WO2006087001A1 - Template-fixed beta-hairpin peptidomimetics with protease inhibitory activity - Google Patents

Template-fixed beta-hairpin peptidomimetics with protease inhibitory activity Download PDF

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Publication number
WO2006087001A1
WO2006087001A1 PCT/EP2005/001622 EP2005001622W WO2006087001A1 WO 2006087001 A1 WO2006087001 A1 WO 2006087001A1 EP 2005001622 W EP2005001622 W EP 2005001622W WO 2006087001 A1 WO2006087001 A1 WO 2006087001A1
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Prior art keywords
lower alkyl
chr
alkenyl
alkyl
pro
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PCT/EP2005/001622
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French (fr)
Inventor
Steven J. Demarco
Kerstin Moehle
Heiko Henze
Odile Sellier
Françoise JUNG
Frank Gombert
Daniel Obrecht
Christian Ludin
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Polyphor Ltd.
Universität Zürich
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Priority to AU2005327825A priority Critical patent/AU2005327825B2/en
Priority to KR1020137000610A priority patent/KR101420198B1/en
Priority to DK05707463.5T priority patent/DK1856140T3/en
Priority to KR1020077021071A priority patent/KR101272633B1/en
Priority to JP2007555456A priority patent/JP4896892B2/en
Priority to US11/816,589 priority patent/US8658604B2/en
Priority to MX2007009873A priority patent/MX2007009873A/en
Priority to ES05707463.5T priority patent/ES2453540T3/en
Priority to PT57074635T priority patent/PT1856140E/en
Priority to NZ556747A priority patent/NZ556747A/en
Priority to EP05707463.5A priority patent/EP1856140B1/en
Priority to CA3035941A priority patent/CA3035941C/en
Priority to PL05707463T priority patent/PL1856140T3/en
Priority to CA2598360A priority patent/CA2598360C/en
Application filed by Polyphor Ltd., Universität Zürich filed Critical Polyphor Ltd.
Priority to CNA2005800491411A priority patent/CN101142228A/en
Priority to BRPI0520016-4A priority patent/BRPI0520016B1/en
Priority to KR1020127025871A priority patent/KR101443171B1/en
Priority to CA2915175A priority patent/CA2915175C/en
Priority to EA200701727A priority patent/EA013814B1/en
Priority to SI200531837T priority patent/SI1856140T1/en
Priority to PCT/EP2005/001622 priority patent/WO2006087001A1/en
Publication of WO2006087001A1 publication Critical patent/WO2006087001A1/en
Priority to IL184804A priority patent/IL184804A/en
Priority to NO20074674A priority patent/NO342041B1/en
Priority to US14/100,878 priority patent/US20140213531A1/en
Priority to CY20141100192T priority patent/CY1115084T1/en
Priority to HK16101770.6A priority patent/HK1213914A1/en
Priority to US15/170,233 priority patent/US10100084B2/en
Priority to US16/130,520 priority patent/US10562933B2/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/06—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length using protecting groups or activating agents
    • C07K1/061—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length using protecting groups or activating agents using protecting groups
    • 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
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
    • A61K38/08—Peptides having 5 to 11 amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
    • A61K38/10—Peptides having 12 to 20 amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • 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 ⁇ -hairpin peptidomimetics incorporating a template-fixed chain of 11 ⁇ -amino acid residues which, depending on their position in the chain, are GIy, or Pro, or Pro(4NHCOPhe), or are 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 10 such as human cathepsin G, elastase, or tryptase.
  • the present invention provides an efficient process by which these compounds can, if desired, be made in library-format.
  • the ⁇ -hairpin peptidomimetics of the invention show improved efficacy, oral 15 bioavailability, improved half-life and most importantly a high selectivity ratio among different serine proteases which depends on the proper choice of certain types of ⁇ -amino acid residues and their position in said chain.
  • these ⁇ -hairpin peptidomimetics show a low hemolysis on red blood cells and low cytotoxicity.
  • ⁇ -Hairpin mimetics are thus ideally suited to lock peptide sequences in an extended conformation.
  • Serine proteases constitute important therapeutic targets. Serine proteases are classified by their substrate specificity, particularly by the type of residue found at Pl, as either trypsin-like (positively charged residues Lys/Arg preferred at Pl), elastase-like (small hydrophobic residues Ala/Val at Pl), or chymotrypsin-like (large hydrophobic residues Phe/Tyr/Leu at Pl).
  • Serine proteases for which protease-inhibitor X-ray crystal data is available on the PDB data base include trypsin, ⁇ -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 IXA, 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.
  • cathepsin G 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).
  • inhibitors of elastase and cathepsin G constitute valuable targets for novel drug candidates particularly for chronic obstructive pulmonary disease (Ohbayashi H, Epert Opin. Investig. Drugs 2002, 11, 965-980).
  • sunflower trypsin inhibitor S. Luckett, R. Santiago Garcia, J. J. Barker, A. V. Konarev, P. R. Shewry, A. R. Clarke, R. L. Brady, J. 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-I sunflower trypsin inhibitor
  • SFTI- 1 inhibited ⁇ -trypsin (K ; ⁇ 0.1nM), cathepsin G (K 1 ⁇ 0.15nM), elastase (Kj ⁇ 105 ⁇ M), chymotrypsin (Kj ⁇ 7.4 ⁇ M) and thrombin (Kj-136mM).
  • ⁇ -hairpin peptidomimetics of the present invention are compounds of the general formula
  • s* is a group of one of the formulae
  • R 1 is H; lower alkyl; or aryl-lower alkyl
  • R 2 is H; alkyl; alkenyl; -(CH 2 ) m (CHR 61 ) s OR 55 ; -(CH 2 ) m (CHR 61 ) s SR :>1
  • R 5 is alkyl; alkenyl; -(CH 2 ) o (CHR 61 ) s OR 55 ; -(CH 2 ) o (CHR 61 ) s SR 56 ; - (CH 2 ) o (CHR 61 ) s NR 33 R 34 ;
  • R 6 is H; alkyl; alkenyl; -(CH 2 ) o (CHR 61 ) s OR 55 ; -(CH 2 ) o (CHR 61 ) s SR 56 ; -
  • R 7 is alkyl; alkenyl; -(CH 2 ) q (CHR 61 ) s OR 55 ; -(CH 2 ) q (CHR 61 ) s NR 33 R 34 ; -(CH 2 ) q (CHR 61 ) s OCONR 33 R 75 ; -(CH 2 ) q (CHR 61 ) s NR 20 CONR 33 R 82 ;
  • R 8 is H; Cl; F; CF 3 ; NO 2 ; lower alkyl; lower alkenyl; aryl; aryl-lower alkyl; -(CH 2 ) o (CHR 61 ) s OR 55 ; -(CH 2 ) o (CHR 61 ) s SR 56 ; -(CH 2 ) o (CHR 61 )NR 33 R 34 ;
  • R u is H; alkyl; alkenyl; -(CH 2 ) m (CHR 61 ) s OR 55 ; -(CH 2 ) m (CHR 61 ) s NR 33 R 34 ; -(CH 2 ) m (CHR 61 ) s OCONR 33 R 75 ; -(CH 2 ) m (CHR 61 ) s NR 20 CONR 33 R 82 ; -(CH 2 ) 0 (CHR 61 ) S COOR 57 ; -(CH 2 ) 0 (CHR 61 ) S CONR 58 R 59 ; - (CH 2 ) o (CHR 61 ) s PO(OR 60 ) 2 ;
  • R 12 is H; alkyl; alkenyl; -(CH 2 ) m (CHR 61 ) s OR 55 ; -(CH 2 ) m (CHR 61 ) s SR 56 ; -(CH 2 ) m (CHR 61 ) s NR 33 R 34 ; -(CH 2 ) m (CHR 61 ) s OCONR 33 R 75 ; -(CH 2 ) m (CHR 61 ) NR 20 CONR 33 R 82 ; -(CH 2 ) r (CHR 61 ) s COOR 57 ; - (CH 2 ) r (CHR 61 ) s CONR 58 R 59 ; -(CH 2 ) r (CHR 61 ) s CONR 58 R 59 ; -(CH 2 ) r (CHR 61 ) s CONR 58 R 59 ; -(CH 2 ) r (CHR
  • R 13 is alkyl; alkenyl; -(CH 2 ) q (CHR 61 ) s OR 55 ; -(CH 2 ) q (CHR 61 ) s SR 56 ; - (CH 2 ) q (CHR 61 ) s NR 33 R 34 ;
  • R 14 is H; alkyl; alkenyl; -(CH 2 ) m (CHR ⁇ l ) s OR 55 ; -(CH 2 ) m (CHR 61 ) s NR 33 R 34 ; -(CH 2 ) m (CHR 61 ) s OCONR 33 R 75 ; -(CH 2 ) m (CHR 61 ) s NR 20 CONR 33 R 82 ; -(CH 2 ) q (CHR 6I ) s COOR 57 ; -(CH 2 ) q (CHR 61 ) s CONR 58 R 59 ; -
  • R 15 is alkyl; alkenyl; -(CH 2 ) o (CHR 61 ) s OR 55 ; -(CH 2 ) O (CHR 61 ) S SR 56 ; -
  • R 16 is alkyl; alkenyl; -(CH 2 ) O (CHR 61 ) S OR 55 ; -(CH 2 ) O (CHR 61 ) S SR 56 ; - (CH 2 ) O (CHR 61 ) S NR 33 R 34 ; -(CH 2 ) o (CHR 61 ) s OCONR 33 R 75 ; -(CH 2 ) o (CHR 61 ) NR 20 CONR 33 R 82 ;
  • R 17 is alkyl; alkenyl; -(CH 2 ) q (CHR 61 ) s OR 55 ; -(CH 2 ) q (CHR 61 ) s SR 56 ; -
  • R 18 is alkyl; alkenyl; -(CH 2 ) P (CHR 61 ) s OR 55 ; -(CH 2 ) P (CHR 61 ), SR 56 ; -
  • R 19 is lower alkyl; -(CH 2 ) P (CHR 61 ) s OR 55 ; -(CH 2 ) P (CHR 61 ) s SR 56 ; -
  • -(CH 2 ) P (CHR 61 ) s SO 2 R 62 ; or -(CH 2 ) o (CHR 61 ⁇ C 6 H 4 R 8 ; or R 18 and R 19 taken together can form: -(CH 2 ) 2 . 6 -; -(CH 2 ) 2 O(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 ) o (CHR 61 ) s OR 55 ; -(CH 2 ) o (CHR 61 ) s SR 56 ; -
  • R 22 is H; alkyl; alkenyl; -(CH 2 ) o (CHR 61 ) s OR 55 ; -(CH 2 ) o (CHR 61 ) s SR 56 ; -
  • R 23 is alkyl; alkenyl; -(CH 2 ) o (CHR 61 ) s OR 55 ; -(CH 2 ) o (CHR 61 ) s SR 56 ; - (CH 2 ) o (CHR 61 ) s NR 33 R 34 ;
  • R 24 is alkyl; alkenyl; -(CH 2 ) o (CHR 61 ) s OR 55 ; -(CH 2 ) o (CHR 61 ) s SR 56 ; -
  • R 25 is H; alkyl; alkenyl; -(CH 2 ) m (CHR 61 ) s OR 55 ; -(CH 2 ) m (CHR 61 ) s SR 56 ;
  • R 26 is H; alkyl; alkenyl; -(CH 2 ) m (CHR 61 ) s OR 55 ; -(CH 2 ) m (CHR 61 ) s SR 56 ;
  • R 25 and R 26 taken together can form: -(CH 2 ) 2 . 6 -; -(CH 2 ),.O(CH 2 ) r ; -(CH 2 ) r S(CH 2 ) r ; or
  • R 27 is H; alkyl; alkenyl; -(CH 2 ) o (CHR 61 ) s OR 55 ; -(CH 2 ) o (CHR 61 ) s SR 56 ; -
  • R 28 is alkyl; alkenyl; -(CH 2 ) o (CHR 61 ) s -OR 55 ; -(CH 2 ) o (CHR 61 ), SR 56 ; -(CH 2 ) o (CHR 61 ) s NR 33 R 34 ; -(CH 2 ) o (CHR 61 ) s OCONR 33 R 75 ; -(CH 2 ) o (CHR 61 ) NR 20 CONR 33 R 82 ; -(CH 2 ) o (CHR 61 ) s COOR 57 ; -(CH 2 ) o (CHR 61 ) s CONR 58 R 59 ; -(CH 2 ) o (CHR 61 ) s PO(OR 60 ) 2 ;
  • R 29 is alkyl; alkenyl; -(CH 2 ) o (CHR 61 ) s OR 55 ; -(CH 2 ) o (CHR 61 ) s SR 56 ; - (CH 2 ) o (CHR 61 ) s NR 33 R 34 ;
  • R 30 is H; alkyl; alkenyl; or aryl-lower alkyl;
  • R 31 is H; alkyl; alkenyl; -(CH 2 ) P (CHR 61 ) s OR 55 ; -(CH 2 ) P (CHR 61 ) s NR 33 R 34 ; -(CH 2 ) p (CHR 61 ) s OCONR 33 R 75 ; -(CH 2 ) p (CHR 61 ) s NR 20 CONR 33 R 82 ; -(CH 2 ) o (CHR 61 ) s COOR 57 ; -(CH 2 ) o (CHR 61 ) s CONR 58 R 59 ; - (CH 2 ) o (CHR 61 ) s PO(OR 60 ) 2 ;
  • R 32 is H; lower alkyl; or aryl-lower alkyl;
  • R 33 is H; alkyl, alkenyl; -(CH 2 ) m (CHR 61 ) s OR 55 ; -(CH 2 ) m (CHR 61 ) s NR 34 R 63 ; -(CH 2 ) m (CHR 61 ) s OCONR 75 R 82 ; -(CH 2 ) m (CHR 61 ) s NR 20 CONR 78 R 82 ; -(CH 2 ) o (CHR 61 ) s COR 64 ; -(CH 2 ) o (CHR 61 ) s -CONR 5S R 59 , -
  • R 34 is H; lower alkyl; aryl, or aryl-lower alkyl;
  • R 33 and R 34 taken together can form: -(CH 2 ) 2 . 6 -; -(CH 2 ) 2 O(CH 2 ) 2 -; -(CH 2 ) 2 S(CH 2 ) 2 -; or -(CH 2 ) 2 NR 57 (CH 2 ) 2 -;
  • R 35 is H; alkyl; alkenyl; -(CH 2 ) m (CHR 61 ) s OR 55 ; -(CH 2 ) m (CHR 61 ) s NR 33 R 34 ; -(CH 2 ) m (CHR 61 ) s OCONR 33 R 75 ; -(CH 2 ) m (CHR 61 ) s NR 20 CONR 33 R 82 ; -(CH 2 ) P (CHR 61 ) s COOR 57 ; -(CH 2 ) P (CHR 61 ) s CONR 58 R 59 ; - (CH 2 ) P (CHR 61 ) s PO(OR 60 ) 2 ; -(CH 2 ) p (CHR 61 ) s SO 2 R 62 ; or -(CH 2 ) P (CHR 61 ) s C 6 H 4 R 8 ;
  • R 36 is H, alkyl; alkenyl; -(CH 2 ) o (CHR 61 ) s OR 55 ; -(CH 2 ) P (CHR 61 ) s NR 33 R 34 ; -(CH 2 ) p (CHR 61 ) s OCONR 33 R 75 ; -(CH 2 ) p (CHR 61 ) s NR 20 CONR 33 R 82 ; -(CH 2 ) p (CHR 61 ) s COOR 57 ; -(CH 2 ) P (CHR 61 ) s CONR 58 R 59 ; - (CH 2 )p(CHR 61 ) s PO(OR 60 ) 2 ; -(CH 2 ) P (CHR 61 ) s SO 2 R 62 ; or -(CH 2 ) o (CHR 61 ) s C 6 H 4 R 8 ; R 37 is H; F; Br; Cl; NO
  • R 38 is H; F; Br; Cl; NO 2 ; CF 3 ; alkyl; alkenyl; -(CH 2 ) P (CHR 61 ) s OR 55 ; - (CH 2 ) P (CHR 61 ) 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; NO 2 ; CF 3 ; alkyl; alkenyl; -(CH 2 ) P (CHR 61 ) s OR 55 ; - (CH 2 ) p (CHR 61 ) s NR 33 R 34 ;
  • R 42 is H; F; Br; Cl; NO 2 ; CF 3 ; alkyl; alkenyl; -(CH 2 ) P (CHR 61 ) s OR 55 ; - (CH 2 ) p (CHR 61 ) s NR 33 R 34 ;
  • R 43 is H; alkyl; alkenyl; -(CH 2 ) m (CHR 61 ) s OR 55 ; -(CH 2 ) m (CHR 61 ) s NR 33 R 34 ; -(CH 2 ) m (CHR 61 ) s OCONR 33 R 75 ; -(CH 2 ) m (CHR 61 ) s NR 20 CONR 33 R 82 ; -(CH 2 ) o (CHR 61 ) s COOR 57 ; -(CH 2 ) o (CHR 61 ) s CONR 58 R 59 ; - (CH 2 ) o (CHR 61 ) s PO(OR 60 ) 2 ;
  • R 44 is alkyl; alkenyl; -(CH 2 ) r (CHR 61 ) s OR 55 ; -(CH 2 ) r (CHR 61 ) s SR 56 ; - (CH 2 ) r (CHR 61 ) s NR 33 R 34 ; -(CH 2 ) r (CHR 61 ) s OCONR 33 R 75 ; -(CH 2 ) r (CHR 51 ) NR 20 CONR 33 R 82 ; 5 001622
  • R 45 is H; alkyl; alkenyl; -(CH 2 ) o (CHR 61 ) s OR 55 ; -(CH 2 ) o (CHR 61 ) s SR 56 ; - (CH 2 ) o (CHR 61 ) s NR 33 R 34 ;
  • R 46 is H; alkyl; alkenyl; or -(CH 2 ) o (CHR 61 ) P C 6 H 4 R 8 ;
  • R 47 is H; alkyl; alkenyl; or -(CH 2 ) o (CHR 61 ) s OR 55 ;
  • R 48 is H; lower alkyl; lower alkenyl; or aryl-lower alkyl;
  • R 49 is H; alkyl; alkenyl; -(CHR 61 ) s COOR 57 ; (CHR 61 ) s CONR 58 R 59 ; (CHR 61 ) 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 61 ) s OR 55 ; -(CH 2 ) m (CHR 61 ) s SR 56 ; -(CH 2 ) m (CHR 61 ) s NR 33 R 34 ; -(CH 2 ) m (CHR 61 ) s OCONR 33 R 75 ; -(CH 2 ) m (CHR 61 ) NR 20 CONR 33 R 82 ; -(CH 2 ) o (CHR 61 ) s COOR 57 ; -(CH 2 ) o (CHR 61 ) s CONR 58 R 59 ; -(CH 2 ) o (CHR 61 ) p PO(OR 60 ) 2 ; -(CH 2 ) p (CHR 61 ) s SO 2 R 62 ; or -(CH 2 ) P (CHR )
  • R 52 is H; alkyl; alkenyl; -(CH 2 ) m (CHR 61 ) s OR 55 ; -(CH 2 ) m (CHR 61 ) s SR 56 ; -(CH 2 ) m (CHR 61 ) s NR 33 R 34 ; -(CH 2 ) m (CHR 61 ) s OCONR 33 R 75 ; -(CH 2 ) m (CHR 61 ) NR 20 CONR 33 R 82 ; -(CH 2 ) o (CHR 61 ) s COOR 57 ; -(CH 2 ) o (CHR 61 ) s CONR 58 R 59 ; -(CH 2 ) o (CHR 61 ) p PO(OR 60 ) 2 ; -(CH 2 ) p (CHR 61 ) s SO 2 R 62 ; or -(CH 2 ) P (CHR 6
  • R 53 is H; alkyl; alkenyl; -(CH 2 ) m (CHR 61 ) s OR 55 ; -(CH 2 ) m (CHR 61 ) s SR 56 ; - (CH 2 ) m (CHR 61 ) s NR 33 R 34 ; -(CH 2 ) m (CHR 61 ) s OCONR 33 R 75 ; -(CH 2 ) m (CHR 61 ) NR 20 CONR 33 R 82 ; -(CH 2 ) o (CHR 61 ) s COOR 57 ; -(CH 2 ) o (CHR 61 ) s CONR 58 R 59 ; -(CH 2 ) o (CHR 61 ) p PO(OR 60 ) 2 ; -(CH 2 ) P (CHR 61 ) s SO 2 R 62 ; or -(CH 2 ) P (CHR 61
  • R 54 is H; alkyl; alkenyl; -(CH 2 ) m (CHR 61 ) s OR 55 ; -(CH 2 ) m (CHR 61 ) s NR 33 R 34 ; -(CH 2 ) m (CHR 61 ) s OCONR 33 R 75 ; -(CH 2 ) m (CHR 61 ) s NR 20 CONR 33 R 82 ; -(CH 2 ) o (CHR 61 )COOR 57 ; -(CH 2 ) o (CHR 61 ) s CONR 58 R 59 ; or -(CH 2 ) o (CHR 61 ) s C ⁇ H 4 R ; R 55 is H; lower alkyl; lower alkenyl; aryl-lower alkyl; -(CH 2 ) m (CHR 61 ) s OR 57 ; -(CH 2 ) m (CHR 61
  • 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 5 is H; lower alkyl; lower alkenyl; aryl; heteroaryl; aryl-lower alkyl; or heteroaryl- lower alkyl; or
  • R 58 and R 59 taken together can form: -(CH 2 ) 2 . 6 -; -(CH 2 ) 2 O(CH 2 ) 2 -; -(CH 2 ) 2 S(CH 2 ) 2 ⁇ ; or -(CH 2 ) 2 NR 57 (CH 2 ) 2 -;
  • R 60 is H; lower alkyl; lower alkenyl; aryl; or aryl-lower alkyl;
  • R 61 is alkyl; alkenyl; aryl; heteroaryl; aryl-lower alkyl; heteroaryl-lower alkyl; -
  • 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; -COR 64 ; -COOR 57 ; -CONR 58 R 59 ; -SO 2 R 62 ; or -PO(OR 60 ) 2 ;
  • R 34 and R 63 taken together can form: -(CH 2 ) 2-6 -; -(CH 2 ) 2 O(CH 2 ) 2 -; -(CH 2 ) 2 S(CH 2 ) 2 -; or
  • R 64 is H; lower alkyl; lower alkenyl; aryl; heteroaryl; aryl-lower alkyl; heteroaryl-lower alkyl; -(CH 2 ) P (CHR 61 ) s OR 65 ; -(CH 2 ) P (CHR 61 ) s SR 66 ; or -(CH 2 ) P (CHR 61 ) s NR 34 R 63 ;
  • 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 66 is H; lower alkyl; lower alkenyl; aryl; aryl-lower alkyl; heteroaryl-lower alkyl; or
  • Z is a chain of 11 ⁇ -amino acid residues, 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, GIy, Pro, Pro(4NHCOPhe) or of formula -A-CO-, or of formula -B-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 H: -NR 20 -CH(CO-)-(CH 2 ) 4 - 7 -CH(CO-)-NR 20 -; -NR 20 -CH(CO-)-(CH 2 ) p SS(CH 2 ) p -CH(CO-)-NR 20 -;
  • R 71 is lower alkyl; lower alkenyl; -(CH 2 ) P (CHR 61 ) s OR 75 ; -(CH 2 ) P (CHR 61 ) s SR 75 ;
  • R 72 is H, lower alkyl; lower alkenyl; -(CH 2 ) P (CHR 61 ) s OR 85 ; or -(CH 2 ) P (CHR 61 ) s SR 85 ;
  • R 73 is -(CR 86 R 87 ) O R 77 ; -(CH 2 ) r O(CH 2 ) o R 77 ; -(CH 2 ) r S(CH 2 ) o R 77 ; or -(CH 2 ) r NR 20 (CH 2 ) o R 77 ;
  • N C(NR 78 R 80 )NR 79 R 80 ; -(CH 2 ) r O(CH 2 ) ra NR 78 R 79 ; - (CH 2 ) r O(CH 2 ) m NR 77 R 80 ;
  • R 75 is lower alkyl; lower alkenyl; or aryl-lower alkyl;
  • R 33 and R 75 taken together can form: -(CHz) 2-6 -; -(CH 2 ) 2 O(CH 2 ) 2 -; -(CH 2 ) 2 S(CH 2 ) 2 -; or
  • -(CH 2 ) 2 NR 57 (CH 2 ) 2 -; R 75 and R 82 taken together can form: -(CH 2 ) 2-6 -; -(CH 2 ) 2 O(CH 2 ) 2 -; -(CH 2 ) 2 S(CH 2 ) 2 -; or
  • R 76 is H; lower alkyl; lower alkenyl; aryl-lower alkyl; -(CH 2 ) o OR 72 ; -(CH 2 ) o SR 72 ;
  • R 77 is -C 6 R 67 R 68 R 69 R 70 R 76 ; or a heteroaryl group of one of the formulae
  • R is H; lower alkyl; aryl; or aryl-lower alkyl;
  • R 78 and R 82 taken together can form: -(CH 2 ) 2 . 6 -; -(CH 2 ) 2 O(CH 2 ) 2 -; -(CH 2 ) 2 S(CH 2 ) 2 -; or
  • 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 O(CH 2 ) 2 -; or -(CH 2 ) 2 NR 57 (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 33 and R 82 taken together can form: -(CH 2 ) 2 . 6 -; -(CH 2 ) 2 O(CH 2 ) 2 -; -(CH 2 ) 2 S(CH 2 ) 2 -; or
  • R 83 is H; lower alkyl; aryl; or -NR 78 R 79 ; R 84 is -(CH 2 ) m (CHR 61 ) s OH; -(CR 86 R 87 )pOR 80 ; -(CR 86 R 87 )pCOOR 80 ; -(CH 2 ) m (CHR 61 ) s SH; -
  • R 85 is lower alkyl; or lower alkenyl
  • R 86 is H; lower alkyl, where H is maybe substituted by halogen; or halogen;
  • R 87 is H; lower alkyl, where H is maybe substituted by halogen; or halogen; with the proviso that in said chain of 1 1 ⁇ -amino acid residues Z if n is 11, the amino acid residues in positions 1 to 11 are:
  • P2 of type C or of Type D or of type E, or of type F;
  • P5 of type E, or of type C, or of type F, or the residue is GIy or Pro;
  • GIy or Pro GIy or Pro
  • - P7 of type C, or of type E, or of type F, or of formula -A-CO-, or the residue is GIy or Pro;
  • P8 of type D, or of type C, or of type F, or of formula -A-CO, or the residue is GIy or Pro or Pro(4NHCOPhe);
  • P9 of type C, or of type D, or of type E, or of type F
  • - PlO of type D, or of type C, or of type F, or of type E
  • PI l of type C, or of type D, or of type E, or of type F; or
  • P2 and PlO taken together, can form a group of type H; and with the further proviso that if the template is D Pro L Pro, the amino acid residues in positions Pl to Pl 1 are other than - Pl : Arg
  • these ⁇ -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 5, 6 or 7, any functional group which may be present in said N-protected amino acid derivative being likewise appropriately protected;
  • X is as defined above and X is an N-protecting group or, alternatively, if
  • step (fa 1 ) coupling the product obtained in step (e) with an appropriately N- protected derivative of an amino acid of the above general formula III, any functional group which may be present in said N-protected amino acid derivative being likewise appropriately protected;
  • step (g) removing the N-protecting group from the product obtained in step (f) or (fc) or (fc 1 );
  • the peptidomimetics of the present invention can be prepared by
  • X is as defined above and X is an N-protecting group or, alternatively, if is to be group (al) or (a2), above,
  • the peptidomimetics of the present invention can also be enantiomers of the compounds of formula I. These enantiomers can be prepared by a modification of the above processes in which enantiomers of all chiral starting materials are used.
