waldmeier, "Solid-supported synthesis of highly substituted xanthene-derived templates for the synthesis of β-turn stabilized cyclic peptide libraries", PhD-thesis, University of Zurich, 1996. For cleaving the phthalimide group 166 is conveniently submitted to hydrazinolysis, e.g. by treatment with hydrazine hydrate in a suitable solvent such as ethanol at an elevated temperature, e.g. at about 80° C. ii: The intermediate aminonitrile is saponified, conveniently under basic conditions, e.g. with aqueous sodium hydroxide in a suitable solvent such as ethanol at an elevated temperature, conveniently under reflux, to yield 167. iii: The intermediate free amino acid formed is conveniently protected with reagents such as 9-fluorenylmethoxcarbonyl chloride or 9-fluorenylmethoxcarbonyl succinimide using a base such as sodium carbonate or triethylamine in a suitable solvent or mixture of solvents such as dioxane and water, or dichloromethane to yield 168 as described by P. Waldmeier, "Solid-supported synthesis of highly substituted xanthene-derived templates for the synthesis of β-turn stabilized cyclic peptide libraries", PhD-thesis, University of Zurich, 1996.
iv: Regioselective bromination of 167 is performed preferably with bromine in acetic acid and dichloromethane. In a similar fashion R37 = NO2 can be introduced by treatment with HNO3 in acetic acid and R37 = CH2-NPht by treatment with hydroxymethyl phthalimide in H2SO4. v: The amino group is conveniently Cbz-protected with reagents such as benzyloxycarbonyl chloride or succinimide in a suitable solvent such as dioxane in presence of a base such as aqueous sodium hydroxide. vi: The carboxylic acid group is esterified, preferably with DBU and methyl iodide in DMF to yield 169. vii: Introduction of lower alkyl, substituted lower alkyl and aryl substituents (R37), conveniently by palladium(O)- catalyzed Stille- (Stifle, J.K. Angew. Chem.1986, 68, 504) and Suzuki- couplings (Oh-e, T.; Mijaura, N.; Suzuki, A. J. Org. Chem. 1993, 58, 2201). Any other functionalization known for aryl bromides can be employed for introduction of substituents R37. viii: Removal of the Cbz-group, e.g. by hydrogenation using H2 and a catalyst such as palladium on charcoal in a suitable solvent such as ethanol, DMF and ethyl acetate, ix: Hydrolysis of the ester group, conveniently under acidic conditions, e.g. with 25% aqueous hydrochloric acid in a suitable solvent such as dioxane at an elevated temperature, preferably at about 100° C. x: The intermediate free amino acid formed is conveniently protected with reagents such as 9-fluorenylmethoxcarbonyl chloride or 9-fluorenylmethoxcarbonyl succinimide using a base such as sodium carbonate or triethylamine in a suitable solvent or mixture of solvents such as dioxane and water, or dichloromethane to yield 170. Scheme 35
Figure imgf000102_0001
i: Double ortho- bromination of 171 is performed preferably with excess bromine in acetic acid and dichloromethane. In a similar fashion R37 = R38 = NO2 can be introduced by treatment with HNO3 in acetic acid and R37 = R38 = CH2-NPht by treatment with hydroxymethyl phthalimide in H2S04. ii: The amino group is protected, conveniently Cbz-protected, with reagents such as benzyloxycarbonyl chloride or succinimide in a suitable solvent such as dioxane in the presence of a base such as aqueous sodium hydroxide. iii: The carboxylic acid group is esterifϊed, preferably with DBU and methyl iodide in DMF to yield 172. iv: Introduction of lower alkyl, substituted lower alkyl and aryl substituents (R37 =
R38), e.g. by palladium(O)- catalyzed Stille- (Stille, J.K. Angew. Chem.1986, 68, 504) and
Suzuki- couplings (Oh-e, T.; Mijaura, N.; Suzuki, A. J. Org. Chem. 1993, 58, 2201). Any other functionalization known for aryl bromides can be employed for introduction of substituents R37 and R38. v: Removal of the Cbz-group of 173, e.g. by hydrogenation using H2 and a catalyst such as palladium on charcoal in a suitable solvent such as ethanol, DMF or ethyl acetate. vi: Hydrolysis of the ester group, conveniently under acidic conditions, e.g. with
25% aqueous hydrochloric acid in a suitable solvent such as dioxane at an elevated temperature, conveniently at about 100° C. vii: The intermediate free amino acid formed is conveniently protected with reagents such as 9-fluorenylmethoxcarbonyl chloride or 9-fluorenylmethoxcarbonyl succinimide using a base such as sodium carbonate or triethylamine in a suitable solvent or mixture of solvents such as dioxane and water, or dichloromethane to yield 174.
Scheme 36
Figure imgf000104_0001
Vlll-X
Figure imgf000104_0002
i: Cleavage of the methoxy groups of 166, preferably by treatment with an excess of boron tribromide in a suitable solvent such as dichloromethane. ii: Hydrolysis of the cyano group under acidic conditions, preferably with 25% aqueous hydrochloric acid in a suitable solvent such as dioxane at an elevated temperature, conveniently at about 100° C. iii: The resulting acid is treated with a dehydrating agent such as thionyl chloride in a suitable solvent such as dioxane to yield 175. iv: Treatment of 175 with an appropriate triflating reagent, preferably trifluoromethanesulfonic acid anhydride in the presence of a base such as 2,6-di-tert- butyl-pyridine in a suitable solvent such as dichloromethane. v: Heating of the intermediate, conveniently in a suitable solvent such as methanol. vi: Introduction of lower alkyl or aryl-lower alkyl (R35) by alkylation to yield 177.
Any other functionalization known for phenol groups can be employed for introduction of substituents R35. vii: Introduction of lower alkyl or aryl (R36), conveniently by palladium(O)- catalyzed Suzuki- coupling (Oh-e, T.; Mijaura, N.; Suzuki, A. J. Org. Chem. 1993, 58, 2201) to yield 178. Any other functionalization known for aryl bromides can be employed for introduction of substituents R36. viii: Hydrolysis of the ester group under acidic conditions, conveniently with 25% aqueous hydrochloric acid in a suitable solvent such as dioxane at an elevated temperature, e.g. at about 100° C. ix: Cleavage of the phthalimido group, conveniently by hydrazinolysis, e.g. with hydrazine hydrate in a suitable solvent such as ethanol. x: The intermediate free amino acid formed is conveniently protected with reagents such as 9-fluorenylmethoxcarbonyl chloride or 9-fluorenylmethoxcarbonyl succinimide using a base such as sodium carbonate or triethylamine in a suitable solvent or mixture of solvents such as dioxane and water, or dichloromethane to yield 179.
Scheme 37
Figure imgf000106_0001
Bromination of 175 using reagents such as bromine in a mixture of acetic acid and dichloromethane at temperatures ranging from about 0° C to about room temperature. n: Benzoylation of the hydroxy group using an appropriate acylating agent such as benzoyl chloride or benzoic acid anhydride, a base such as pyridine or triethylamine and a suitable solvent such as dichloromethane to yield 180. in: 180 is treated with methanol and a catalytic amount of an acidic catalyst such as camphor sulfonic acid under heating. iv: Introduction of lower alkyl or aryl-lower alkyl (R35) by alkylation using a base such as sodium hydride or potassium tert.-butoxide in a solvent such as tetrahydrofuran, dimethoxyethane or DMF gives 181. v: Lower alkyl, substituted lower alkyl and aryl substituents (R38) are introduced, e.g. by palladium(O)- catalyzed Stille- (Stille, J.K. Angew. Chem.1986, 68, 504) and
Suzuki- couplings (Oh-e, T.; Mijaura, N.; Suzuki, A. J. Org. Chem. 1993, 58, 2201). Any other functionalization known for aryl bromides can be employed for introduction of substituents R38. vi: For cleaving the benzyloxy group the intermediate is conveniently heated with sodium cyanide adsorbed on aluminum oxide and methanol. vii: Treatment with an appropriate triflating reagent, preferably trifluoromethanesulfonic acid anhydride, in the presence of a base such as 2,6-di-tert.-butyl-pyridine in a suitable solvent such as dichloromethane. viii: Introduction of lower alkyl and aryl substituents (R36), e.g. by palladium(O)- catalyzed Stille- (Stille, J.K. Angew. Chem.1986, 68, 504) and Suzuki- couplings (Oh-e,
T.; Mijaura, N.; Suzuki, A. J. Org. Chem. 1993, 58, 2201) yields 182. Any other functionalization known for aryl bromides can be employed for introduction of substituents R36. ix: Bromination under standard conditions such as using bromine in acetic acid and dichloromethane at temperatures ranging from about 0° C to about room temperature. x: Lower alkyl, substituted lower alkyl and aryl substituents (R37) are introduced, e.g. by palladium(O)- catalyzed Stille- (Stille, J.K. Angew. Chem.1986, 68, 504) and Suzuki- couplings (Oh-e, T.; Mijaura, N.; Suzuki, A. J. Org. Chem. 1993, 58, 2201) to yield 184. Any other functionalization known for aryl bromides can be employed for introduction of substituents R37. xi: The ester group is hydrolyzed under acidic conditions, conveniently with 25% aqueous hydrochloric acid in a suitable solvent such as dioxane at an elevated temperature, e.g. at about 100° C. xii: The phthali ido group is cleaved, e.g. by hydrazinolysis, conveniently with hydrazine hydrate in a suitable solvent such as ethanol. xiii: The intermediate free amino acid formed is conveniently protected with reagents such as
9-fluorenylmethoxcarbonyl chloride or 9-fluorenylmethoxcarbonyl succinimide using a base such as sodium carbonate or triethylamine in a suitable solvent or mixture of solvents such as dioxane and water, or dichloromethane to yield 185.
Templates of type (c2) can be prepared as shown in Schemes 38 and 39. Scheme 38
Figure imgf000108_0001
3,7-Dimethoxyphenothiazine 186 is prepared and converted into 187 according to Mϋller, K.; Obrecht, D.; Knierzinger, A.; Spiegler, C; Bannwarth, W.; Trzeciak, A.; Englert, G; Labhardt, A.; Schonholzer, P. Perspectives in Medicinal Chemistry, Editor Testa, B.; Kyburz, E.; Fuhrer, W.; Giger, R., Weinheim, New York, Basel, Cambridge: Verlag Helvetica Chimica Acta, 1993, 513-531; Bannwarth, W.; Gerber, F.; Grieder, A.; Knierzinger, A.; Mϋller, K.; Obrecht. D.; Trzeciak, A. Can. Pat. Appl. CA2101599(131 pages). The benzyl group is cleaved off from 187 conveniently by hydrogenation, e.g. with H2 and a catalyst such as palladium on charcoal in a suitable solvent such as ethanol, DMF or ethyl acetate. ii: Introduction of lower alkyl (R43) by alkylation using an appropriate alkylating agent (R43-X'; X - OTf, Br, I) and strong bases such as sodium amide in liquid ammonia or sodium hydride in tetrahydrofuran, dioxan or DMF in the presence of a phase transfer catalyst such as TDA-I. In a similar manner substituted lower alkyl (R43) can be introduced; thus, for example R43 = CH2COOR55 and CH2CH2COOR55 can be introduced by treatment with the appropriate 2-halo acetic and, respectively, 3 -halo propionic acid derivatives. Any other functionalization known for diarylamines can be employed for introduction of substituents R43. iii: Cleavage of the methoxy groups of 188, conveniently by treatment with an excess of boron tribromide in a suitable solvent such as dichloromethane at temperatures ranging from about -20° C to about room temperature, iv: For the introduction of lower alkyl, substituted lower alkyl or aryl-lower alkyl substituents (R39 and R40) the intermediate bis-phenol derivative is conveniently reacted with a reagent of the formula R39-and R40-X' (X' = OTf, Br, I) in the presence of strong bases such as sodium hydride in tetrahydrofuran, dioxan or DMF in the presence of a phase transfer catalyst such as TDA-I. Any other functionalization known for phenol groups can be employed for introduction of substituents R39 and R40. v: The cyano group of 188 and, respectively, 189 is hydrolyzed, conveniently under acidic conditions, e.g. with 25% aqueous hydrochloric acid in a suitable solvent such as dioxane at an elevated temperature, e.g. at about 100° C. vi: The phthalimide group of the intermediate is cleaved, conveniently by hydrazinolysis, e.g. with hydrazine hydrate in a suitable solvent such as ethanol. vii: The free amino group is conveniently protected with reagents such as 9- fluorenylmethoxcarbonyl chloride or 9-fluorenylmethoxcarbonyl succinimide using a base such as sodium carbonate or triethylamine in a suitable solvent or mixture of solvents such as dioxane and water, or dichloromethane to yield 190 and, respectively, 191.
Scheme 39
Figure imgf000110_0001
IX-XII
IX-XII
Figure imgf000110_0002
i: The cyano group of 188 is hydrolyzed, conveniently under acidic conditions, e.g. with
25% aqueous hydrochloric acid in a suitable solvent such as dioxane at an elevated temperature, e.g. at about 100° C. ii: The phthalimide group of the intermediate is cleaved, conveniently by hydrazinolysis, e.g. with hydrazine hydrate in a suitable solvent such as ethanol to yield 192. iii: Double ortho- bromination of 192 is performed preferably with excess bromine in acetic acid and dichloromethane. In a similar fashion R41 = R42 = NO2 can be introduced by treatment with HNO3 in acetic acid and R41 = R42 = CH2-NPht by treatment with hydroxymethyl phthalimide in H2S0 . Any other functionalization by electrophilic aromatic substitution known can be employed for introduction of substituents R41 and R42. iv: The amino group is protected, conveniently Cbz-protected, with reagents such as benzyloxycarbonyl chloride or succinimide in a suitable solvent such as dioxane in the presence of a base such as aqueous sodium hydroxide. v: The carboxylic acid group is esterified, preferably with DBU and methyl iodide in DMF to yield 193. vi: Regioselective bromination of 192 is performed preferably with bromine in acetic acid and dichloromethane. In a similar fashion R41 = N02 can be introduced by treatment with HN03 in acetic acid and R41 = CH2-NPt by treatment with hydroxymethyl phthalimide in H2SO4. Any other functionalization by electrophilic aromatic substitution known can be employed for introduction of substituents R41. vii: The amino group is conveniently Cbz-protected with reagents such as benzyloxycarbonyl chloride or succinimide in a suitable solvent such as dioxane in presence of a base such as aqueous sodium hydroxide. viii: The carboxylic acid group is esterified, preferably with DBU and methyl iodide in DMF to yield 194. ix: Introduction of lower alkyl, substituted lower alkyl and aryl substituents (R41)for
194 and (R41 and R42) for 193, conveniently by ρalladium(O)- catalyzed Stille- (Stille, J.K. Angew. Chem.1986, 68, 504) and Suzuki- couplings (Oh-e, T.; Mijaura, N.; Suzuki, A. J.
Org. Chem. 1993, 58, 2201). Any other functionalization known for aryl bromides can be employed for introduction of substituents R41 and R42. x: Removal of the Cbz-group, e.g. by hydrogenation using H2 and a catalyst such as palladium on charcoal in a suitable solvent such as ethanol, DMF and ethyl acetate. xi: Hydrolysis of the ester group, conveniently under acidic conditions, e.g. with
25% aqueous hydrochloric acid in a suitable solvent such as dioxane at an elevated temperature, preferably at about 100° C. xii: The intermediate free amino acid formed is conveniently protected with reagents such as 9-fluorenylmethoxcarbonyl chloride or 9-fluorenylmethoxcarbonyl succinimide using a base such as sodium carbonate or triethylamine in a suitable solvent or mixture of solvents such as dioxane and water, or dichloromethane to yield 195 and 196.
Templates of type (c3) can be prepared as shown in Schemes 40 and 41. ill
Scheme 40
Resorufin
Figure imgf000112_0001
V-VII
V-VII
Figure imgf000112_0002
i: 197 can be prepared from commercial resorufin and coverted into 198 according to Mϋller, K.; Obrecht, D.; Knierzinger, A.; Spiegler, C; Bannwarth, W.; Trzeciak, A.; Englert, G.; Labhardt, A.; Schonholzer, P. Perspectives in Medicinal Chemistry, Editor Testa, B.; Kyburz, E.; Fuhrer, W.; Giger, R, Weinheim, New York, Basel, Cambridge: Verlag Helvetica Chimica Acta, 1993, 513-531; Bannwarth, W.; Gerber, F.; Grieder, A.; Knierzinger, A.; Mϋller, K.; Obrecht. D.; Trzeciak, A. Can. Pat. Appl. CA2101599(131 pages). For splitting off the benzyl group 198 is conveniently hydrogenated e.g. with H2 and a catalyst such as palladium on charcoal in a suitable solvent such as ethanol, DMF or ethyl acetate. ii: Introduction of lower alkyl (R ") by alkyiation with R43-X' (X* = OTf, Br, I) using strong bases such as sodium amide in liquid ammonia or sodium hydride in tetrahydrofuran, dioxan or DMF in the presence of a phase transfer catalyst such as TDA- I to yield 199. In a similar manner substituted lower alkyl (R43) can be introduced; thus, for example, R43 = CH2COOR55 and CH2CH2COOR55 can be introduced by treatment with the appropriate 2-halo acetic and, respectively, 3-halo propionic acid derivatives. Any other functionalization of diarylamino groups known can be employed for introduction of substituents R43. iii: Cleavage of the methoxy groups of 199, conveniently by treatment with excess boron tribromide in dichloromethane at temperatures ranging from about -20° to about room temperature. iv: The intermediate bis-phenol derivative is preferably reacted with R39 and R 0-X' (X- OTf, Br, I) in the presence of strong bases such as sodium hydride in tetrahydrofuran, dioxan or DMF in the presence of a phase transfer catalyst such as TDA- I. Any other functionalization for phenol groups can be employed for introduction of substituents R39 and R40. v: The cyano group of 199 and, respectively, 200 is hydrolyzed under acidic conditions, e.g. with 25% aqueous hydrochloric acid in a suitable solvent such as dioxane at an elevated temperature, conveniently at about 100° C. vi: The phthalimide group is cleaved, conveniently by hydrazinolysis, e.g. with hydrazine hydrate in suitable solvent such as ethanol. vii: The free amino group is conveniently protected with reagents such as 9- fluorenylmethoxcarbonyl chloride or 9-fluorenylmethoxcarbonyl succinimide using a base such as sodium carbonate or triethylamine in suitable solvent or mixture of solvents such as dioxane and water, or dichloromethane to yield 201 and, respectively, 202.