  • 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 , NO 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).
  • the structural element -B-CO- forms in combination with another structural element -B-CO- template (a3).
  • the template (a3) is less preferred in formula I.
  • Templates (a) through (p) constitute building blocks which have an N-terminus and a C-terminus oriented in space in such a way that the distance between those two groups may lie between 4.0-5.5A.
  • the peptide chain Z 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 serine protease inhibitory activity 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 the selective inhibitory activity but also for the synthesis process defined hereinabove, as incorporation of the templates at the beginning or near the middle of the linear protected peptide precursors enhances cyclization yields significantly.
  • 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- L B-CO- to D A69-CO- L B-CO-.
  • D Pro- L Pro constitutes the prototype of templates (al).
  • Less preferred, but possible are combinations
  • L Pro- D Pro constitutes the prototype of template (a2).
  • building blocks -Al-CO- to -A69-CO- in which A has (D)- conf ⁇ guration 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. Examples of preferred values for R 2 to R 17 are:
  • R 2 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 : 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 33 and R 34 taken together form:
  • 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 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; or R 33 and R 34 taken together form: -(CH 2 ) 2 .
  • R 1 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 : 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; or R 33 and R 34 taken together form: -(CH 2 ) 2-6 -; -(CH 2 ) 2 O(CH 2 ) 2 -; -(CH 2 ) 2 S(CH 2 ) 2 -; or -(CH 2 ) 2 NR 57 (CH 2 ) 2 -; where R 57 : H; or lower alkyl); -(CH 2 ) m OCONR 33 R 75 (where R 33 : H; or lower alkyl; or lower alkenyl; R 75 : lower
  • R 5 lower alkyl; lower alkenyl; -(CHz) 0 OR 55 (where R 55 : lower alkyl; or lower alkenyl); -(CH 2 ) o SR 56 (where R 56 : lower alkyl; or lower alkenyl); -(CH 2 ) o NR 33 R 34 (where R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl; or R 33 and R 34 taken together form: -(CH 2 ) Z-6 -; -(CHz) 2 O(CHz) 2 -; -(CH 2 ) 2 S(CH 2 ) 2 -; or -(CH 2 ) 2 NR 57 (CH 2 ) 2 -; where R 57 : H; or lower alkyl); -(CH 2 ) o OCONR 33 R 75 (where R 33 : H; or lower alkyl; or lower alkenyl; R 75 : lower alkyl; or
  • R 6 H; lower alkyl; lower alkenyl; -(CH 2 ) o OR 55 (where R 55 : lower alkyl; or lower alkenyl); -(CH 2 ) o SR 56 (where R 56 : lower alkyl; or lower alkenyl); -(CH 2 ) o NR 33 R 34 (where R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl; or R 33 and R 34 taken together form: -(CH 2 ) 2 .
  • 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 : 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; or R 33 and R 34 taken together form:
  • R 82 (where R 20 : H; or lower lower alkyl; R 33 : H; or lower alkyl; or lower alkenyl; R 82 : H; or lower alkyl; or R 33 and R 82 taken together form: -(CH 2 ) 2 . 6 -; -(CH 2 ) 2 O(CH 2 ) 2 -; -(CH 2 ) 2 S(CH 2 ) 2 -; or -(CH 2 ) 2 NR 57 (CH 2 ) 2 -; where R 57 : H; or lower alkyl);
  • R 20 H; or lower alkyl
  • R 64 lower alkyl; or lower alkenyl
  • R 58 lower alkyl; or lower alkenyl; and R 59 : H; or lower alkyl; or R 58 and R 59 taken together form:
  • R 33 lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl; or R 33 and R 34 taken together form:
  • R 33 H; or lower lower alkyl;
  • R 33 H; or lower alkyl; or lower alkenyl;
  • R 82 H; or lower alkyl; or
  • R 33 and R 82 taken together form: -(CH 2 W; -(CH 2 ) 2 O(CH 2 ) 2 -; -(CH 2 ) 2 S(CH 2 ) 2 -; or
  • R 1 lower alkyl; lower alkenyl; -(CH 2 ) o OR 55 (where R 55 : lower alkyl; or lower alkenyl); -(CH 2 ) o SR 56 (where R 56 : lower alkyl; or lower alkenyl); -(CH 2 ) o NR 33 R 34 (where R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl; or R 33 and R 34 taken together form:
  • R 11 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 : 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; or R 33 and R 34 taken together form: -(CH 2 W; -(CH 2 ) 2 O(CH 2 ) 2 -; -(CH 2 ) 2 S(CH 2 ) 2 -; or -(CH 2 ) 2 NR 57 (CH 2 ) r ; where R 57 : H; or lower alkyl); -(CH 2 ) m OCONR 33 R 75 (where R 33 : H; or lower alkyl; or lower alkenyl; R 75 : lower alkyl; or
  • R 12 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 : lower alkyl; or lower alkenyl); -(CH 2 )JNfR 33 R 34 (where
  • R 33 lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl; or R 33 and R 34 taken together form: -(CH 2 ) 2 . 6 -; -(CH 2 ) 2 O(CH 2 ) 2 -; -(CH 2 ) 2 S(CH 2 ) 2 -; or -(CH 2 ) 2 NR 57 (CH 2 ) 2 -; where R 57 :
  • R 33 H; or lower alkyl
  • R 75 lower alkyl; or R 33 and R 75 taken together form: -(CH 2 ) 2 . 6 -; -(CH 2 ) 2 O(CH 2 ) 2 -; -
  • 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 : 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; or R 33 and R 34 taken together form: -(CH 2 W; -(CH 2 ) 2 O(CH 2 ) 2 -; -(CH 2 ) 2 S(CH 2 ) r ; or -(CH 2 ) 2 NR 57 (CH 2 ) 2 -; where R 57 : H; or lower alkyl); -(CH 2 ) q OCONR 33 R 75 (where R 33 : H; or lower alkyl; or lower alkenyl; R 75 : lower alkyl; or R 33 and R 75 taken together form: -(CH 2 ) 2 . 6 -; -(CH 2 ) 2 O(CH 2 ) 2 -; -
  • R 8 (where R 8 : H; F; Cl; CF 3 ; lower alkyl; lower alkenyl; or lower alkoxy).
  • - 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 : 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; or R 33 and R 34 taken together form: -(CH 2 ) 2-6 -; -(CH 2 ) 2 O(CH 2 ) 2 -; -(CH 2 ) 2 S(CH 2 ) 2 -; or -(CH 2 ) 2 NR 57 (CH 2 ) 2 -; where R 57
  • R 15 lower alkyl; lower alkenyl; -(CH 2 ) o OR 55 (where R 55 : lower alkyl; or lower alkenyl); -(CH 2 ) o SR 56 (where R 56 : lower alkyl; or lower alkenyl); -(CH 2 ) o NR 33 R 34 (where R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl; or R 33 and R 34 taken together form:
  • R 58 and R 59 taken together form: -(CH 2 ) 2 -6-; -(CH 2 ) 2 O(CH 2 ) 2 -; -(CH 2 ) 2 S(CH 2 ) 2 -; or -(CH 2 ) 2 NR 57 (CH 2 ) 2 -; where R 57 : H; or lower alkyl); -(CH 2 ) o PO(OR 60 ) 2 (where R 60 : lower alkyl; or lower alkenyl); -(CH 2 ) o SO 2 R 62 (where R 62 : lower alkyl; or lower alkenyl); or
  • 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 ) o OR 55 (where R 55 : lower alkyl; or lower alkenyl); -(CH 2 ) o SR 56 (where R 56 : lower alkyl; or lower alkenyl); -(CH 2 ) o NR 33 R 34 (where R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl; or R 33 and R 34 taken together form:
  • R 33 H; or lower lower alkyl;
  • R 33 H; or lower alkyl; or lower alkenyl;
  • R 82 H; or lower alkyl; or
  • R 33 and R 82 taken together form: -(CH 2 W; -(CH 2 ) 2 O(CH 2 ) 2 -; -(CH 2 ) 2 S(CH 2 ) 2 -; or
  • R 57 H; or lower alkyl
  • R 57 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 ) o COOR 57 (where R 57 : lower alkyl; or lower alkenyl); -(CH 2 ) o CONR 58 R 59 (where R 58 : lower alkyl; or lower alkenyl; and R 59 : H; or lower alkyl; or R 58 and R 59 taken together form: -(CH 2 ) 2 . 6 -; -
  • R 17 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 : 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; or R 33 and R 34 taken together form:
  • building blocks Al to A69 the following are preferred: A5 with R 2 being H, A8, A22, A25, A38 with R 2 being H, A42, A47 and A50. Most preferred are building blocks of type A8':
  • R 20 is H or lower alkyl; and R 64 is alkyl; alkenyl; [(CH 2 ) u -X] r CH 3 , wherein X is -
  • O-, -NR 20 or -S-, u is 1-3 and t is 1-6; aryl; aryl-lower alkyl; or heteroaryl-lower alkyl; especially those wherein R 64 is n-hexyl (A8'-l); n-heptyl (A8'-2); 4-(phenyl)benzyl (A8'- 3); diphenylmethyl (A8'-4); 3 -amino-propyl (A8'-5); 5-amino-pentyl (A8'-6); methyl
  • Building block A70 belongs to the class of open-chain ⁇ -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.
  • 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 21 to R 29 in building blocks A70 to A104 are the following:
  • R 18 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 : 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; or R 33 and R 34 taken together form: -(CH 2 W; -(CH 2 ) 2 O(CH 2 ) 2 -; -(CH 2 ) 2 S(CH 2 ) 2 -; or -(CH 2 ) 2 NR 57 (CH 2 ) 2 -; where R 57 : H; or lower alkyl); -(CH 2 ) P OCONR 33 R 75 (where R 33 : H; or lower alkyl; or lower alkenyl; R 75 : lower alkyl; or R 33 and R 75 taken
  • R 2 ' H; lower alkyl; lower alkenyl; -(CH 2 ) o OR 55 (where R 55 : lower alkyl; or lower alkenyl); -(CH 2 ) o SR 56 (where R 56 : lower alkyl; or lower alkenyl); -(CH 2 ) o NR 33 R 34 (where R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl; or R 33 and R 34 taken together form:
  • R 22 lower alkyl; lower alkenyl; -(CH 2 ) o OR 55 (where R 55 : lower alkyl; or lower alkenyl); -(CH 2 ) o SR 56 (where R 56 : lower alkyl; or lower alkenyl); -(CH 2 ) o NR 33 R 34 (where R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl; or R 33 and R 34 taken together form: -(CH 2 ) 2 .
  • R 23 H; lower alkyl; lower alkenyl; -(CH 2 ) o OR 55 (where R 55 : lower alkyl; or lower alkenyl); -(CH 2 ) o SR 56 (where R 56 : lower alkyl; or lower alkenyl); -(CH 2 ) o NR 33 R 34 (where R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl; or R 33 and R 34 taken together, form: -(CH 2 W; -(CH 2 ) 2 O(CH 2 ) 2 -; -(CH 2 ) 2 S(CH 2 ) 2 -; or -(CH 2 ) 2 NR 57 (CH 2 ) 2 -; where R 57 : H; or lower alkyl); -(CH 2 ) o OCONR 33 R 75 (where R 33 : H; or lower alkyl; or lower alkenyl; R 75 : lower alky
  • R 82 (where R 20 : H; or lower lower alkyl; R 33 : H; or lower alkyl; or lower alkenyl; R 82 : H; or lower alkyl; or R 33 and R 82 taken together form: -(CH 2 ) 2-6 -; -(CH 2 ) 2 O(CH 2 ) 2 -; -(CH 2 ) 2 S(CH 2 ) 2 -; or -(CH 2 ) 2 NR 57 (CH 2 ) r ; where R 57 : H; or lower alkyl);
  • 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 34 (where R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl; or R 33 and R 34 taken together form: -(CH 2 ) 2-6 -; -(CH 2 ) 2 O(CH 2 ) 2 -; -(CH 2 ) 2 S(CH 2 ) 2 -; or -(CH 2 ) 2 NR 57 (CH 2 ) 2 -; where R 57 : H; or lower alkyl); -(CH 2 ) ra OCONR 33 R 75 (where R 33 : H; or lower alkyl; or lower alkenyl; R 75 : lower alkyl; or R 33 and R 75 taken together form: -
  • R 82 (where R 20 : H; or lower lower alkyl; R 33 : H; or lower alkyl; or lower alkenyl; R 82 : H; or lower alkyl; or R 33 and R 82 taken together form: -(CH 2 ) 2 . 6 -;
  • R 20 H; or lower alkyl
  • R 64 lower alkyl; or lower alkenyl
  • R 25 and R 26 taken together can be -(CH 2 W; -(CH 2 ) 2 O(CH 2 ) 2 -;
  • R 27 H; lower alkyl; lower alkenyl; -(CH 2 ) o OR 55 (where R 55 : lower alkyl; or lower alkenyl); -(CH 2 ) o SR 56 (where R 56 : lower alkyl; or lower alkenyl); -(CH 2 ) o NR 33 R 34 (where R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl; or R 33 and R 34 taken together form:
  • R 2li lower alkyl; lower alkenyl; -(CH 2 ) o OR 55 (where R 55 : lower alkyl; or lower alkenyl); -(CH 2 ) o SR 56 (where R 56 : lower alkyl; or lower alkenyl); -(CH 2 ) o NR 33 R 34 (where R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl; or R 33 and R 34 taken together form:
  • R 29 lower alkyl; lower alkenyl; -(CH 2 ) o OR 55 (where R 55 : lower alkyl; or lower alkenyl); -(CH 2 ) o SR 56 (where R 56 : lower alkyl; or lower alkenyl); -(CH 2 ) o NR 33 R 34 (where R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl; or R 33 and R 34 taken together form: -(CH 2 )M-; -(CH 2 ) 2 O(CH 2 ) r ; -(CH 2 ) 2 S(CH 2 ) r ; or -(CH 2 ) 2 NR 57 (CH 2 ) 2 -; where R 57 : H; or lower alkyl); -(CH 2 ) o OCONR 33 R 75 (where R 33 : H; or lower alkyl; or lower alkenyl; R 75 : lower alkyl;
  • R 23 , R 24 and R 29 is -NR 20 -CO-lower alkyl where R 20 is H or lower alkyl.
  • R 1 H; or lower Alkyl
  • R 8 H; F; Cl; CF 3 ; lower alkyl; lower alkenyl; -(CH 2 ) o OR 55 (where R 55 : lower alkyl; or lower alkenyl); -(CH 2 ) o SR 56 (where R 56 : lower alkyl; or lower alkenyl); - (CH 2 ) o NR 33 R 34 (where R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl; or R 33 and R 34 taken together form: -(CH 2 ) 2 .
  • R 62 lower alkyl; or lower alkenyl
  • R 8 -(CH 2 ) q C 6 H 4
  • R 20 H; or lower alkyl.
  • R 30 H, methyl.
  • R 31 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; or R 33 and R 34 taken together form: -(CH 2 ) 2 . 6 -; -(CH 2 ) 2 O(CH 2 ) r ; -(CH 2 ) 2 S(CH 2 ) 2 -; or
  • R 82 (where R 20 : H; or lower lower alkyl; R 33 : H; or lower alkyl; or lower alkenyl; R 82 : H; or lower alkyl; or R 33 and R 82 taken together form: -(CH 2 ) 2 . 6 -; -(CH 2 ) 2 O(CH 2 ) 2 -; -(CH 2 ) 2 S(CH 2 ) 2 -; or -(CH 2 ) 2 NR 57 (CH 2 ) 2 -; where R 57 : H; or lower alkyl);
  • R 33 lower alkyl; lower alkenyl; -(CH 2 ) m OR 55 (where R 55 : lower alkyl; or lower alkenyl); -(CH 2 ) m NR 34 R 63 (where R 34 : lower alkyl; or lower alkenyl; R 63 : H; or lower alkyl; or R 34 and R 63 taken together form: -(CH 2 )M-; -(CH 2 )2 ⁇ (CH 2 ) 2 -; -(CH 2 ) 2 S(CH 2 ) 2 -; or
  • R 58 lower alkyl; or lower alkenyl
  • R 59 H; lower alkyl; or R 58 and R 59 taken together form:
  • R 3S 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; or R 33 and R 34 taken together form: -(CH 2 W; -(CH 2 ) 2 O(CH 2 ) 2 -; -(CH 2 ) 2 S(CH 2 ) 2 -; or -(CH 2 ) 2 NR 57 (CH 2 ) 2 -; where R 57 : H; or lower alkyl); -(CH 2 ) m OCONR 33 R 75 (where R 33 : H; or lower alkyl; or lower alkenyl; R 75 : lower alkyl; or R 33 and R 75 taken together form: -
  • 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; or R 33 and R 34 taken together form: -(CH 2 ) 2 .
  • R 82 (where R 20 : H; or lower alkyl; R 33 : H; or lower alkyl; or lower alkenyl; R 82 : H; or lower alkyl; or R 33 and R 82 taken together form: -(CH 2 ) 2-6 -;
  • R 20 H; or lower alkyl
  • R 64 lower alkyl; or lower alkenyl
  • R 38 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; or R 33 and R 34 taken together form: -(CH 2 ) M S -(CH 2 ) 2 O(CH 2 ) 2 -; -(CH 2 ) 2 S(CH 2 ) 2 -; or -(CH 2 ) 2 NR 57 (CH 2 ) 2 -; where R 57 : H; or lower alkyl); -(CH 2 ) P OCONR 33 R 75 (where R 33 : H; or lower alkyl; or lower alkenyl; R 75 : lower alkyl; or R 33 and R 78 taken together form: -
  • R 82 (where R 20 : H; or lower lower alkyl; R 33 : H; or lower alkyl; or lower alkenyl; R 82 : H; or lower alkyl; or R 33 and R 82 taken together form: -(CH 2 ) 2 . 6 -; -(CH 2 ) 2 O(CH 2 ) 2 -; -(CH 2 ) 2 S(CH 2 ) 2 -; or -(CH 2 ) 2 NR 57 (CH 2 ) 2 -; where R 57 : H; or lower alkyl);
  • R 20 H; or lower alkyl
  • R 64 lower alkyl; or lower alkenyl
  • 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 ) o COOR 57 (where R 57 : lower alkyl; or lower alkenyl); -(CH 2 ) o CONR 58 R 59
  • R 58 lower alkyl; or lower alkenyl
  • R 59 H; lower alkyl; or R 58 and R 59 taken together form: -(CH 2 ) 2-6 s -(CH 2 ) 2 O(CH 2 ) 2 -; -(CH 2 ) 2 S(CH 2 ) 2 -; or -(CH 2 ) 2 NR 57 (CH 2 ) 2 -; where R 57 : H; or lower alkyl).
  • R 40 lower alkyl; lower alkenyl; or aryl-lower alkyl.
  • R 82 (where R 20 : H; or lower lower alkyl; R 33 : H; or lower alkyl; or lower alkenyl; R 82 : H; or lower alkyl; or R 33 and R 82 taken together form: -(CH 2 ) 2 . 6 -;
  • R 20 H; or lower alkyl
  • R 64 lower alkyl; or lower alkenyl
  • R 58 lower alkyl, or lower alkenyl
  • R 59 H; lower alky; or R 58 and R 59 taken together form: -(CH 2 ) 2 . 6 s -(CH 2 ) 2 O(CH 2 ) 2 -; -(CH 2 ) 2 S(CH 2 ) 2 -; or -(CH 2 ) 2 NR 57 (CH 2 ) 2 -; where R 57 : H; or lower alkyl); -(CH 2 ) o PO(OR 60 ) 2 (where R 60 : lower alkyl; or lower alkenyl); -
  • 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; or R 33 and R 34 taken together form: -(CH 2 ) 2 . fi -; -(CH 2 ) 2 O(CH 2 ) 2 -; -(CH 2 ) 2 S(CH 2 ) 2 -; or
  • R 82 (where R 20 : H; or lower lower alkyl; R 33 : H; or lower alkyl; or lower alkenyl; R 82 : H; or lower alkyl; or R 33 and R 82 taken together form: -(CH 2 ) 2 . 6 -; -(CH 2 ) 2 O(CH 2 ) 2 -; -(CH 2 ) 2 S(CH 2 ) 2 -; or -(CH 2 ) 2 NR 57 (CH 2 ) 2 -; where R 57 : H; or lower alkyl);
  • R 20 H; or lower alkyl
  • R 64 lower alkyl; or lower alkenyl
  • R 58 lower alkyl, or lower alkenyl
  • R 59 H; lower alkyl; or R 58 and R 59 taken together form:
  • R 33 H; or lower lower alkyl; R 33 : H; or lower alkyl; or lower alkenyl; R 82 : H; or lower alkyl; or R 33 and R 82 taken together form: -(CH 2 ) 2 . 6 -; -(CH 2 ) 2 O(CH 2 ) r ; -(CH 2 ) 2 S(CH 2 ) 2 -; or -(CH 2 ) 2 NR 57 (CH 2 ) 2 -; where R 57 : 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 ) o COOR 57 (where R 57 : lower alkyl; or lower alkenyl); -(CH 2 ) o CONR 58 R 59 (where R 58 : lower alkyl; or lower alkeny
  • R 1'1 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 : 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; or R 33 and R 34 taken together form:
  • R 33 H; or lower lower alkyl; R 33 : H; or lower alkyl; or lower alkenyl; R 82 : H; or lower alkyl; or R 33 and R 82 taken together form: -(CH 2 W; -(CH 2 ) 2 O(CH 2 ) 2 -; -(CH 2 ) 2 S(CH 2 ) r ; or -(CH 2 ) 2 NR 57 (CH 2 ) 2 -; where R 57 : 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;
  • K 15 H; lower alkyl; lower alkenyl; -(CH 2 ) o OR 55 (where R 55 : lower alkyl; or lower alkenyl); -(CH 2 ) o SR 56 (where R 56 : lower alkyl; or lower alkenyl); -(CH 2 ) o NR 33 R 34 (where
  • R 33 lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl; or R 33 and R 34 taken together form: -(CH 2 W; -(CH 2 ) 2 O(CH 2 ) r ; -(CH 2 ) 2 S(CH 2 ) 2 -; or -(CH 2 ) 2 NR 57 (CH 2 ) 2 -; where R 57 : H; or lower alkyl); -(CH 2 ) s OCONR 33 R 75 (where R 33 : H; or lower alkyl; or lower alkenyl; R 75 : lower alkyl; or R 33 and R 75 taken together form: -(CH 2 W; -(CH 2 ) 2 O(CH 2 ) 2 -; -
  • R 16 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 : lower alkyl; or lower alkenyl); -(CH 2 ) s NR 33 R 34 (where R 33 : lower alkyl; or lower alkenyl; R 34 : H; or lower alkyl; or R 33 and R 34 taken together form:
  • R 48 H; or lower alkyl.
  • R 49 H;lower alkyl; -(CH 2 ) o COOR 57 (where R 57 : lower alkyl; or lower alkenyl); -(CH 2 ) o CONR 58 R 59 (where R 58 : lower alkyl; or lower alkenyl; and R 59 : H; lower alkyl; or R 58 and R 59 taken together form: -(CH 2 ) 2 .
  • R 51 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; or R 33 and R 34 taken together form: -(CH 2 ) 2 . 6 -; -(CH 2 ) 2 O(CH 2 ) 2 -; -(CH 2 ) 2 S(CH 2 ) 2 -; or
  • R 82 (where R 20 : H; or lower lower alkyl; R 33 : H; or lower alkyl; or lower alkenyl; R 82 : H; or lower alkyl; or R 33 and R 82 taken together form: -(CH 2 ) 2 . 6 -;
  • R 20 H; or lower alkyl
  • R 64 lower alkyl; or lower alkenyl
  • R 58 lower alkyl; or lower alkenyl
  • R 59 H; lower alkyl; or R 58 and R 59 taken together form: -(CH 2 ) M -; -(CH 2 ) 2 O(CH 2 ) 2 -; -(CH 2 ) 2 S(CH 2 ) 2 -; or -(CH 2 ) 2 NR 57 (CH 2 ) 2 -; where R 57 : H; or lower alkyl); or -(CH 2 )AH 4 R 8 (where R 8 : H; F; Cl; CF 3 ; lower alkyl; lower alkenyl; or lower alkoxy).
  • 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; or R 33 and R 34 taken together form: -(CH 2 ) 2-6 -; -(CH 2 ) 2 O(CH 2 ) 2 -; -(CH 2 ) 2 S(CH 2 ) 2 -; or
  • R 82 (where R 20 : H; or lower lower alkyl; R 33 : H; or lower alkyl; or lower alkenyl; R 82 : H; or lower alkyl; or R 33 and R 82 taken together form: -(CH 2 ) 2 . 6 -;
  • R 58 lower alkyl; or lower alkenyl
  • R 59 H; lower alkyl; or R 58 and R 59 taken together form:
  • 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; or R 33 and R 34 taken together form: -(CH 2 ) 2 . 6 -; -(CH 2 ) 2 O(CH 2 ) 2 -; -(CH 2 ) 2 S(CH 2 ) 2 -; or
  • R 82 (where R 20 : H; or lower lower alkyl; R 33 : H; or lower alkyl; or lower alkenyl; R 82 : H; or lower alkyl; or R 33 and R 82 taken together form: -(CH 2 ) 2 . 6 -;
  • R 20 H; or lower alkyl
  • R 64 lower alkyl; or lower alkenyl
  • R 58 lower alkyl; or lower alkenyl
  • R 59 H; lower alkyl; or R 58 and R 59 taken together form: -(CH 2 ) 2 . 6 -; -(CH 2 ) 2 O(CH 2 ) 2 -; -(CH 2 ) 2 S(CH 2 ) 2 -; or -(CH 2 ) 2 NR 57 (CH 2 ) 2 -; where R 57 : H; or lower alkyl); or -(CH 2 )AH 4 R 8 (where R 8 : H; F; Cl; CF 3 ; lower alkyl; lower alkenyl; or lower alkoxy).
  • R 54 lower alkyl; lower alkenyl; or aryl-lower alkyl.
  • R 1 is H; R 20 is H; R 30 is H; R 31 is carboxymethyl; or lower alkoxycarbonylmethyl; R 32 is H; R 35 is methyl; R 36 is methoxy; R 37 is H and R 38 is H.
  • the building block -B-CO- within templates (al), (a2) and (a3) 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 64 is alkyl; alkenyl; -[(CH 2 ) u -X]rCH 3 (where X is
  • R 64 is n-hexyl (A8"-21); n-heptyl (A8"-22); 4- (phenyl)benzyl (A8"-23); diphenylmethyl (A8"-24); 3-amino-propyl (A8"-25); 5- amino-pentyl (A8"-26); methyl (A8"-27); ethyl (A8"-28); isopropyl (A8"-29); isobutyl (A8"-30); n-propyl (A8"-31); cyclohexyl (A8"-32); cyclohexylmethyl (A8"-33); n-butyl (A8"-34); phenyl (A8 M -35);
  • the peptidic chain Z of the ⁇ -hairpin mimetics described 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- or of formula -B-CO- wherein A and B are as defined above.
  • GIy can also be an amino acid residue in chain Z
  • Pro and Pro(4-NHCOPhe) can be amino acid residues in chain Z, too, with the exception of positions where an interstrand linkage (H) is possible.
  • 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 .
  • residues 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 and
  • Group E comprises amino acids containing side chains with polar-cationic, acylamino- and urea-derived residues according to the general definition for substituent 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 urea derived amino acid 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 being 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, carboxyclic 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 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. , Solid Phase Peptide Synthesis, 2d Ed., Pierce Chemical Company, III., 1984; Ahmed et al. J. Biol. Chem.
  • disulfide linkages can be prepared 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 5 N- carbonylimidazole to form cyclic ureas.
  • positions for an interstrand linkage are positions P2 and 10, taken together.
  • 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 abbreviations corresponding to generally adopted usual practice:
  • residues for group C are: Ala L-Alanine
  • residues for group D are:
  • residues for group F are Asn L-Asparagine
  • the peptidic chain Z within the ⁇ -hairpin mimetics of the invention comprises 11 amino acid residues.
  • the positions Pl to Pl 1 of each amino acid residue in the chain Z are unequivocally defined as follows: Pl 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), or of the group -B-CO- forming the C-terminus of template (a3); and Pl 1 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), or of the group - B-CO- forming the N-terminus of template (a3).
  • Each of the positions Pl to Pl 1 will preferably contain an amino acid residue belonging to one of the above types C, D, E, F, H, or of formula -A-CO- or of formula -B-CO-, or being GIy, Pro or Pro(4NHCOPhe) as follows:
  • ⁇ -amino acid residues in positions 1 to 11 of the chain Z are preferably: - Pl : of type C, or of type D, or of type E, or of type F;
  • P2 of type E, or of type F, or of type C;
  • P4 of type C, or of type E, or of type F, or the residue is GIy or Pro;
  • P5 of type E, or of type F, or the residue is GIy or Pro
  • - P6 of type C, or of type D, or of type F, or the residue is GIy or Pro;
  • P7 of type F or of formula -A-CO-or the residue is GIy or Pro;
  • P8 of type D, or of type C, or of formula -A-CO or the residue is GIy or Pro or Pro(4NHCOPhe);
  • P9 of type C, or of type D, or of type E, or of type F
  • - PlO of type F, or of type C, or type E
  • PI l of type E, or of type F, or of type C or of type D; or
  • ⁇ -amino acid residues in positions 1 to 11 are most preferably:
  • Pl NIe, He, Aoc, hLeu, Chg, OctG, hPhe, 4AmPhe 3 Cha, Phe, Tyr, 2C1-Phe, Trp, 1 -NaI, Leu, Cha, or Arg;
  • P2 Cys, GIu, NIe, Thr, or GIn ;
  • P3 Thr, Ala or Abu;
  • P4 Lys, NIe, Ala, Abu, or Thr;
  • P5 Ser, AlloThr, or Dpr;
  • - P6 He, c5al, Leu, NIe, Aoc, OctG, Cha, hLeu, hPhe, Chg, t-BuA, GIu 5 or Asp;
  • P7 Pro;
  • P8 Pro, Ala, or Pro(4NHCOPhe);
  • P9 Tyr, Phe, He, NIe, Cha, GIn, Arg, Lys, His, Thr, or Ala; - PlO: Cys, Arg, NIe, GIn, Lys, Met, Thr, or Ser;
  • PI l Tyr, GIn, Arg, Ser, NIe, 2-Nal, 2C1-Phe, Cha, Phg, Tyr, Phe, Asp,
  • Cys if present at P2 and PlO, may form a disulfide bridge.
  • ⁇ -amino acid residues in positions 1 to 1 1 of the chain Z are preferably:
  • Pl of type C, or of type D, or of type E;
  • P2 of type F, or of type C
  • - P3 of type F
  • P4 of type C, or of type E;
  • P5 of type E, or of type F;
  • P7 of type F, or of formula -A-CO-, or the residue is GIy or Pro
  • - P8 of type C, or of formula -A-CO-, or the residue is GIy or Pro or
  • P9 of type C, or of type D, or of type F;
  • PlO of type F, or of type C, or type E;
  • PI l of type E, or of type D, or of type F; or - P2 and PlO, taken together, form a group of type H.
  • the ⁇ -amino acid residues in positions 1 to 11 are most preferably
  • Pl Phe, hPhe, 4AmPhe, NIe, Chg, He, Tyr, Arg, Trp, 2C1-Phe, Arg, 1 - NaI, or Cha;
  • P2 Cys, GIu, or NIe;
  • P4 Lys, or NIe
  • P5 Ser, AlloThr, or Dpr
  • - P6 Asp, or GIu
  • P9 lie, NIe, Cha, GIn, Tyr, or Ala;
  • PlO Cys, Arg, or NIe
  • - PI l Thr, Asp, Ser, Tyr, Phe, Asn, or Arg
  • Cys if present at P2 and PlO, may form a disulfide bridge.
  • the ⁇ -amino acid residues in positions 1 to 11 of the chain Z are preferably - Pl : of type C, or of type D;
  • P3 of type F or of type C;
  • P4 of type C or of type F;
  • P7 of formula -A-CO-or the residue is GIy or Pro;
  • P8 of formula -A-CO or the residue is GIy or Pro or
  • P9 of type D, or of type F or of type C
  • - PlO of type F, or of type C, or type E
  • Pl 1 of type E, or of type F, or of type D; or
  • ⁇ -amino acid residues in positions 1 to 1 1 are most preferably:
  • P2 Cys, GIu, Thr, or GIn;
  • P3 Thr, Ala, or Abu
  • P4 Ala, Thr, or Abu
  • - P5 Ser
  • P6 OctG, He, Cha, Leu, c5al, NIe, Aoc, Chg, tBuA, or hLeu;
  • P8 Pro, or Pro(4NHCOPhe);
  • P9 GIn, Tyr, ILe, or Phe;
  • PlO Cys, Lys, GIn, Thr, Met, or Arg
  • PI l Tyr, Ser, Arg, GIn, NIe, 2-Nal, 2C1-Phe, Phe, Cha, or Phg;
  • Cys if present at P2 and PlO, may form a disulfide bridge.
  • ⁇ -amino acid residues in positions 1 to 11 of the chain Z are preferably:
  • Pl of type C, or of type D, or of type E;
  • P6 of type C, or of type D;
  • P7 of type F, or of formula -A-CO-, or the residue is GIy or Pro;
  • P8 of type C, or of formula -A-CO-, or the residue is GIy or Pro;
  • - P9 of type C, or of type E, or of type F;
  • ⁇ -amino acid residues in positions 1 to 11 of the chain Z are most preferably:
  • Particularly preferred ⁇ -peptidomimetics of the invention include those described in Examples 5, 19, 20, 22, 23, 38, 39, 40, and 75 as inhibitors of cathepsin G; Examples 91, 121, 153, 154, 155, 156, 157, 158, 159, 160, 161 177, and 178 as inhibitors of elastase; and Examples 193, 194, and 195 as inhibitors of Tryptase.
  • the processes of the invention can advantageously be carried out as parallel array syntheses to yield libraries of template-fixed ⁇ -hairpin peptidomimetics of the above general formula I.
  • Such parallel syntheses allow one to obtain arrays of numerous
  • 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 Synthesis of Small-Molecular-Weight Compound Libraries", Tetrahedron Organic Chemistry Series, 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.
  • 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.
  • 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.
  • Type 1 linkers are designed to release the amide group under acidic conditions (Rink H, Tetrahedron Lett. 1987, 28, 3783-3790).
  • Linkers of this kind form amides of the carboxyl group of the amino acids; examples of resins functionalized by such linker structures include 4-[(((2,4-dimethoxyphenyl)Fmoc-aminomethyl)phenoxyacetamido) aminomethyl] PS resin, 4-[(((2,4-dimethoxyphenyl)Fmoc- aminomethyl)phenoxyacetamido) aminomethyl] -4-methylbenzydrylamine PS resin (Rink amide MBHA PS Resin), and 4-[(((2,4-dimethoxyphenyl)Fmoc- aminomethyl)phenoxyacetamido) aminomethyl] benzhydrylamine PS-resin (Rink amide BHA PS resin).
  • the support is derived from polystyrene crosslinked with, most preferably 1-5%, divinylbenzene and functionalized by means of the 4- (((2,4- dimethoxyphenyl)Fmoc-aminomethyl)phenoxyacetamido) linker.
  • Type 2 linkers are designed to eventually release the carboxyl group under acidic conditions.
  • Linkers of this kind form acid-labile esters with the carboxyl group of the amino acids, usually acid-labile benzyl, benzhydryl and trityl esters; examples of such linker structures 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.
  • 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
  • the appropriate functionalized solid support which is preferably derived from polystyrene cross-linked with 1 to 3% of divinylbenzene, or from 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) 3 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
  • 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
  • guanidino group (as is present e. g. in the side-chain of arginine)
  • Ts tosyl i. e. p-toluenesulfonyl
  • Trt trityl and for the mercapto group (as is present e. g. in the side-chain of cysteine) Acm acetamidomethyl tBu tert.-butyl
  • the 9-fluorenylmethoxycarbonyl- (Fmoc)-protected amino acid derivatives are preferably used as the building blocks for the construction of the template-fixed ⁇ -hairpin 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 acid derivative, is usually 1 to 20 equivalents based on the milliequivalents per gram (meq/g) loading of the functional ized 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 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), the resulting dicyclohexylurea and diisopropylurea is insoluble and, respectively, soluble in the solvents generally used.
  • 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 (DPPA) or O-(7-aza-benzotriazol-l-yl)-N,N,N',N ! - tetramethyluronium tetrafluoroborate (TATU) or O-(7-aza-benzotriazol-l-yl)-N,N,N',N'- tetramethyluronium hexafiuorophosphate (HATU)/7-aza-l -hydroxy benzotriazole (HOAt, Carpino et al., Tetrahedron Lett. 1994, 35, 2279-2281) have also been used as coupling reagents.
  • DPPA diphenoxyphosphoryl azide
  • TATU O-(7-aza-benzotriazol-l-yl)-N,N,N',N ! - 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; 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.
  • 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 by-product removal by methods such as TLC, GC, or inspection of the washings.
  • an amino group can be acylated by means of an acylating agent corresponding to the acyl substituent to be introduced.
  • pegylated amino acids such as IPegK, or SPegK
  • HATU N-[(dimethylamino)-lH-l ,2,3-triazolo[4,5- ⁇ >]pyridin-l- ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide
  • DIPEA Diisopropyl ethylamine
  • 2-[2-(2-methoxyethoxy)ethoxy] acetic acid (IPeg) and, respectively, 2-(2- methoxyethoxy)acetic acid (sPeg) is applied to the liberated amino group of the appropiate amino acid side chain for 3 h.
  • 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 cysteine and homocysteine residues at opposite positions of the ⁇ -strand; or lactam bridges formed by glutamic and aspartic acid residues linking ornithine and, respectively, lysine residues, or by glutamic acid residues linking 2,4- diaminobutyric acid residues located at opposite ⁇ -strand positions by amide bond formation.
  • groups of the type H which can, for example, be disulfide bridges formed by cysteine and homocysteine residues at opposite positions of the ⁇ -strand; or lactam bridges formed by glutamic and aspartic acid residues linking ornithine and, respectively, lysine residues, or by glutamic acid residues linking 2,4- diaminobutyric acid residues located at opposite ⁇ -strand positions by amide bond formation.
  • a solution of 10 equivalents of iodine solution is applied in DMF or in a mixture Of CH 2 Cb /MeOH for 1.5 h which is repeated for another 3h with a fresh iodine solution after filtering of the iodine solution, or in a mixture of DMSO and acetic acid solution, buffered with 5% with NaHCO 3 to pH 5-6 for 4h, or in water adjusted to pH 8 with ammonium hydroxide solution by stirring for 24h, or in ammonium acetate buffer adjusted to pH 8 in the presence of air, or in a solution of NMP and tri-n- butylphosphine (preferably 50 eq.).
  • 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.
  • Cyclization is then effected in solution using solvents such as DCM, DMF, dioxane, THF and the like.
  • 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 16 hours.
  • the progress of the reaction is followed, e. g. by RP-HPLC (Reverse Phase High Performance Liquid Chromatography).
  • the solvent is removed by evaporation, the fully protected cyclic peptide derivative is dissolved in a solvent which is not miscible with water, such as DCM, and the solution is extracted with water or a mixture of water- miscible solvents, in order to remove any excess of the coupling reagent.
  • the fully protected peptide derivative is treated with 95% TFA, 2.5% H 2 O, 2.5% TIS or another combination of scavengers for effecting the cleavage of protecting groups.
  • the cleavage reaction time is commonly 30 minutes to 12 hours, preferably about 2.5 hours.
  • the volatiles are evaporated to dryness and the crude peptide is dissolved in 20% AcOH in water and extracted with isopropyl ether or other solvents which are suitable therefor.
  • the aqueous layer is collected and evaporated to dryness, and the fully deprotected cyclic peptide derivative of formula I is obtained as end-product.
  • the detachment, cyclization and complete deprotection of the fully protected peptide from the solid support can be achieved manually in glass vessels.
  • this peptide derivative can be used directly for biological assays, or it has to be further purified, for example by preparative HPLC.
  • the ⁇ -hairpin peptidomimetics of the invention can be used in a wide range of applications where inflammatory diseases or pulmanory diseases or infections or immunological diseases or cardiovascular diseases or neurodegenerative diseases are mediated or resulting from serine protease activity, or where cancer is mediated or resulting from serine protease activity.
  • the ⁇ -hairpin peptidomimetics may be administered per se or may be applied as an appropriate formulation together with carriers, diluents or excipients well known in the art.
  • the ⁇ - hairpin peptidomimetics can be administered singly, as mixtures of several ⁇ -hairpin peptidomimetics, in combination with other anti-inflammatory agents, or antimicrobial agents or anti-cancer agents and/or in combination with other pharmaceutically active agents.
  • the ⁇ -hairpin peptidomimetics can be administered per se or as pharmaceutical compositions.
  • compositions comprising ⁇ -hairpin 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 ⁇ -hairpin peptidomimetics into preparations which can be used pharmaceutically. Proper formulation depends upon the method of administration chosen.
  • the ⁇ -hairpin peptidomimetics of the invention may be formulated as solutions, gels, ointments, creams, suspensions, etc. as are well-known in the art.
  • 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.
  • the ⁇ -hairpin peptidomimetics of the invention may be formulated in adequate solutions, preferably in physiologically compatible buffers such as Hink's solution, Ringer's solution, or physiological saline buffer.
  • the solutions may contain formulatory agents such as suspending, stabilizing and/or dispersing agents.
  • the ⁇ -hairpin peptidomimetics of the invention may be in powder form for combination with a suitable vehicle, e.g., sterile pyrogen-free water, before use.
  • a suitable vehicle e.g., sterile pyrogen-free water
  • penetrants appropriate to the barrier to be permeated are used in the formulation as known in the art.
  • the ⁇ -hairpin peptidomimetics of the invention can be readily formulated by combining them with pharmaceutically acceptable carriers well known in the art.
  • Such carriers enable the ⁇ -hairpin peptidomimetics of the invention to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions etc., for oral ingestion by a patient to be treated.
  • suitable excipients include fillers such as sugars, e. g.
  • 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.
  • des integrating agents may be added, such as cross-linked polyvinylpyrrolidones, agar, or alginic acid or a salt thereof, such as sodium alginate.
  • solid dosage forms may be sugar-coated or enteric-coated using standard techniques.
  • suitable carriers, excipients or diluents include water, glycols, oils, alcohols, etc.
  • flavoring agents, preservatives, coloring agents and the like may be added.
  • the composition may take the form of tablets, lozenges, etc., formulated as usual.
  • the ⁇ -hairpin peptidomimetics 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.
  • a suitable propellant e.g. dichlorodifluoromethane, trichlorofluromethane, carbon dioxide or another suitable gas.
  • 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 ⁇ -hairpin 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.
  • the ⁇ -hairpin 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.
  • the ⁇ -hairpin 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.
  • ⁇ -hairpin peptidomimetics of the invention may be delivered using a sustained-release system, such as semipermeable matrices of solid polymers containing the therapeutic agent.
  • 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.
  • ⁇ -hairpin pepdidomimetics of the invention may contain charged residues, they may be included in any of the above-described formulations as such or as pharmaceutically acceptable salts.
  • Pharmaceutically acceptable salts tend to be more soluble in aqueous and other protic solvents than are the corresponding free forms.
  • ⁇ -hairpin peptidomimetics of the invention will generally be used in an amount effective to achieve the intended purpose. It is to be understood that the amount used will depend on a particular application.
  • a therapeutically effective dose can be determined using, for example, the in vitro assays provided in the examples.
  • the treatment may be applied while the disease is visible, or even when it is not visible.
  • An ordinary skilled expert will be able to determine therapeutically effective amounts to treat topical diseases without undue experimentation.
  • a therapeutically effective dose can be estimated initially from in vitro assays.
  • a dose can be formulated in animal models to achieve a circulating ⁇ -hairpin peptidomimetic concentration range that includes the IC 50 as determined in the cell culture. Such information can be used to more accurately determine useful doses in humans.
  • 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 skill in the art could readily optimize administration to humans based on animal data.
  • Dosage amounts for applications as serine protease inhibitory agents may be adjusted individually to provide plasma levels of the ⁇ -hairpin peptidomimetics of the invention which are sufficient to maintain the therapeutic effect.
  • Therapeutically effective serum levels may be achieved by administering multiple doses each day.
  • the effective local concentration of the ⁇ -hairpin peptidomimetics of the invention may not be related to plasma concentration.
  • One having the ordinary skill in the art will be able to optimize therapeutically effective local dosages without undue experimentation.
  • the amount of ⁇ -hairpin 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.
  • a therapeutically effective dose of the ⁇ -hairpin peptidomimetics described herein will provide therapeutic benefit without causing substantial toxicity.
  • Toxicity of the ⁇ -hairpin peptidomimetics of the invention can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., by determining the LD 50 (the dose lethal to 50% of the population) or the LD 100 (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 ⁇ -hairpin 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.1, p.l).