Scheme 41
Figure imgf000114_0001
IX-XII
IX-XII
Figure imgf000114_0002
The cyano group of 199 is hydrolyzed, conveniently under acidic conditions, e.g. with
25% aqueous hydrochloric acid in a suitable solvent such as dioxane at an elevated temperature, e.g. at about 100° C. The phthalimide group of the intermediate is cleaved, conveniently by hydrazinolysis, e.g. with hydrazine hydrate in a suitable solvent such as ethanol to yield 203. iii: Double ortho- bromination of 203 is performed preferably with excess bromine in acetic acid and dichloromethane. In a similar fashion R41 = R42 = NO2 can be introduced by treatment with HNO3 in acetic acid and R41 = R42 = CH2-NPht by treatment with hydroxyrnethyl phthalimide in H2S04. Any other functionalization by electrophilic aromatic substitution can be employed for introduction of substituents R41 and R42. iv: The amino group is protected, conveniently Cbz-protected, with reagents such as benzyloxycarbonyl chloride or succinimide in a suitable solvent such as dioxane in the presence of a base such as aqueous sodium hydroxide. v: The carboxylic acid group is esterified, preferably with DBU and methyl iodide in DMF to yield 204. vi: Regioselective bromination of 203 is performed preferably with bromine in acetic acid and dichloromethane. In a similar fashion R41 = NO2 can be introduced by treatment with HNO3 in acetic acid and R4I=CH2-NPht by treatment with hydroxymethyl phthalimide in H2S04. vii: The amino group is conveniently Cbz-protected with reagents such as benzyloxycarbonyl chloride or succinimide in a suitable solvent such as dioxane in presence of a base such as aqueous sodium hydroxide. viii: The carboxylic acid group is esterified, preferably with DBU and methyl iodide in DMF to yield 205. ix: Introduction of lower alkyl, substituted lower alkyl and aryl substituents (R41)for
205 and (R41 and R42) for 204, conveniently by ρalladium(O)- catalyzed Stille- (Stille, J.K.
Angew. Chem.1986, 68, 504) and Suzuki- couplings (Oh-e, T.; Mijaura, N.; Suzuki, A. J. Org. Chem. 1993, 58, 2201). Any other functionalization known for aryl bromides can be employed for introduction of substituents R41 and R42. x: Removal of the Cbz-group, e.g. by hydrogenation using H2 and a catalyst such as palladium on charcoal in a suitable solvent such as ethanol, DMF and ethyl acetate. xi: Hydrolysis of the ester group, conveniently under acidic conditions, e.g. with 25% aqueous hydrochloric acid in a suitable solvent such as dioxane at an elevated temperature, preferably at about 100° C. xii: The intermediate free amino acid formed is conveniently protected with reagents such as 9-fluorenylmethoxcarbonyl chloride or 9-fluorenylmethoxcarbonyl succinimide using a base such as sodium carbonate or triethylamine in a suitable solvent or mixture of solvents such as dioxane and water, or dichloromethane to yield 206 and 207.
Templates(d) can be prepared according to D. Obrecht, U. Bohdal, C. Lehmann, P. Schonholzer, K. Mϋller, Tetrahedron 1995, 51, 10883; D. Obrecht, C. Abrecht, M. Altorfer, U. Bohdal, A. Grieder, M. Kleber, P. Pfyffer, K. Mϋller, Helv. Chim. Acta 1996, 79, 1315-1337.
Templates (el) and (e2): See R. Mueller, L. Revesz, Tetrahedron Lett. 1994, 35, 4091; H.-G. Lubell, W. D. Lubell, J. Org. Chem. 1996, 61, 9437; L. Colombo, M. DiGiacomo, G. Papeo, O. Carugo, C. Scolastico, L. Manzoni, Tetrahedron Lett. 1994, 35, 4031. Templates (e3): See S. Hanessian, B. Ronan, A. Laoui, Bioorg. Med. Chem. Lett. 1994, 4, 1397.
Templates (e4): See S. Hanessian, G. McNaughton-Smith, Bioorg. Med. Chem. Lett. 1996, 6, 1567.
Templates (f): See T.P. Curran, P. M. McEnay, Tetrahedron Lett. 1995, 36, 191-194.
Templates (g): See D. Gramberg, C. Weber, R. Beeli, J. Inglis, C. Bruns, J. A. Robinson, Helv. Chem. Acta 1995, 78, 1588-1606; K. H. Kim, J. P. Dumas, J. P. Germanas, J. Org. Chem. 1996, (57, 3138-3144.
Templates (h): See S. de Lombart, L. Blanchard, L. B. Stamford, D. M. Sperbeck, M. D. Grim, T. M. Jenson, H. R. Rodriguez, Tetrahedron Lett. 1994, 35, 7513-7516.
Templates (il): See J. A. Robl, D. S. Karanewski, M. M. Asaad, Tetrahedron Lett. 1995, 5, 773- 758.
Templates (i2): See T. P. Burkholder, T.-B. Le, E. L. Giroux, G. A. Flynn, Bioorg. Med. Chem. Lett. 1992, 2, 579.
Templates (i3) and (i4): See L. M. Simpkins, J. A. Robl, M. P. Cimarusti, D. E. Ryono, J. Stevenson, C.-Q. Sun, E. W. Petiillo, D. S. Karanewski, M. M. Asaad, J. E. Bird, T. R. Schaeffer, N. C. Trippodo, Abstracts of papers, 210th Am. Chem. Soc Meeting, Chicago, II 1, MEDI 064 (1995).
Templates (k): See D. Benlshai, A. R. McMurray, Tetrahedron 1993, 49, 6399.
Templates (I): See E. G. von Roedern, H. Kessler, Angew. Chem. Int. Ed. Engl. 1994, 33, 687- 689.
Templates (m): See R. Gonzalez-Muniz, M. J. Dominguez, M. T. Garcia-Lopez, Tetrahedron 1992, 45, 5191-5198. Templates (n): See F. Esser, A. Carpy, H. Briem, -H. όppen, K.-H. Pook, Int. J. Pept. Res. 1995, 45, 540-546.
Templates (o): See N. De la Figuera, I. Alkorta, T. Garcia-Lopez, R. Herranz, R. Gonzalez- Muniz, Tetrahedron 1995, 51, 7841.
Templates (p): See U. Slomcynska, D. K. Chalmers, F. Cornille, M. L. Smythe, D. D. Benson, K. D. Moeller, G. R. Marshall, J. Org. Chem. 1996, 61, 1198-1204.
Medicaments containing a β-hairpin mimetis of general formula I , a solvate or a salt thereof are likewise objects of the present invention, as is a process for the manufacture of such medicaments which comprises bringing one or more of said compounds and , where desired, one or more additional therapeutically valuable substances into a galenical dosage form.
For the control or prevention of a given illness amenable to treatment with protease inhibitors, the β-haiφin mimetics of the invention can be administered singly, as mixtures of several β-haiφin mimetics, in combination with other inhibitors of protease enzymes or in combination with other pharmaceutically active agents. The β-haiφin mimetics of the invention can be administered per se or as pharmaceutical compositions. The dosage of the active substance, that is, a compound of formula I, can vary within wide limits and will, of course, be fitted to the individual requirements in each particular case. In general, in the case of oral or parenteral, for example, intravenous or subcutaneous, administration a dosage of about 0.1-29mg/kg, preferably of about 0.5-5mg/kg, per day should be appropriate for adults, although the upper limit just given can also be increased or lowered, when this is shown to be indicated.
Pharmaceutical compositions comprising β-haiφin peptidomimetics of the invention may be manufactured by means of conventional mixing, dissolving, granulating, coated tablet-making, levigating, emulsifying, encapsulating, entrapping or lyophilizing processes. Pharmaceutical compositions may be formulated in conventional manner using one or more physiologically acceptable carriers, diluents, excipients or auxilliaries which facilitate processing of the active β- haiφin peptidomimetics into preparations which can be used pharmaceutically. Proper formulation depends upon the method of administration chosen. Systemic formulations include those designed for administration by injection, e.g. subcutaneous, intravenous, intramuscular, intrathecal or intraperitoneal injection, as well as those designed for transdermal, transmucosal, oral or pulmonary administration.
For injections, the β-hairpin peptidomimetics of the invention may be formulated in aqueous solutions, preferably in physiologically compatible buffers such as Hink's solution, Ringer's solution, or physiological saline buffer. The solution may contain formulatory agents such as suspending, stabilizing and/or dispersing agents. Alternatively, the β-haiφin peptidomimetics of the invention may be in powder form for combination with a suitable vehicle, e.g., sterile pyrogen-free water, before use.
For transmucosal administration, penetrants appropriate to the barrier to be permeated are used in the formulation as known in the art.
For oral administration, the compounds can be readily formulated by combining the active β- haiφin peptidomimetics of the invention with pharmaceutically acceptable carriers well known in the art. Such carriers enable the β-haiφin peptidomimetics of the invention to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions etc., for oral ingestion of a patient to be treated. For oral formulations such as, for example, powders, capsules and tablets, suitable excipients include fillers such as sugars, such as lactose, sucrose, mannitol and sorbitol; cellulose preparations such as maize starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methyl cellulose, hydroxypropylmethyl cellulose, sodium carboxymethylcellulose, and/or polyvinylpyrrolidone (PVP); granulating agents; and binding agents. If desired, desintegrating agents maybe added, such as cross-linked polyvinylpyrrolidones, agar, or alginic acid or a salt thereof, such as sodium alginate. If desired, solid dosage forms may be sugar-coated or enteric-coated using standard techniques.
For oral liquid preparations such as, for example, suspensions, elixirs and solutions, suitable carriers, excipients or diluents include water, glycols, oils, alcohols, etc. In addition, flavoring agents, preservatives, coloring agents and the like may be added.
For buccal administration, the composition may take the form of tablets, lozenges, etc. formulated as usual. For administration by inhalation, the β-haiφin pcμuuυimmetics of the invention are conveniently delivered in form of an aeorosol spray from pressurized packs or a nebulizer, with the use of a suitable propellant, e.g. dichlorodifluoromethane, trichlorofluromethane, carbon dioxide or another suitable gas. In the case of a pressurized aerosol the dose unit may be determined by providing a valve to deliver a metered amount. Capsules and cartridges of e.g. gelatin for use in an inhaler or insufflator may be formulated containing a powder mix of the β-haiφin peptidomimetics of the invention and a suitable powder base such as lactose or starch.
The compounds may also be formulated in rectal or vaginal compositions such as suppositories together with appropriate suppository bases such as cocoa butter or other glycerides.
In addition to the formulation described previously, the β-haiφin peptidomimetics of the invention may also be formulated as depot preparations. Such long acting formulations may be administered by implantation (e.g. subcutaneously or intramuscularly) or by intramuscular injection. For the manufacture of such depot preparations the β-haiφin peptidomimetics of the invention may be formulated with suitable polymeric or hydrophobic materials (e.g. as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble salts.
In addition, other pharmaceutical delivery systems may be employed such as liposomes and emulsions well known in the art. Certain organic solvents such as dimethylsulfoxide also may be employed. Additionally, the β-haiφin peptidomimetics of the invention may be delivered using a sustained-release system, such as semipermeable matrices of solid polymers containing the therapeutic agent. Various sustained-release materials have been established and are well known by those skilled in the art. Sustained-release capsules may, depending on their chemical nature, release the compounds for a few weeks up to over 100 days. Depending on the chemical nature and the biological stability of the therapeutic agent, additional strategies for protein stabilization may be employed.
As the β-haiφin pepdidomimetics of the invention may contain charged residues, they may be included in any of the above-described formulations as free bases or as pharmaceutically acceptable salts. Pharmaceutically acceptable salts tend to be more soluble in aqueous and other protic solvents than are the corresponding free base forms. The β-haiφin peptidomimetics of the invention , or compositions thereof, will generally be used in an amount effective to achieve the intended puφose. It is to be understood that the amount used will depend on a particular application.
For use to treat or prevent diseases amenable to treatment with protease inhibitors, the β-haiφin pepidomimetics of the invention, or compositions thereof, are administered or applied in a therapeutically effective amount. By therapeutically effective amount is meant an amount effective in ameliorating the symptoms of, or ameliorate, treat or prevent diseases related to protease activity. Determination of a therapeutically effective amount is well within the capacities of those skilled in the art, especially in view of the detailed disclosure provided herein.
Initial dosages can also be determined from in vivo data, e.g. animal models, using techniques that are well known in the art. One having ordinary skills in the art could readily optimize administration to humans based on animal data.
Dosage amount and interval may be adjusted individually to provide plasma levels of the β- haiφin peptidomimetics of the invention which are sufficient to maintain the therapeutic effect. Usual patient dosages for administration by injection range from about 0.1-5mg/kg day, preferably from about 0.5 to 1 mg/kg/day. Therapeutically effective serum levels may be achieved by administering multiple doses each day.
The amount of β-haiφin peptidomimetics administered will, of course, be dependent on the subject being treated, on the subject's weight, the severity of the affliction, the manner of administration and the judgement of the prescribing physician.
Normally, a therapeutically effective dose of the β-haiφin peptidomimetics described herein will provide therapeutic benefit without causing substantial toxicity.
Toxicity of the β-haiφin peptidomimetics of the invention herein can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., by determining the LD50 (the dose lethal to 50% of the population) or the LDι00(the dose lethal to 100% of the population). The dose ratio between toxic and therapeutic effect is the therapeutic index. Compounds which exhibit high therapeutic indices are preferred. The data obtained from these cell culture assays and animal studies can be used in formulating a dosage range that is not toxic for use in humans. The dosage of the β-haiφin peptidomimetics of the invention lies preferably within a range of circulating concentrations that include the effective dose with little or no toxicity. The dosage may vary within the range depending upon the dosage form employed and the route of administration utilized. The exact formulation, route of administration and dose can be chosen by the individual physician in view of the patient's condition (see, e.g. Fingl et al. 1975, In : The Pharmacological Basis of Therapeutics, Ch.l, p.l).
Compounds of formula I containing a free thiol group, i.e. compounds containing as R2-R6, R8- R10, R12, R13, R15-R19, R2,-R29, R33 or R64 a residue of the formula -(CH2)m(CHR61)sSR56 in which R56 is H, can be immobilized on gold-coated wawers, and interactions with ligands can be determined by means of the so-called surface plasmon resonance (SPR) biosensor analysis (cf. M. Fivash, E.M. Towler and R.J. Fisher, Curr. Opin. in Biotechnol. 1998. 9, 97-101; and R.L. Rich and D.G. Myszka, Curr. Opin. in Biotechnol. 2000. 11, 54-61).
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:
HBTU : 1-benzotriazol-l-yl-tetramethylurounium hexafluorophosphate 5 (Knorr et al. Tetrahedron Lett. 1989, 30, 1927-1930) HOBt : 1-hydroxybenzotriazole DIEA : diisopropylethylamine HO AT: 7-aza- 1-hydroxybenzotriazole
HATU: O-(7-aza-benzotriazole-l -yl)-N,N,N',N'-tetramethyluronoium hexafluorophosphate 0 Caφino et al. Tetrahedron Lett. 1994, 35, 2279-2281)
Examples
1. Peptide synthesis 5
Coupling of the first protected amino acid residue
0.5 g of 2-chlorotritylchloride resin (Barlos et al. Tetrahedron Lett. 1989, 30, 3943-3946) (0.83 mMol/g, 0.415 mmol) was filled into a dried flask. The resin was suspended in CH2C12 (2.5 ml) 0 and allowed to swell at room temperature under constant stirring. The resin was treated with
0.415 mMol (leq) of the first suitably protected amino acid residue (see below) and 284 μl (4eq) of diisopropylethylamine (DIEA) in CH2C12 (2.5 ml), the mixture was shaken at 25°C for 15 minutes, poured onto the pre-swollen resin and stirred at 25°C for 18 hours. The resin colour changed to puφle and the solution remained yellowish. The resin was washed extensively
!5 (CH2C12 /MeOH/DIEA : 17/2/1 ; CH2C12, DMF; CH2C12; Et2O, 3 times each) and dried under vacuum for 6 hours. Loading was typically 0.6-0.7 mMol/g
The following preloaded resins were prepared: Fmoc-Ser(tBu)O-chlorotritylresin and Fmoc-AlaO-chlorotritylresin. 1.1. Procedure 1
The synthesis was carried out using a Syro-peptide synthesizer (Multisyntech) using 24 to 96 reaction vessels. In each vessel was placed 60 mg (weight of the resin before loading) of the above resin. The following reaction cycles were programmed and carried out:
Step Reagent Time
1 CH2C12, wash and swell (manual) 3 x 1 min.
2 DMF, wash and swell 1 x 5 min
3 40 % piperidine/DMF 1 5 min.
4 DMF, wash 5 2 min.
5 5 equiv. Fmoc amino acid/DMF
+ 5 eq. HBTU
+ 5 eq. HOBt
+ 5 eq. DIEA 1 x 120 min.
6 DMF, wash 4 x 2 min.
7 CH2C12, wash (at the end of the synthesis) 3 x 2 min.
Steps 3 to 6 are repeated to add each amino-acid.
Cleavage of the fully protected peptide fragment
After completion of the synthesis, the resin was suspended in 1 ml (0.39 mMol) of 1% TFA in CH2C12 (v/v) for 3 minutes, filtered and the filtrate was neutralized with lml (1.17 mMol, 3eq.) of 20% DIEA in CH2C12 (v/v). This procedure was repeated twice to ensure completion of the cleavage. The filtrate was evaporated to dryness and the product was fully deprotected to be analyzed by reverse phase-HPLC (column .) to monitor the efficiency of the linear peptide synthesis.
Cyclization of the linear peptide
100 mg of the fully protected linear peptide were dissolved in DMF (9 ml, cone. 10 mg/ml). Then 41.8 mg (0.110 mMol, 3 eq.) of HATU, 14.9 mg (0.110 mMol, 3 eq) of HOAt and 1 ml (0.584 mMol) of 10% DIEA in DMF (v/v) were added and the mixture vortexed at 20°C for 16 hours and subsequently concentrated under high vacuum. The residue was partitioned between CH2C12 and H20/CH3CN (90/10: v/v). The CH2C12 phase was evaporated to yield the fully protected cyclic peptide.