  • HBTU 1-benzotriazol-l-yl-tetramethylurounium hexafluorophosphate
  • HOBt 1-hydroxybenzotriazole
  • DIEA diisopropylethylamine
  • HOAT 7-aza-l-hydroxybenzotriazole
  • HATU O-(7-aza-benzotriazo Ie- 1 -y 1)-N,N,N ' ,N ' -tetramethy lurono i um hexafluorophosphate (Carpino et al. Tetrahedron Lett. 1994, 35, 2279-2281).
  • the resin was shaken (CH 2 Cl 2 /MeOH/DIEA : 17/2/1), 30 ml for 30 min; then washed in the following order with CH 2 Cl 2 (Ix), DMF (Ix), CH 2 Cl 2 (Ix), MeOH (Ix), CH 2 Cl 2 (Ix), MeOH (Ix), CH 2 Cl 2 (2x), Et 2 O (2x) and dried under vacuum for 6 hours. Loading was typically 0.6-0.7 mMol/g.
  • the following preloaded resins were prepared: Fmoc-Pro-2-chlorotritylresin, Fmoc-Asp (OtBu)-2-chlorotritylresin, Fmoc-Pro(5RPhe)-2-chlorotritylresin, Fmoc-Leu-2-chlorotritylresin, Fmoc-Glu(OtBu)-2-chlorotritylresin, Fmoc- Asp(OtBu) -2-chlorotritylresin,.
  • Fmoc-Phe-2-chlorotritylresin Fmoc-Gln(Trt)-2- chlorotritylresin, Fmoc-Ser (OtBu) -2-chlorotritylresin, Fmoc-Val-2- chlorotritylresin, Fmoc-Thr(OtBu) -2-chlorotritylresin and Fmoc-Ile-2- chlorotritylresin.
  • Steps 3 to 6 are repeated to add each amino-acid.
  • Procedure A i. e. disulfide ⁇ -strand linkages
  • the fully protected linear peptide was dissolved in DMF (8 ml, cone. 10 mg/ml). Two eq. of HATU (0.72 mMol) in ImI of DMF and 4 eq. of DIEA (1.44 mMol) in I mI of DMF were added, and the mixture was stirred at room temperature for 16 h. The volatile was evaporated to dryness.
  • the crude cyclised peptide was dissolved in 7 ml Of CH 2 Cl 2 and extracted with 10% acetonitrile in water (4.5 ml) three times. The CHbCl 2 layer was evaporated to dryness.
  • cyclic peptide obtained was dissolved in 3 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 2O 0 C for 2.5 hours and then concentrated under vacuum. The crude peptide was dissolved in 20% AcOH in water (7 ml) and extracted with diisopropylether (4 ml) three times. The aqueous layer was collected and evaporated to dryness, and the residue was purified by preparative reverse phase LC-MS.
  • TFA trifluoroacetic acid
  • TIS triisopropylsilane

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Abstract

Templates-fixed β-hairpin peptidomimetics of the general formulae (I), wherein Z is a chain of 11 α-amino acid residues which, depending on their positions in the chain (counted starting from the N-terminal amino acid) are Gly, or Pro, or Pro(4NHCOPhe), or of certain types which , as the remaining symbols in the above formula, are defined in the description and the claims, and salts thereof, have the property to inhibit proteases, in particular serine proteases, especially Cathepsin G or Elastase or Tryptase. These β-hairpin peptidomimetics can be manufactured by processes which are based on a mixed solid- and solution phase synthetic strategy.

Description

TEMPLATE-FIXED BETA-HAIRPIN PEPTIDOMIMETICS WITH PROTEASE INHIBITORY ACTIVITY
5 The present invention provides template-fixed β-hairpin peptidomimetics incorporating a template-fixed chain of 11 α-amino acid residues which, depending on their position in the chain, are GIy, or Pro, or Pro(4NHCOPhe), or are 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 10 such as human cathepsin G, elastase, or tryptase. In addition the present invention provides an efficient process by which these compounds can, if desired, be made in library-format.
The β-hairpin peptidomimetics of the invention show improved efficacy, oral 15 bioavailability, improved half-life and most importantly a high selectivity ratio among different serine proteases which depends on the proper choice of certain types of α-amino acid residues and their position in said chain. In addition these β-hairpin peptidomimetics show a low hemolysis on red blood cells and low cytotoxicity.
20 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, T. Rockway, Expert Opin. Ther. Patents 2003, 13, 773-786), parasitic,
25 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 ); 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.
30 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. Wickersham, Z. Hostomsky, N. Habuka, E. W. Moomaw, T. Adachi, Z. Hostomska, Cell, 1996, 87, 331 -342), herpes (W.
35 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. E. Hugli, Trends Biotechnol. 1996, 14, 409-12, ), cardiovascular (M. T. Stubbs, W. A. Bode, Thromb. Res. 1993, 69, 1-58; H. Fukami et al, Current Pharmaceutical Design 1998, 4, 439-453), and neurodegenerative defects including Alzheimer's disease (R. Vassar, B. D. Bennett, S. Babu-Kahn, S. Kahn, E. A. Mendiaz, Science, 1999, 286, 735-41), angiogenesis (Kaatinen M et al, Atherosklerosis 1996, 123 1-2, 123-131) and multiple sclerosis (Ibrahim MZ et al, J. Neuroimmunol 1996, 70, 131-138.
As most proteases bind their substrates in extended or β-strand 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 Pl, as either trypsin-like (positively charged residues Lys/Arg preferred at Pl), elastase-like (small hydrophobic residues Ala/Val at Pl), or chymotrypsin-like (large hydrophobic residues Phe/Tyr/Leu at Pl). Serine proteases for which protease-inhibitor X-ray crystal data is available on the PDB data base (PDB: www.rcsb.org/pdb) include trypsin, α-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 IXA, 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. LiIa5 P. Gloanec, L. Cadet, Y. Herve, J. Fournier, F. Leborgne, T. J. Verbeuren, G. DeNanteuil, Synth. Comm. 1998, 28, 4419-29) and factor Xa (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.
Strimpter, 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), urokinase inhibitors for breast cancer, and chymase inhibitors for heart related diseases. 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 elastase and cathepsin G constitute valuable targets for novel drug candidates particularly for chronic obstructive pulmonary disease (Ohbayashi H, Epert Opin. Investig. Drugs 2002, 11, 965-980).
Of the many occurring proteinaceous serine protease inhibitors, one is a 14 amino acid cyclic peptide from sunflower seeds, termed sunflower trypsin inhibitor (SFTI-I) (S. Luckett, R. Santiago Garcia, J. J. Barker, A. V. Konarev, P. R. Shewry, A. R. Clarke, R. L. Brady, J. 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 trypsin- reactive loop of the Bowman-Birk family of serine protease inhibitors. The inhibitor adopts a β-hairpin conformation when bound to the active site of bovine β-trypsin. SFTI- 1 inhibited β-trypsin (K;<0.1nM), cathepsin G (K1 ~0.15nM), elastase (Kj~105μM), chymotrypsin (Kj~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), and serine proteinase-inhibiting template-fixed peptidomimetics and methods for their synthesis have been described in International Patent Application WO2003/054000 Al and in Descours A, Moehle K., Renard A, Robinson J. ChemBioChem 2002, 3, 318-323 but the previously disclosed molecules do not exhibit high selectivity and particularly high potency. 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, HeIv. CMm. Acta. 2000, 83, 3097-3112).
These methods allow the synthesis and screening of large hairpin mimetic libraries, which in turn considerably facilitates structure-activity studies, and hence the discovery of new molecules with highly potent and selective serine protease inhibitory activity, oral bioavailability, low hemolytic activity to human red blood cells and low cytotoxicity.
The β-hairpin peptidomimetics of the present invention are compounds of the general formula
Figure imgf000005_0001
(I) wherein
Figure imgf000005_0002
is a group of one of the formulae
Figure imgf000005_0003
(a3)
Figure imgf000006_0001
Figure imgf000007_0001
03) (W) ϋ)
Figure imgf000007_0002
(k) (I) (m)
Figure imgf000007_0003
(n) (o) (P)
wherein
Figure imgf000007_0004
is GIy or 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;
s* is a group of one of the formulae
Figure imgf000008_0001
A1 A2 A3 A4
Figure imgf000008_0002
A5 A6 A7 Aδ A9
Figure imgf000008_0003
A15 A16 A17 A18 A19
Figure imgf000008_0004
A25 A26 A27 A28
Figure imgf000009_0001
A38 A39 A40 A41 A42
1
Figure imgf000009_0002
A48 A49 A50 A51 A52
Figure imgf000009_0003
A53 A54 A55 A56 A57
Figure imgf000010_0001
Figure imgf000011_0001
R1 is H; lower alkyl; or aryl-lower alkyl;
R2 is H; alkyl; alkenyl; -(CH2)m(CHR61)sOR55; -(CH2)m(CHR61)sSR:>1
, 6K
-(CH2)m(CHR61)sNR33R34; -(CH2)m(CHR61)sOCONR33R75;
-(CH2)m(CHR61)sNR20CONR33R82; -(CH2)o(CHR61)sCOOR57; -(CH2)o(CHR61)sCONR58R59; -(CH2)o(CHR61)sPO(OR60)2; -(CH2)o(CHR61)s SO2R62; or -(CH2)o(CHR61)sC6H4R8; R3 is H; alkyl; alkenyl; -(CH2)m(CHR61)sOR55; -(CH2)m(CHR61)sSR56; -(CH2)m(CHR61)sNR33R34; -(CH2)m(CHR61)sOCONR33R75; -(CH2)m(CHR61)sNR20CONR33R82; -(CH2)o(CHR61)sCOOR57 ; -(CH2)o(CHR61)sCONR58R59 ; -(CH2)o(CHR61)sPO(OR60)2; -(CH2)o(CHR61)s SO2R62; or -(CH2)o(CHR61)sC6H4R8; R4 is H; alkyl; alkenyl; -(CH2)m(CHR61)sOR55; -(CH2)m(CHR61)sSR56; - (CH2)m(CHR61)sNR33R34;
-(CH2)m(CHR61)sOCONR33R75; -(CH2)ra(CHR61)sNR20CONR33R82; -(CH2)P(CHR61)sCOOR57; -(CH2)P(CHR61)sCONR58R59; - (CH2)P(CHR61)sPO(OR60)2;
-(CH2)P(CHR61)s SO2R62; or -(CH2)o(CHR61)sC6H4R8; R5 is alkyl; alkenyl; -(CH2)o(CHR61)sOR55; -(CH2)o(CHR61)sSR56; - (CH2)o(CHR61)sNR33R34;
-(CH2)o(CHR61)sOCONR33R75; -(CH2)o(CHR61)sNR20CONR33R82; -(CH2)o(CHR61)sCOOR57; -(CH2)o(CHR61)sCONR58R59; -
(CH2)o(CHR61)sPO(OR60)2;
-(CH2)o(CHR61)s SO2R62; or -(CH2)o(CHR61)sC6H4R8; R6 is H; alkyl; alkenyl; -(CH2)o(CHR61)sOR55; -(CH2)o(CHR61)sSR56; -
(CH2)o(CHR6])sNR33R34; -(CH2)o(CHR61)sOCONR33R75; -(CH2)o(CHR61)sNR20CONR33R82;
-(CH2)o(CHR61)sCOOR57; -(CH2)o(CHR61)sCONR58R59; - (CH2)o(CHR61)sPO(OR60)2;
-(CH2)o(CHR61)s SO2R62; or -(CH2)o(CHR61)sC6H4R8; R7 is alkyl; alkenyl; -(CH2)q(CHR61)sOR55; -(CH2)q(CHR61)sNR33R34; -(CH2)q(CHR61)sOCONR33R75; -(CH2)q(CHR61)sNR20CONR33R82;
-(CH2)r(CHR61)sCOOR57; -(CH2)r(CHR61)sCONR58R59; - (CH2)r(CHR61)sPO(OR60)2;
-(CH2)r(CHR61)sSO2R62; or -(CH2)r(CHR61)s C6H4R8;
R8 is H; Cl; F; CF3; NO2; lower alkyl; lower alkenyl; aryl; aryl-lower alkyl; -(CH2)o(CHR61)sOR55; -(CH2)o(CHR61)sSR56; -(CH2)o(CHR61)NR33R34 ;
-(CH2)o(CHR61)sOCONR33R75; -(CH2)o(CHR61)sNR20CONR33R82; -(CH2)o(CHR61)sCOOR57; -(CH2)o(CHR6' )sCONR58R59; - (CH2)o(CHR61)sPO(OR60)2; -(CH2)o(CHR61)sSO2R62; or -(CH2)o(CHR6 ^5COR64; R9 is alkyl; alkenyl; -(CH2)o(CHR61)sOR55; -(CH2)o(CHR61)sSR56; - (CH2)o(CHR61)sNR33R34;
-(CH2)o(CHR61)sOCONR33R75; -(CH2)o(CHR61)sNR20CONR33R82; -(CH2)o(CHR61)sCOOR57; -(CH2)o(CHR61)sCONR58R59; - (CH2)o(CHR61)sPO(OR60)2; -(CH2)o(CHR61)s SO2R62; or -(CH2)o(CHR61)sC6H4RS; R10 is alkyl; alkenyl; -(CH2)O(CHR61)SOR55; -(CH2)O(CHR61)SSR56; - (CH2)O(CHR61)SNR33R34;
-(CH2)o(CHR61)sOCONR33R75; -(CH2)o(CHR51)sNR20CONR33R82; -(CH2)o(CHR61)sCOOR57; -(CH2)0(CHR61)SCONR58R59; - (CH2)o(CHR61)sPO(OR60)2;
-(CH2)O(CHR61)S SO2R62; or -(CH2)O(CHR61^C6H4R8; Ru is H; alkyl; alkenyl; -(CH2)m(CHR61)sOR55; -(CH2)m(CHR61)sNR33R34; -(CH2)m(CHR61)sOCONR33R75; -(CH2)m(CHR61)sNR20CONR33R82; -(CH2)0(CHR61)SCOOR57; -(CH2)0(CHR61)SCONR58R59; - (CH2)o(CHR61)sPO(OR60)2;
-(CH2)O(CHR61)SSO2R62; or -(CH2)O(CHR61)S C6H4R8; R12 is H; alkyl; alkenyl; -(CH2)m(CHR61)sOR55; -(CH2)m(CHR61)sSR56; -(CH2)m(CHR61)sNR33R34; -(CH2)m(CHR61)sOCONR33R75; -(CH2)m(CHR61)sNR20CONR33R82; -(CH2)r(CHR61)sCOOR57; - (CH2)r(CHR61)sCONR58R59; -(CH2)r(CHR61)sPO(OR60)2; -(CH2)r(CHR61)s SO2R62; or -(CH2)r(CHR61)sC6H4R8;
R13 is alkyl; alkenyl; -(CH2)q(CHR61)sOR55; -(CH2)q(CHR61)sSR56; - (CH2)q(CHR61)sNR33R34;
-(CH2)q(CHR61)sOCONR33R75; -(CH2)q(CHR61)sNR20CONR33R82; -(CH2)q(CHR61)sCOOR57; -(CH2)q(CHR61)sCONR58R59; -
(CH2)q(CHR61)sPO(OR60)2;
-(CH2)q(CHR61)s SO2R62; or -(CH2)q(CHR61)sC6H4R8; R14 is H; alkyl; alkenyl; -(CH2)m(CHRδl)sOR55; -(CH2)m(CHR61)sNR33R34; -(CH2)m(CHR61)sOCONR33R75; -(CH2)m(CHR61)sNR20CONR33R82; -(CH2)q(CHR6I)sCOOR57; -(CH2)q(CHR61)sCONR58R59; -
(CH2)q(CHR61)sPO(OR60)2;. -(CH2)q(CHR61)sSOR62; or -(CH2)q(CHR61)s C6H4R8; R15 is alkyl; alkenyl; -(CH2)o(CHR61)sOR55; -(CH2)O(CHR61)SSR56; -
(CH2)O(CHR61)SNR33R34; -(CH2)O(CHR61)SOCONR33R75; -(CH2)o(CHR61)sNR20CONR33R82;
-(CH2)0(CHR61)SCOOR57; -(CH2)o(CHR61)sCONR58R59; - (CH2)o(CHR61)sPO(OR60)2;
-(CH2)o(CHR61)s SO2R62; or -(CH2)o(CHR61)sC5H4R8; R16 is alkyl; alkenyl; -(CH2)O(CHR61)SOR55; -(CH2)O(CHR61)SSR56; - (CH2)O(CHR61)SNR33R34; -(CH2)o(CHR61)sOCONR33R75; -(CH2)o(CHR61)sNR20CONR33R82;
-(CH2)o(CHR61)sCOOR57; -(CH2)o(CHR61)sCONR58R59; -
(CH2)o(CHR61)sPO(OR60)2;
-(CH2)o(CHR61)s SO2R62; or -(CH2)o(CHR61)sC6H4R8; R17 is alkyl; alkenyl; -(CH2)q(CHR61)sOR55; -(CH2)q(CHR61)sSR56; -
(CH2)q(CHR61)sNR33R34;
-(CH2)q(CHR61)sOCONR33R75; -(CH2)q(CHR61)sNR20CONR33R82;
-(CH2)q(CHR61)sCOOR57; -(CH2)q(CHR61)sCONR58R59; -
(CH2)q(CHR61)sPO(OR60)2; -(CH2)q(CHR61)s SO2R62; or -(CH2)q(CHR61)sC6H4R8;
R18 is alkyl; alkenyl; -(CH2)P(CHR61)sOR55; -(CH2)P(CHR61), SR56; -
(CH2)P(CHR61)sNR33R34;
-(CH2)p(CHR61)sOCONR33R75; -(CH2)p(CHR61)sNR20CONR33R82;
-(CH2)P(CHR61)sCOOR57; -(CH2)P(CHR61)sCONR58R59; - (CH2)P(CHR61)sPO(OR60)2;
-(CH2)p(CHR61)s SO2R62; or -(CH2)o(CHR61)sC6H4R8; R19 is lower alkyl; -(CH2)P(CHR61)sOR55; -(CH2)P(CHR61)sSR56; -
(CH2)P(CHR61)sNR33R34;
-(CH2)p(CHR61)sOCONR33R75; -(CH2)p(CHR61)sNR20CONR33R82; -(CH2)P(CHR61)sCOOR57; -(CH2)P(CHR61)sCONR58R59; -
(CH2)P(CHR61)sPO(OR60)2;
-(CH2)P(CHR61)s SO2R62; or -(CH2)o(CHR61^C6H4R8; or R18 and R19 taken together can form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-; R20 is H; alkyl; alkenyl; or aryl-lower alkyl;
R21 is H; alkyl; alkenyl; -(CH2)o(CHR61)sOR55; -(CH2)o(CHR61)sSR56; -
(CH2)o(CHR61)sNR33R34;
-(CH2)o(CHR61)sOCONR33R75; -(CH2)o(CHR61)sNR20CONR33R82;
-(CH2)o(CHR61)sCOOR57; -(CH2)o(CHR61)sCONR58R59; - (CH2)o(CHR61)sPO(OR60)2;
-(CH2)o(CHR61)s SO2R62; or -(CH2)o(CHR61)sC6H4R8; R22 is H; alkyl; alkenyl; -(CH2)o(CHR61)sOR55; -(CH2)o(CHR61)sSR56; -
(CH2)o(CHR61)sNR33R34;
-(CH2)o(CHR61)sOCONR33R75; -(CH2)o(CHR61)sNR20CONR33R82; -(CH2)o(CHR61)sCOOR57; -(CH2)o(CHR61)sCONR58R59; -
(CH2)o(CHR61)sPO(ORδ0)2;
-(CH2)o(CHR61)s SO2R62; or -(CH2)o(CHR61)sC6H4R8; R23 is alkyl; alkenyl; -(CH2)o(CHR61)sOR55; -(CH2)o(CHR61)sSR56; - (CH2)o(CHR61)sNR33R34;
-(CH2)o(CHR61)sOCONR33R75; -(CH2)o(CHR61)sNR20CONR33R82;
-(CH2)o(CHR61)sCOOR57; -(CH2)o(CHR61)sCONR58R59; -
(CH2)o(CHR61)sPO(ORδ0)2;
-(CH2)o(CHR61)s SO2R62; or -(CH2)o(CHR61)sC5H4R8; R24 is alkyl; alkenyl; -(CH2)o(CHR61)sOR55; -(CH2)o(CHR61)sSR56; -
(CH2)o(CHR61)sNR33R34;
-(CH2)o(CHR61)sOCONR33R75; -(CH2)o(CHR61)sNR20CONR33R82;
-(CH2)o(CHR61)sCOOR57; -(CH2)o(CHR61)sCONR58R59; -
(CH2)o(CHR61)sPO(OR60)2; -(CH2)o(CHR6')s SO2R62; or -(CH2)o(CHR61)sC6H4R8;
R25 is H; alkyl; alkenyl; -(CH2)m(CHR61)sOR55; -(CH2)m(CHR61)sSR56;
-(CH2)m(CHR61)sNR33R34; -(CH2)m(CHR61)sOCONR33R75;
-(CH2)m(CHR6l)sNR20CONR33R82; -(CH2)o(CHR61)sCOOR57;
-(CH2)o(CHR61)sCONR58R59; -(CH2)o(CHR61)sPO(OR60)2; -(CH2)o(CHR61)sSO2R62; or -(CH2)o(CHR61)sC6H4R8;
R26 is H; alkyl; alkenyl; -(CH2)m(CHR61)sOR55; -(CH2)m(CHR61)sSR56;
-(CH2)m(CHR61)sNR33R34; -(CH2)m(CHR61)sOCONR33R75;
-(CH2)m(CHR61)sNR20CONR33R82; -(CH2)o(CHR61)sCOOR57; -
(CH2)o(CHR61)sCONR58R59; -(CH2)o(CHR61)sPO(OR60)2; -(CH2)o(CHR61)s SO2R62; or -(CH2)o(CHR61)sC6H4R8; or
R25 and R26 taken together can form: -(CH2)2.6-; -(CH2),.O(CH2)r; -(CH2)rS(CH2)r; or
-(CH2)rNR57(CH2)r-; R27 is H; alkyl; alkenyl; -(CH2)o(CHR61)sOR55; -(CH2)o(CHR61)sSR56; -
(CH2)o(CHR61)sNR33R34; -(CH2)o(CHR61)sCOOR57; -(CH2)o(CHR61)sCONR58R59; -
(CH2)o(CHR61)sOCONR33R75;
-(CH2)o(CHR61)sNR20CONR33R82; -(CH2)o(CHR6')sPO(OR60)2;
-(CH2)o(CHR61)s SO2R62; or -(CH2)o(CHR61)sC6H4R8;
R28 is alkyl; alkenyl; -(CH2)o(CHR61)s-OR55; -(CH2)o(CHR61 ), SR56; -(CH2)o(CHR61)s NR33R34; -(CH2)o(CHR61)sOCONR33R75; -(CH2)o(CHR61)sNR20CONR33R82; -(CH2)o(CHR61)s COOR57; -(CH2)o(CHR61)s CONR58R59; -(CH2)o(CHR61)s PO(OR60)2;
-(CH2)o(CHR61)s SO2R62; or -(CH2)o(CHR61)s C6H4R8; R29 is alkyl; alkenyl; -(CH2)o(CHR61)sOR55; -(CH2)o(CHR61)sSR56; - (CH2)o(CHR61)sNR33R34;
-(CH2)o(CHR61)sOCONR33R75; -(CH2)o(CHR61)sNR20CONR33R82; -(CH2)o(CHR61)sCOOR57; -(CH2)o(CHR61)sCONR58R59; - (CH2)o(CHR61)sPO(OR60)2; -(CH2)o(CHR61)s SO2R62; or -(CH2)o(CHR61^C6H4R8;
R30 is H; alkyl; alkenyl; or aryl-lower alkyl;
R31 is H; alkyl; alkenyl; -(CH2)P(CHR61)sOR55; -(CH2)P(CHR61)sNR33R34; -(CH2)p(CHR61)sOCONR33R75; -(CH2)p(CHR61)sNR20CONR33R82; -(CH2)o(CHR61)sCOOR57; -(CH2)o(CHR61)sCONR58R59; - (CH2)o(CHR61)sPO(OR60)2;
-(CH2)o(CHR61)sSO2R62; or -(CH2)o(CHR61)s C6H4R8; R32 is H; lower alkyl; or aryl-lower alkyl;
R33 is H; alkyl, alkenyl; -(CH2)m(CHR61)sOR55; -(CH2)m(CHR61)sNR34R63; -(CH2)m(CHR61)sOCONR75R82; -(CH2)m(CHR61)sNR20CONR78R82; -(CH2)o(CHR61)sCOR64; -(CH2)o(CHR61)s-CONR5SR59, -
(CH2)o(CHR61)sPO(OR60)2;
-(CH2)o(CHR61)s SO2R62; or -(CH2)o(CHR61^C6H4R8; R34 is H; lower alkyl; aryl, or aryl-lower alkyl;
R33 and R34 taken together can form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-;
R35 is H; alkyl; alkenyl; -(CH2)m(CHR61)sOR55; -(CH2)m(CHR61)sNR33R34; -(CH2)m(CHR61)sOCONR33R75; -(CH2)m(CHR61)sNR20CONR33R82; -(CH2)P(CHR61)sCOOR57; -(CH2)P(CHR61)sCONR58R59; - (CH2)P(CHR61)sPO(OR60)2; -(CH2)p(CHR61)sSO2R62; or -(CH2)P(CHR61)s C6H4R8;
R36 is H, alkyl; alkenyl; -(CH2)o(CHR61)sOR55; -(CH2)P(CHR61)sNR33R34; -(CH2)p(CHR61)sOCONR33R75; -(CH2)p(CHR61)sNR20CONR33R82; -(CH2)p(CHR61)sCOOR57; -(CH2)P(CHR61)sCONR58R59; - (CH2)p(CHR61)sPO(OR60)2; -(CH2)P(CHR61)sSO2R62; or -(CH2)o(CHR61)s C6H4R8; R37 is H; F; Br; Cl; NO2; CF3; lower alkyl; -(CH2)P(CHR61)sOR55; - (CH2)P(CHR61)sNR33R34;
-(CH2)p(CHR61)sOCONR33R75; -(CH2)p(CHR61)sNR20CONR33R82; -(CH2)o(CHR61)sCOOR57; -(CH2)o(CHR61)sCONR58R59; - (CH2)o(CHR61)sPO(OR60)2;
-(CH2)o(CHR61)sSO2R62; or -(CH2)o(CHR61)s C6H4R8; R38 is H; F; Br; Cl; NO2; CF3; alkyl; alkenyl; -(CH2)P(CHR61)sOR55; - (CH2)P(CHR61)sNR33R34;
-(CH2)P(CHR61)sOCONR33R75; -(CH2)P(CHR61)sNR20CONR33R82; -(CH2)o(CHR61 )sCOOR57; -(CH2)o(CHR61)sCONR58R59; -
(CH2)o(CHR61)sPO(OR60)2;
-(CH2)o(CHR61)sSO2R62; or -(CH2)o(CHR61)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; NO2; CF3; alkyl; alkenyl; -(CH2)P(CHR61)sOR55; - (CH2)p(CHR61)sNR33R34;
-(CH2)P(CHR61)sOCONR33R75; -(CH2)P(CHR61)sNR20CONR33R82; -(CH2)o(CHR61)sCOOR57; -(CH2)o(CHR61)sCONR58R59; - (CH2)o(CHR61)sPO(OR60)2; -(CH2)o(CHR61)sSO2R62; or -(CH2)o(CHR61)s C6H4R8;
R42 is H; F; Br; Cl; NO2; CF3; alkyl; alkenyl; -(CH2)P(CHR61)sOR55; - (CH2)p(CHR61)sNR33R34;
-(CH2)p(CHR61)sOCONR33R75; -(CH2)p(CHR61)sNR20CONR33R82; -(CH2)o(CHR61)sCOOR57; -(CH2)o(CHR61)sCONR58R59; - (CH2)o(CHR61)sPO(OR60)2;
-(CH2)o(CHR61)sSO2R62; or -(CH2)o(CHR61)s C6H4R8; R43 is H; alkyl; alkenyl; -(CH2)m(CHR61)sOR55; -(CH2)m(CHR61)sNR33R34; -(CH2)m(CHR61)sOCONR33R75; -(CH2)m(CHR61)sNR20CONR33R82; -(CH2)o(CHR61)sCOOR57; -(CH2)o(CHR61)sCONR58R59; - (CH2)o(CHR61)sPO(OR60)2;