Deprotection and purification of the cyclic peptide:
The cyclic peptide obtained was dissolved in 1 ml of the cleavage mixture containing 95% trifluoroacetic acid (TFA), 2.5% water and 2.5% triisopropylsilane (TIS). The mixture was left to stand at 20°C for 2.5 hours and then concentrated under vacuum. The residue was dissolved in a solution of H2O/acetic acid (75/25: v/v) and the mixture extracted with di-isopropylether.
The water phase was dried under vacuum and then the product purified by preparative reverse phase HPLC.
After lyophilisation products were obtained as a white powder and analysed by ESI-MS. The analytical data comprising HPLC retension times and ESI-MS are shown in table 1.
Examples ex.1-11 (n=7) are shown in table 1. The peptides were synthesized starting with the amino acid at position P3 which was coupled to the resin. Starting resins were Fmoc-Ser(tBu)0- chlorotritylresin and Fmoc-AlaO-chlorotrityl resin, which were prepared as described above. The linear peptides were synthesized on solid support according to procedure 1 in the following sequence: P4-P6-P7 -DPro-Pro-Pl-P2-P3 -resin, cleaved, cyclized, deprotected and purified as indicated.
Examples ex.12 and 13 (n=7) are also shown in table 1. The peptides were synthesized starting with the amino acid at position P3 which was grafted to the resin. Starting resin was Fmoc- Ser(tBu)O-chlorotritylresin, which was prepared as described above. The linear peptides were synthesized on solid support according to procedure 1 in the following sequence: P4-P5-P6-P7- DPro-(A8'-l)-Pl-P2-P3-resin (ex 12) and, respectively, P4-P5-P6-P7-DPro-(A8"-l)-Pl-P2-P3- resin (ex. 13), cleaved, cyclized, deprotected and purified as indicated.
Example ex.14 (n=7) is shown in table 1, too. The peptide was synthesized starting with the amino acid at position P3 which was grafted to the resin. Starting resin was Fmoc-Ser(tBu)0- chlorotritylresin, which was prepared as described above. The linear peptide was synthesized on solid support according to procedure 1 in the following sequence: P4-P5-P6-P7-(cl-l)-Pl-P2- P3 -resin, cleaved, cyclized, deprotected and purified as indicated. Building block (cl-1) is described below.
Example ex.15 (n=l 1) is likewise shown in table 1. The peptide was synthesized starting with the amino acid at position P5 which was coupled to the resin. Starting resin was Fmoc-Ser(tBu)O- chlorotritylresin, which was prepared as described above. The linear peptide was synthesized on solid support according to procedure 3 (see below)in the following sequence: P6-P7-P8-P9-P10- Pl l-DPro-Pro-Pl-P2-P3-P4-P5-resin, cleaved, cyclized, oxidized, deprotected and purified as indicated.
Tablet. Examples ex. 1-15
Figure imgf000126_0001
H20 + 0.1% CF3COOH over 15 min. and 50-100% MeCN/H2O + 0.1% CF3COOH over 4 min. b) Vydac C-18-column; gradient: 5-60% MeCN/H20 + 0.1% CF3COOH over 20 min. c) Nd: not determined
1.2. Procedure 2
Example ex.13 was also synthesized using procedure 2.
The peptide synthesis is carried out by solid phase method using standard Fmoc 5 chemistry on a peptide synthesizer- ABI 433A.
The first amino acid, Fmoc-Ser(tBu)-OH (0.68g, 1.2 equiv.) is coupled to the 2- chlorotritylchloride resin (Barlos et al. Tetrahedron Lett. 1989, 30, 3943-3946) ( 2g, 0.83 mmol/g) in presence of DIEA (1.12mL, 4 equiv.) in CH2C12 (20 mL), with swirling for 3 hr at room temperature. The resin is then washed with 3 x CH2C12 /MeOH/DIEA(l 7:2:1), 3 x CH2C12, 0 2 x DMF, 2 x CH2C12, 2 x MeOH. Finally, the resin is dried under vacuum and the substitution level was measured by weight increase (-0.6 mmol/g)
In case it is desired to use different acylating agents, the resin with the synthesized linear peptide, He-Lys(Boc)-Pro-Pro-Ile-ι:)Pro-212-Thr(tBu)-Lys(Boc)-Ser(tBu)-resin, is preferably divided into equal parts and placed in different reaction vessels in order to carry out the acylation reaction in
15 parallel format. The coupling and deprotection reactions in the following steps are monitored by Kaiser's test (Kaiser et al. Anal. Biochemistry 1970, 43, 595).
Removal of Alloc protecting group:
!0 To the linear peptide resin (100 mg per reaction vessel) is added Pd(PPh3) (15mg, 0.5 equiv.) under argon followed by dry CH2C12 (10 mL) and phenylsilane (17μL, 30 equiv.). The reaction mixture is left for 1 hour in the dark, filtered, and the resin is washed twice with CH2C12, DMF, and CH2C12.
5 Acylation of 4-amino-proline group
To the resin is added the corresponding acylating agent (usually a carboxyxlic acid (R64 COOH, 3 equiv.), HBTU (22.3mg, 4 equiv.), HOBt (8mg, 4 equiv.) and DIEA (125μL, 6 equiv.) in DMF (2mL) for 1.5-2 hrs at room temperature. The resin is filtered, washed with 2 x DMF, 3 x CH2C12, 0 2 x DMF.
Deprotection oflΨ'-Fmoc group: Deprotection of the Fmoc-group is achieved by treating the resin with 20% piperidine in DMF for 20 min. The resin is subsequently filtered and washed three times with DMF, and CH2C12, and twice with DMF, and CH2C12.
Cleavage of peptide from the resin:
The linear side-chain protected peptide is cleaved from the resin using AcOH: TFE: CH2C12 (2:2:6, v/v/v) for 2 hrs at room temperature. The resin is filtered off and washed twice with a mixture of AcOH:TFE:DCM and once with CH2C12. The filtrate is subsequently diluted with hexane (14 times by vol.) and concentrated. Evaporation is repeated twice with hexane to remove traces of AcOH. The residue is dried under vacuum. Yield of the linear protected peptide is generally about 40-50 mg.
Cyclization of the linear protected peptide:
Cyclization is carried out in DMF at a concentration of 5 mg/mL using HATU (13.12 mg, 3 equiv.), HOAT (4.7 mg, 3 equiv.), DIEA (153μL, 6 equiv.). The reaction mixture is stirred for 16 hrs at room temperature and the completion of reaction is monitored by HPLC. After the evaporation of DMF, CH3CN/H2O (90/10, v/v) is added to the residue and extracted with DCM. The organic layer is washed once with water and evaporated to dryness. Dried under vacuum.
Cleavage of side chain protecting groups:
The final deprotection of the side-chain protecting groups is carried out by treating the peptide with TFA:triisopropylsilane:H2O (95:2.5:2.5, v/v/v) at room temperature for 3 hrs. TFA is then evaporated and the residue triturated with cold ether.
Purification:
The crude peptides thus obtained are analyzed and purified by HPLC on a VYDAC Cl 8 preparative column using 5-60% CH3CN H2O+0.1%TFA in 30 min as gradient and a flow rate of lOml/min. The purity of the final peptide is checked by analytical HPLC and by ESI-MS. 1.3. Procedure 3
Procedure 3 describes the synthesis of β-haiφin mimetics having disulfide β-strand linkages.
n=ll: :The peptides are synthesized according to procedure 1 starting with the amino acid at position P5, coupled to the resin. The linear peptides are synthesized on solid support according to procedure 1 in the following sequence: P6-P7-P8-P9-P10-Pll-DPro-Pro-Pl-P2-P3-P4-P5- resin, where at positions P2 and P10 Fmoc-Cys(Acm)OH or Fmoc-Cys(Tr)OH are incoφorated. The linear peptides are cleaved and cyclized as described in procedure 1.
When Cys(Acm) was incoφorated as protected building block, the cyclized side chain protected β-haiφin mimetics are dissolved in methanol (0.5ml) to which is added dropwise a solution of iodine in methanol (IN, 1.5equiv.) at room temperature. The reaction mixture is stirred for 4 hours at room temperature and the solvent evaporated. The crude product is subsequently deprotected and purified as described in procedure 1.
When Cys(Tr) was incoφorated as protected cysteine building block, the cyclic fully protected protected β-haiφin mimetics are treated with a mixture containing trifluoro acetic acid/thioanisol/phenol/H2O/ethane-dithiol/triisopropylsilane (82.5:5:5:2.5:2.5:2.5) at room temperature for 2 hours. The reduced peptide is subjected to air oxidation by stirring for 30 minutes in ammonium acetate buffer and purified as in procedure 1.
2. Synthesis of the templates
2.1. The syntheses of (2S,4S)-4-[(allyloxy)carbonylamino]-l-[(9H-fluoren-9- yl)methoxycarbonyl]-proline (212) and (2S,4R)-4-[(allyloxy)carbonylamino]-l-[(9H-fluoren-9- 5 yl)methoxy-carbonyl]proline (217) are shown in Schemes 42 and 43.
Scheme 42
OOMe
Figure imgf000130_0001
211 212 i: SOCI2, MeOH; ii: Boc20, DMAP, Et3N; iii: pN02C6H4S02CI, Et3N; iv: NaN3, DMF; v: SnCI2,dioxane/H20; vi: CICOOCH2CH=CH2, aq.NaHCO3, dioxane: vii: LiOH, MeOH, H2O; viii: TFA, CH2CI2; ix: Fmoc-OSu, DIEA
I0 (2S,4S)-4-[(Allyloxy)carbonylamino]-l-[( H-fluoren-9-yl)methoxycarbonyl]- proline (212)
i,ii: To a solution of (2S,4R)-4-hydroxyproline (30 g, 0.18 mol) in abs. methanol (300 ml) at 0 °C thionyl chloride (38 ml, 2.5 eq, 0.45 mol) was added dropwise. The solution was heated to
5 reflux and stirred for 3 h under nitrogen. Then the solution was concentrated by rotary evaporation and the ester precipitated by adding diethylether. After filtration the white solid was washed with diethylether, then dried at ΗV: (2S,4R)-4-hydroxyproline-methylester-hydrochloride as a white solid (29.9 g, 90 %). %). TLC (CΗ2Cl2/Me0Η/water 70:28:2): Rf 0.82. [CC]D = -24.5 (c = 1.01, MeOH). IR (KBr): 3378s (br.), 2950m, 2863w, 1745s, 1700s, 1590m, 1450s, 1415-?,
0 1360s, 1215s, 1185s, 1080m, 700m. Η-NMR (300MHz, MeOH-d4) 4.66-4.55 (m, 2H, H-C(4), H- C(2)); 3.85 (s, 3H, H3C-O); 3.45 (dd, J= 12.2, 3.8, IH, H-C(5)); 3.37-3.25 (m, IH, H-C(5)); 2.44- 2.34 (m, IH, H-C(3)), 2.27-2.12 (m, IH, H-C(3)). I3C-NMR (75MHz, MeOH-d4): 170.8 (s, COOMe); 70.8 (d, C(4)); 59.6 (d, C(2)); 55.2 (t, C(5)); 54.2 (q, Me); 38.7 (t, C(3)). CI-MS (NH3): 146.1 ([ -C1]+).
(30 g, 0.17 mmol) of the above intermediate was dissolved in CH2C12 (300 ml), cooled to 0 °C and triethylamine (45 ml, 1.5 eq, 0.25 mol) was added dropwise. Then di-tert.-butyldicarbonate (54.0 g, 1.5 eq, 0.25 mmol) in CH2C12 (15 ml) and 4-NN-dimethylaminopyridine (2.50 g, 0.1 eq, 17 mmol) was added and the solution stirred at room temperature overnight. Then the solution was washed with IN aq. citric acid solution, sat. aq. NaHC03 solution, dried (Na2S04), evaporated and the residue dried at high vaccum: (2S,4R)-4-Hydroxy-l-[(tert- butoxy)carbonyl]proline-methylester (209) as a white solid (39.6 g, 78 %). TLC (CH2Cl2 MeOH 9:1): R_ 0.55. [α]* = -55.9 (c = 0.983, CHC13). IR (KBr): 3615w, 3440w (br.), 2980m, 2950m, 2880m, 1750s, 1705s, 1680s, 1480m, 1410s, 1370s, 1340m, 1200s, 1160s, 1130m, 1090m, 1055w, 960w, 915w, 895w, 855m, 715m. Η-NMR (300MHz, CDC13): 4.47-4.37 (m, 2H, H-C(4), H-C(2)); 3.73 (s, 3H, H3C-O)); 3.62 (dd, J= 11.8, 4.1, IH, H-C(5)); 3.54-3.44 (m, IH, H-C(5)); 2.32-2.25 ( , IH, H-C(3)); 2.10-2.03 ( , IH, H-C(3)); 1.46+1.41 (2s, 9H, tBu). 13C-NMR (75 MHz, CDC13): 173.6 (s, COOMe); 154.3+153.9 (2s, COOtBu); 80.3 (s, C-tBu); 70.0+69.3 (2d, C(4)); 57.9+57.4 (2d, C(2)); 54.6 (t, C(5)); 51.9 (q, Me); 39.0+38.4 (2t, C(3)); 28.1+27.6 (2q, tBu). CI-MS: 246.2 ([ +H]+); 190.1 ([ -tBu+H]+); 146.1 ([ -BOC+H]+). iii, iv: (39 g, 0.16 mol) of 209 was dissolved in CH2C12 (300 ml) followed by addition of 4- nitrobenzenesulfonyl chloride (46 g, 1.3 eq, 0.21 mol) and Et3N (33 ml, 1.5 eq, 0.24 mol) at 0 °C. Then the solution was stirred overnight and brought gradually to room temperature, washed with IN hydrochloric acid, sat. aq. NaHCO3 solution and dried (Na2SO ). The solvents were evaporated and the crude product was purified by filtration on silica gel with (2:1) hexane/ AcOEt. The product was crystallized from hexane/AcOEt: (2S,4S)-4-[(p-nitrobenzyl)sulfonyloxy]-l- [(tert-butoxy)carbonyl]proline methylester as white crystals (46.4 g, 65 %). TLC (hexane/AcOEt 1:1): Rf 0.78. M.p.: 93-95 °C. [α] g = -32.3 ° (c = 0.907, CHC13). IR (KBr): 31 lOw, 3071ιv, 2971w, 1745s, 1696s, 1609s, 1532s, 1414s, 1365s, 1348m, 1289m, 1190m, 1173m, 1122w, 1097w, 1043w, 954 , 912w, 755w, 578w. 'H-NMR (600MHz, CDC13): 8.42-8.34 (m, 2H, H- C(Nos)); 8.11-8.04 (m, 2H, H-C(Nos)); 5.14 (s, IH, H-C(4)); 4.39-4.28 (m, IH, H-C(2)); 3.70- 3.56 (m, 5H, H2-C(5), H3C-O); 2.58-2.38 ( , IH, H-C(3)); 2.25-2.11 (m, IH, H-C(3)); 1.37+1.33 (2s, 9H, tBu). 13C-NMR (150 MHz, CDC13): 172.4+172.2 (2s, COOMe); 153.6+153.0 (2s,
COOtBu); 150.8+142.0 (2s, C(Nos)); 129.0+124.6 (2d, C(Nos)); 80.4 (s, C-tBu); 80.8+79.9 (2d, C(4)); 57.1+56.9 (2d, C(2)); 52.2+51.7 (2t, C(5)); 52.3 (q, Me); 37.1+35.9 (2t, C(3)); 28.0 (q, tBu). ESI-MS (MeOH + Nai): 453.0 ([ +Na]+). (38 g, 88 mmol) of the above intermediate was dissolved in DMF (450 ml) then heated to 40 °C when sodium azide (34 g, 6 eq, 0.53 mol) was added and the solution stirred overnight. DMF was evaporated and the solid suspended in diethylether. The suspension was washed with water and dried (Na2S04). The solvent was evaporated and the product dried at high vacuum: (2S,4S)-4- Azido-l-[(tert-butoxy)carbonyl]proline methylester (210) yellow oil (21.1 g, 88 %). [α] D = -36.9 (c = 0.965, CHC13). !H-NMR (600MHz, CDC13): 4.46-4.25 (2m, IH, H-C(2)); 4.20-4.10 (m, IH, H-C(4)); 3.80-3.65 (m, 4H, H-C(5), H3C-O); 3.53-3.41 (m, IH, H-C(5)); 2.54-2.39 (m, IH, H- C(3)); 2.21-2.12 (m, IH, H-C(3)); 1.47+1.41 (2s, 9H, tBu). 13C-NMR (150 MHz, CDC13): 172.2+171.9 (2s, COOMe); 153.9+153.4 (2s, COOtBu); 80.5 (s, C-tBu); 59.2+58.2 (2d, C(4)); 57.7+57.3 (2d, C(2)); 52.4+52.2 (2q, Me); 51.2+50.7 (2t, C(5)); 36.0+35.0 (2t, C(3)); 28.3+28.2 (2q, tBu). EI-MS (70ev): 270.1 ([M]+); 22 '.1 ([ -C02+H]+); 169.1 ([ -BOC+H]+);. v, vz: (21.1 g, 78 mmol) of the above intermediate was dissolved in a (3:l)-mixture of dioxane/water (500 ml) and SnCl2 (59.2 g, 4 eq, 0.31 mol) was added at 0° and the solution stirred for 30 min. and graduallly brought to room temperature and stirred for another 5h. After adjusting the pH to 8 with solid NaHCO3, allyl chloroformate (41.5 ml, 5 eq, 0.39 mol) was added and the solution stirred at room temperature overnight. The reaction mixture was evaporated and extracted with AcOEt. The organic phase was washed with brine, dried (Na2SO4), the solvent evaporated and the product was dried at high vacuum: (2S,4S)-4- [(Allyloxy)carbonylamino]-l-[(tert-butoxy)carbonyl]proline methylester (211) as a clear thick oil (22.3 g, 87 %). [α] $ = -30.2 ° (c = 1.25, CHC13). Η-NMR (300MHz, CDC13): 5.98-5.77 ( , IH, H-C(β)(Alloc)); 5.32-5.12 (m, 2H, H2-C(γ)(Alloc); 4.59-4.46 (m, 2H, H2-C(α)(Alloc)); 4.40-4.16 ( , 2H, H-C(4), H-C(2)); 3.80-3.53 (m, 4H, H-C(5), H3C-O); 3.53-3.31 (m, IH, H-C(5)); 2.54- 2.17 (m, IH, H-C(3)); 1.98-1.84 (m, IH, H-C(3)); 1.41+1.37 (2s, 9H, tBu). ESI-MS (MeOH+ CH2C12): 351.2 ([/W+Na]+); 229.0 ([ -BOC+H]+). vii-ix : 22 g, 67 mmol) of 211 was dissolved in a (4:l)-mixtøe of methanol/water (100 ml) and LiOH (5 g, 2 eq, 134 mmol) was added at room temperature and the solution stirred for 3.5 h. The reaction mixture was evaporated and extracted with IN hydrochloric acid (100 ml) and AcOEt. The solvent was removed and the resulting solid dissolved in 1:1 TFA/ CH2C12 (200ml) and stirred for 2 h. The solvents were evaporated and the product dried at high vacuum: (2S,4S)-4- [(Allyloxy)carbonylamino]proline as a clear oil (21 g, 96 %) Η-ΝMR (600MHz, MeOH-d ): 5.98-5.85 (m, IH, H-C(β)(AUoc)); 5.30 (dd, J=17.1, 1.5 Hz, IH, H-C(γ)(Alloc)); 5.12 (d, J=10.7 Hz, IH, H-C(γ)(Alloc)); 4.54 (d, J=4.4 Hz, 2H, H2-C(α)(Alloc)); 4.44 (t, J=8.9 Hz, IH, H-C(2)); 4.36-4.27 ( , IH, H-C(4)); 3.58 (dd, J=12.2, 7.3 Hz, IH, H-C(5)); 3.34-3.32 ( , IH, H-C(5)); 2.73 (ddd, J=13.6, 8.7, 7.2 Hz, IH, H-C(3)); 2.23-2.15 (m, IH, H-C(3)). I3C-NMR (150 MHz, MeOH-d4): 171.3 (s, COOMe); 158.3 (s, COOAllyl); 134.1 (d, C(β)(Alloc)); 118.0 (t, C(γ)(Alloc)); 66.8 (t, C(α)(Alloc)); 59.7 (d, C(2)); 51.3 (d, C(4)); 51.1 (t, C(5)); 34.9 (t, C(3)). • ESI-MS (DCM+MeOH): 237.0 ([ +Na]+); 215.0 ([ +H]+). (15 g, 70 mmol) of the above intermediate and 9-fluorenylmethoxycarbonylsuccinimid (28 g, 1.2 eq, 84 mmol) were dissolved in DCM (700 ml) and DIEA (48 ml, 6 eq, 0.42 mol) was added and the solution stirred overnight at room temperature. The solvent was removed and the residue dissolved in AcOEt and washed with 17V hydrochloric acid and dried (Na2S0 ). After evaporation, the crude product was purified by filtration on silica gel with a gradient of (3:1) hexane/AcOEt to AcOEt. The solvent was evaporated and the residue crystallized from hexane at -20 °C. The product was dried at high vacuum: (2S,4S)-4-[(Allyloxy)carbonylamino]-l-[(PH- fluoren-9-yl)methoxycarbonyl proline (212) as a white solid (23.8 mg, 78 %) [CC] D = -27.0 (c = 1.1, CHC13). IR (KBr): 3321w (br.), 3066w, 2953u>, 1707s, 1530m, 1451s, 1422s, 1354m, 1250m, 1205m, 1173m, 1118m, 1033m, 977m, 936m, 759m, 739s, 621m, 597w, 571 , 545s. 'H-NMR (300MHz, MeOH-d4): 7.88-7.78 ( , 2H, H-C(4')(Fmoc)); 7.71-7.61 (m, 2H, H-C(l')(Fmoc)); 7.49-7.29 (m, 4H, H-C(3')(Fmoc), H-C(2')(Fmoc)); 6.08-5.68 ( , IH, H-C(β)(Alloc)); 5.41-5.17 (m, 2H, H2-C(γ)(Alloc); 4.58 (s, 2H, H2-C(α)(Alloc)); 4.74-4.17 ( , 5H, H2-C(10')(Fmoc), H- C(9')(Fmoc), H-C(4), H-C(2)); 3.94-3.73 (m, IH, H-C(5)); 3.41-3.26 (m, IH, H-C(5)); 2.74-2.54 (m, IH, H-C(3)); 2.12-1.92 ( , IH, H-C(3)). ESI-MS (DCM+MeOH): 459.3 ([ +Na]+); 437.3 ([ +H]4).