-(CH2)o(CHR61)sSO2R62; or -(CH2)o(CHR61)s C6H4R8; R44 is alkyl; alkenyl; -(CH2)r(CHR61)sOR55; -(CH2)r(CHR61)sSR56; - (CH2)r(CHR61)sNR33R34; -(CH2)r(CHR61)sOCONR33R75; -(CH2)r(CHR51)sNR20CONR33R82; 5 001622
17
-(CH2)r(CHR61)sCOOR57; -(CH2)r(CHR61)sCONR58R59; - (CH2)r(CHR61)sPO(OR60)2;
-(CH2)r(CHR61)s SO2R62; or -(CH2)r(CHR61)sC6H4R8; R45 is H; alkyl; alkenyl; -(CH2)o(CHR61)sOR55; -(CH2)o(CHR61)sSR56; - (CH2)o(CHR61)sNR33R34;
-(CH2)o(CHR61)sOCONR33R75; -(CH2)o(CHR61)sNR20CONR33R82; -(CH2)o(CHR61)sCOOR57; -(CH2)s(CHR61)sCONR58R59; - (CH2)s(CHR61)sPO(OR60)2;
-(CH2)s(CHR61)s SO2R62; or -(CH2)s(CHR61)sC6H4R8; R46 is H; alkyl; alkenyl; or -(CH2)o(CHR61)PC6H4R8; R47 is H; alkyl; alkenyl; or -(CH2)o(CHR61)sOR55; R48 is H; lower alkyl; lower alkenyl; or aryl-lower alkyl; R49 is H; alkyl; alkenyl; -(CHR61)sCOOR57; (CHR61)sCONR58R59; (CHR61)sPO(OR60)2;
-(CHR61)sSOR62; or -(CHR61^C6H4R8; R50 is H; lower alkyl; or aryl-lower alkyl;
R51 is H; alkyl; alkenyl; -(CH2)m(CHR61)sOR55; -(CH2)m(CHR61)sSR56; -(CH2)m(CHR61)sNR33R34; -(CH2)m(CHR61)sOCONR33R75; -(CH2)m(CHR61)sNR20CONR33R82; -(CH2)o(CHR61)sCOOR57; -(CH2)o(CHR61)sCONR58R59; -(CH2)o(CHR61)pPO(OR60)2; -(CH2)p(CHR61)s SO2R62; or -(CH2)P(CHR61)sC6H4R8;
R52 is H; alkyl; alkenyl; -(CH2)m(CHR61)sOR55; -(CH2)m(CHR61)sSR56; -(CH2)m(CHR61)sNR33R34; -(CH2)m(CHR61)sOCONR33R75; -(CH2)m(CHR61)sNR20CONR33R82; -(CH2)o(CHR61)sCOOR57; -(CH2)o(CHR61)sCONR58R59; -(CH2)o(CHR61)pPO(OR60)2; -(CH2)p(CHR61)s SO2R62; or -(CH2)P(CHR6 ^5C6H4R8;
R53 is H; alkyl; alkenyl; -(CH2)m(CHR61)sOR55; -(CH2)m(CHR61)sSR56; - (CH2)m(CHR61)sNR33R34; -(CH2)m(CHR61)sOCONR33R75; -(CH2)m(CHR61)sNR20CONR33R82; -(CH2)o(CHR61)sCOOR57; -(CH2)o(CHR61)sCONR58R59; -(CH2)o(CHR61)pPO(OR60)2; -(CH2)P(CHR61)s SO2R62; or -(CH2)P(CHR61)sC6H4R8;
R54 is H; alkyl; alkenyl; -(CH2)m(CHR61)sOR55; -(CH2)m(CHR61)sNR33R34; -(CH2)m(CHR61)sOCONR33R75; -(CH2)m(CHR61)sNR20CONR33R82; -(CH2)o(CHR61)COOR57; -(CH2)o(CHR61)sCONR58R59; or -(CH2)o(CHR61)s C^H4R ; R55 is H; lower alkyl; lower alkenyl; aryl-lower alkyl; -(CH2)m(CHR61)sOR57; -(CH2)m(CHR61)sNR34R63; -(CH2)m(CHR61)sOCONR75R82; -(CH2)m(CHR61)sNR20CONR78R82; -(CH2)o(CHR61)s-COR64; - (CH2)o(CHR61)COOR57; or -(CH2)o(CHR61)sCONR58R59; R56 is H; lower alkyl; lower alkenyl; aryl-lower alkyl; -(CH2)m(CHR61)sOR57; -(CH2)m(CHR61)sNR34R63; -(CH2)m(CHR61)sOCONR75R82; -(CH2)m(CHR61)sNR20CONR78R82; -(CH2)o(CHR61)s-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; R5 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)2O(CH2)2-; -(CH2)2S(CH2)2~; or -(CH2)2NR57(CH2)2-;
R60 is H; lower alkyl; lower alkenyl; aryl; or aryl-lower alkyl;
R61 is alkyl; alkenyl; aryl; heteroaryl; aryl-lower alkyl; heteroaryl-lower alkyl; -
(CH2)mOR55;
-(CH2)mNR33R34; -(CH2)mOCONR75R82; -(CH2)mNR20CONR78R82; - (CH2)oCOOR37;
-(CH2)oNR58R59; or -(CH2)oPO(COR60)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; -SO2R62; or -PO(OR60)2;
R34and R63 taken together can form: -(CH2)2-6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-;
R64 is H; lower alkyl; lower alkenyl; aryl; heteroaryl; aryl-lower alkyl; heteroaryl-lower alkyl; -(CH2)P(CHR61)sOR65; -(CH2)P(CHR61)sSR66; or -(CH2)P(CHR61)sNR34R63;
-(CH2)P(CHR61)sOCONR75R82; -(CH2)P(CHR61)sNR20CONR78R82; R65 is H; lower alkyl; lower alkenyl; aryl, aryl-lower alkyl; heteroaryl-lower alkyl; - COR57; -COOR57; or -CONR58R59; R66 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 11 α-amino acid residues, 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, GIy, Pro, Pro(4NHCOPhe) or of formula -A-CO-, or of formula -B-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-;
-NR20-CH(CO-)-(-(CH2)pNR20CO(CH2)p-CH(CO-)-NR20-; and -NR20-CH(CO-)-(-(CH2)pNR20CONR20(CH2)p-CH(CO-)-NR20-; R71 is lower alkyl; lower alkenyl; -(CH2)P(CHR61)sOR75; -(CH2)P(CHR61)sSR75;
-(CH2)P(CHR61)sNR33R34; -(CH2)P(CHR61)sOCONR33R75; - (CH2)p(CHR61)sNR20CONR33R82;
-(CH2)o(CHR61)sCOOR75; -(CH2)PCONR58R59; -(CH2)PPO(OR62)2; -(CH2)PSO2R62; or -(CH2)o-C6R67R68R69R70R76;
R72 is H, lower alkyl; lower alkenyl; -(CH2)P(CHR61)sOR85; or -(CH2)P(CHR61)sSR85; R73 is -(CR86R87)OR77; -(CH2)rO(CH2)oR77; -(CH2)rS(CH2)oR77; or -(CH2)rNR20(CH2)oR77; R74 is -(CH2)PNR78R79; -(CH2)PNR77R80; -(CH2)pC(=NR80)NR78R79; - (CH2)PC(=NOR50)NR78R79;
-(CH2)pC(=NNR78R79)NR78R79; -(CH2)pNR80C(=NR80)NR78R79; -(CH2)pN=C(NR78R80)NR79R80;-(CH2)pC6H4NR78R79; -(CH2)pC6H4NR77R80; -(CH2)PC6H4C(=NR80)NR78R79; -(CH2)PC6H4C(=NOR50)NR78R79; -(CH2)pC6H4C(=NNR78R79)NR78R79; -(CH2)pC6H4NR80C(=NR80)NR78R79;
-(CH2)pC6H4N=C(NR78R80)NR79R80; -(CH2)rO(CH2)raNR78R79; - (CH2)rO(CH2)mNR77R80;
-(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)inN=C(NR78R80)NR79R80; -(CH2)rO(CH2)pC6H4CNR78R79; EP2005/001622
20
-(CH2)rO(CH2)pC6H4C(=NR80)NR78R79; -(CH2)rO(CH2)pC6H4C(=NOR50)NR78R79;
-(CH2)rO(CH2)pC6H4C(=NNR78R79)NR78R79;
-(CH2)rO(CH2)pCδH4NR80C(=NR80)NR78R79; -(CH2)rS(CH2)raNR78R79;
-(CH2)rS(CH2)mNR77R80;-(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)mN=C(NR78R80)NR79R80;
-(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)pC6H4NRS0C(=NR80)NR78R79; -(CH2)PNR80COR64; -
(CH2)pNR80COR77;
-(CH2)PNR80CONR78R79; -(CH2)PC6H4NR80CONR78R79; or
-(CH2)pNR20CO-[(CH2)u-X]rCH3 where X is -O-; -NR20-, or -S-; u is 1-3, and t is 1-
6; R75 is lower alkyl; lower alkenyl; or aryl-lower alkyl;
R33 and R75 taken together can form: -(CHz)2-6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-; R75 and R82 taken together can form: -(CH2)2-6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-; R76 is H; lower alkyl; lower alkenyl; aryl-lower alkyl; -(CH2)oOR72; -(CH2)oSR72;
-(CH2)oNR33R34; -(CH2)oOCONR33R75; -(CH2)oNR20CONR33R82;
-(CH2)oCOOR75; -(CH2)oCONR58R59; -(CH2)oPO(OR60)2; -(CH2)PSO2R62; or
-(CH2)oCOR64;
R77 is -C6R67R68R69R70R76; or a heteroaryl group of one of the formulae
Figure imgf000021_0001
H1 H2 H3 H4 H5
Figure imgf000021_0002
H6 H7 H8 H9 H10
Figure imgf000022_0001
H11 H12 H13 H14 H15
Figure imgf000022_0002
H21 H22 H23 H24 H25
Figure imgf000022_0003
H26 H27 H28 H29
Figure imgf000022_0004
Figure imgf000022_0005
Figure imgf000022_0007
Figure imgf000022_0006
H38 H39 H40 H41
Figure imgf000022_0008
H42 H43 H44 H45
Figure imgf000023_0001
H50 H51 H52 H53 H54
Figure imgf000023_0002
H46 H47 H48 H49
Figure imgf000023_0003
,78
R is H; lower alkyl; aryl; or aryl-lower alkyl;
R78 and R82 taken together can form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-;
R79 is H; lower alkyl; aryl; or aryl-lower alkyl; or
R78 and R79, taken together, can be -(CH2)2.7-; -(CH2)2O(CH2)2-; or -(CH2)2NR57(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; R33 and R82 taken together can form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-; R83 is H; lower alkyl; aryl; or -NR78R79; R84 is -(CH2)m(CHR61)sOH; -(CR86R87)pOR80; -(CR86R87)pCOOR80; -(CH2)m(CHR61)sSH; -
> 80.
(CR86R87)pSR80;-(CH2)pCONR78R79; -(CH2)PNR80CONR78R79; -
(CH2)pC5H4CON
Figure imgf000023_0004
-(CR86R87)OPO(OR60)2; - (CR86R87)PSO2R60; -(CR86R87)PSOR60; -(CH2)m(CHR61)s OPO(OR60)2,; or - (CH2)m(CHR61)s OSO2R60 ,;
R85 is lower alkyl; or lower alkenyl;
R86 is H; lower alkyl, where H is maybe substituted by halogen; or halogen;
R87 is H; lower alkyl, where H is maybe substituted by halogen; or halogen; with the proviso that in said chain of 1 1 α-amino acid residues Z if n is 11, the amino acid residues in positions 1 to 11 are:
Pl : of type C or of type D or of type E or of type F;
P2: of type C or of Type D or of type E, or of type F;
P3: or of type C, of type F, or the residue is GIy; - P4: of type C, or of type D, or of type F, or of type E, or the residue is
GIy or Pro;
P5: of type E, or of type C, or of type F, or the residue is GIy or Pro;
P6: of type D, or of type F, or of type E or of type C, or the residue is
GIy or Pro; - P7: of type C, or of type E, or of type F, or of formula -A-CO-, or the residue is GIy or Pro;
P8: of type D, or of type C, or of type F, or of formula -A-CO, or the residue is GIy or Pro or Pro(4NHCOPhe);
P9: of type C, or of type D, or of type E, or of type F; - PlO: of type D, or of type C, or of type F, or of type E; and
PI l : of type C, or of type D, or of type E, or of type F; or
P2 and PlO, taken together, can form a group of type H; and with the further proviso that if the template is DProLPro, the amino acid residues in positions Pl to Pl 1 are other than - Pl : Arg
P2: Cys, linked with Cys in position PlO by a disulfide bridge
P3: Thr
P4: Lys
P5: Ser - P6: He
P7: Pro
P8: Pro
P9: He
PlO: Cys, linked with Cys in position PlO by a disulfide bridge; and - PI l: Phe
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 5, 6 or 7, 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 steps (c) and (d) until the N-terminal amino acid residue has been introduced;
(f) coupling the product thus obtained with a compound of the general formula
Figure imgf000025_0001
II wherein
Figure imgf000025_0002
is as defined above and X is an N-protecting group or, alternatively, if
Figure imgf000025_0003
is to be group (al) or (a2), above,
(fa) coupling the product obtained in step (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, III, any functional group which may be present in said N-protected amino acid derivative being likewise appropriately protected; and, respectively, if
Figure imgf000026_0001
is to be group (a3), above,
(fa1) coupling the product obtained in step (e) with an appropriately N- protected derivative of an amino acid of the above general formula III, any functional group which may be present in said N-protected amino acid derivative being likewise appropriately protected;
(fb1) removing the N-protecting group from the product thus obtained; and (fc1) coupling the product thus obtained with an appropriately N-protected derivative of an amino acid of the above general formula III, 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) or (fc1);
(h) coupling the product thus obtained with an appropriately N-protected derivative of that amino acid which in the desired end-product is in position 11 , 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 with an appropriately N-protected derivative of that amino acid which in the desired end-product is one position farther away from position 11, 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 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;
(n) if desired, forming an interstrand linkage between side-chains of appropriate amino acid residues at positions 2 and 10;
(o) detaching the product thus obtained from the solid support;
(p) cyclizing the product cleaved from the solid support;
(q) 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.
Alternatively, the peptidomimetics of the present invention can be prepared by
(aτ) coupling an appropriately functionalized solid support with a compound of the general formula
Figure imgf000027_0001
II wherein
Figure imgf000027_0002
is as defined above and X is an N-protecting group or, alternatively, if
Figure imgf000028_0001
is to be group (al) or (a2), above,
(a'a) coupling said appropriately functionalized solid support 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;
. (a'b) removing the N-protecting group from the product thus obtained; and (a'c) coupling the product thus obtained with an appropriately N-protected derivative of an amino acid of the above general formula IV and, respectively, III, any functional group which may be present in said N-protected amino acid derivative being likewise appropriately protected; and, respectively, if
Figure imgf000028_0002
is to be group (a3), above,
(a'a1) coupling said appropriately functionalized solid support with an appropriately N-protected derivative of an amino acid of the above general formula III, any functional group which may be present in said N-protected amino acid derivative being likewise appropriately protected; (a'b1) removing the N-protecting group from the product thus obtained; and (a'c1) coupling the product thus obtained with an appropriately N-protected derivative of an amino acid of the above general formula III, any functional group which may be present in said N-protected amino acid derivative being likewise appropriately protected; (b1) removing the N-protecting group from the product obtained in step (a1), (a'c) or
(a'c1);
(c1) coupling the product thus obtained with an appropriately N-protected derivative of that amino acid which in the desired end-product is in position 11, any functional group which may be present in said N-protected amino acid derivative being likewise appropriately protected;
(d1) removing the N-protecting group from the product thus obtained;
(e1) coupling the product thus obtained with an appropriately N-protected derivative of that amino acid which in the desired end-product is one position farther away from position 1 1, any functional group which may be present in said N-protected amino acid derivative being likewise appropriately protected;
(f ) removing the N-protecting group from the product thus obtained;
(g1) repeating steps (e1) and (f ) until all amino acid residues have been introduced; (h1) if desired, selectively deprotecting one or several protected functional group(s) ' present in the molecule and appropriately substituting the reactive group(s) thus liberated;
(i1) if desired forming an interstrand linkage between side-chains of appropriate amino acid residues at positions 2 and 10;
(j1) detaching the product thus obtained from the solid support; (kτ) cyclizing the product cleaved from the solid support;
(1') 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
(m1) 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.
The peptidomimetics of the present invention can also be enantiomers of the compounds of formula I. These enantiomers can be prepared by a modification of the above processes in which enantiomers of all chiral starting materials are used.
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, NO2, 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). The structural element -B-CO- forms in combination with another structural element -B-CO- template (a3). The template (a3) is less preferred in formula I. Templates (a) through (p) constitute building blocks which have an N-terminus and a C-terminus oriented in space in such a way that the distance between those two groups may lie between 4.0-5.5A. The peptide chain Z 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 serine protease inhibitory activity 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 the selective inhibitory activity but also for the synthesis process defined hereinabove, as incorporation of the templates at the beginning or near the middle of the linear protected peptide precursors enhances cyclization yields significantly.
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-LB-CO- to DA69-CO-LB-CO-. Thus, for example, DPro-LPro constitutes the prototype of templates (al). Less preferred, but possible are combinations
-LA1-CO-DB-CO- to -LA69-CO-DB-CO- forming templates (a2). Thus, for example, LPro- DPro constitutes the prototype of template (a2).
It will be appreciated that building blocks -Al-CO- to -A69-CO- in which A has (D)- confϊguration, 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)mOR55 (where R55: lower alkyl; or lower alkenyl); (CH2)mSR56 (where R56: lower alkyl; or lower alkenyl); (CH2)mNR33R34 (where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl; R33 and R34 taken together form:
-(CH2)M-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; R57: H; or lower alkyl); (CH2)mOCONR33R75 (where R33: H; or lower alkyl; or lower alkenyl; R75: lower alkyl; or R33 and R75 taken together form: -(CH2)MS -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)mNR20CONR33R82 (where R20: H; or lower lower alkyl; R33: H; or lower alkyl; or lower alkenyl; R82: H; or lower alkyl; or R33 and R82 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oN(R20)COR64(where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); -(CH2)oCOOR57 (where R57: lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; or lower alkyl; or R58 and R59 taken together form: -(CH2)M-; - (CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); - (CH2)oPO(OR60)2 (where R60 : lower alkyl; or lower alkenyl); -(CH2)oSO2R62 (where R62: lower alkyl; or lower alkenyl); or -(CH2)C1C6H4R8 (where Rs: 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 R56: lower alkyl; or lower alkenyl); -(CH2)mNR33R34 (where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl; or R33 and R34 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)mOCONR33R75 (where R33: H; or lower alkyl; or lower alkenyl; R75: lower alkyl; or R33 and R75 taken together form: -(CH2)2-6-; -(CH2)2O(CH2)2-; - (CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)mNR20CONR33R82 (where R20: H; or lower lower alkyl; R33: H; or lower alkyl; or lower alkenyl; R82: H; or lower alkyl; or R33 and R82 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); -(CH2)oCOOR57 (where R57: lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; lower alkyl; or R58 and R59 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); (CH2)oPO(OR60)2 (where R60: lower alkyl; or lower alkenyl); -(CH2)oSO2R62 (where R62: lower alkyl; or lower alkenyl); or -(CH2)qC6H4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
R1: H; lower alkyl; lower alkenyl; -(CH2)mOR55 (where R55: lower alkyl; or lower alkenyl); -(CH2)mSR56 (where R56: lower alkyl; or lower alkenyl); -(CH2)mNR33R34 (where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl; or R33 and R34 taken together form: -(CH2)2-6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)mOCONR33R75 (where R33: H; or lower alkyl; or lower alkenyl; R75: lower alkyl; or R33 and R75 taken together form: -(CH2)2.5-; -(CH2)2O(CH2)2-; - (CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)mNR20CONR33R82 (where R20: H; or lower lower alkyl; R33: H; or lower alkyl; or lower alkenyl; R82: H; or lower alkyl; or R33 and R82 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)mN(R20)COR64(where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); -(CH2)oCOOR57 (where R57: lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; or lower alkyl; or R58 and R59 taken together form: -(CH2)2-6-; - (CH2)2O(CH2)r;
-(CHz)2S(CH2)I-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); - (CH2)oPO(OR60)2 (where R60: lower alkyl; or lower alkenyl); -(CH2)oSO2R62 (where R62: lower alkyl; or lower alkenyl); or -(CHz)11C6H4R8 (where Rs: H; F; Cl; CF3; lower alkyl; lower alkenyl;or lower alkoxy).
R5: lower alkyl; lower alkenyl; -(CHz)0OR55 (where R55: lower alkyl; or lower alkenyl); -(CH2)oSR56 (where R56: lower alkyl; or lower alkenyl); -(CH2)oNR33R34 (where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl; or R33 and R34 taken together form: -(CH2)Z-6-; -(CHz)2O(CHz)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oOCONR33R75 (where R33: H; or lower alkyl; or lower alkenyl; R75: lower alkyl; or R33 and R75 taken together form: -(CH2)2-6-; -(CH2)2O(CH2)2-; - (CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; R57: where H; or lower alkyl); (CH2)oNR20CONR33R82 (where R20: H; or lower lower alkyl; R33: H; or lower alkyl; or lower alkenyl; R82: H; or lower alkyl; or R33 and R82 taken together form: -(CH2)2-6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)r; or -(CH2)zNR57(CH2)2-; where R57: H; or lower alkyl); (CH2)oN(R20)COR64(where: R20: H; or lower alkyl; R64: alkyl; alkenyl; aryl; and aryl-lower alkyl; heteroaryl-lower alkyl); -(CH2)oCOOR57 (where R57: lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; or lower alkyl; or R58 and R59 taken together form: -(CH2)2-6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oPO(OR60)2 (where R60: lower alkyl; or lower alkenyl); -(CH2)oSO2R62 (where R62: lower alkyl; or 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)oOR55 (where R55: lower alkyl; or lower alkenyl); -(CH2)oSR56 (where R56 : lower alkyl; or lower alkenyl); -(CH2)oNR33R34 (where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl; or R33 and R34 taken together form: -(CH2)2.6-; -(CHj)2O(CHz)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oOCONR33R75 (where R33: H; or lower alkyl; or lower alkenyl; R75: lower alkyl; or R33 and R75 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; - (CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oNR20CONR33R82 (where R20: H; or lower lower alkyl; R33: H; or lower alkyl; or lower alkenyl; R82: H; or lower alkyl; or R33 and R82 taken together form: -(CHz)2-6-; -(CH2)z0(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); -(CH2)oCOOR57 (where R57: lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; or lower alkyl; or R58 and R59 taken together form: -(CH2)2-6-; - (CH2)2O(CH2)2-;
-(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); - (CH2)oPO(OR60)2 (where R60 : lower alkyl; or lower alkenyl); -(CH2)oSO2R62 (where R62: lower alkyl; or lower alkenyl); or -(CH2)C1C6H4R8 (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 : lower alkyl; or lower alkenyl); -(CH2)qNR33R34 (where
R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl; or R33 and R34 taken together form:
-(CH2W; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)qOCONR33R75 (where R33: H; or lower alkyl; or lower alkenyl; R75:
> 33 __ . r.75 . lower alkyl; or R" and R" taken together form: -(CH2W; -(CH2)2O(CH2)2-; - (CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); (CH2)qNR20CONR33R82 (where R20: H; or lower lower alkyl; R33: H; or lower alkyl; or lower alkenyl; R82: H; or lower alkyl; or R33 and R82 taken together form: -(CH2)M-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)qN(R20)COR64(where: R20: H; or lower alkyl; R54: 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; or R58 and R59 taken together form: -(CH2)2.6-; - (CH2)2O(CH2)2-;
-(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); - (CH2)rPO(OR60)2 (where R60 : lower alkyl; or lower alkenyl); (CH2)rSO2R62 (where R62: lower alkyl; or lower alkenyl); or -(CH2)qC6H4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl;or lower alkoxy). - Rs; H; F; Cl; CF3; lower alkyl; lower alkenyl; -(CH2)oOR55 (where R55: lower alkyl; or lower alkenyl); (CH2)oSR56 (where R56: lower alkyl; or lower alkenyl); - (CH2)oNR33R34 (where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl; or R33 and R34 taken together form: -(CH2W; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or - (CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oOCONR33R75 (where R33: H; or lower alkyl; or lower alkenyl; R75: lower alkyl; or R33 and R75 taken together form: - (CH2W; -(CH2)2O(CH2)2-;
-(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)r; where R57: H; or lower alkyl);
-(CH2)oNR20CONR33R82 (where R20: H; or lower lower alkyl; R33: H; or lower alkyl; or lower alkenyl; R82: H; or lower alkyl; or R33 and R82 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl);
-(CH2)oN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl);
-(CH2)oCOOR57 (where R57 : lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where •
R58: lower alkyl; or lower alkenyl; and R59: H; or lower alkyl; or R58 and R59 taken together form:
-(CH2)M-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oPO(OR60)2 (where R60: lower alkyl; or lower alkenyl); -
(CH2)oSO2R62 (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)oOR55 (where R55: lower alkyl; or lower alkenyl); -(CH2)oSR56 (where R56: lower alkyl; or lower alkenyl); -(CH2)oNR33R34 (where
R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl; or R33 and R34 taken together form:
-(CH2)M-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oOCONR33R75 (where R33: H; or lower alkyl; or lower alkenyl; R75: lower alkyl; or R33 and R75 taken together form: -(CH2W; -(CH2)2O(CH2)2-; -
(CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)mNR20CONR33R82 (where R20:
H; or lower lower alkyl; R33: H; or lower alkyl; or lower alkenyl; R82: H; or lower alkyl; or R33 and R82 taken together form: -(CH2W; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oN(R20)COR64(where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); -(CH2)oCOOR57 (where R57: lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where R58 : lower alkyl; or lower alkenyl; and R59: H; or lower alkyl; or R58 and R59 taken together form: -(CH2W; - (CH2)2O(CH2>;
-(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -
(CH2)oPO(OR60)2 (where R60: lower alkyl; or lower alkenyl); -(CH2)oSO2R62 (where R62: lower alkyl; or lower alkenyl); or -(CH2^C6H4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy). - R1": lower alkyl; lower alkenyl; -(CH2)oOR55 (where R55: lower alkyl; or lower alkenyl); -(CH2)oSR56 (where R56: lower alkyl; or lower alkenyl); -(CH2)oNR33R34 (where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl; or R33 and R34 taken together form:
-(CH2)MS -(CH2)2O(CH2)2-; -(CH2)2S(CH2)r; or -(CH2)2NR57(CH2)2s where R57: H; or lower alkyl); -(CH2)oOCONR33R75 (where R33: H; or lower alkyl; or lower alkenyl; R75: lower alkyl; or R33 and R75 taken together form: -(CH2W; -(CH2)2O(CH2)2-; - (CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oNR20CONR33R82 (where R20: H; or lower lower alkyl; R33: H; or lower alkyl; or lower alkenyl; R82: H; or lower alkyl; or R33 and R82 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)r; where R57: H; or lower alkyl); -(CH2)oN(R20)COR64(where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); -(CH2)oCOOR57 (where R57: lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; lower alkyl; or R58 and R59 taken together form: -(CH2W; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -
(CH2)oPO(OR60)2 (where R60: lower alkyl; or lower alkenyl); -(CH2)oSO2R62 (where R62: lower alkyl; or lower alkenyl); or -(CH2)C1C6H4R8 (where R8: H; F; Cl; CF3; lower alkyi; lower alkenyl; or lower alkoxy).
R11: H; lower alkyl; lower alkenyl; -(CH2)mOR55 (where R55: lower alkyl; or lower alkenyl); -(CH2)mSR56 (where R56: lower alkyl; or lower alkenyl); -(CH2)mNR33R34 (where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl; or R33 and R34 taken together form: -(CH2W; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)r; where R57: H; or lower alkyl); -(CH2)mOCONR33R75 (where R33: H; or lower alkyl; or lower alkenyl; R75: lower alkyl; or R33 and R75 taken together form: -(CH2W; -(CH2)2O(CH2)r; - (CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)mNR20CONR33R82 (where R20: H; or lower lower alkyl; R33: H; or lower alkyl; or lower alkenyl; R82: H; or lower alkyl; or R33 and R82 taken together form: -(CH2W; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)mN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); -(CH2)oCOOR57 (where R57: lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; lower alkyl; or R58 and R59 taken together form: -(CH2W; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)r; where R57: H; or lower alkyl); - (CH2)oPO(OR60)2 (where R60: lower alkyl; or lower alkenyl); -(CH2)oSO2R62 (where R62: lower alkyl; or lower alkenyl); or -(CH2)qC6H4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
R12: H; lower alkyl; lower alkenyl; -(CH2)mOR55 (where R55: lower alkyl; or lower alkenyl); -(CH2)mSR56 (where R56: lower alkyl; or lower alkenyl); -(CH2)JNfR33R34 (where
R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl; or R33 and R34 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 :
H; or lower alkyl); -(CH2)mOCONR33R75 (where R33: H; or lower alkyl; or lower alkenyl;
R75: lower alkyl; or R33 and R75taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -
(CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)mNR20CONR33R82 (where R20: H; or lower lower alkyl; R33: H; or lower alkyl; or lower alkenyl; R82: H; or lower alkyl; or R33 and R82 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-; where R57: 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; or R58 and R59 taken together form: -(CH2)2.6-; -
(CH2)2O(CH2)2-;
-(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -
(CH2)rPO(OR60)2 (where R60: lower alkyl; or lower alkenyl); -(CH2)o SO2R62 (where R62: lower alkyl; or lower alkenyl); or -(CH2)qC6H4R8 (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: lower alkyl; or lower alkenyl); -(CH2)qNR33R34 (where
R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl; or R33 and R34 taken together form: -(CH2W; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)r; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)qOCONR33R75 (where R33: H; or lower alkyl; or lower alkenyl; R75: lower alkyl; or R33 and R75 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -
(CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)qNR20CONR33R82 (where R20: H; or lower lower alkyl; R33: H; or lower alkyl; or lower alkenyl; R82: H; or lower alkyl; or R33 and R82 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)qN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); -(CH2)rCOO57 (where R57: lower alkyl; or lower alkenyl); -(CH2)qCONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; or lower alkyl;or R58 and R59 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; - (CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)rPO(OR60)2 (where R60: lower alkyl; or lower alkenyl); -(CH2)rSO2R62 (where R62: lower alkyl; or lower alkenyl); or
-(CH2)C1C6H4R8 (where R8: 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: lower alkyl; or lower alkenyl); -(CH2)mNR33R34 (where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl; or R33 and R34 taken together form: -(CH2)2-6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)mOCONR33R75 (where R33: H; or lower alkyl; or lower alkenyl; R75: lower alkyl; or R33 and R75 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; - (CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)mNR20CONR33R82 (where R20: H; or lower lower alkyl; R33: H; or lower alkyl; or lower alkenyl; R82: H; or lower alkyl; or R33 and R82 taken together form: -(CH2)2-6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)mN(R20)COR64 (where: R20: H; lower alkyl; R54: lower alkyl; or lower alkenyl); -(CH2)oCOOR57 (where R57: lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where RS8: lower alkyl; or lower alkenyl; and R59: H; or lower alkyl; or R58 and R59 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); - (CH2)oPO(OR60)2 (where R50: lower alkyl; or lower alkenyl); -(CH2)oSO2R62 (where R62: lower alkyl; or lower alkenyl); -(CH^qC6H4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
R15: lower alkyl; lower alkenyl; -(CH2)oOR55 (where R55: lower alkyl; or lower alkenyl); -(CH2)oSR56 (where R56: lower alkyl; or lower alkenyl); -(CH2)oNR33R34 (where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl; or R33 and R34 taken together form:
-(CH2)2-6-; -(CH2)2O(CH2)2S -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oOCONR33R75 (where R33: H; or lower alkyl; or lower alkenyl; R75: lower alkyl; or R33 and R75 taken together form: -(CH2)2-6-; -(CH2)2O(CH2)2-; - (CH2)2S(CH2)r; or
-(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oNR20CONR33R82 (where R20: H; or lower lower alkyl; R33: H; or lower alkyl; or lower alkenyl; R82: H; or lower alkyl; or R33 and R82 taken together form: -(CH2)2-6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: 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)oCOOR57 (where R57: lower alkyl; or lower alkenyl);
-(CH2)oCONR58R59 (where R58: lower alkyl, or lower alkenyl; and R59: H; lower alkyl; or
R58 and R59 taken together form: -(CH2)2-6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oPO(OR60)2 (where R60 : lower alkyl; or lower alkenyl); -(CH2)oSO2R62 (where R62: lower alkyl; or lower alkenyl); or
-(CH2)C1C6H4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy). R16: lower alkyl; lower alkenyl; -(CH2)oOR55 (where R55: lower alkyl; or lower alkenyl); -(CH2)oSR56 (where R56: lower alkyl; or lower alkenyl); -(CH2)oNR33R34 (where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl; or R33 and R34 taken together form:
-(CH2)M-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oOCONR33R75 (where R33: H; or lower alkyl; or lower alkenyl; R75: lower alkyl; or R33 and R75 taken together form: -(CH2W; -(CH2)2O(CH2)2-; - (CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oNR20CONR33R82 (where R20:
H; or lower lower alkyl; R33: H; or lower alkyl; or lower alkenyl; R82: H; or lower alkyl; or R33 and R82 taken together form: -(CH2W; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or
-(CH2)2NR"(CH2)2-; where R57: H; or lower alkyl); -(CH2)oN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); -(CH2)oCOOR57 (where R57: lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; or lower alkyl; or R58 and R59 taken together form: -(CH2)2.6-; -
(CH2)2O(CH2)2-;
-(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); - (CH2)oPO(OR60)2 (where R60: lower alkyl; or lower alkenyl); -(CH2)oSO2R62 (where R62: lower alkyl; or lower alkenyl); or -(CH2)qC6H4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
R17: lower alkyl; lower alkenyl; -(CH2)qOR55 (where R55: lower alkyl; or lower alkenyl); -(CH2)qSR56 (where R56: lower alkyl; or lower alkenyl); -(CH2)qNR33R34 (where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl; or R33 and R34 taken together form:
-(CH2)M-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)qOCONR33R75 (where R33: H; or lower alkyl; or lower alkenyl; R75: lower alkyl; or R33 and R75 taken together form: -(CH2)M-; -(CH2)2O(CH2)2-; - (CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)CNR20CONR33R82 (where R20: H; or lower lower alkyl; R33: H; or lower alkyl; or lower alkenyl; R82: H; or lower alkyl; or R33 and R82 taken together form: -(CH2)M-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: 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; lower alkyl; or R58 and R59 taken together form: -(CH2W; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); - (CH2)rPO(OR60)2 (where R60: lower alkyl; or lower alkenyl); -(CH2)rSO2R62 (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':
Figure imgf000040_0001
A81 wherein R20 is H or lower alkyl; and R64 is alkyl; alkenyl; [(CH2)u-X]rCH3, wherein X is -
O-, -NR20 or -S-, u is 1-3 and t is 1-6; aryl; aryl-lower alkyl; or heteroaryl-lower alkyl; especially those wherein R64 is n-hexyl (A8'-l); n-heptyl (A8'-2); 4-(phenyl)benzyl (A8'- 3); diphenylmethyl (A8'-4); 3 -amino-propyl (A8'-5); 5-amino-pentyl (A8'-6); methyl
(A8'-7); ethyl (A8'-8); isopropyl (A8'-9); isobutyl (A8'-10); n-propyl (A8'-ll); cyclohexyl (A8'-12); cyclohexylmethyl (A8'-13); n-butyl (A8'-14); phenyl (A8'-15); benzyl (A8'-16); (3-indolyl)methyl (A8'-17); 2-(3-indolyl)ethyl (A8'-18); (4- phenyl)phenyl (A8'-19); n-nonyl (A8'-20); CH3-OCH2CH2-OCH2- and CH3- (OCH2CH2)S-OCH2-.
Building block A70 belongs to the class of open-chain 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, VoU, 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, VoU, 1-68; D. Obrecht, C. Spiegler, P. Schδnholzer, K. Muller, H. Heimgartner, F. Stierli, HeIv. CUm. Acta 1992, 75, 1666-1696; D. Obrecht, U. Bohdal, J. Daly, C. Lehmann, P. Schδnholzer, K. Muller, Tetrahedron 1995, 51, 10883-10900; D. Obrecht, C. Lehmann, C. Ruffieux, P. Schϋnholzer, K. Muller, HeIv. CMm. Acta 1995, 78, 1567- 1587; D. Obrecht, U. Bohdal, C. Broger, D. Bur, C. Lehmann, R. Ruffieux, P. Schδnholzer, C. Spiegler, HeIv. CMm. Acta 1995, 78, 563-580; D. Obrecht, H.
Karajiannis, C. Lehmann, P. Schδnholzer, C. Spiegler, HeIv. CMm. 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 R21 to R29 in building blocks A70 to A104 are the following:
R18: lower alkyl.
R19: lower alkyl; lower alkenyl; -(CH2)POR55 (where R55: lower alkyl; or lower alkenyl); -(CH2)PSR56 (where R56: lower alkyl; or lower alkenyl); -(CH2)PNR33R34 (where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl; or R33 and R34 taken together form: -(CH2W; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)POCONR33R75 (where R33: H; or lower alkyl; or lower alkenyl; R75: lower alkyl; or R33 and R75 taken together form: -(CH2)2-6-; -(CH2)2O(CH2)2-; - (CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)PNR20CONR33R82 (where R20: H; or lower lower alkyl; R33: H; or lower alkyl; or lower alkenyl; R82: H; or lower alkyl; or R33 and R82 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: 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; or R58 and R59 taken together form: -(CH2)2.6-; - (CH2)2O(CH2)2-;
-(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); - (CH2)oPO(OR60)2 (where R50: lower alkyl; or lower alkenyl); -(CH2)PSO2R62 (where R62: lower alkyl; or lower alkenyl); or -(CH2)oC6H4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
R2': H; lower alkyl; lower alkenyl; -(CH2)oOR55 (where R55: lower alkyl; or lower alkenyl); -(CH2)oSR56 (where R56: lower alkyl; or lower alkenyl); -(CH2)oNR33R34 (where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl; or R33 and R34 taken together form:
-(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oOCONR33R75 (where R33: H; or lower alkyl; or lower alkenyl; R75: lower alkyl; or R33 and R75 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; - (CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oNR20CONR33R82 (where R20: H; or lower lower alkyl; R33: H; or lower alkyl; or lower alkenyl; R82: H; or lower alkyl; or R33 and R82 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)r; where R57: H; or lower alkyl); -(CH2)oN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); -(CH2)oCOOR57 (where R57: lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where R58: lower alkyl, or lower alkenyl; and R59: H; lower alkyl; or R58 and R59 taken together form: -(CH2)2-6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); - (CH2)oPO(OR60)2 (where R60: lower alkyl; or lower alkenyl); (CH2)oSO2R62 (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)oOR55 (where R55: lower alkyl; or lower alkenyl); -(CH2)oSR56 (where R56: lower alkyl; or lower alkenyl); -(CH2)oNR33R34 (where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl; or R33 and R34 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; Or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oOCONR33R75 (where R33: H; or lower alkyl; or lower alkenyl; R75: lower alkyl; or R33 and R75 taken together form: -(CH2)2-6-; -(CH2)2O(CH2)r; - (CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oNR20CONR33R82 (where R20: H; or lower lower alkyl; R33: H; or lower alkyl; or lower alkenyl; R82: H; or lower alkyl; or R33 and R82 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oN(R20)COR64(where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); -(CH2)oCOOR57 (where R57: lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where R58: lower alkyl, or lower alkenyl; and R59: H; lower alkyl; or R58 and R59 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); - (CH2)oPO(OR60)2 (where R60: lower alkyl; or lower alkenyl); -(CH2)oSO2R62 (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)oOR55 (where R55: lower alkyl; or lower alkenyl); -(CH2)oSR56 (where R56: lower alkyl; or lower alkenyl); -(CH2)oNR33R34 (where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl; or R33 and R34 taken together, form: -(CH2W; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oOCONR33R75 (where R33: H; or lower alkyl; or lower alkenyl; R75: lower alkyl; or R33 and R75 taken together form: -(CH2W; -(CH2)2O(CH2)2-; - (CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oNR20CONR33R82 (where R20: H; or lower lower alkyl; R33: H; or lower alkyl; or lower alkenyl; R82: H; or lower alkyl; or R33 and R82 taken together form: -(CH2W; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-; where R57: 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)oCOOR57 (where R57: lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where R58: lower alkyl, or lower alkenyl; and R59: H; lower alkyl; or R58 and R59 taken together form: -(CH2W; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oPO(OR60)2 (where R60: lower alkyl; or lower alkenyl); -(CH2)oSO2R62 (where R62: lower alkyl; or lower alkenyl); or -(CH2)C1C6H4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy); - R24: lower alkyl; lower alkenyl; -(CH2)oOR55 (where R55: lower alkyl; or lower alkenyl); -(CH2)oSR56 (where R56: lower alkyl; or lower alkenyl); -(CH2)oNR33R34 (where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl; or R33 and R34 taken together form: -(CH2)MS -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oOCONR33R75 (where R33: H; or lower alkyl; or lower alkenyl; R75: lower alkyl; or R33 and R75 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; - (CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oNR20CONR33R82 (where R20: H; or lower lower alkyl; R33: H; or lower alkyl; or lower alkenyl; R82: H; or lower alkyl; or R33 and R82 taken together form: -(CH2)2-6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-; where R57: 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)oCOOR57 (where R57: lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where R58: lower alkyl, or lower alkenyl; and R59: H; lower alkyl; or R58 and R59 taken together form: -(CH2)2-6s -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oPO(OR60)2 (where R60: lower alkyl; or lower alkenyl); ~(CH2)oSO2R62 (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); -(CHo)111NR33R34 (where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl; or R33 and R34 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)mOCONR33R75 (where R33: H; or lower alkyl; or lower alkenyl; R75: lower alkyl; or R33 and R75 taken together form: - (CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl);
-(CH2)mNR20CONR33R82 (where R20: H; or lower lower alkyl; R33: H; or lower alkyl; or lower alkenyl; R82: H; or lower alkyl; or R33 and R82 taken together form: -(CH2)2-6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)r; where R57: H; or lower alkyl);
-(CH2)mN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); -(CH2)oCOOR57 (where R57: lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; lower alkyl; or R58 and R59 taken together form:
-(CH2)M-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oPO(OR60)2 (where R60: lower alkyl; or lower alkenyl); - (CH2)oSO2R62 (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)mNR33R34 (where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl; or R33 and R34 taken together form: -(CH2)2-6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)raOCONR33R75 (where R33: H; or lower alkyl; or lower alkenyl; R75: lower alkyl; or R33 and R75 taken together form: -
(CH2W;
-(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl);
-(CH2)mNR20CONR33R82 (where R20: H; or lower lower alkyl; R33: H; or lower alkyl; or lower alkenyl; R82: H; or lower alkyl; or R33 and R82 taken together form: -(CH2)2.6-;
-(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl);
-(CH2)mN(R20)COR64(where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl);
-(CH2)oCOOR57 (where R57: lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; lower alkyl; or R58 and R59 taken together form:
-(CH2W; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oPO(ORδ0)2 (where R60: lower alkyl; or lower alkenyl); -
(CH2)oSO2R62 (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 together can be -(CH2W; -(CH2)2O(CH2)2-;
-(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl).