Scheme 43
OOMe
Figure imgf000134_0001
208 213 214
Figure imgf000134_0002
215 216 217 i: Ac20, AcOH; ii: SOCI2, MeOH; iii: Boc2O, DMAP, Et3N; vi: pN02C6H4SO2CI, Et3N; v: NaN3, DMF; vi: SnCI2,dioxane/H2O; vii: CICOOCH2CH=CH2, aq.NaHC03, dioxane: viii: LiOH, MeOH, H20; ix: TFA, CH2CI2; x: Fmoc-OSu, DIEA
2.2. (2R,4S)-4-[(Allyloxy)carbonylamino]-l -[(9H-fluoren-9-yl)methoxycarbonyl]- proline (217)
i: A solution of acetic anhydride (1.02 kg, 5.3eq, 10 mol) in glacial acetic acid (3 1) was heated to 50 °C and (2S,4R)-4-hydroxyproline (208) (247 g, 1.88 mol) was added in one portion. The solution was refluxed for 5.5 h., cooled to room temperature and the solvent was removed under reduced pressure giving a thick oil. The oil was then dissolved in 2N hydrochloric acid (3.5 1) and heated to reflux for 4 h and treated with charcoal and filtered through Celite. As the solution was evaporated, white needles formed, which were filtered. The product was dried at high vacuum: (2R,4R)-4-hydroxyproline-hydrochloride (213) white cryst. needles (220.9 g, 70 %). M.p.: 117 °C. [α] D = +19.3 ° (c = 1.04, water). IR (KBr): 3238s 3017s, 2569m, 1712s, 1584m, 1376s, 1332m, 1255s, 1204m, 1181w, 1091w, \066w, 994vμ, 725m, 499s. Η-NMR (600MΗz, MeOH-dt): 9.64 (s, IH, H-N); 8.89 (s, IH, H-N); 4.55-4.53 (m, IH, H-C(4)); 4.51 (dd, J= 10.4, 3.6 Hz, IH, H-C(2)); 3.44-3.35 (m, 2H, H2-C(5)); 2.54-2.48 (m, IH, H-C(3)); 2.40-2.34 (m, IH, H-C(3)). 13C-NMR (150MHz, MeOH-d4): 171.9 (s, COOH); 70.3 (d, C(4)); 59.6 (d, C(2)); 55.0 (t, C(5)); 38.5 (t, C(3)). EI-MS (NH3): 132.1 ([ -C1]+). The filtrate was further concentrated to give an additional 59.5 g (19 %). ii.iii: To a solution of 213 (30 g, 0.18 mol) in abs. methanol (550 ml) was added dropwise at 0 °C thionyl chloride (38 ml, 2.5 eq, 0.45 mol). The solution refluxed for 3 h under nitrogen atmosphere. The solution was evaporated and the ester hydrochloride precipitated by adding diethylether. After filtration the white solid was washed with diethylether and dried at high vacuum: (2R,4R)-4-hydroxyproline methylester -hydrochloride white solid (29 g, 89 %). [<X] D = +8.6 ° (c = 0.873, MeOH). IR (KBr): 3388s (br.), 2980s (br.), 1730s, 1634m, 1586s, 1384s, 1248s, 1095s, 1064s, 1030m, 877m. 'H-NMR (300MHz, MeOH-d4): 4.59-4.44 ( , 2H, H- C(4), H-C(2)); 3.81 (s, 3H, H3C-O); 3.37-3.31 (m, 2H, H2-C(5)); 2.50-2.37 (m, IH, H-C(3)), 2.37- 2.27 ( , IH, H-C(3)). 13C-NMR (75MHz, MeOH-d4): 170.9 (s, COOMe); 70.2 (d, C(4)); 59.8 (d, C(2)); 55.1 (t, C(5)); )); 54.1 (q, C(Me)); 38.4 (t, C(3)). EI-MS (NH3): 146.1 ([Λf-Clf).
(10 g, 55 mmol) of the above intermediate was dissolved in CH2C12 (100 ml), cooled to 0 °C and triethylamine (15.2 ml, 2 eq, 0.11 mol) was added dropwise. Then di-tert.-butyldicarbonate (18.0 g, 1.5 eq, 83 mmol) in CH2C12 (10 ml) and 4-NN-dimethylaminopyridine (0.67 g, 0.1 eq, 5 mmol) were added and the solution was stirred at RT overnight. The solution was washed with IMaq. citric acid solution and sat. aqueous NaHC03 solution, dried (Na2SO ), the solvents evaporated and dried at high vaccum: (2R,4R)-4-hydroxy-l-[(tert-butoxy)- carbonyl]prolinemethylester (214) as a white solid (13 g, 97 %). [α] o =+13.0 ° (c = 1.06, CHC13). IR (KBr): 3466s (br.), 2985s, 2930m, 1729s, 1679s, 1424s, 1283m, 1262m, 1122s, 1089s, 969m, 770m. 'H-NMR (300MHz, CDC13): 4.43-4.26 (m, 2H, H-C(4), H-C(2)); 3.80+3.79 (2s, 3H, H3C-O)); 3.76-3.47 ( , 2H, H2-C(5)); 2.44-2.24 (m, IH, H-C(3)); 2.16-2.03 (m, IH, H- C(3)); 1.47+1.43 (2s, 9H, tBu). ESI-MS: 268.1 ([ +Na]+). tv,v: 214 (12.2 g, 50 mmol) was dissolved in CH2C12 (130 ml), cooled to 0 °C and 4- nitrobenzenesulfonyl chloride (14.3 g, 1.3 eq, 65 mmol) and Et3N (10.3 ml, 1.5 eq, 75 mmol) were added. The reaction mixture was stirred overnight and gradually brought to room temperature. The solution was washed with INhydrochloric acid and saturated aqueous ΝaHC03 solution, dried (Na2S0 ), the solvents were evaporated and the crude product was purified by filtration on silica gel with (2:l)-mixture of hexane/AcOEt: 18 g (84 %). The product was then recrystallized from hexane/AcOEt: (2R,4R)-4-[(^-nitrobenzyl)sulfonyloxy]-l-[(tert- butoxy)carbonyl]proline-methylester as white crystals (13.7 g, 64 %). TLC (hexane/AcOEt 1:1): Rf 0.76. M.p.: 113-115 °C. [α] D = +21.6 ° (c = 0.924, CHC13). IR (KBr): 3112s (br.), 2981s, 2955s, 2882m, 1755s, 1683s, 1532s, 1413s, 1375s, 1348s, 1192s, 928s, 911s, 759m, 745s, 610s. 'H-NMR (600MHz, CDC13): 8.45-8.35 (m, 2H, H-C(Nos)); 8.15-8.06 (m, 2H, H-C(Nos)); 5.27- 5.16 ( , IH, H-C(4)); 4.53-4.32 (m, IH, H-C(2)); 3.75-3.60 (m, 5H, H2-C(5), H3C-O); 2.59-2.35 (m, 2H, H2-C(3)); 1.42+1.39 (2s, 9H, tBu). 13C-NMR (150 MHz, CDC13): 171.8 + 171.6 (s, COOMe); 153.8+153.4 (s, COOtBu); 151.0+142.6 (s, C(Nos)); 129.2+124.7 (d, C(Nos)); 81.0 (s, C-tBu); 80.8+79.7 (d, C(4)); 57.4+57.1 (d, C(2)); 52.6+52.5+52.3+51.8 (t, C(5), q, Me); 37.2+36.3 (t, C(3)); 28.5+28.3 (q, tBu). ESI-MS (DCM + MeOH + Nai): 453.2 ([Λf+Na] ). (13 g, 30 mmol) of the above intermediate was dissolved in DMF (200 ml), heated to 40 °C and sodium azide (14.3 g, 6 eq, 180 mmol) was added and the reaction mixture stirred over- night. The reaction mixture was evaporated and the residue suspended in diethylether. The suspension was filtered, the filtrate washed with water and the organic phase dried(Na2S0 ). The solvent was evaporated and the product dried at high vacuum: (2R,4S)-4-azido-l-[(tert- butoxy)carbonyl]prolinemethylester (215) as a yellow oil (8.15 g, 99 %). [α] D = +42.8 ° (c = 1.05, CHC13). 'H-NMR (300MHz, CDC13): 4.58-4.37 (m, IH, H-C(2)); 4.34-4.23 (m, IH, H- C(4)); 3.92-3.51 (m, 5H, H2-C(5), H3C-0); 2.52-2.33 (m, IH, H-C(3)); 2.33-2.20 (m, IH, H- C(3)); 1.56+1.51 (2s, 9H, tBu). CI-MS (NH3): 288.2 ([ +NHJ÷); 271.1 ([M+Ef). vi.vii: 215 (8 g, 30 mmol) was dissolved in a (3:l)-mixture of dioxane/water (400 ml), cooled to 0 °C and SnCl2 (22.4 g, 4 eq, 120 mmol) was added and the reaction mixture stirred for 30 min. at 0°, gradually warmed to room temperature and stirred for another 5h. After adjusting the pH of the solution to 8 with solid NaHC03, allyl chloroformate (15.7 ml, 5 eq, 150 mmol) was added. The reaction mixture was stirred overnight at room temperature, evaporated and extracted with AcOEt and the organic phase washed with brine. After drying the organic phase (Na2SO4), the solvent was evaporated and the product dried at high vacuum: (2R,4S)-4-
[(Allyloxy)carbonylamino]-l-[(tert-butoxy)carbonyl] proline-methylester as a clear thick oil (216) (8.7 g, 89 %). [a] r? = +41.9° (c = 0.928, CHC13). Η-NMR (300MHz, CDC13): 5.98-5.87 (m, IH, H-C(β)(AUoc)); 5.34-5.02 (m, 2H, H2-C(γ)(Alloc); 4.62-4.49 (TO, 2H, H2-C(a)(Alloc)); 4.41-4.23 (m, 2H, H-C(4), H-C(2)); 3.82-3.66 (m, 4H, H-C(5), H3C-0); 3.43-3.20 ( , IH, H- C(5)); 2.33-2.07 (m, 2H, H2-C(3)); 1.43+1.39 (2s, 9H, tBu). CI-MS (NH3): 329.1 ([M+Ef). vii-x: 216 (8.4 g, 25 mmol) was dissolved in (4:l)-mixture of methanol/water (100 ml) at room temperature, LiOH (2.2 g, 2 eq, 50 mmol) added and the solution stirred overnight. Methanol was evaporated and the residue poured onto INhydrochloric acid (100 ml) and extracted with AcOEt. The solvent was removed and the residue dissolved in (1 : l)-mixture of TFA/ CH2C12 (200ml) and stirred for 2h. The solvents were evaporated and the product dried at high vaccum: (2R,4R)- 4-[(Allyloxy)carbonylamino]proline as a clear oil (5.2 g, 96 %) Η-ΝMR (300MHz, MeOH-d,): 6.04-5.88 (m, IH, H2-C(β)(Alloc)); 5.38-5.19 ( , 2H, H2-C(γ)(Alloc); 4.64-4.54 ( , 3H, H2- C(α)(Alloc), H-C(4)); 4.39-4.22 (m, IH, H-C(2)); 3.71-3.60 (m, IH, H-C(5)); 3.45-3.32 ( , IH, H-C(5)); 2.51-2.41 (m, 2H, H2-C(3)). CI-MS (NH3): 215.1 ([M+Ef). (200 mg, 0.86 mmol) of the above intermediate and 9-fluorenylmethoxycarbonylsuccinimide (440 mg, 1.5 eq, 1.3 mmol) were dissolved in CH2C12 (10 ml) and DIEA (466 μl, 4 eq, 3.44 mmol) was added, and the solution stirred overnight at room temperature. The solvent was removed and the residue dissolved in AcOEt, washed with INhydrochloric acid dried (Νa2S04). After evaporation, the crude product was purified by filtration over silica gel with first a gradient of (3:1) hexane/AcOEt to AcOEt. The solvent was coevaporated with CH2C12 and the product dried at high vacuum: (2R,4S)-4-[(Allyloxy)carbonylamino]-l-[(PH-fluoren-9-yl)methoxy- carbonyl]- proline (217) white solid (90 mg, 33 %) [α] r? = +29.3 ° (c = 1.08, CHC13). IR (KBr): 3314s (br.), 3066s (br.), 2952s (br.), 1708s (br.), 1536m, 1424s, 1353s, 1126m, 1030m, 909m, 759m, 738s, 620m. Η-NMR (300MHz, CDC13): 8.74 (s, IH, H-N); 7J9-7.66 (m, 2H, H- C(4')(fmoc)); 7.62-7.49 (m, 2H, H-C(l')(fhιoc)); 7.44-7.22 (m, 4H, H-C(3')(frnoc), H- C(2')(fmoc)); 6.03-5.74 (TO, IH, H-C(β)(Alloc)); 5.41-5.07 (m, 2H, H2-C(γ)(Alloc); 4.74-4.17 ( , 7H, H2-C(10')(fmoc), H-C(9')(fhιoc), H-C(4), H-C(2), H2-C(α)(Alloc)); 3.91-3.76 (m, IH, H- C(5)); 3.48-3.25 (m, IH, H-C(5)); 2.45-2.08 (m, 2H, H2-C(3)). ESI-MS (MeOH): 437.3 ([M+Ef); ESI-MS (MeOH+Na): 459.1 ([ +Na]+).
2.3. Starting from derivatives 210 and 215 the key precursors 219a and 221a can be prepared according to Scheme 44.
RX n-hexyl (219a, 221a).
Figure imgf000138_0001
220a 221a
i: SnCI2, dioxane/H20; ii: R64COCi, diisopropylethylamine, CH2CI2; iii: LiOHxl H20, MeOH,
HzO; iv. TFA, CH2CI2; v: FmocOSu, Na2C03 aq., dioxane Scheme 44
i, ii: Typical procedures:
To a solution of 78 mmol of azides 210 and 215 in a (3:l)-mixture of dioxane/water (500 ml) was added at 0 °C SnCl2 (59.2 g, 4 eq, 0.31 mol) and the solution was stirred for 30 minutes. The reaction mixture was gradually warmed up to room temperature and stirred for another 5 hours. After adjusting the pH to 8 with solid NaHCO3, the reaction mixture was extracted with CH2C12, the organic fraction dried (MgSO4), evaporated and the residue dried under reduced pressure. The residue was dissolved in CH2C12 (300ml), cooled to 4° with an ice bath, followed by addition of DIEA (20.0ml, 117mmol) and a solution of the appropriate acid chloride R64 COCl (lOl.Ommol) in CH2C12 (50ml) at 4°C. The reaction mixture was stirred for 1 hour at 4° and for 18 hours at room temperature and extracted with HCl aq. (0.5N, 200ml) and CH2C12. The organic fraction was dried (MgSO ), evaporated and the residue chromatographed on SiO 2 with gradients of ethylacetate/hexane yielding 218a and 220a, which were converted into the final products 219a and 221a as described for the conversion of 216 into 217. The overall yields were 50-60%.