R27: H; lower alkyl; lower alkenyl; -(CH2)oOR55 (where R55: lower alkyl; or lower alkenyl); -(CH2)oSR56 (where R56: lower alkyl; or lower alkenyl); -(CH2)oNR33R34 (where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl; or R33 and R34 taken together form:
-(CH2)MS -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oOCONR33R75 (where R33: H; or lower alkyl; or lower alkenyl; R75: lower alkyl; or R33 and R75 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; - (CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oNR20CONR33R82 (where R20: H; or lower lower alkyl; R33: H; or lower alkyl; or lower alkenyl; R82: H; or lower alkyl; or R33 and R82 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)r; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); -(CH2)oCOOR57 (where R57: lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where R58: lower alkyl, or lower alkenyl; and R59: H; lower alkyl; or R58 and R59 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2s where R57: H; or lower alkyl); - (CH2)oPO(OR60)2 (where R60: lower alkyl; or lower alkenyl); -(CH2)oSO2R62 (where R62: lower alkyl; or lower alkenyl); or -(CH2)qC6H4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
R2li : lower alkyl; lower alkenyl; -(CH2)oOR55 (where R55: lower alkyl; or lower alkenyl); -(CH2)oSR56 (where R56: lower alkyl; or lower alkenyl); -(CH2)oNR33R34 (where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl; or R33 and R34 taken together form:
-(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R37: H; or lower alkyl); -(CH2)oOCONR33R75 (where R33: H; or lower alkyl; or lower alkenyl; R75: lower alkyl; or R33 and R75 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; - (CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oNR20CONR33R82 (where R20: H; or lower lower alkyl; R33: H; or lower alkyl; or lower alkenyl; R82: H; or lower alkyl; or R33 and R82 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)r; Or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oN(R20)COR64(where: R20 : H; or lower alkyl; R64: lower alkyl; or lower alkenyl); -(CH2)oCOOR57 (where R57: lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where R58: lower alkyl, or lower alkenyl; and R59: H; lower alkyl; or R58 and R59 taken together form: -(CH2)2-6s -(CH2)2O(CH2)2-; -(CH2)2S(CH2)r; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); - (CH2)oPO(OR60)2 (where R60: lower alkyl; or lower alkenyl); -(CH2)oSO2R62 (where R62: lower alkyl; or lower alkenyl); or -(CH2^C6H4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy). R29: lower alkyl; lower alkenyl; -(CH2)oOR55 (where R55: lower alkyl; or lower alkenyl); -(CH2)oSR56 (where R56: lower alkyl; or lower alkenyl); -(CH2)oNR33R34 (where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl; or R33 and R34 taken together form: -(CH2)M-; -(CH2)2O(CH2)r; -(CH2)2S(CH2)r; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oOCONR33R75 (where R33: H; or lower alkyl; or lower alkenyl; R75: lower alkyl; or R33 and R75 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)r; - (CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oNR20CONR33R82 (where R20: H; or lower lower alkyl; R33: H; or lower alkyl; or lower alkenyl; R82: H; or lower alkyl; or R33 and R82 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oN(R20)COR64(where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); particularly favored are NR20COlower-alkyl (R20=H; or lower alkyl); -(CH2)oCOOR57 (where R57: lower alkyl; or lower alkenyl);
-(CH2)oCONR58R59 (where R58: lower alkyl, or lower alkenyl; and R59: H; lower alkyl; or R58 and R59 taken together form: -(CH2)MS -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oPO(OR60)2 (where R60: lower alkyl; or lower alkenyl); -(CH2)oSO2R62 (where R62: lower alkyl; or lower alkenyl); or -(CH2)qC6H4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
The preferred value for R23, R24 and R29 is -NR20-CO-lower alkyl where R20 is H or lower alkyl.
For templates (b) to (p), such as (bl) and (I), the preferred values for the various symbols are the following:
R1: H; or lower Alkyl;
R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; -(CH2)oOR55 (where R55: lower alkyl; or lower alkenyl); -(CH2)oSR56 (where R56: lower alkyl; or lower alkenyl); - (CH2)oNR33R34 (where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl; or R33 and R34 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or - (CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oOCONR33R75 (where R33: H; or lower alkyl; or lower alkenyl; R75: lower alkyl; or R33 and R75 taken together form: - (CH2)MS -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oNR20CONR33R82 (where R20: H; or lower lower alkyl; R33: H; or lower alkyl; or lower alkenyl; R82: H; or lower alkyl; or R33 and R82 taken together form: -(CH2)2-6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); -(CH2)oCOOR57 (where R57: lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; or lower alkyl; or R58 and R59 taken together form:
-(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oPO(OR60)2 (where R60: lower alkyl; or lower alkenyl); -
(CH2)oSO2R62 (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. - R31: 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; or R33 and R34 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)r; -(CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)POCONR33R75 (where R33: H; or lower alkyl; or lower alkenyl; R75: lower alkyl; or R33 and R75 taken together form: - (CH2W; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl);
-(CH2)pNR20CONR33R82 (where R20: H; or lower lower alkyl; R33: H; or lower alkyl; or lower alkenyl; R82: H; or lower alkyl; or R33 and R82 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl);
-(CH2)pN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); -(CH2)oCOOR57 (where R57: lower alkyl; or lower alkenyl); (-CH2)oCONR58R59 (where R5S: lower alkyl, or lower alkenyl; and R59: H; lower alkyl; or R58 and R59 taken together form:
-(CH2)MS -(CH2)2O(CH2)2-; -(CH2)2S(CH2)r; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oPO(OR60)2 (where R60: lower alkyl; or lower alkenyl); - (CH2)oSO2R62 (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 is - 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)mNR34R63 (where R34: lower alkyl; or lower alkenyl; R63: H; or lower alkyl; or R34 and R63 taken together form: -(CH2)M-; -(CH2)2θ(CH2)2-; -(CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl) ; (CH2)mOCONR75R82(where R75: lower alkyl; or lower alkenyl; R82: H; or lower alkyl; or R75 and R82 taken together form: -
(CH2)M-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2VNR20CONR78R82 (where R20: H; or lower lower alkyl; R78: H; or lower alkyl; or lower alkenyl; R82: H; or lower alkyl; or R78 and R82 taken together form: -(CH2)2.6-;
-(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl);
-(CH2)mN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); -(CH2)oCOOR57 (where R57: lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where
R58: lower alkyl; or lower alkenyl; and R59: H; lower alkyl; or R58 and R59 taken together form:
-(CH2)M-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl). - R3-1: H; or lower alkyl.
R3S: 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; or R33 and R34 taken together form: -(CH2W; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)mOCONR33R75 (where R33: H; or lower alkyl; or lower alkenyl; R75: lower alkyl; or R33 and R75 taken together form: -
(CH2)2.6-;
-(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CHz)1nNR20CONR33R82 (where R20: H; or lower lower alkyl; R33: H; or lower alkyl; or lower alkenyl; R82: H; or lower alkyl; or R33 and R82 taken together form: -(CH2)2.6-;
-(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl);
-(CH2)mN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); -(CH2)oCOOR57 (where R57: lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; lower alkyl; or R58 and R59 taken together form:
-(CH2)M-; -(CH2)2O(CH2)r; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or 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; or R33 and R34 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)POCONR33R75 (where R33: H; or lower alkyl; or lower alkenyl; R75: lower alkyl; or R33 and R75 taken together form: -
(CH2)M-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl);
-(CH2)PNR20CONR33R82 (where R20: H; or lower alkyl; R33: H; or lower alkyl; or lower alkenyl; R82: H; or lower alkyl; or R33 and R82 taken together form: -(CH2)2-6-;
-(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl);
-(CH2)PN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl);
-(CH2)oCOOR57 (where R57 : lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where R58: lower alkyl, or lower alkenyl; and R59: H; lower alkyl; or R58 and R59 taken together form:
-(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 : H; or lower alkyl); -(CH2)oPO(OR60)2 (where R60: lower alkyl; or lower alkenyl); -
(CH2)oSO2R62 (where R62: lower alky; or lower alkenyl); or -(CH2)qC6H4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
R38: 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; or R33 and R34 taken together form: -(CH2)MS -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)POCONR33R75 (where R33: H; or lower alkyl; or lower alkenyl; R75: lower alkyl; or R33 and R78 taken together form: -
(CH2)MS -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl);
-(CH2)pNR20CONR33R82 (where R20: H; or lower lower alkyl; R33: H; or lower alkyl; or lower alkenyl; R82: H; or lower alkyl; or R33 and R82 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl);
-(CH2)PN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl);
-(CH2)oCOOR57 (where R57: lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where R58: lower alkyl, or lower alkenyl; and R59: H; lower alkyl; or R58 and R59 taken together form:
-(CH2)MS -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2s where R57: H; or lower alkyl); -(CH2)oPO(OR60)2 (where R60: lower alkyl; or lower alkenyl); -
(CH2)oSO2R62 (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)oCOOR57 (where R57: lower alkyl; or lower alkenyl); -(CH2)oCONR58R59
(where R58: lower alkyl; or lower alkenyl; and R59: H; lower alkyl; or R58 and R59 taken together form: -(CH2)2-6s -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl).
R40: lower alkyl; lower alkenyl; or aryl-lower alkyl.
R'": 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; or R33 and R34 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)r; where R57: H; or lower alkyl); -(CH2)POCONR33R75 (where R33: H; or lower alkyl; or lower alkenyl; R75: lower alkyl; or R33 and R75 taken together form: -
(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl);
-(CH2)PNR20CONR33R82 (where R20: H; or lower lower alkyl; R33: H; or lower alkyl; or lower alkenyl; R82: H; or lower alkyl; or R33 and R82 taken together form: -(CH2)2.6-;
-(CH2)2O(CH2)2-; -(CH2)2S(CH2)r; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl);
-(CH2)PN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl);
-(CH2)oCOOR57 (where R57: lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where
R58: lower alkyl, or lower alkenyl; and R59: H; lower alky; or R58 and R59 taken together form: -(CH2)2.6s -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oPO(OR60)2 (where R60: lower alkyl; or lower alkenyl); -
(CH2)oSO2R62 (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; or R33 and R34 taken together form: -(CH2)2.fi-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)POCONR33R75 (where R33: H; or lower alkyl; or lower alkenyl; R75: lower alkyl; or R33 and R75 taken together form: - (CH2W;
-(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl);
-(CH2)pNR20CONR33R82 (where R20: H; or lower lower alkyl; R33: H; or lower alkyl; or lower alkenyl; R82: H; or lower alkyl; or R33 and R82 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl);
-(CH2)pN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl);
-(CH2)oCOOR57 (where R57: lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where
R58: lower alkyl, or lower alkenyl; and R59: H; lower alkyl; or R58 and R59 taken together form:
-(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oPO(OR60)2 (where R60: lower alkyl; or lower alkenyl); -
(CH2)oSO2R62 (where R62: lower alkyl; or lower alkenyl); or -(CH2)qC6H4R8 (where R8: H;
F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy). - R13: H; lower alkyl; lower alkenyl; -(CH2)mOR55 (where R55: lower alkyl; or lower alkenyl); -(CH2)mSR56 (where R56 : lower alkyl; or lower alkenyl); -(CH2)mNR33R34
(where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl; or R33 and R34 taken together form:
-(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)mOCONR33R75 (where R33: H; or lower alkyl; or lower alkenyl; R75: lower alkyl; or R33 and R75 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -
(CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)r; where R57: H; or lower alkyl); -(CH2)mNR20CONR33R82 (where R20:
H; or lower lower alkyl; R33: H; or lower alkyl; or lower alkenyl; R82: H; or lower alkyl; or R33 and R82 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)r; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)mN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); -(CH2)oCOOR57 (where R57: lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; lower alkyl; or R58 and R59 taken together form: -(CH2)2.5-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -
(CH2)oPO(OR60)2 (where R60: lower alkyl; or lower alkenyl); -(CH2)oSO2R62 (where R62: lower alkyl; or lower alkenyl); or -(CH2)qC6H4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
R1'1: lower alkyl; lower alkenyl; -(CH2)POR55 (where R55: lower alkyl; or lower alkenyl); -(CH2)PSR56 (where R56: lower alkyl; or lower alkenyl); -(CH2)PNR33R34 (where
R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl; or R33 and R34 taken together form:
-(CH2W; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)POCONR33R75 (where R33: H; or lower alkyl; or lower alkenyl; R75: lower alkyl; or R33 and R78 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -
(CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)PNR20CONR33R82 (where R20:
H; or lower lower alkyl; R33: H; or lower alkyl; or lower alkenyl; R82: H; or lower alkyl; or R33 and R82 taken together form: -(CH2W; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)r; or -(CH2)2NR57(CH2)2-; where R57: 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 R58 and R59 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-;
-(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); or - (CH2)oC6H4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
K15: H; lower alkyl; lower alkenyl; -(CH2)oOR55 (where R55: lower alkyl; or lower alkenyl); -(CH2)oSR56 (where R56: lower alkyl; or lower alkenyl); -(CH2)oNR33R34 (where
R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl; or R33 and R34 taken together form: -(CH2W; -(CH2)2O(CH2)r; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)sOCONR33R75 (where R33: H; or lower alkyl; or lower alkenyl; R75: lower alkyl; or R33 and R75 taken together form: -(CH2W; -(CH2)2O(CH2)2-; -
(CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oNR20CONR33R82 (where R20: H; or lower lower alkyl; R33: H; or lower alkyl; or lower alkenyl; R82: H; or lower alkyl; or R33 and R82 taken together form: -(CH2)2-6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)r; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)oN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); -(CH2)oCOOR57 (where R57: lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; lower alkyl; or R58 and R59 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); or - (CH2)AH4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
R16: H; lower alkyl; lower alkenyl; -(CH2)sOR55 (where R55: lower alkyl; or lower alkenyl); -(CH2)sSR56 (where R56: lower alkyl; or lower alkenyl); -(CH2)sNR33R34 (where R33: lower alkyl; or lower alkenyl; R34: H; or lower alkyl; or R33 and R34 taken together form:
-(CH2)2-6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)sOCONR33R75 (where R33: H; or lower alkyl; or lower alkenyl; R75: lower alkyl; or R33 and R75 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; - (CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)sNR20CONR33R82 (where R20: H; or lower lower alkyl; R33: H; or lower alkyl; or lower alkenyl; R82: H; or lower alkyl; or R33 and R82 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)sN(R20)COR64 (where: R20: H; or lower alkyl; R64: lower alkyl; or lower alkenyl); -(CH2)oCOOR57 (where R57: lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; lower alkyl; or R58 and R59 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); or - (CH2)sC6H4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy). - R47: H; or OR55 (where R55: lower alkyl; or lower alkenyl).
R48: H; or lower alkyl.
R49: H;lower alkyl; -(CH2)oCOOR57 (where R57 : lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where R58: lower alkyl; or lower alkenyl; and R59: H; lower alkyl; or R58 and R59 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); or (CH2)sC6H4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy). R5": H; methyl.
R51: 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; or R33 and R34 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); (CH2)mOCONR33R75 (where R33: H; or lower alkyl; or lower alkenyl; R75: lower alkyl; or R33 and R75 taken together form: -
-(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl);
-(CH2)mNR20CONR33R82 (where R20: H; or lower lower alkyl; R33: H; or lower alkyl; or lower alkenyl; R82: H; or lower alkyl; or R33 and R82 taken together form: -(CH2)2.6-;
-(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: 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 R58 and R59 taken together form: -(CH2)M-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); or -(CH2)AH4R8 (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; or R33 and R34 taken together form: -(CH2)2-6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)mOCONR33R75 (where R33: H; or lower alkyl; or lower alkenyl; R75: lower alkyl; or R33 and R75 taken together form: -
(CH2)M-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl);
-(CH2)mNR20CONR33R82 (where R20: H; or lower lower alkyl; R33: H; or lower alkyl; or lower alkenyl; R82: H; or lower alkyl; or R33 and R82 taken together form: -(CH2)2.6-;
-(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; R57: 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 R58 and R59 taken together form:
-(CH2W; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); or -(CH2)rC6H4R8 (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; or R33 and R34 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); -(CH2)mOCONR33R75 (where R33: H; or lower alkyl; or lower alkenyl; R75: lower alkyl; or R33 and R75 taken together form: -
(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl);
-(CH2)mNR20CONR33R82 (where R20: H; or lower lower alkyl; R33: H; or lower alkyl; or lower alkenyl; R82: H; or lower alkyl; or R33 and R82 taken together form: -(CH2)2.6-;
-(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: 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 R58 and R59 taken together form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl); or -(CH2)AH4R8 (where R8: H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
R54: lower alkyl; lower alkenyl; or aryl-lower alkyl.
Most preferably 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.
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 templates (al), (a2) and (a3) 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
GIy Glycine
His L-Histidine
He L-Isoleucine
Leu L-Leucine
Lys L-Lysine
Met L-Methionine
Phe L-Phenylalanine
Pro L-Proline
Pro(5RPhe) (2S,5R)-5-phenylpyrrrolidine-2-carbocyclic acid
Ser L-Serine
Thr L-Threonine
Trp L-Tryptophan
Tyr L-Tyrosine
VaI L-Valine
Cit L-Citrulline
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 c lopenty lalan ine
NIe L-Norleucine
2-Nal L-2-Naphthylalanine 1-Nal L-I -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 -Fluoropheny lalanine 2F-Phe L-2-Fluorophenylalanine Tic L-l,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-Aminobutyric acid (GABA) Aha ε-Aminohexanoic acid Aib α-Aminoisobutyric acid
Y(BzI) L-O-Benzyltyrosine Bip L-Biphenylalanine S(BzI) L-O-Benzylserine T(BzI) 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 MeNIe L-N-Methylnorleucine MeAIa L-N-Methy IaI anine MeIIe L-N-Methylisoleucine
MeVaI 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 imgf000059_0001
A8" wherein R20 is H or lower alkyl and R64 is alkyl; alkenyl; -[(CH2)u-X]rCH3 (where X is
-O-; -NR20-, or -S-, u is 1 -3 and t is 1-6), aryl; aryl-lower alkyl; or heteroaryl-lower alkyl; especially those wherein R64 is n-hexyl (A8"-21); n-heptyl (A8"-22); 4- (phenyl)benzyl (A8"-23); diphenylmethyl (A8"-24); 3-amino-propyl (A8"-25); 5- amino-pentyl (A8"-26); methyl (A8"-27); ethyl (A8"-28); isopropyl (A8"-29); isobutyl (A8"-30); n-propyl (A8"-31); cyclohexyl (A8"-32); cyclohexylmethyl (A8"-33); n-butyl (A8"-34); phenyl (A8M-35); benzyl (A8M-36); (3-indolyl)methyl (A8"-37); 2-(3-indolyl)ethyl (A8"-38); (4-phenyl)phenyl (A8"-39); n-nonyl (A8"- 40); CH3-OCH2CH2-OCH2- (A8"-41) and CH3-(OCH2CH2)2-OCH2- (A8"-42).
The peptidic chain Z of the β-hairpin mimetics described 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 or 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-M-(CH2)pNR20CONR20(CH2)p-CH(CO-)-NR20-; "interstrand linkage" Furthermore, the amino acid residues in chain Z can also be of formula -A-CO- or of formula -B-CO- wherein A and B are as defined above. Finally, GIy can also be an amino acid residue in chain Z, and Pro and Pro(4-NHCOPhe) can be amino acid residues in chain Z, too, with the exception of positions where an interstrand linkage (H) is possible.
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 such residues 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, acylamino- and urea-derived residues according to the general definition for substituent 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 urea derived amino acid 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 being 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, carboxyclic 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 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. , Solid Phase Peptide 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 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 N5N- carbonylimidazole to form cyclic ureas.
As mentioned earlier, positions for an interstrand linkage are positions P2 and 10, 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 abbreviations corresponding to generally adopted usual practice:
three letter code one letter code
Ala L-Alanine A
Arg L-Arginine R
Asn L-Asparagine N
Asp L-Aspartic acid D
Cys L-Cysteine C
GIu L-Glutamic acid E
GIn L-Glutamine Q
GIy Glycine G
His L-Histidine H
He 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
VaI 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
NIe L-Norleucine
2-Nal L-2-Naphthylalanine
1-Nal L-I -Naphthylalanine
4C1-Phe L-4-Chlorophenylalanine
3C1-Phe L-3-Chlorophenylalanine
2C1-Phe L-2-Chlorophenylalanine
3,4C12-Phe L-3 ,4-Dichloropheny lalanine
4F-Phe L-4-Fluorophenylalanine
3F-Phe L-3-Fluorophenylalanine
2F-Phe L-2-Fluorophenylalanine
Tic l,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(BzI) L-O-Benzyltyrosine
Bip L-(4-phenyl)phenylalanine
S(BzI) L-O-Benzylserine
T(BzI) 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-pyrrolidine-L-carboxylic acid
4-AmPyrr2 (2S,4R)-4-Amino-pyrrolidine-L-carboxylic acid
4-PhePyrrl (2S,5R)-4-Phenyl-pyrrolidine-L-carboxylic acid
4-PhePyrr2 (2S,5 S)-4-Pheny 1-pyrrolidine-L-carboxylic acid
5-PhePyrrl (2S,5R)-5-Phenyl-pyrrolidine-L-carboxylic acid
5-PhePyrr2 (2S,5S)-5-Phenyl-pyrrolidine-L-carboxylic acid
Pro(4-OH)1 (4S)-L-Hydroxyproline
Pro(4-OH)2 (4R)-L-Hydroxyproline
Pip L-Pipecolic acid
DPiP D-Pipecolic acid
OctG L-Octylglycine
NGIy N-Methylglycine
MePhe L-N-Methylphenylalanine
MeNIe L-N-Methylnorleucine
MeAIa L-N-Methylalanine MeIIe L-N-Methylisoleucine
MeVaI L-N-Methylvaline
MeLeu L-N-Methylleucine
DimK L-(N' ,NΛ Dimethyl)-lysine
Lpzp L-Piperazinic acid
Dpzp D-Piperazinic acid
Isorn L-(N' ,Nλ -diisobuty l)-ornithine
PipAla L-2-(4λ -piperidiny l)-alanine
PirrAla L-2-(3"-pyrrolidinyl)-alanine
Ampc 4-Amino-piperidine-4-carboxylic acid
NMeR L-N-Methylarginine
NMeK L-N-Methyllysine
NMePhe L-N-Methylphenylalanine
IPegK L-2-Amino-6-{2-[2-(2-methoxy- ethoxy)ethoxy]acety lamino } -hexanoic acid
SPegK L-2-Amino-6-[2-(2methoxy-ethoxy)- acetylamino]-hexanoic acid
Dab L-2,4-Diamino-butyric acid IPegDab L-2-Araino-4{2-[2-(2-methoxy-ethoxy)-ethoxy]- acetylamino} -butyric acid
SPegDab L-2-Amino-4[2-(2-methoxy-ethoxy)-acetylamino] butyric acid
4-PyrAla L-2-(4Tyridyl)-alanine OrnPyr L-2-Amino-5-[(2'carbonylpyrazine)]amino- pentanoic acid
BnG N-Benzylglycine
AlIoT Allo-Threonin
Pro(4NHCOPhe) (2S)-4-benzamidino-pyrrolidine-2-carboxylic acid
Aoc 2-(S)-Aminooctanoic acid
Particularly preferred residues for group C are: Ala L-Alanine
He L-Isoleucine
Leu L-Leucine
Met L-Methionine VaI L-Valine tBuA L-t-Butylalanine t-BuG L-tert.-Butylglycine
Cha L-Cyclohexylalanine C4al L-3-Cyclobutylalanine
C5al L-3-Cyclopentylalanine
NIe L-Norleucine hCha L-Homo-cyclohexylalanine
OctG L-Octylglycine MePhe L-N-Methylphenylalanine
MeNIe L-N-Methylnorleucine
MeAIa L-N-Methylalanine
MeIIe L-N-Methylisoleucine
MeVaI L-N-Methylvaline MeLeu L-N-Methylleucine
Aoc 2-(S)-Aminooctanoic acid
Particularly 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-NaphthylaIanine 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(BzI) L-O-Benzyltyrosine Bip L-Biphenylalanine
S(BzI) L-O-Benzylserine
T(BzI) L-O-Benzylthreonine hPhe L-Homo-phenylalanine
Bpa L-4-Benzoylphenylalanine
PirrAla L-2-(3 ' -pyrrolidiny l)-alanine
NMePhe L-N-Methylphenylalanine
4-PyrAla L-2-(4Λ Pyridy l)-alanine
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(mNH2) L-meta-Aminophenylalanine
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
DimK L-(N\N'Dimethyl)-lysine
Isorn L-(N\N*-diisobutyl)-ornithine
NMeR L-N-Methylarginine
NMeK L-N-Methyllysine
IPegK L-2-Amino-6-{2-[2-(2-methoxy- ethoxy)ethoxy]acetylamino}-hexanoic acid
SPegK L-2-Amino-6-[2-(2methoxy-ethoxy)- acetylamino]-hexanoic acid
Dab L-2,4-Diamino-butyric acid
IPegDab L-2-Amino-4 { 2-[2-(2-methoxy-ethoxy)-ethoxy] acetylamino}-butyric acid SPegDab L-2-Amino-4[2-(2-methoxy-ethoxy)-acetylamino] butyric acid
OrnPyr L-2-Amino-5-[(2Λ carbonylpyrazine)]amino- pentanoic
PipAla L-2-(4* -piperidinyl)-alanine
Particularly preferred residues for group F are Asn L-Asparagine
Asp L-Aspartic acid
Cys L-Cysteine
GIn L-Glutamine
GIu L-Glutamic acid Ser L-Serine
Thr L-Threonine
AlloThr AlIo 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 11 amino acid residues. The positions Pl to Pl 1 of each amino acid residue in the chain Z are unequivocally defined as follows: Pl 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), or of the group -B-CO- forming the C-terminus of template (a3); and Pl 1 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), or of the group - B-CO- forming the N-terminus of template (a3). Each of the positions Pl to Pl 1 will preferably contain an amino acid residue belonging to one of the above types C, D, E, F, H, or of formula -A-CO- or of formula -B-CO-, or being GIy, Pro or Pro(4NHCOPhe) as follows:
In general the α-amino acid residues in positions 1 to 11 of the chain Z are preferably: - Pl : of type C, or of type D, or of type E, or of type F;
P2: of type E, or of type F, or of type C;
P3: or of type C, of type F or the residue is GIy;
P4: of type C, or of type E, or of type F, or the residue is GIy or Pro;
P5: of type E, or of type F, or the residue is GIy or Pro; - P6: of type C, or of type D, or of type F, or the residue is GIy or Pro;
P7: of type F or of formula -A-CO-or the residue is GIy or Pro;
P8: of type D, or of type C, or of formula -A-CO or the residue is GIy or Pro or Pro(4NHCOPhe);
P9: of type C, or of type D, or of type E, or of type F; - PlO: of type F, or of type C, or type E;
PI l: of type E, or of type F, or of type C or of type D; or
P2 and PlO, taken together, form a group of type H;
with the proviso that if template is DPro-LProthe amino acid residues in positions Pl to Pl 1 are other than
Pl : Arg
P2: Cys, linked with Cys in position PlO by a disulfide bridge
P3: Thr
P4 Lys - P5 Ser
P6 He
P7 Pro
P8 Pro
P9 He - PlO Cys, linked with Cys in position Pl 0 by a disulfide bridge; and
PI l Phe.