Templates (bio¬
synthesis of (2S,6S,8aR)-8a-{[(tert.-butyl)oxycarbonyl]met^^ fluoren-9-yl)methoxycarbonyl]amino}-pyrrolo[l,2-a]pyrazine-6-acetic acid (222):
Figure imgf000140_0001
To a stirred solution of 250mg (0.414mmol) of allyl {(2S,6S,8aR)-8a-[(tert.-butyl)oxycarbonyl] methyl}perhydro-5,8-dioxo-{[(9H-fluoren-9-yl)methoxycarbonyl]amino} -pyrrolo[l,2-a]pyrazin- 6-acetate in a degassed mixture of dichloromethane/methanol (9:1, 3ml) were added under argon 25mg (0.0216mmol) of tetrakis(triphenylphosphine)palladium, 0.05ml of acetic acid and 0.025ml of N-methylmorpholine. The reaction mixture was stirred for 48 hours at room temperature and poured onto water and dichloromethane. The organic phase was dried (MgS04), evaporated and the residue chromatographed on SiO2 with dichloromethane/methanol (9:1) to yield 180mg (77%) of(2S,6S,8aE)-8a-{[(tert.-butyl)oxycarbonyl]methyl}ρerhydro-5,8-dioxo-{[(9H-fluoren-9-yl)- methoxycarbonyljamino} -pyrrolo[l,2-a]pyrazine-6-acetic acid (222) as a white powder.
Η-NMR(300MHz, DMSO-d6): 8.30 (s, IH); 7.88 (d, J- 7.2, 2H); 7.67 (d, J=7.4, 2H); 7.62 (br.s, IH); 7.41 (t, J= 7.2, 2H); 7.33 (t, J=7.4, 2H); 4.35-4.2 (m, 5H); 3.55 (br.d, J= 6.3, 2H); 2.8-2.55 (m, 3H); 2.45-2.25 (m, 2H); 2.1-1.95 (m, IH); 1.35 (s, 9H); MS(ESD: 586.1 (M+Na) +, 564.1 (M+H)+.
Templates (cl):
Figure imgf000140_0002
(d-1)(168) Experimental procedure described in W. Bannwarth, A. Knierzinger, K. Mϋller, D. Obrecht, A. Trzeciak, EP 0 592 791 A2, 1993.
3. Biological methods 3.1. Enzymatic assays
The active enzyme concentrations were calculated using the equation described by Hendersen (P. J. F. Hendersen, Biochem. J. 1972, 127, 321-333). The inhibitor concentrations were determined by quantitative amino acid analysis. All assays were repeated in quadruplicate.
Determination of antitrypsin activity
A solution of trypsin was incubated for 5 minutes with increasing concentrations of inhibitor. The assays were carried out at 20°C in Tris-HCI buffer (pH 7.8, lOOmM) containing lOmM CaCl2. The substrate was N-α-benzoyl-L-arginine-4-nitroanilide (3.2 mM) and the initial reaction rate was monitored over 30 minutes at 405nm.
Determination of antielastase activity
As above, exept the substrate was N-succinyl-L-alanyl-L-alanyl-L-ρrolyl-L-phenylalanin-4- nitroanilide (1.6mM).
Apparent Ki values were calculated by fitting the initial rate data to the following equation, which assumes competitive tight-binding inhibition (J. F. Williams, J. F. Morrison, Methods Enzymol. 1979, 63, 437-467):
Figure imgf000142_0001
Determination of anti cathepsin G activity
10ml of a solution of cathepsin G (0.2 U/mL, corresponding to around 2μM, purchased from Calbiochem) were incubated for 15 minutes with increasing concentrations of inhibitor. The assays were carried out at 37°C in a total volume of 700μl of HEPES buffer (pH 7.5; 0.1 mol/L) containing 0.05mol/L CaCl2. Then, 70μl of substrate (N-succinyl-L-alanyl-L-alanyl-L-prolyl — L- phenylalanin-4-nitroanilide, 20mM in DMSO) were added. The release of of p-nitroanilide was monitored at 405 nm to determine the initial velocities of the reactions. Each measurement was reproduced three times (A. J. Barrett, Methods in Enzymology 1981, 80, 561-565).
3.2. Results
Figure imgf000143_0001
Nd: not determined

Claims

1. Compounds of the general formula
Figure imgf000144_0001
(I)
wherein
Figure imgf000144_0002
is a group of one of the formulae
Figure imgf000144_0003
(a1) (a2)
Figure imgf000144_0004
Figure imgf000145_0001
03) (14) 0)
Figure imgf000146_0001
(n) (o) (P)
wherein
Figure imgf000146_0002
is the residue of an L-α-amino acid with B being a residue of formula -NR20CH(R71)- or the enantiomer of one of the groups Al to A69 as defined hereinafter;
I
X
is a group of one of the formulae
Figure imgf000147_0001
A1 A2 A3 A4
Figure imgf000147_0002
A5 A6 A7 A8 A9
Figure imgf000147_0003
A10 A11 A12 A13 A14
Figure imgf000147_0004
Figure imgf000147_0005
A20 A21 A22 A23 A24
Figure imgf000148_0001
A38 A39 A40 A41 A42
Figure imgf000148_0002
Figure imgf000149_0001
A63 A64 A65 A66
Figure imgf000149_0002
A70 A71 A72 A73 A74
Figure imgf000149_0003
A75 A76 A77 A78 A79
Figure imgf000150_0001
A80 A81 A82 A83 A84
Figure imgf000150_0002
R1 is H; lower alkyl; or aryl-lower alkyl;
R2 is H; alkyl; alkenyl; -(CH2)m(CHR61)s0R5S; -(CH2)m(CHR61)sSR56;
(CH2)m(CHR61)sNR33R34; -(CH2)m(CHR61)sOCNR33R78; -
(CH2)m(CHR61)sNR20CONR33R78; -(CH2)0(CHR61)sCOOR57; -(CH2)0(CHR6,)sCONR5sR59; -(CH2)o(CHR6,)sPO(OR6t,)2;
-(CH2)0(CHRδ,)s S02R62; or -(CH2)0(CHR61)SC6H4R8; R3 is H; alkyl; alkenyl; -(CH2)m(CHR6,)sOR55; -(CH2)m(CHR61)sSR56; -
(CH2)m(CHR61)sNR33R34; -(CH2)0(CHR6I)sCOOR57 ; -(CH2)0(CHR6I)sCONR58R59 ; -(CH2)o(CHR61)sPO(OR60)2;
-(CH2)0(CHR61)S SO2R62; or -(CH2)0(CHR61)SC6H4R8; R4 is H; alkyl; alkenyl; -(CH2)m(CHR61)sOR55; -(CH2)m(CHR6I)sSR56; -
(CH2)m(CHR61)sNR33R34;
-(CH2)p(CHR61)sCOOR57; -(CH2)p(CHR6I)sCONR58R59; -(CH2)p(CHR6I)sPO(ORδ0)2; -(CH2)P(CHR61)S S02R62; or -(CH2)0(CHR6I)SC6H4R8;
R5 is alkyl; alkenyl; -(CH2)0(CHR61)sOR55; -(CH2)0(CHR61)SSR56; -(CH2)0(CHR61)SNR33R34; - (CH2)m(CHR61)sOCNR33R78; -(CH2)m(CHR6I)sNR20CONR33R78; -(CH2)o(CHR6I)sCOOR57; -(CH2)o(CHR61)sCONR58R59; -(CH2)o(CHR61)sPO(OR60)2; -(CH2)0(CHR61)S SO2R62; or -(CH2)0(CHR61)SC6H4R8; R6 is H; alkyl; alkenyl; -(CH2)0(CHR6I)sOR5S; -(CH2)0(CHR6I)SSR56; -(CH2)0(CHR61)SNR33R34; -(CH2)o(CHR6I)sCOOR57; -(CH2)o(CHR61)sCONRS8R59; -(CH2)o(CHR6I)sPO(OR60)2; -(CH2)0(CHR61)S SO2R62; or -(CH2)0(CHR61)SC6H4R8; R7 is alkyl; alkenyl; -(CH2)q(CHR61)sOR55; -(CH2)q(CHR61)sNR33R34; -(CH2)r(CHR61)sCOOR57; -(CH2)r(CHR61)sCONR58R59; -(CH2)r(CHR6I)sPO(OR60)2; -(CH2)r(CHR61)sS02R62; or -(CH2)r(CHR61)s Cg-tttR8;
R8 is H; Cl; F; CF3; NO2; lower alkyl; lower alkenyl; aryl; aryl-lower alkyl; - (CH2)o(CHR61)sOR55;
-(CH2)o(CHR61)sSR56; -(CH2)0(CHR61)NR33R3 ; -(CH2)0(CHR6I)sOCNR33R78; - (CH2)o(CHR6I)sNR20CONR33R78; -(CH2)o(CHR61)sCOOR57; -(CH2)o(CHR61)sCONR58R59; -(CH2)„(CHR61)SPO(OR60)2; -(CH2)0(CHR61)sS02R62; or -(CH2)0(CHR6J)SCOR64;
R9 is alkyl; alkenyl; -(CH2)0(CHR61)sOR55; -(CH2)0(CHR61)SSR56; -(CH2)0(CHR61)SNR33R34; -(CH2)0(CHR61)sCOOR57; -(CH2)o(CHR6I)sCONR58R59; -(CH2)o(CHR61)sPO(OR60)2; -(CH2)0(CHR61)S S02R62; or -(CH2)0(CHR61)SC6H4R8; R10 is alkyl; alkenyl; -(CH2)0(CHR61)sORS5; -(CH2)0(CHR6I)SSR56; -(CH2)0(CHR61)SNR33R34; -(CH2)o(CHR61)sCOOR57; -(CH2)0(CHR61)sCONR58R59; -(CH2)0(CHR61)SPO(OR60)2;
-(CH2)0(CHR61)S SO2R62; or -(CH2)0(CHR61)SC6H4R8; R11 is H; alkyl; alkenyl; -(CH2)m(CHR61)sOR55; -(CH2)m(CHR61)sSR56; - (CH2)m(CHR61)sNR33R34; -(CH2)m(CHR61)sOCNR33R78; - (CH2)m(CHR6I)sNR20CONR33R78; -(CH2)o(CHR61)sCOOR57; -(CH2)0(CHR61)SC0NR58R59; -(CH2)o(CHR61)sPO(OR60)2; -(CH2)„(CHR61)sS02R62; or
Figure imgf000152_0001
R12 is H; alkyl; alkenyl; -(CH2)m(CHR61)sOR55; -(CH2)m(CHR61)sSR56; -
(CH2)m(CHR61)sNR33R34; -(CH2)r(CHRδl)sCOOR57; -(CH2)r(CHR61)sCONR58R59; -(CH2)r(CHR61)sPO(OR60)2;
-(CH2)r(CHR6,)s S02R62; or -(CH2)r(CHRδI)sC6H4R8; R13 is alkyl; alkenyl; -(CH2)q(CHR61)sOR55; -(CH2)q(CHR61)sSR56; -(CH2)q(CHR61)sNR33R34;
-(CH2)q(CHR61)sCOOR57; -(CH2)q(CHR6I)sCONR58RS9; -(CH2)q(CHR6I)sPO(OR60)2;
-(CH2)q(CHR6I)s S02R62; or -(CH2)q(CHR6,)sC6H4R8; R14 is H; alkyl; alkenyl; -(CH2)m(CHR61)sOR55; -(CH2)m(CHRδI)sNR33R34; -
(CH2)q(CHR61)sCOOR57;
-(CH2)q(CHR61)sCONR58R59; -(CH2)q(CHR61)sPO(OR60)2; -(CH2)q(CHRδl)sSOR62; or
Figure imgf000152_0002
R15 is alkyl; alkenyl; -(CH2)0(CHRδl)sOR55; -(CH2)0(CHRδl)sSR56; -(CH2)0(CHRδI)sNR33R34; -(CH2)0(CHRδl)sCOOR57; -(CH2)0(CHRδl)sCONR58R59; -(CH2)0(CHRδl)sPO(OR60)2;
-(CH2)0(CHRδl)s SO2R62; or -(CH2)0(CHRδI)sC6H4R8; RIδ is alkyl; alkenyl; -(CH2)0(CHRδl)sOR55; -(CH2)0(CHRδl)sSRSδ; -(CH2)0(CHRδl)sNR33R34;
-(CH2)o(CHR61)sCOORS7; -(CH2)o(CHR61)sCONRS8R59; -(CH2)o(CHR6l)sPO(OR60)2;
-(CH2)0(CHRδI)s SO2R62; or -(CH2)0(CHRδI)sC6H4R8; R17 is alkyl; alkenyl; -(CH2)m(CHRδI)sOR5S; -(CH2)m(CHR61)sSR56; -(CH2)m(CHRδI)sNR33R34;
-(CH2)q(CHRδI)sCOOR57; -(CH2)q(CHRδI)sCONR58R59; -(CH2)q(CHR6I)sPO(OR60)2;
-(CH2)q(CHR61)s SO2R62; or -(CH2)q(CHR61)sC6H4R8; R18 is alkyl; alkenyl; -(CH2)p(CHRδI)sORss; -(CH2)p(CHRδl)sSR5δ; -(CH2)p(CHRδl)sNR33R34;
-(CH2)p(CHRδl)sCOOR57; -(CH2)p(CHRδl)sCONR58R59; -(CH2)p(CHRδl)sPO(ORδ0)2; -(CH2)p(CHRδI)s SO2R62; or -(CH2)0(CHRδI)sC6H4R8;
R19 is lower alkyl; -(CH2)p(CHR61)sOR55; -(CH2)P(CHR61)SSR56; -(CH2)P(CHR61)SNR33R34;
-(CH2)p(CHRδI)sCOOR57; -(CH2)p(CHRδl)sCONR58R59; -(CH2)p(CHR61)sPO(OR60)2;
-(CH2)P(CHR61)S S02R62; or -(CH2)0(CHRδl)sC6H4R8; or R18 and R19 taken together can form: -(CH2)2.6-; -(CH2)20(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR34(CH2)2-;
R20 is H; alkyl; alkenyl; or aryl-lower alkyl;
R21 is H; alkyl; alkenyl; -(CH2)0(CHRδl)sOR55; -(CH2)0(CHRδl)sSR5δ; -(CH2)0(CHRδl)sNR33R34;
-(CH2)0(CHRδl)sCOOR57; -(CH2)0(CHR61)sCONR58R59; -(CH2)0(CHR61)sPO(ORδ0)2;
-(CH2)0(CHRδl)s SO2R62; or -(CH2)0(CHRδl)sC6H4R8; R22 is H; alkyl; alkenyl; -(CH2)0(CHRδl)sOR55; -(CH2)0(CHR61)SSR55; -(CH2)0(CHR61)SNR33R34;
-(CH2)o(CHRδl)sCOOR57; -(CH2)o(CHRδl)sCONR58R59; -(CH2)o(CHRδ,)sPO(ORδ0)2;
-(CH2)0(CHR6I)S SO2R62; or -(CH2)0(CHR61)SC6H4R8; R23 is alkyl; alkenyl; -(CH2)0(CHR61)SOR55; -(CH2)0(CHRδl)sSR56; -(CH2)0(CHRδl)sNR33R34; -(CH2)0(CHR61)sCOOR57; -(CH2)0(CHR61)sCONR58R59; -(CH2)o(CHRδl)sPO(ORδ0)2;
-(CH2)0(CHRδl)s SO2R62; or -(CH2)0(CHRδl)sC6H4R8; R24 is alkyl; alkenyl; -(CH2)0(CHRδI)sOR55; -(CH2)0(CHRδl)sSR5δ; -(CH2)0(CHR61)SNR33R34;
-(CH2)o(CHRδl)sCOOR57; -(CH2)o(CHRδI)sCONR58R59; -(CH2)o(CHRδ,)sPO(ORδ0)2;
-(CH2)0(CHRδI)s SO2R62; or -(CH2)0(CHRδI)sC6H4R8; R25 is H; alkyl; alkenyl; -(CH2)m(CHR61)sOR55; -(CH2)m(CHRδl)sSR56; -
(CH2)ra(CHRδl)sNR33R34;
-(CH2)o(CHRδl)sCOOR57; -(CH2)o(CHRδ,)sCONR58R59; -(CH2)o(CHRδl)sPO(ORδ0)2;
-(CH2)o(CHRδl)sSO2R62; or -(CH2)0(CHR61)SC6H4R8; R2δ is H; alkyl; alkenyl; -(CH2)m(CHRδl)sOR55; -(CH2)m(CHRδl)sSRSδ; - (CH2)m(CHRδl)sNR33R34;
-(CH2)o(CHRδl)sCOOR57; -(CH2)o(CHR6I)sCONRS8R59; -(CH2)o(CHRδl)sPO(ORδ0)2;
-(CH2)o(CHRδl)s SO2R62; or -(CH2)0(CHRδI)sC6H4R8; or R25 and R26 taken together can form: -(CH2)2.6-; -(CH2)rO(CH2)r-; -(CH2)rS(CH2)r-; or
-(CH2)rNR34(CH2)r-; R27 is H; alkyl; alkenyl; -(CH2)0(CHRδI)sOR55; -(CH2)0(CHRδI)sSR5δ; -(CH2)0(CHR61)SNR33R34;
-(CH2)o(CHRδl)sCOOR57; -(CH2)o(CHR61)sCONR58R59; -(CH2)o(CHR61)sPO(OR60)2;
-(CH2)0(CHR61)s SO2R62; or -(CH2)0(CHRδl)sC6H4R8; R28 is alkyl; alkenyl; -(CH2)0(CHRδl)s-OR55; -(CH2)0(CHR6I)S SR5δ; -(CH2)0(CHRδl)s NR33R34;
-(CH2)o(CHRδl)s COOR57; -(CH2)0(CHRδI)s CONR58R59; -(CH2)0(CHR6I)S PO(OR60)2; -(CH2)0(CHRδl)s SO2R62; or -(CH2)0(CHR61)S G^8;
R29 is alkyl; alkenyl; -(CH2)0(CHRδl)sOR55; -(CH2)0(CHR61)SSR56; -(CH2)0(CHRδl)sNR33R34;
-(CH2)o(CHR61)sCOOR57; -(CH2)o(CHR61)sCONR58R59; -(CH2)o(CHR61)sPO(OR60)2;
-(CH2)o(CHRδI)s SO2R62; or -(CH2)0(CHRδl)sC6H4R8; R30 is H; alkyl; alkenyl; or aryl-lower alkyl; R31 is H; alkyl; alkenyl; -(CH2)P(CHR61)S0R55; -(CH2)P(CHRδl)sNR33R34; -
(CH2)o(CHRδI)sCOOR57;
-(CH2)0(CHRδl)sCONR58R59; -(CH2)0(CHR61)SPO(OR60)2; -(CH2)0(CHR61)sS02R62; or
-(CH2)o(CHRδl)s C6H4R8; R32 is H; lower alkyl; or aryl-lower alkyl; R33 is H; alkyl, alkenyl; -(CH2)m(CHRδl)s0R55; -(CH2)m(CHR6,)sSR56; -
(CH2)m(CHR61)sNR34R63;-(CH2)m(CHR61)sOCNR33R78; -(CH2)m(CHRδl)sNR20CONR33R78;
-(CH2)0(CHRδl)sCOR64;
-(CH2)0(CHR61)S-CONR58R59, -(CH2)o(CHR6I)sPO(OR60)2; -(CH2)0(CHRδl)s S02R62; or -(CH2)0(CHRδl)sC6H4R8;
R34 is H; lower alkyl; aryl, or aryl-lower alkyl;
R35 is H; alkyl; alkenyl; -(CH2)m(CHRδI)sOR55; -(CH2)m(CHRδl)sNR33R34; -
(CH2)p(CHRδl)sCOOR57;
-(CH2)p(CHRδ,)sCONR58R59; -(CH2)p(CHRδl)sPO(OR60)2; -(CH2)p(CHR61)sS02R62; or -(CH2)p(CHRδI)s C6H4R8;
R3δ is H, alkyl; alkenyl; -(CH2)0(CHRδl)s0R55; -(CH2)p(CHRδI)sNR33R34; -
(CH2)p(CHR61)sC00R57;
-(CH2)p(CHRδl)sCONR58R59; -(CH2)p(CHRδl)sPO(ORδ0)2; -(CH2)p(CHRδl)sS02R62; or
-(CH2)0(CHR61)S C6H4R8; R37 is H; F; Br; Cl; N02; CF3; lower alkyl; -(CH2)p(CHR61)s0R55; -(CH2)p(CHRδl)sNR33R34;
-(CH2)o(CHR61)sCOOR57; -(CH2)o(CHRδl)sCONR58R59; -(CH2)o(CHR6I)sPO(ORδ0)2;
-(CH2)0(CHRδl)sSO2R62; or -(CH2)0(CHR6I)S
Figure imgf000154_0001
R38 is H; F; Br; Cl; NO2; CF3; alkyl; alkenyl; -(CH2)p(CHRδl)sOR55; -(CH2)p(CHRδl)sNR33R34;
-(CH2)o(CHR61)sCOOR57; -(CH2)o(CHRδl)sCONR58R59; -(CH2)o(CHRδl)sPO(OR60)2; -(CH2)0(CHR61)sSO2R62; or -(CH2)0(CHRδl)sC6H4R8;
R39 is H; alkyl; alkenyl; or aryl-lower alkyl; R40 is H; alkyl; alkenyl; or aryl-lower alkyl; R41 is H; F; Br; Cl; N02; CF3; alkyl; alkenyl; -(CH2)p(CHR61)sOR5S; -(CH2)p(CHRδl)sNR33R34;
-(CH2)0(CHRδl)sCOOR57; -(CH2)o(CHRδI)sCONR58R59; -(CH2)o(CHRδl)sPO(OR60)2; -(CH2)0(CHR61)SSO2R62; or -(CH2)0(CHRδl)s Ce-H4R8;
R42 is H; F; Br; Cl; NO2; CF3; alkyl; alkenyl; -(CH2)p(CHR61)sOR55; -(CH2)p(CHRδl)sNR33R34;
-(CH2)o(CHRδl)sCOOR57; -(CH2)o(CHRδl)sCONRS8R59; -(CH2)o(CHRδl)sPO(ORδ0)2;
-(CH2)0(CHR61)sS02R62; or -(CH2)0(CHRδI)s C^; R43 is H; alkyl; alkenyl; -(CH2)m(CHRδl)sOR5S; -(CH2)m(CHRδl)sNR33R34; - (CH2)0(CHRδl)sCOOR57;
-(CH2)0(CHRδl)sCONR58R59; -(CH2)o(CHRδl)sPO(ORδ0)2; -(CH2)0(CHRδl)sS02R62; or
-(CH2)0(CHRδl)s CeH4R8; R44 is alkyl; alkenyl; -(CH2)r(CHR61)sOR55; -(CH2)r(CHRδl)sSR5δ; -(CH2)r(CHRδl)sNR33R34;
-(CH2)r(CHRδl)sCOOR57; -(CH2)r(CHRδl)sCONR58R59; -(CH2)r(CHRδl)sPO(ORδ0)2; -(CH2)r(CHRδl)s S02R62; or -(CH2)r(CHR6I)sC6H4R8; R45 is H; alkyl; alkenyl; -(CH2)0(CHRδl)sOR55; -(CH2)0(CHRδl)sSR5δ; -(CH2)0(CHRδl)sNR33R34;
-(CH2)o(CHR6I)sCOOR57; -(CH2)s(CHRδl)sCONR58R59; -(CH2)s(CHR61)sPO(ORδ0)2;
-(CH2)S(CHR6I)S SO2R62; or -(CH2)s(CHRδl)sC6H4R8; R46 is H; alkyl; alkenyl; or -(CH2)0(CHR61)PC6H4R8; R47 is H; alkyl; alkenyl; or -(CH2)0(CHRδI)sOR55; R48 is H; lower alkyl; lower alkenyl; or aryl-lower alkyl; R49 is H; alkyl; alkenyl; -(CHRδl)sCOOR57; (CHR6I)sCONRS8R59; (CHR61)SPO(OR60)2;
-(CHRδl)sSOR62; or -(CHRδI)sC6H4R8; R50 is H; lower alkyl; or aryl-lower alkyl;
R51 is H; alkyl; alkenyl; -(CH2)m(CHRδl)sOR55; -(CH2)m(CHRδI)sSR5δ; -
(CH2)m(CHRδl)sNR33R34;
-(CH2)o(CHR6I)sCOOR57; -(CH2)o(CHR61)sCONR58R59; -(CH2)o(CHR61)pPO(OR60)2;
-(CH2)P(CHR6I)S SO2R62; or -(CH2)p(CHRδl)sC6H4R8; R52 is H; alkyl; alkenyl; -(CH2)m(CHRδl)sOR55; -(CH2)m(CHRδl)sSR56; -
(CH2)m(CHRδl)sNR33R34;
-(CH2)o(CHRδl)sCOOR57; -(CH2)o(CHRδl)sCONR58R59; -(CH2)o(CHRδl)pPO(ORδ0)2;
-(CH2)p(CHR61)s SO2R62; or -(CH2)P(CHR61)SC6H4R8; R53 is H; alkyl; alkenyl; -(CH2)m(CHRδl)sOR55; -(CH2)ra(CHR6I)sSR56; - (CH2)m(CHRδl)sNR33R34;
-(CH2)o(CHR6I)sCOOR57; -(CH2)o(CHRδl)sCONRS8R59; -(CH2)o(CHRδl)pPO(OR60)2;
-(CH2)P(CHR61)S SO2R62; or -(CH2)P(CHR61)SC6H4R8; R54 is H; alkyl; alkenyl; -(CH2)m(CHRδl)sOR55; -(CH2)m(CHRδl)sNR33R34; -
(CH2)0(CHRδl)COOR57; -(CH2)0(CHRδl)sCONR58R59; or -(CH2)0(CHRδl)s C6H4R8;
R55 is H; lower alkyl; lower alkenyl; aryl-lower alkyl; -(CH2)m(CHRδl)sOR57;
-(CH2)m(CHR61)sNR34R63; -(CH2)0(CHRδl)s-COR64; -(CH2)0(CHR61)COOR57; or
-(CH2)0(CHRδl)sCONR58R59; R56 is H; lower alkyl; lower alkenyl; aryl-lower alkyl; -(CH2)m(CHRδI)sOR57; -(CH2)m(CHRδl)sNR34R63; -(CHzMC-HR' COR64; or -(CH2)o(CHR61)sCONR58R59;
R57 is H; lower alkyl; lower alkenyl; aryl lower alkyl; or heteroaryl lower alkyl; R58 is H; lower alkyl; lower alkenyl; aryl; heteroaryl; aryl-lower alkyl; or heteroaryl-lower alkyl; R59 is H; lower alkyl; lower alkenyl; aryl; heteroaryl; aryl-lower alkyl; or heteroaryl-lower alkyl; or
R58 and R59 taken together can form: -(CH2)2.6-; -(CH2)20(CH2)2-; -(CH2)2S(CH2)2-; or
-(CH2)2NR34(CH2)2-; Rδ0 is H; lower alkyl; lower alkenyl; aryl; or aryl-lower alkyl; Rδl is alkyl; alkenyl; aryl; heteroaryl; aryl-lower alkyl; heteroaryl-lower alkyl; -(CH2)m0R >555; .
-(CH2)mNR33R34; -(CH2)0COOR37; -(CH2)0NR58R59; or -(CH2)oPO(CORδ0)2; R62 is lower alkyl; lower alkenyl; aryl, heteroaryl; or aryl-lower alkyl; R63 is H; lower alkyl; lower alkenyl; aryl, heteroaryl; aryl-lower alkyl; heteroaryl-lower alkyl;
-COR64; -COOR57; -C0NR58R59; -S02R62; or -PO(ORδ0)2; R64 is H; lower alkyl; lower alkenyl; aryl; heteroaryl; aryl-lower alkyl; heteroaryl-lower alkyl;
-(CH2)p(CHRδl)sOR65; -(CH2)p(CHRδl)sSR6δ; or -(CH2)p(CHRδl)sNR34R63;
R is H; lower alkyl; lower alkenyl; aryl, aryl-lower alkyl; heteroaryl-lower alkyl; -COR 57.
-COOR57; or -CONR58R59; R is H; lower alkyl; lower alkenyl; aryl; aryl-lower alkyl; heteroaryl-lower alkyl; or ■ CONR58R59; m is 2-4; o is 0-4; p is 1-4; q is 0-2; r is 1 or 2; s is 0 or 1;
Z is a chain of n α-amino acid residues, n being the integer 7 or 11, the positions of said amino acid residues in said chain being counted starting from the N-terminal amino acid, whereby these amino acid residues are, depending on their position in the chain, Gly, or Pro, or of formula -A- CO-,or of one of the types C: -NR20CH(R72)CO-
D -NR20CH(R73)CO-: E -NR20CH(R74)CO F -NR20CH(R84)CO and H; -NR20-CH(CO-)-(CH2)4.7-CH(CO-)-NR20-; -NR20-CH(CO-)-(CH2)pSS(CH2)p-CH(CO-)-NR20-;
-NR2 -CH(CO-)-(-(CH2)pNR'uCO(CH2)P-CH(CO-)-NRzu-; and
•NR20-CH(CO-)-(-(CH2 ))PpNNRR2200CCOONNRR2200((CCHH22V)p-CH(CO-)-NR20-;
R71 is H; lower alkyl; lower alkenyl; -(CH2)p(CHRδI)sOR75; -(CH2)p(CHRδl)sSR75; -(CH2)PNR78R79 ;
-(CH2)0(CHRδI)sCOOR75; -(CH2)PC0NR78R79; -(CH2)pPO(OR62)2; -(CH2)pS02R62; or
-(CH2)o-C6R67R68R69R70R76; R72 is H; lower alkyl; lower alkenyl; -(CH2)p(CHRδl)sOR85; or -(CH2)p(CHRδl)sSR85; R73 is -(CH2)0R77; -(CH2)rO(CH2)0R77; -(CH2)rS(CH2)0R77; or -(CH2)rNR20(CH2)0R77; R74 is -(CH2)PNR78R79; -(CH2)PC(=NR80)NR78R79; -(CH2)pC(=NOR50)NR78R79;
-(CH2)pC(=NNR78R79)NR78R79; -(CH2)pNR80C(=NR80)NR78R79; -(CH2)pC6H4NR78R79;
Figure imgf000157_0001
-(CH2)pC6H4C(=NNR78R79)NR78R79; -(CH2)PC6H4NR80C(=NR80)NR78R79;
-(CH2)rO(CH2)mNR78R79; -(CH2)rO(CH2)pC(=NR80)NR78R79; -
(CH2)rO(CH2)pC(=NOR50)NR78R79;
-(CH2)rO(CH2)pC(=NNR78R79)NR78R79; -(CH2)rO(CH2)mNR80C(=NR80)NR78R79;
-(CH2)rO(CH2)pC6H4CNR78R79; -(CH2)rO(CH2)pC6H4C(=NR80)NR78R79; -(CH2)rO(CH2)pC6H4C(=NOR50)NR78R79;- (CH2)rO(CH2)pC6H4C(=NNR78R79)NR78R79;
-(CH2)rO(CH2)pC6H4NR80C(=NR80)NR78R79; -(CH2)rS(CH2)mNR78R79;
-(CH2)rS(CH2)pC(=NR80)NR78R79; -(CH2)rS(CH2)pC(=NOR50)NR78R79;
-(CH2)rS(CH2)pC(=NNR78R79)NR78R79; -(CH2)rS(CH2)mNR80C(=NR80)NR78R79;
-(CH2)rS(CH2)pC6H4CNR78R79; -(CH2)rS(CH2)PC6H4C(=NR80)NR78R79; -(CH2)tS(CH2)pCeΗ4C(=NOR50)NR78R79; -(CH2)rS(CH2)pC6H4C(=NNR78R79)NR78R79;
-(CH2)rS(CH2)pC6H4NR80C-(=NR80)NR78R79; -(CH2)PNR80CONR78R79; or
-(CH2)pC6H4NR80CONR7 R79; R75 is lower alkyl; lower alkenyl; or aryl-lower alkyl;
R7δ is H; lower alkyl; lower alkenyl; aryl-lower alkyl; -(CH2)0OR72; -(CH2)0SR72; - (CH2)oNR33R34;
-(CH2)oCOOR7S; -(CH2)0CONR58R59; -(CH2)0PO(ORδ0)2; -(CH2)pSO2R62; or -
(CH2)0COR64; R77 is -C6R67Rδ8Rδ9R70R7δ; or a heteroaryl group of one of the formulae
Figure imgf000157_0002
H1 H2 H3 H4 H5
Figure imgf000157_0003
H6 H7 H8 H9 H10
Figure imgf000158_0001
H11 H12 H13 H14 H15
Figure imgf000158_0002
H16 H17 H18 H19 H20
Figure imgf000158_0003
H21 H22 H23 H24 H25
Figure imgf000158_0004
H26 H27 H28 H29
Figure imgf000158_0005
H30 H31 H32 H33
Figure imgf000158_0006
H34 H35 H36 H37
Figure imgf000158_0007
H38 H39 H40 H41
Figure imgf000159_0001
H42 H43 H44 H45
Figure imgf000159_0002
H46 H47 H48 H49
Figure imgf000159_0003
H50 H51
R78 is H; lower alkyl; aryl; or aryl-lower alkyl;
R79 is H; lower alkyl; aryl; or aryl-lower alkyl; or
R78 and R79, taken together, can be -(CH2)2-7-; -(CH2)20(CH2)2-; or -(CH2)2NR33(CH2)2-;
R80 is H; or lower alkyl;
R81 is H; lower alkyl; or aryl-lower alkyl;
R82 is H; lower alkyl; aryl; heteroaryl; or aryl-lower alkyl;
R83 is H; lower alkyl; aryl; or -NR78R79;
R84 is -(CH2)ra(CHRδl)sOH; -(CH2)pCONR78R79; -(CH2)pNR80CONR78R79; -(CH2)pC6H4CONR78R79;
(CH2)pCOOR80 or -(CH2)PC6H4NR80CONR78R79; R85 is lower alkyl; or lower alkenyl;
with the proviso that in said chain of n α-amino acid residues Z if n is 7, the amino acid residues in positions 1 to 7 are: - PI: of type C or of type F or of type D;
P2: of type E or of type C or of type D or of type F;
P3 : of type F or of type C, or the residue is Gly or Pro;
P4: of type C or type D or of type F, or the residue is Gly or Pro;
P5 : of type F or of formula -A-CO-, or the residue is Gly or Pro; P6: of type C or of type E or of formula -A-CO-, or the residue is Pro; P7: of type C or of type F or of type D;
if n is 11, the amino acid residues in positions 1 to 11 are: - PI: of type E or oftype F or of type C;
P2: of type C or of type F or of type E;
P3: of type C or of type F;
P4: of type E or of type C or of type D or of type F, or the residue is Gly or Pro;
P5: of type F or of type C, or the residue is Gly or Pro; - P6: of type C or of type D or of type F, or the residue is Gly or Pro;
P7: of type F or of formula -A-CO-, or the residue is Gly or Pro;
P8: of type C or of type E or of formula -A-CO-, or the residue is Gly or Pro;
P9: of type C or of type F;
P10: of type F or of type C; - Pl l: of type D or of type E or of type F or of type C; or
P2 and P 10, taken together, can form a group of type H;
and pharmaceutically acceptable salts thereof.
Compounds according to claim 1 wherein
Figure imgf000161_0001
is a group of formula (al) or (a2).
3. Compounds according to claim 2 wherein A is a group of one of the formulae Al to A69;
R1 is hydrogen or lower alkyl;
R2 is H; lower alkyl; lower alkenyl; -(CH2)mOR55 (where R55 is lower alkyl; or lower alkenyl);
-(CH2)mSR5δ (where R56 is H; or lower alkyl; or lower alkenyl); -(CH2)mNR33R34 (where R33 is lower alkyl; or lower alkenyl; and R34 is H; or lower alkyl) ; -(CH2)mOCONR33R78 (where R33: lower alkyl; or lower alkenyl; R78: H; or lower alkyl); -(CH2)mNR20CONR33R78 (where R20: H or lower alkyl; R33: lower alkyl; or lower alkenyl; R78: H; or lower alkyl); -
(CH2)oN(R20)CORδ (where R20 is H; or lower alkyl; and R64 is lower alkyl; or lower alkenyl); -
(CH2)0COOR57 (where R57 is lower; or lower alkenyl); -(CH2)0CONR58R59 (where R58 is lower alkyl; or lower alkenyl; and R59 is H; or lower alkyl); -(CH2)oPO(ORδ0)2 (where Rδ0 is lower alkyl; or lower alkenyl); -(CH2)0SO2R62 (where R62 is lower alkyl; or lower alkenyl); or -(CH2)qC6H4R8 (where R8 is H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy);
R3 is H; lower alkyl; lower alkenyl; -(CH2)mOR55 (where R55 is lower alkyl; or lower alkenyl);
-(CH2)mSR5δ (where R5δ is H; or lower alkyl; or lower alkenyl); -(CH2)mNR33R34( where R33 is lower alkyl; or lower alkenyl; and R34 is H; or lower alkyl) ; -(CH2)0N(R20)COR64 (where R20 is
H; or lower alkyl; and R64 is lower alkyl; or lower alkenyl); -(CH2)0COOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)0CONR58R59 (where R58 is lower alkyl; or lower alkenyl; and R59 is H; or lower alkyl);
-(CH2)oPO(ORδ0)2 (where Rδ0 is lower alkyl; or lower alkenyl); -(CH2)0SO2R62 (where R62 is lower alkyl; or lower alkenyl); or -(CH^qCsH^8 (where R8 is H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy);
R4 is H; lower alkyl; lower alkenyl; -(CH2)mOR5S (where R55 is lower alkyl; or lower alkenyl);
-(CH2)mSR5δ (where R56 is H; or lower alkyl; or lower alkenyl); -(CH2)mNR33R34( where R33 is lower alkyl; or lower alkenyl; and R34is H or lower alkyl);
Figure imgf000161_0002
R20 is H; or lower alkyl; and R64 is lower alkyl; or lower alkenyl); -(CH2)0COOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)0CONR58R59 (where R58 is lower alkyl; or lower alkenyl; and R59 is H; or lower alkyl);
-(CH2)0PO(ORδ0)2 (where R60 is lower alkyl; or lower alkenyl); -(CH2)0S02R62 (where R52 is lower alkyl; or lower alkenyl); or -(CH2)qC6H4R8 (where R8 is H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy); R5 is lower alkyl; lower alkenyl; -(CH2)0OR55 (where R55 is lower alkyl; or lower alkenyl); -(CH2)0SR5δ (where R56 is H; or lower alkyl; or lower alkenyl); -(CH2)0NR33R34 (where R33 is lower alkyl; or lower alkenyl; and R34 is H; or lower alkyl); -(CH2)mOCONR33R78 (where R33: lower alkyl; or lower alkenyl; R78: H; or lower alkyl); -(CH2)mNR20CONR33R78 (where R20: H or lower alkyl; R33: lower alkyl; or lower alkenyl; R78: H; or lower alkyl); - (CH2)0N(R20)CORδ4(where R20 is H; or lower alkyl; and R64 is alkyl; alkenyl; aryl; aryl-lower alkyl; or heteroaryl-lower alkyl); -(CH2)0COOR57 (where R57 is lower alkyl; or lower alkenyl); - (CH2)0CONR58R59 (where R58 is lower alkyl; or lower alkenyl; and R59 is H; or lower alkyl); - (CH2)oPO(OR60)2 (where R60 is lower alkyl; or lower alkenyl); -(CH2)0S02R62 (where R62 is lower alkyl; or lower alkenyl); or -(CH2)qC6H4R8 (where R8 is H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy);
Rδ is H; lower alkyl; lower alkenyl; -(CH2)0OR55 (where R5S is lower alkyl; or lower alkenyl); -(CH2)0SR56 (where R56 is H; or lower alkyl; or lower alkenyl); -(CH2)0NR33R34 (where R33 is lower alkyl; or lower alkenyl; and R34 is H; or lower alkyl) ; -(CH2)oN(R20)COR64 (where R20 is H; or lower alkyl; and R64 is lower alkyl; or lower alkenyl); -(CH2)0COOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)0CONR58R59 (where R58 is lower alkyl; or lower alkenyl; and R59 is H; or lower alkyl);
-(CH2)0PO(ORδo)2 (where R60 is lower alkyl; or lower alkenyl); -(CH2)0SO2R62 (where R62 is lower alkyl; or lower alkenyl); or
Figure imgf000162_0001
(where R8 is H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy); R7 is lower alkyl; lower alkenyl; -(CH2)qOR55 (where R55 is lower alkyl; or lower alkenyl); -(CH2)qSR56 (where R56 is H; or lower alkyl; or lower alkenyl); -(CH2)qNR33R34(where R33 is lower alkyl; or lower alkenyl; and R34 is H; or lower alkyl); -(CH2)qN(R20)COR64(where R20 is H; or lower alkyl; and R64 is lower alkyl; or lower alkenyl); -(CH2)rCOOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)qCONR58R59 (where R58 is lower alkyl; or lower alkenyl; and R59 is H; or lower alkyl);
-(CH2)rPO(OR60)2 (where R60 is lower alkyl; or lower alkenyl); -(CH2)rSO2R62 (where R62 is lower alkyl; or lower alkenyl); or -(CH2)qC6H4R8 (where R8 is H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy); R8 is H; F; Cl; CF3; lower alkyl; lower alkenyl; -(CH2)00R55 (where R55 is lower alkyl; or lower alkenyl); -(CH2)0SR5δ (where R56 is H; or lower alkyl; or lower alkenyl); -(CH2)0NR33R34 (where R33 is lower alkyl; or lower alkenyl; and R34 is H or lower alkyl); -(CH2)0OCONR33R78 (where R33: lower alkyl; or lower alkenyl; R78: H; or lower alkyl); -(CH2)oNR20CONR33R78 (where R20: H or lower alkyl; R33: lower alkyl; or lower alkenyl; R78: H; or lower alkyl); -(CH2)oN(R20)COR64 (where is R20: H; or lower alkyl; and R64 is lower alkyl; or lower alkenyl); -(CH2)0COOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)0CONR58R59 (where R58 is lower alkyl; or lower alkenyl; and R59 is H; or lower alkyl); -(CH2)0PO(OR60)2 (where R60 is lower alkyl; or lower alkenyl); -(CH2)0S02R62 (where R62 is lower alkyl; or lower alkenyl); or
Figure imgf000163_0001
(where R8 is H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy);
R9 is lower alkyl; lower alkenyl; -(CH2)0OR55 (where R55 is lower alkyl; or lower alkenyl); -(CH2)0SR5δ (where R56 is H; or lower alkyl; or lower alkenyl); -(CH2)0NR33R34( where R33 is lower alkyl; or lower alkenyl; and R34 is H; or lower alkyl); -(CH2)oN(R20)COR64 (where R20 is H; or lower alkyl; and R64 is lower alkyl; or lower alkenyl); -(CH2)0COOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)0CONR58R59 (where R58 is lower alkyl; or lower alkenyl; and R59 is H; or lower alkyl);
-(CH2)oPO(OR60)2 (where Rδ0 is lower alkyl; or lower alkenyl); -(CH2)0S02R62 (where R62 is lower alkyl; or lower alkenyl); or -(CH2)qC„-H R8 (where R8 is H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy); R10 is lower alkyl; lower alkenyl; -(CH2)0OR55 (where R55 is lower alkyl; or lower alkenyl); -(CH2)0SR5δ (where R56 is H; or lower alkyl; or lower alkenyl); -(CH2)0NR33R34 (where R33 is lower alkyl; or lower alkenyl; and R34 is lower alkenyl); -(CH2)0COOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)0CONR58R59 (where R58 is lower alkyl; or lower alkenyl; and R59 is H; lower alkyl); -(CH2)oPO(ORδ0)2 (where R60 is lower alkyl; or lower alkenyl); -(CH2)0SO2R62 (where R62 is lower alkyl; or lower alkenyl); or -(CH2)qC6H R8 (where R8 is H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy) ;
R11 is H; lower alkyl; lower alkenyl; -(CH2)mOR55 (where R55 is lower alkyl; or lower alkenyl); -(CH2)mSR5δ (where R5δ is H; or lower alkyl; or lower alkenyl); -(CH2)m R33R34(where R33 is lower alkyl; or lower alkenyl; and R34 is H; or lower alkyl); -(CH2)mOCONR33R78 (where R33: lower alkyl; or lower alkenyl; R78: H; or lower alkyl); -(CH2)mNR20CONR33R78 (where R20: H or lower alkyl; R33: lower alkyl; or lower alkenyl; R78: H; or lower alkyl); -(CH2)mN(R20)COR64 (where R20 is H; or lower alkyl; and R64 is lower alkyl; or lower alkenyl); -(CH2)0COOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)0CONR58R59 (where R58 is lower alkyl; or lower alkenyl; and R59 is H; or lower alkyl);
-(CH2)oPO(OR60)2 (where Rδ0 is lower alkyl; or lower alkenyl); -(CH2)0S02R62 (where R62 is lower alkyl; or lower alkenyl); or
Figure imgf000164_0001
(where R8 is H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy); R12 is H; lower alkyl; lower alkenyl; -(CH2)mOR55 (where R55 is lower alkyl; or lower alkenyl);
-(CH2)mSR56 (where R56 is H; or lower alkyl; or lower alkenyl); -(CH2)mNR33R34 (where R33 is lower alkyl; or lower alkenyl; and R34 is H; or lower alkyl); -(CH2)mN(R20)COR64 (where: R20 is
H; or lower alkyl; and R64 is lower alkyl; or lower alkenyl); -(CH2)rCOOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)rCONR58R59 (where R58 is lower alkyl; or lower alkenyl; and R59 is H; or lower alkyl);
-(CH2)rPO(OR60)2 (where R60 is lower alkyl; or lower alkenyl); -(CH2)0S02R62 (where R62 is lower alkyl; or lower alkenyl); or -(CH2)qC6H4R8 (where R8 is H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy);
R13 is lower alkyl; lower alkenyl; -(CH2)q0R55 (where R55 is lower alkyl; or lower alkenyl); -(CH2)qSR56 (where R56 is H; or lower alkyl; or lower alkenyl); -(CH2)qNR33R34 (where R33 is lower alkyl; or lower alkenyl; and R34 is H; or lower alkyl); -(CH2)qN(R20)COR64 (where R20 is H; or lower alkyl; and R64 is lower alkyl; or lower alkenyl); -(CH2)rCOO57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)qC0NR58R59 (where R58 is lower alkyl; or lower alkenyl; and R59 is H; or lower alkyl); -(CH2)rPO(OR60)2 (where R60 is lower alkyl; or lower alkenyl); -(CH2)rS02R62 (where R62 is lower alkyl; or lower alkenyl); or -(CH2)qC6H4R8 (where R8 is H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy);
R14 is H; lower alkyl; lower alkenyl; -(CH2)mOR55 (where R55 is lower alkyl; or lower alkenyl);
-(CH2)mSR56 (where R56 is H; or lower alkyl; or lower alkenyl); -(CH2)mNR33R34 (where R33 is lower alkyl; or lower alkenyl; and R34 is H; or lower alkyl); -(CH2)raN(R20)COR64 (where R20 is
H; lower alkyl; and R64 is lower alkyl; or lower alkenyl); -(CH2)0COOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)0CONR58R59 (where R58 is lower alkyl; or lower alkenyl; and R59 is H; or lower alkyl); -(CH2)oPO(OR60)2 (where R60 is lower alkyl; or lower alkenyl); -
(CH2)0S02R62 (where R62 is lower alkyl; or lower alkenyl); or -(CH2)qC6H4R8 (where R8 is H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy);
R15 is lower alkyl; lower alkenyl; -(CH2)0OR5S (where R55 is lower alkyl; or lower alkenyl);
-(CH2)0SR56 (where R56 is H; or lower alkyl; or lower alkenyl); -(CH2)0NR33R34 (where R33 is lower alkyl; or lower alkenyl; and R34 is H; or lower alkyl); -(CH^o ^COR64 (where R20 is H; or lower alkyl; and R64 is lower alkyl; or lower alkenyl); -(CH2)0COOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)0CONR58R59 (where R58 is lower alkyl, or lower alkenyl; and R59 is H; lower alkyl);
-(CH2)oPO(OR60)2 (where Rδ0 is lower alkyl; or lower alkenyl); -(CH2)0SO2R62 (where R62 is lower alkyl; or lower alkenyl); or -(CH2)qC6H4R8 (where R8 is H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy);
RIδ is lower alkyl; lower alkenyl; -(CH2)0OR55 (where R55 is lower alkyl; or lower alkenyl); -(CH2)oSR5δ (where R5δ is H; or lower alkyl; or lower alkenyl); -(CH2)0NR33R34(where R33 is lower alkyl; or lower alkenyl; and R34 is H; or lower alkyl) ; -(CH2)oN(R20)COR64 (where R20 is H; or lower alkyl; and R64 is lower alkyl; or lower alkenyl); -(CH2)0COOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)0CONR58R59 (where R58 is lower alkyl; or lower alkenyl; and R59 is H; or lower alkyl);
-(CH2)oPO(OR60)2 (where R60 is lower alkyl; or lower alkenyl); -(CH2)0S02R62 (where R62 is lower alkyl; or lower alkenyl); or -(CH2)qC6H4R8 (where R8 is H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy); and R17 is lower alkyl; lower alkenyl; -(CH2)qOR55 (where R55 is lower alkyl; or lower alkenyl); -(CH2)qSR56 (where R56 is H; or lower alkyl; or lower alkenyl); -(CH2)qNR33R34(where R33 is lower alkyl; or lower alkenyl; and R34 is H; or lower alkyl); -(CH2)qN(R20)COR64 (where R20 is H; or lower alkyl; and R64 is lower alkyl; or lower alkenyl); -(CH2)rCOOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)qCONRS8R59 (where R58 is lower alkyl; or lower alkenyl; and R59 is H; or lower alkyl);
-(CH2)rPO(ORδ0)2 (where R60 is lower alkyl; or lower alkenyl); -(CH2)rS02R62 (where R62 is lower alkyl; or lower alkenyl); or -(CH2)qC6H4R8 (where R8 is H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
4. Compounds according to claim 2 or 3 wherein A is a group of one of the formulae A5
(with R2 being H); A8; A22; A25; A38 (with R2 being H); A42; and A50. 5. Compounds according to claim 4 wherein A is a group of formula
5.
Figure imgf000166_0001
A8" A8"
wherein R20 is H or lower alkyl; and R64 is alkyl; alkenyl; aryl; aryl-lower alkyl; or heteroaryl- lower alkyl and R75 is lower alkyl; lower alkenyl; or aryl-lower-alkyl.
6. Compounds according to claim 5 wherein R34 is H and R64 is n-hexyl.
7. Compounds according to claim 2 wherein A is a group of one of the formulae A70 to
A104;
R20 is H; or lower alkyl; R18 is lower alkyl;
R19 is lower alkyl; lower alkenyl; -(CH2)pOR55 (where R55 is lower alkyl; or lower alkenyl);
-(CH2)pSR5δ (where R5δ is H; or lower alkyl; or lower alkenyl); -(CH2)PNR33R34 ( here R33 is lower alkyl; or lower alkenyl; and R34 is H; or lower alkyl); -(CH2)PN(R20)COR64 (where R20 is H; or lower alkyl; and R64 is lower alkyl; or lower alkenyl); -(CH2)pCOOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)pCONR58R59 (where R58 is lower alkyl; or lower alkenyl; and R59 is H; or lower alkyl);
-(CH2)oPO(OR60)2 (where R60 is lower alkyl; or lower alkenyl); -(CH2)pSO2R62 (where R62 is lower alkyl; or lower alkenyl); or -(CH2)0C6H4R8 (where R8 is H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy); R21 is H; lower alkyl; lower alkenyl; -(CH2)0OR55 (where R55 is lower alkyl; or lower alkenyl);
-(CH2)0SR5δ (where R56 is H; or lower alkyl; or lower alkenyl); -(CH2)0NR33R34 (where R33 is lower alkyl; or lower alkenyl; and R34 is H; or lower alkyl); -(CH^^R^COR64 (where R20 is H; or lower alkyl; and R64 is lower alkyl; or lower alkenyl); -(CH2)0COOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)0CONR58R59 (where R58 is lower alkyl, or lower alkenyl; and R59 is H; lower alkyl);
-(CH2)oPO(OR60)2 (where R60 is lower alkyl; or lower alkenyl); -(CH2)0SO2R62 (where R62 is lower alkyl; or lower alkenyl); or
Figure imgf000166_0002
(where R8 is H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy); R22 is lower alkyl; lower alkenyl; -(CH2)0OR55 (where R55 is lower alkyl; or lower alkenyl);
-(CH2)0SR5δ (where R5δ is H; or lower alkyl; or lower alkenyl); -(CH2)0NR33R34 (where R33 is lower alkyl; or lower alkenyl; and R34 is H; or lower alkyl); -(CH2)oN(R20)CORδ4(where R20 is H; or lower alkyl; and R64 is lower alkyl; or lower alkenyl); -(CH2)0COOR57 (where R57 is lower alkyl; or lower alkenyl);
-(CH2)0CONR58R59 (where R58 is lower alkyl, or lower alkenyl; and R59 is H; or lower alkyl);
-(CH2)oPO(OR60)2 (where Rδ0 is lower alkyl; or lower alkenyl); -(CH2)0SO2R62 (where R62 is lower alkyl; or lower alkenyl); or -(CH2)qC6H R8 (where R8 is H; F; Cl; CF; lower alkyl; lower alkenyl; or lower alkoxy); R23 is H; lower alkyl; lower alkenyl; -(CH2)0OR55 (where R55 is lower alkyl; or lower alkenyl);
-(CH2)0SR56 (where R56 is H; or lower alkyl; or lower alkenyl); -(CH2)0NR33R34 (where R33 is lower alkyl; or lower alkenyl; and R34 is H; or lower alkyl); -(CH2)0N(R20)COR64 (where R20 is H; or lower alkyl; and R64 is lower alkyl; or lower alkenyl); -(CH2)0COOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)0CONR58R59 (where R58 is lower alkyl, or lower alkenyl; and R59 is H; lower alkyl);
-(CH2)oPO(ORδ0)2 (where R60 is lower alkyl; or lower alkenyl); -(CH2)0S02R62 (where R62 is lower alkyl; or lower alkenyl); or
Figure imgf000167_0001
(where R8 is H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy);
R24 is lower alkyl; lower alkenyl; -(CH2)0OR55 (where R55 is lower alkyl; or lower alkenyl); -(CH2)0SR5δ (where R5δ is H; or lower alkyl; or lower alkenyl); -(CH2)0NR33R34 (where R33 is lower alkyl; or lower alkenyl; and R34 is H; or lower alkyl); -(CH2)oN(R20)COR64 (where R20 is H; or lower alkyl; and R64 is lower alkyl; or lower alkenyl); -(CH2)0COOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)0CONR58R59 (where R58 is lower alkyl, or lower alkenyl; and R59 is H; or lower alkyl); -(CH2)0PO(ORδo)2 (where Rδ0 is lower alkyl; or lower alkenyl); -(CH2)0SO2R62 (where R62 is ' lower alkyl; or lower alkenyl); or -(CH2)qC6H4R8 (where R8 is H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy);
R25 is H; lower alkyl; lower alkenyl; -(CH2)mOR55 (where R55 is lower alkyl; or lower alkenyl);
-(CH2)mNR33R34(where R33 is lower alkyl; or lower alkenyl; and R34 is H; or lower alkyl); -(CH^n-N R^COR64 (where R20 is H; or lower alkyl; and R64 is lower alkyl; or lower alkenyl);
-(CH2)0COOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)0CONR58R59 (where R58 is lower alkyl; or lower alkenyl; and R59 is H; lower alkyl);
Figure imgf000167_0002
(where R60 is lower alkyl; or lower alkenyl); -(CH2)0SO2R62 (where R62 is lower alkyl; or lower alkenyl); or -
(CEyqCβH-tR8 (where R8 is H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy); R26 is H; lower alkyl; lower alkenyl; -(CH2)mOR55 (where R55 is lower alkyl; or lower alkenyl);
-(CH2)mNR33R34(where R33 is lower alkyl; or lower alkenyl; and R34 is H; or lower alkyl);
-(CH2)mN(R20)CORδ4 (where R20 is H; or lower alkyl; and R64 is lower alkyl; or lower alkenyl);
-(CH2)0COOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)0CONR58R59 (where R58 is lower alkyl; or lower alkenyl; and R59 is H; or lower alkyl); -(CH2)oPO(OR60)2 (where R60 is lower alkyl; or lower alkenyl); -(CH2)0S02R62 (where R62 is lower alkyl; or lower alkenyl); or -
(CH2)qC6H4R8 (where R8 is H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy); or
R25 and R2δ taken together are -(CH2)2.6-; -(CH2)20(CH2)2-; -(CH2)2S(CH2)2-; or -
(CH2)2NR34(CH2)2-; R27 is H; lower alkyl; lower alkenyl; -(CH2)0OR55 (where R55 is lower alkyl; or lower alkenyl);
-(CH2)0SR5δ (where R5δ is lower alkyl; or lower alkenyl); -(CH2)0NR33R34 (where R33 is lower alkyl; or lower alkenyl; and R34 is H; or lower alkyl); -(CH2)0N(R20)COR64 (where: R20 is H; or lower alkyl; and R64 is lower alkyl; or lower alkenyl); -(CH2)0COOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)0CONR58R59 (where R58 is lower alkyl, or lower alkenyl; and R59 is H; or lower alkyl);
-(CH2)oPO(ORδ0)2 (where Rδ0 is lower alkyl; or lower alkenyl); -(CH2)0S02R62 (where R62 is lower alkyl; or lower alkenyl); or -(CH2)qC6H4R8 (where R8 is H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy);
R28 is lower alkyl; lower alkenyl; -(CH2)0OR55 (where RS5 is lower alkyl; or lower alkenyl); -(CH2)0SR56 (where R5δ is H; or lower alkyl; or lower alkenyl); -(CH2)0NR33R34 (where R33 is lower alkyl; or lower alkenyl; and R34 is H; or lower alkyl); -(CH2)oN(R20)COR64 (where R20 is H; or lower alkyl; and R64 is lower alkyl; or lower alkenyl); -(CH2)0COOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)0CONR58R59 (where R58 is lower alkyl, or lower alkenyl; and R59 is H; or lower alkyl); -(CH2)oPO(ORδ0)2 (where Rδ0 is lower alkyl; or lower alkenyl); -(CH2)0S02R62 (where R62 is lower alkyl; or lower alkenyl); or
Figure imgf000168_0001
(where R8 is H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy); and
R29 is lower alkyl; lower alkenyl; -(CH2)00R55 (where R55 is lower alkyl; or lower alkenyl);
-(CH2)0SR56 (where R5δ is H; or lower alkyl; or lower alkenyl); -(CH2)0NR33R34 (where R33 is lower alkyl; or lower alkenyl; and R34 is H; or lower alkyl); -(CH2)0N(R20)COR64 (where R20 is H; or lower alkyl; and R64 is lower alkyl; or lower alkenyl); -(CH2)0COOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)0C0NR5 R59 (where R58 is lower alkyl, or lower alkenyl; and R59 is H; or lower alkyl);
-(CH2)oPO(ORδ0)2 (where Rδ0 is lower alkyl; or lower alkenyl); -(CH2)0S02R62 (where R62 is lower alkyl; or lower alkenyl); or -(CH2)qC6H4R (where R8 is H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
8. Compounds according to claim 7 wherein R23, R24 and R29 are -NR20-CO-lower alkyl where R20 is H; or lower alkyl.
9. Compounds according to claim 7 or 8 wherein A is a group of one of the formulae A74 (with R22 being H); A75; A76; A77 (with R22 being H); A78; and A79.
10. Compounds according to any one of claims 2 to 9 wherein B is a group of formula
-NR20CH(R71)- or an enantiomer of one of the groups A5 (with R2 being H); A8; A22; A25; A38 (with R2 being H); A42; A47; and A50.
11. Compounds according to claim 10 wherein B-CO is Ala; Arg; Asn; Cys; Gin; Gly; His; He; Leu; Lys; Met; Phe; Pro; Ser; Thr; Trp; Tyr; Val; Cit; Orn; tBuA; Sar; t-BuG; 4AmPhe;
3AmPhe; 2AmPhe; Phe(mC(NH2)=NH; Phe(pC(NH2)=NH; Phe(mNHC (NH2)=NH; Phe(pNHC (NH2)=NH; Phg; Cha; C al; C5al; Nle; 2-Nal; 1-Nal; 4C1-Phe; 3C1-Phe; 2C1-Phe; 3,4Cl2Phe; 4F- Phe; 3F-Phe; 2F-Phe; Tic; Thi; Tza; Mso; AcLys; Dpr; A2Bu; Dbu; Abu; Aha; Aib; Y(Bzl); Bip; S(Bzl); T(Bzl); hCha; hCys; hSer, hArg; hPhe; Bpa; Pip; OctG; MePhe; MeNle; MeAla; Melle; MeVal; or MeLeu.
12. Compounds according to claim 10 or 11 wherein B is a group, having (L)-confιguration, of formula
Figure imgf000169_0001
A8'
wherein R20 is H; or lower alkyl; and R64 is alkyl; alkenyl; aryl; aryl-lower alkyl; or heteroaryl- lower alkyl.
13. Compounds according to claim 11 wherein R64 is n-hexyl.
14. Compounds according to claim 1 wherein
Figure imgf000170_0001
is a group of formula (bl) or (cl); R1 is H; or lower alkyl; R20 is H; or lower alkyl; R30 is H; or methyl;
R31 is H; lower alkyl; lower alkenyl; -(CH2)pOR55 (where R55 is lower alkyl; or lower alkenyl); -(CH2)PNR33R34 (where R33 is lower alkyl; or lower alkenyl; and R34 is H; or lower alkyl); -CH2)PN(R20)COR64 (where is R20: H; or lower alkyl; and R64 is lower alkyl; or lower alkenyl); -(CH2)0COOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)0CONR58R59 (where R58 is lower alkyl, or lower alkenyl; and R59 is H; or lower alkyl); -(CH2)0PO(ORδ0)2 (where R60 is lower alkyl; or lower alkenyl); -(CH2)0SO2R62 (where R62 is lower alkyl; or lower alkenyl); or - (CH2)rC6H4R8 (where R8 is H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy); R32 is H; or methyl;
R35 is H; lower alkyl; lower alkenyl; -(CH2)mOR55 (where R55 is lower alkyl; or lower alkenyl); -(CH^NζR^COR64 (where: R20 is H; or lower alkyl; and R64 is lower alkyl; or lower alkenyl); -(CH2)0COOR57 (where R57 is lower alkyl; or lower alkenyl); or -(CH2)0CONR58R59 (where R58 is lower alkyl; or lower alkenyl; and R59 is H; lower alkyl); R3δ is lower alkyl; lower alkenyl; or aryl-lower alkyl;
R37 is H; lower alkyl; lower alkenyl; -(CH2)pOR55 (where R55 is lower alkyl; or lower alkenyl); -(CH2)PNR33R34 (where R33 is lower alkyl; or lower alkenyl; and R34 is H; or lower alkyl); -(CH2)PN(R20)COR64 (where R20 is H; or lower alkyl; and R64 is lower alkyl; or lower alkenyl); -(CH2)0COOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)0CONR58R59 (where R58 is lower alkyl, or lower alkenyl; and R59 is H; or lower alkyl); -(CH2)oPO(ORδ0)2 (where R60 is lower alkyl; or lower alkenyl); -(CH2)0SO2R62 (where R62 is lower alky; or lower alkenyl); or - (CH2)qC6H4R8 (where R8 is H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy); and R38 is H; lower alkyl; lower alkenyl; -(CH2)pOR55 (where R55 is lower alkyl; or lower alkenyl); -(CH2)PNR33R34 (where R33 is lower alkyl; or lower alkenyl; and R34 is H; or lower alkyl); -(CH2)PN(R20)COR64 (where R20 is H; or lower alkyl; and R64 is lower alkyl; or lower alkenyl); -(CH2)0COOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)0CONR58R59 (where R58 is lower alkyl, or lower alkenyl; and R59 is H; or lower alkyl); -(CH2)oPO(ORδ0)2 (where R60 is lower alkyl; or lower alkenyl); -(CH2)oS02R62 (where R62 is lower alkyl; or lower alkenyl); or - (CH2)qC6H4R8 (where R8 is H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
15. Compounds according to claim 14 wherein R1 is H; R20 is H; R30 is H; R31 is carboxymethyl; or lower alkoxycarbonylmethyl; R32 is H; R35 is methyl; R36 is methoxy; R37 is H and R38 is H.
16. Compounds according to any one of claims 1 to 15 wherein in the chain of α-amino acid residues Z - if n is 7, the amino acid residues in positions 1 to 7 are:
PI : of type C or of type F;
P2: of type E or of type C or of type D;
P3: of type F or of type C;
P4: of type C or of type F or of type D; - P5: of type F, or the residue is Pro;
P6: of type C or of type E, or the residue is Pro;
P7: of type C or of type F;
if n is 11, the amino acid residues in positions 1 to 11 are: - PI : of type E or of type F;
P2: of type C or of type F;
P3: of type C or of type F;
P4: of type E or of type C or of type D;
P5: of type F or of type C; - P6: of type C or of type D;
P7: of type F, or the residue is Pro;
P8: of type C or of type E, or the residue is Pro;
P9: of type C or of type F;
P10: of type F or of type C; - PI 1 : of type D or of type E; or
P2 and P 10, taken together, can form a group of type H.
17. A compound according to claim 1 wherein the template is ^ro^Pro; n is 7; and the amino acid residues in position 1 -7 are: PI Thr;
P2 Lys;
P3 Ser;
P4 lie;
P5 Pro;
P6 Pro;
P7 He.
18. A compound according to claim 1 wherein the template is DPro- Pro; n is 7; and the amino acid residues in position 1 - 7 are:
PI Thr;
P2 Lys;
P3 Ala;
P4 lie;
P5 Pro;
P6 Pro;
P7 lie.
19. A compound according to claim 1 wherein the template is ^ro^Pro; n is 7; and the amino acid residues in position 1 - 7 are:
PI Thr;
P2 Lys;
P3 Ser;
P4 lie;
P5 Pro;
P6 Ala;
P7 lie.
20. A compound according to claim 1 wherein the template is "•'Pro^Pro; n is 7; and the amino acid residues in position 1 -7 are:
PI Thr;
P2 Lys;
P3 Ser;
P4 He; P5: Pro;
P6: Pro;
P7: Ala.
21. A compound according to claim 1 wherein the template is DPro-LPro; n is 11; and the amino acid residues in position 1 - 11 are:
PI Arg;
P2 Cys;
P3 Thr;
P4 Lys;
P5 Ser;
P6 He;
P7 Pro;
P8 Pro;
P9 He;
PI s): Cys;
PI 1: Phe, the two Cys residues forming a disulfide bridge.
22. A compound according to claim 1 wherein the template is DPro-(2R,4S)-4-[n- hexylcarbonylamino]-LPro; n is 7; and the amino acid residues in position 1 - 7 are:
PI Thr;
P2 Lys;
P3 Ser;
P4 He;
P5 Pro;
P6 Pro;
P7 He.
23. A compound according to claim 1 wherein the template is DPro-(2R,4S)-4- allyloxycarbonylamino-LPro; n is 7; and the amino acid residues in position 1 - 7 are:
PI ;Thr
P2 Lys;
P3 Ser; P4 He;
P5: Pro;
P6 Pro;
P7 He.
24. A compound according to claim 1 wherein the template is of formula (cl) wherein R20 is H; R35 is methyl; R3 is methoxy; R37 is H and R38 is H; n is 7; and the amino acid residues in position
- 7 are:
- PI Thr;
- P2 Lys;
- P3 Ser;
- P4 He;
- P5 Pro;
- P6 Pro;
_ P7 He.
25. Compounds according to any one of claims 1 to 24 for use as therapeutically active substances.
26. A pharmaceutical composition containing a compound according to any one of claims 1 to 24 and a pharmaceutically inert carrier.
27. The use of compounds according to any one of claims 1 to 24 for the manufacture of a medicament for use as an inhibitor of protease enzymes.
28. A process for the manufacture of compounds according to any one of claims 1 to 24 which process comprises
(a) coupling an appropriately functionalized solid support with an appropriately N-protected derivative of that amino acid which in the desired end-product is in position 2+V2 or 72-V2, any functional group which may be present in said N-protected amino acid derivative being likewise appropriately protected;
(b) removing the N-protecting group from the product thus obtained;
(c) coupling the product thus obtained with an appropriately N-protected derivative of that amino acid which in the desired end-product is one position nearer the N-terminal amino acid residue, any functional group which may be present in said N-protected amino acid derivative being likewise appropriately protected;
(d) removing the N-protecting group from the product thus obtained;
(e) repeating, if necessary, steps (c) and (d) until the N-terminal amino acid residue has been introduced; (f) coupling the product thus obtained to a compound of the general formula
Figure imgf000175_0001
wherein
Figure imgf000175_0002
is as defined above and X is an N-protecting group or, if
Figure imgf000175_0003
is to be group (al) or (a2), above, alternatively
(fa) coupling the product obtained in step (d) or (e) with an appropriately N-protected derivative of an amino acid of the general formula
HOOC-B-H III or HOOC-A-H IV wherein A and B are as defined above , any functional group which may be present in said N-protected amino acid derivative being likewise appropriately protected; (fb) removing the N-protecting group from the product thus obtained; and (fc) coupling the product thus obtained with an appropriately N-protected derivative of an amino acid of the above general formula IV and, respectively, IH, any functional group which may be present in said N-protected amino acid derivative being likewise appropriately protected;
(g) removing the N-protecting group from the product obtained in step (f) or (fc); (h) coupling the product thus obtained to an appropriately N-protected derivative of that amino acid which in the desired end-product is in position n, any functional group which may be present in said N-protected amino acid derivative being likewise appropriately protected;
(i) removing the N-protecting group from the product thus obtained;
(j) coupling the product thus obtained to an appropriately N-protected derivative of that amino acid which in the desired end-product is one position farther away from position n, any functional group which may be present in said N-protected amino acid derivative being likewise appropriately protected;
(k) removing the N-protecting group from the product thus obtained;
(1) repeating, if necessary, steps (j) and (k) until all amino acid residues have been introduced; J*~
(m) if desired, selectively deprotecting one or several protected functional group(s) present in the molecule and appropriately substituting the reactive group(s) thus liberated;
(o) detaching the product thus obtained from the solid support;
(p) cyclizing the product cleaved from the solid support; (q) if desired, forming an interstrand linkage between side-chains of appropriate amino acid residues at opposite positions of the β-strand region;
(r) removing any protecting groups present on functional groups of any members of the chain of amino acid residues and, if desired, any protecting group(s) which may in addition be present in the molecule; and (r) if desired, converting the product thus obtained into a pharmaceutically acceptable salt or converting a pharmaceutically acceptable, or unacceptable, salt thus obtained into the ' corresponding free compound of formula I or into a different, pharmaceutically acceptable, salt.
PCT/EP2001/014528 2001-12-11 2001-12-11 Template-fixed peptidomimeticsas inhibitors of serine proteases WO2003054000A1 (en)

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CNA2007101543999A CN101157924A (en) 2001-12-11 1996-10-25 Neutrophil leukocyte gene alpha
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PCT/EP2001/014528 WO2003054000A1 (en) 2001-12-11 2001-12-11 Template-fixed peptidomimeticsas inhibitors of serine proteases
CN2007101537108A CN101157720B (en) 2001-12-11 2001-12-11 Template fixed peptide analogue as serine protease inhibitor
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DE60137654T DE60137654D1 (en) 2001-12-11 2001-12-11 FROM SERIN PROTEASES
AT01985390T ATE422501T1 (en) 2001-12-11 2001-12-11 TEMPLATE-FIXED PEPTIDOMIMETICS AS INHIBITORS OF SERINE PROTEASES
CNA018238807A CN1582296A (en) 2001-12-11 2001-12-11 Template-fixed peptidomimeticsas inhibitors of serine proteases
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CA2466591A CA2466591C (en) 2001-12-11 2001-12-11 Template-fixed peptidomimetics as inhibitors of serine proteases
JP2003554716A JP4098244B2 (en) 2001-12-11 2001-12-11 Template-bound peptide mimetics as serine protease inhibitors
EP01985390A EP1458747B1 (en) 2001-12-11 2001-12-11 Template-fixed peptidomimetics as inhibitors of serine proteases
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