The α-amino acid residues in positions 1 to 11 are most preferably:
Pl : NIe, He, Aoc, hLeu, Chg, OctG, hPhe, 4AmPhe3 Cha, Phe, Tyr, 2C1-Phe, Trp, 1 -NaI, Leu, Cha, or Arg; P2: Cys, GIu, NIe, Thr, or GIn ; P3: Thr, Ala or Abu; P4: Lys, NIe, Ala, Abu, or Thr; P5: Ser, AlloThr, or Dpr; - P6: He, c5al, Leu, NIe, Aoc, OctG, Cha, hLeu, hPhe, Chg, t-BuA, GIu5 or Asp; P7: Pro;
P8: Pro, Ala, or Pro(4NHCOPhe);
P9: Tyr, Phe, He, NIe, Cha, GIn, Arg, Lys, His, Thr, or Ala; - PlO: Cys, Arg, NIe, GIn, Lys, Met, Thr, or Ser;
PI l: Tyr, GIn, Arg, Ser, NIe, 2-Nal, 2C1-Phe, Cha, Phg, Tyr, Phe, Asp,
Asn, or Thr; and
Cys, if present at P2 and PlO, may form a disulfide bridge.
For inhibitors of Cathepsin G the α-amino acid residues in positions 1 to 1 1 of the chain Z are preferably:
Pl : of type C, or of type D, or of type E;
P2: of type F, or of type C; - P3: of type F;
P4: of type C, or of type E;
P5: of type E, or of type F;
P6: of type F;
P7: of type F, or of formula -A-CO-, or the residue is GIy or Pro; - P8: of type C, or of formula -A-CO-, or the residue is GIy or Pro or
Pro(4NHCOPhe);
P9: of type C, or of type D, or of type F;
PlO: of type F, or of type C, or type E;
PI l : of type E, or of type D, or of type F; or - P2 and PlO, taken together, form a group of type H.
For inhibitors of Cathepsin G, the α-amino acid residues in positions 1 to 11 are most preferably
Pl : Phe, hPhe, 4AmPhe, NIe, Chg, He, Tyr, Arg, Trp, 2C1-Phe, Arg, 1 - NaI, or Cha; P2: Cys, GIu, or NIe;
P3: Thr;
P4: Lys, or NIe;
P5: Ser, AlloThr, or Dpr; - P6: Asp, or GIu;
P7: Pro;
P8: Pro;
P9: lie, NIe, Cha, GIn, Tyr, or Ala;
PlO: Cys, Arg, or NIe; - PI l : Thr, Asp, Ser, Tyr, Phe, Asn, or Arg; and
Cys, if present at P2 and PlO, may form a disulfide bridge.
For inhibitors of Elastase the α-amino acid residues in positions 1 to 11 of the chain Z are preferably - Pl : of type C, or of type D;
P2: of type F;
P3: of type F or of type C;
P4: of type C or of type F;
P5: of type F; - P6: of type C;
P7 : of formula -A-CO-or the residue is GIy or Pro;
P8: of formula -A-CO or the residue is GIy or Pro or
Pro(4NHCOPhe);
P9: of type D, or of type F or of type C; - PlO: of type F, or of type C, or type E;
Pl 1 : of type E, or of type F, or of type D; or
P2 and PlO, taken together, form a group of type H.
For inhibitors of Elastase, the α-amino acid residues in positions 1 to 1 1 are most preferably:
Pl : He, NIe, Aoc, hLeu, Chg, OctG, or hPhe;
P2: Cys, GIu, Thr, or GIn;
P3: Thr, Ala, or Abu;
P4: Ala, Thr, or Abu; - P5: Ser; P6: OctG, He, Cha, Leu, c5al, NIe, Aoc, Chg, tBuA, or hLeu;
P7: Pro;
P8: Pro, or Pro(4NHCOPhe);
P9: GIn, Tyr, ILe, or Phe;
PlO: Cys, Lys, GIn, Thr, Met, or Arg;
PI l : Tyr, Ser, Arg, GIn, NIe, 2-Nal, 2C1-Phe, Phe, Cha, or Phg; and
Cys, if present at P2 and PlO, may form a disulfide bridge.
For inhibitors of Tryptase the α-amino acid residues in positions 1 to 11 of the chain Z are preferably:
Pl : of type C, or of type D, or of type E;
P2: of type F;
P3: oftype F; - P4: oftype E;
P5: oftype F;
P6: of type C, or of type D;
P7: of type F, or of formula -A-CO-, or the residue is GIy or Pro;
P8: of type C, or of formula -A-CO-, or the residue is GIy or Pro; - P9: of type C, or of type E, or of type F;
PlO: of type F;
PI l : of type E, or of type D; or
P2 and PlO, taken together, form a group of type H; with the proviso that if the template is DPro-LPro, the amino acid residues in positions Pl to Pl 1 are other than
Pl : Arg
P2: Cys, linked with Cys in position PlO by a disulfide bridge
P3: Thr
P4 Lys - P5 Ser
P6 He
P7 Pro
P8 Pro
P9 He - PlO Cys, linked with Cys in position PlO by a disulfide bridge; and PIl Phe.
For inhibitors of Tryptase the α-amino acid residues in positions 1 to 11 of the chain Z are most preferably:
Pl : Cha, Tyr , or Trp
P2: Cys
P3: Thr
P4: Lys
P5: Ser
P6: Leu
P7: Pro
P8: Pro
P9: Lys
PlO: Cys
PI l: Arg; and the Cys residues present at P2 and PlO may form a disulfide bridge.
Particularly preferred β-peptidomimetics of the invention include those described in Examples 5, 19, 20, 22, 23, 38, 39, 40, and 75 as inhibitors of cathepsin G; Examples 91, 121, 153, 154, 155, 156, 157, 158, 159, 160, 161 177, and 178 as inhibitors of elastase; and Examples 193, 194, and 195 as inhibitors of Tryptase.
The processes of the invention can advantageously be carried out as parallel array syntheses to yield libraries of template-fixed β-hairpin peptidomimetics of the above general formula I. Such parallel syntheses allow 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 Synthesis of Small-Molecular-Weight Compound Libraries", Tetrahedron Organic Chemistry Series, 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 two types of linkers are used:
Type 1 linkers are designed to release the amide group under acidic conditions (Rink H, Tetrahedron Lett. 1987, 28, 3783-3790). Linkers of this kind form amides of the carboxyl group of the amino acids; examples of resins functionalized by such linker structures include 4-[(((2,4-dimethoxyphenyl)Fmoc-aminomethyl)phenoxyacetamido) aminomethyl] PS resin, 4-[(((2,4-dimethoxyphenyl)Fmoc- aminomethyl)phenoxyacetamido) aminomethyl] -4-methylbenzydrylamine PS resin (Rink amide MBHA PS Resin), and 4-[(((2,4-dimethoxyphenyl)Fmoc- aminomethyl)phenoxyacetamido) aminomethyl] benzhydrylamine PS-resin (Rink amide BHA PS resin). Preferably, the support is derived from polystyrene crosslinked with, most preferably 1-5%, divinylbenzene and functionalized by means of the 4- (((2,4- dimethoxyphenyl)Fmoc-aminomethyl)phenoxyacetamido) linker.
Type 2 linkers are designed to eventually release the carboxyl group under acidic conditions. Linkers of this kind form acid-labile esters with the carboxyl group of the amino acids, usually acid-labile benzyl, benzhydryl and trityl esters; examples of such linker structures 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 parallel array syntheses the processes of the invention can be advantageously carried out as described herein below but it will be immediately apparent to those skilled in the art how these procedures 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 which is preferably derived from polystyrene cross-linked with 1 to 3% of divinylbenzene, or from 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)3 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, 29 4005-4008) 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 processes • 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 triniethylsilylethoxycarbonyl
Tec trichloroethoxycarbonyl
Nps o-nitrophenylsulfonyl;
Trt triphenyraethyl 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
AUyI
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
TYt 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-hairpin 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 acid derivative, is usually 1 to 20 equivalents based on the milliequivalents per gram (meq/g) loading of the functional ized 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 and diisopropylurea is insoluble and, respectively, 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 hexafiuorophosphate (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 (DPPA) or O-(7-aza-benzotriazol-l-yl)-N,N,N',N!- tetramethyluronium tetrafluoroborate (TATU) or O-(7-aza-benzotriazol-l-yl)-N,N,N',N'- tetramethyluronium hexafiuorophosphate (HATU)/7-aza-l -hydroxy benzotriazole (HOAt, Carpino et al., Tetrahedron 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 by-product 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 has been obtained.
Before this fully protected linear peptide is detached from 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 an amino protecting group which can be selectively removed, e.g. by means of Pd° and phenylsilane in
CH2Cl2, 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. For the formation of pegylated amino acids such as IPegK, or SPegK, preferably a solution of 5 equivalents of HATU (N-[(dimethylamino)-lH-l ,2,3-triazolo[4,5-έ>]pyridin-l- ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide) in dry DMF and a solution of 10 equivalents of DIPEA (Diisopropyl ethylamine) in dry DMF and 5 equivalents of 2-[2-(2-methoxyethoxy)ethoxy] acetic acid (IPeg) and, respectively, 2-(2- methoxyethoxy)acetic acid (sPeg), is applied to the liberated amino group of the appropiate amino acid side chain for 3 h. The procedure is thereafter repeated for another 3h with a fresh solution of reagents after filtering and washing the resin. Before this fully protected linear peptide is detached from the solid support , it is also possible, if desired, to form an interstrand linkages between side-chains of appropriate amino acid residues at positions 2 and 10.
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 cysteine and homocysteine residues at opposite positions of the β-strand; or lactam bridges formed by glutamic and aspartic acid residues linking ornithine and, respectively, lysine residues, or by glutamic acid residues linking 2,4- diaminobutyric acid residues 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.
For the formation of disulfide bridges preferably a solution of 10 equivalents of iodine solution is applied in DMF or in a mixture Of CH2Cb /MeOH for 1.5 h which is repeated for another 3h with a fresh iodine solution after filtering of the iodine solution, or in a mixture of DMSO and acetic acid solution, buffered with 5% with NaHCO3 to pH 5-6 for 4h, or in water adjusted to pH 8 with ammonium hydroxide solution by stirring for 24h, or in ammonium acetate buffer adjusted to pH 8 in the presence of air, or in a solution of NMP and tri-n- butylphosphine (preferably 50 eq.).
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 16 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, such as DCM, and the solution is extracted with water or a mixture of water- miscible solvents, in order to remove any excess of the coupling reagent.
Finally, the fully protected peptide derivative is treated with 95% TFA, 2.5% H2O, 2.5% TIS or another combination of scavengers for effecting the cleavage of protecting groups. The cleavage reaction time is commonly 30 minutes to 12 hours, preferably about 2.5 hours. The volatiles are evaporated to dryness and the crude peptide is dissolved in 20% AcOH in water and extracted with isopropyl ether or other solvents which are suitable therefor. The aqueous layer is collected and evaporated to dryness, and the fully deprotected cyclic peptide derivative of formula I is obtained as end-product.
Alternatively the detachment, cyclization and complete deprotection of the fully protected peptide from the solid support can be achieved manually in glass vessels.
Depending on its 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 of formula I 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 template starting materials of formula II used in the processes of the invention, pre- starting materials therefor, and the preparation of these starting and pre-starting materials are described in International Application PCT/EP02/01711 of the same applicants, published as WO 02/070547 Al .
The β -hairpin peptidomimetics of the invention can be used in a wide range of applications where inflammatory diseases or pulmanory diseases or infections or immunological diseases or cardiovascular diseases or neurodegenerative diseases are mediated or resulting from serine protease activity, or where cancer is mediated or resulting from serine protease activity. For the control or prevention of a given illness or disease amenable to treatment with protease inhibitors, the β-hairpin peptidomimetics may be administered per se or may be applied as an appropriate formulation together with carriers, diluents or excipients well known in the art.
When used to treat or prevent diseases such as pulmonary emphysema, rheumatoid arthritis, osteoarthritis, atherosklerosis, psoriaris, cystic fibrosis, multiple sclerosis, adult respiratory distress syndrome, pancreatitis, asthma, allergic rhinitis, inflammatory dermatoses, postangioplasty restenosis, cardiac hypertrophy, heart failure or cancer such as, but not limited to, breast cancer, or cancer related to angiogenesis or metastasis, the β- hairpin peptidomimetics can be administered singly, as mixtures of several β-hairpin peptidomimetics, in combination with other anti-inflammatory agents, or antimicrobial agents or anti-cancer agents and/or in combination with other pharmaceutically active agents. The β-hairpin peptidomimetics can be administered per se or as pharmaceutical compositions.
Pharmaceutical compositions comprising β-hairpin 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-hairpin peptidomimetics into preparations which can be used pharmaceutically. Proper formulation depends upon the method of administration chosen. For topical administration the β-hairpin peptidomimetics of the invention may be formulated as solutions, gels, ointments, creams, suspensions, etc. as are well-known in the art.
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 adequate solutions, preferably in physiologically compatible buffers such as Hink's solution, Ringer's solution, or physiological saline buffer. The solutions may contain formulatory agents such as suspending, stabilizing and/or dispersing agents.
Alternatively, the β-hairpin 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 β-hairpin peptidomimetics of the invention can be readily formulated by combining them with pharmaceutically acceptable carriers well known in the art. Such carriers enable the β-hairpin peptidomimetics of the invention to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions etc., for oral ingestion by a patient to be treated. For oral formulations such as, for example, powders, capsules and tablets, suitable excipients include fillers such as sugars, e. g. 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, des integrating agents may be 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-hairpin peptidomimetics 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-hairpin 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 formulations described previously, the β-hairpin 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-hairpin 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 may also be employed. Additionally, the β-hairpin 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-hairpin pepdidomimetics of the invention may contain charged residues, they may be included in any of the above-described formulations as such or as pharmaceutically acceptable salts. Pharmaceutically acceptable salts tend to be more soluble in aqueous and other protic solvents than are the corresponding free forms.
The β-hairpin peptidomimetics of the invention, or compositions thereof, will generally be used in an amount effective to achieve the intended purpose. It is to be understood that the amount used will depend on a particular application.
For topical administration to treat or prevent diseases amenable to treatment with beta hairpin mimetics a therapeutically effective dose can be determined using, for example, the in vitro assays provided in the examples. The treatment may be applied while the disease is visible, or even when it is not visible. An ordinary skilled expert will be able to determine therapeutically effective amounts to treat topical diseases without undue experimentation.
For systemic administration, a therapeutically effective dose can be estimated initially from in vitro assays. For example, a dose can be formulated in animal models to achieve a circulating β-hairpin peptidomimetic concentration range that includes the IC50 as determined in the cell culture. Such information can be used to more accurately determine useful doses in humans.
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 skill in the art could readily optimize administration to humans based on animal data.
Dosage amounts for applications as serine protease inhibitory agents may be adjusted individually to provide plasma levels of the β-hairpin peptidomimetics of the invention which are sufficient to maintain the therapeutic effect. Therapeutically effective serum levels may be achieved by administering multiple doses each day.
In cases of local administration or selective uptake, the effective local concentration of the β-hairpin peptidomimetics of the invention may not be related to plasma concentration. One having the ordinary skill in the art will be able to optimize therapeutically effective local dosages without undue experimentation. The amount of β-hairpin 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-hairpin peptidomimetics described herein will provide therapeutic benefit without causing substantial toxicity.
Toxicity of the β-hairpin peptidomimetics of the invention 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 LD100 (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-hairpin 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.1, p.l).
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
(Knorr et al. Tetrahedron Lett, 1989, 30, 1927-1930); HOBt: 1-hydroxybenzotriazole; DIEA: diisopropylethylamine; HOAT: 7-aza-l-hydroxybenzotriazole;
HATU : O-(7-aza-benzotriazo Ie- 1 -y 1)-N,N,N ' ,N ' -tetramethy lurono i um hexafluorophosphate (Carpino et al. Tetrahedron Lett. 1994, 35, 2279-2281). Examples
1. Peptide synthesis
Coupling of the first protected amino acid residue to the resin
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 CH2CI2 (2.5 ml) and allowed to swell at room temperature under constant stirring for 30 min. 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 CH2Cl2 (2.5 ml), the mixture was shaken at 25°C for 4 hours. The resin colour changed to purple and the solution remained yellowish. The resin was shaken (CH2Cl2 /MeOH/DIEA : 17/2/1), 30 ml for 30 min; then washed in the following order with CH2Cl2 (Ix), DMF (Ix), CH2Cl2 (Ix), MeOH (Ix), CH2Cl2(Ix), MeOH (Ix), CH2Cl2 (2x), Et2O (2x) and dried under vacuum for 6 hours. Loading was typically 0.6-0.7 mMol/g.
The following preloaded resins were prepared: Fmoc-Pro-2-chlorotritylresin, Fmoc-Asp (OtBu)-2-chlorotritylresin, Fmoc-Pro(5RPhe)-2-chlorotritylresin, Fmoc-Leu-2-chlorotritylresin, Fmoc-Glu(OtBu)-2-chlorotritylresin, Fmoc- Asp(OtBu) -2-chlorotritylresin,. Fmoc-Phe-2-chlorotritylresin, Fmoc-Gln(Trt)-2- chlorotritylresin, Fmoc-Ser (OtBu) -2-chlorotritylresin, Fmoc-Val-2- chlorotritylresin, Fmoc-Thr(OtBu) -2-chlorotritylresin and Fmoc-Ile-2- chlorotritylresin.
Synthesis of the fully protected peptide fragment
The synthesis was carried out using a Syro-peptide synthesizer (Multisyntech) using 24 to 96 reaction vessels. In each vessel were 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 CH2Cl2, wash and swell (manual) 3 x 1 min.
2 DMF, wash and swell 1 x 5 min
3 40 % piperidine/DMF 1 x 5 min. 4 DMF, wash 5 x 2 min.
5 5 equiv. Fmoc amino acid/DMF
+ 5 eq. HBTU + 5 eq. HOBt
+ 5 eq. DIEA I x 120 min. ■ 6 DMF, wash 4 x 2 min.
7 CH2Cl2, wash (at the end of the synthesis) 3 x 2 min.
Steps 3 to 6 are repeated to add each amino-acid.
After the synthesis of the fully protected peptide fragment had been terminated, then subsequently either Procedure A or Procedure B , as described hereinbelow, was adopted, depending on whether not interstrand linkages (i. e. disulfide β-strand linkages) were to be formed.
Procedure A: Cyclization and work up of backbone cyclized peptides
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 CH2Cl2 (v/v) for 3 minutes, filtered and the filtrate was neutralized with 1 ml ( 1.17 mMol, 3 eq.) of 20% DIEA in CH2Cl2 (v/v). This procedure was repeated twice to ensure completion of the cleavage. An aliquot (200 μL) of the filtrate was fully deprotected with 0.5ml of the cleavage mixture containing 95% trifluoroacetic acid (TFA), 2.5% water and 2.5% triisopropylsilane (TIS) and analysed by reverse phase-LC MS to monitor the efficiency of the linear peptide synthesis.
Cyclization of the linear peptide
The fully protected linear peptide was dissolved in DMF (8 ml, cone. 10 mg/ml). Two eq. of HATU (0.72 mMol) in ImI of DMF and 4 eq. of DIEA (1.44 mMol) in I mI of DMF were added, and the mixture was stirred at room temperature for 16 h. The volatile was evaporated to dryness. The crude cyclised peptide was dissolved in 7 ml Of CH2Cl2 and extracted with 10% acetonitrile in water (4.5 ml) three times. The CHbCl2 layer was evaporated to dryness.
Deprotection and purification of the cyclic peptide The cyclic peptide obtained was dissolved in 3 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 2O0C for 2.5 hours and then concentrated under vacuum. The crude peptide was dissolved in 20% AcOH in water (7 ml) and extracted with diisopropylether (4 ml) three times. The aqueous layer was collected and evaporated to dryness, and the residue was purified by preparative reverse phase LC-MS.
After lyophilisation the products were obtained as white powders and analysed by LC- MS. The analytical data comprising purity after preparative HPLC and ESI-MS are shown in Table 1.
Analytical method:
Analytical HPLC retention times (RT, in minutes) were determined using an Jupiter Proteo 9OA, 150 x 2.0 mm, (cod. 00F4396-B0 - Phenomenex) with the following solvents A (H2O + 0.1% TFA) and B (CH3CN + 0.1% TFA) and the gradient: 0 min: 95%A, 5%B; 20 min: 40%A 60%B; 21-23 min: 0%A, 100%B; 23.1-30 min: 95% A, 5%B.
Procedure B: Cyclization and work up of backbone cyclized peptides having disulfide β- strand linkages
Formation of disulfide β-strand linkage
After completion of the synthesis, the resin was swelled in 3 ml of dry DMF for 1 h. Then 10 eq. of iodine solution in DMF (6 ml) were added to the reactor, followed by stirring for 1.5 h. The resin was filtered and a fresh solution of iodine (10 eq.) in DMF (6 ml) was added, followed by stirring for another 3 h. The resin was filtered and washed with DMF (3x) and CH2Cl2 (3x).
Backbone cyclization, cleavage and purification of the peptide
After formation of the disulfide β-strand linkage, the resin was suspended in 1 ml (0.39 mMol) of 1% TFA in CH2Cl2 (v/v) for 3 minutes and filtered, and the filtrate was neutralized with ImI (1.17 mMol, 3eq.) of 20% DIEA in CH2Cl2 (v/v). This procedure was repeated twice to ensure completion of the cleavage. The resin was washed with 2ml Of CH2Cl2. The CH2Cl2 layer was evaporated to dryness.
The fully protected linear peptide was solubilised in 8 ml of dry DMF. Then 2 eq. of HATU in dry DMF (ImI) and 4 eq. of DIPEA in dry DMF (1 ml) were added to the peptide, followed by stirring for 16 h. The volatiles were evaporated to dryness. The crude cyclised peptide was dissolved in 7 ml Of CH2Cl2 and extracted with 10% acetonitrile in water (4.5 ml) three times. The CH2Cl2 layer was evaporated to dryness. To deprotect the peptide fully, 3 ml of cleavage cocktail TFA:TIS:H2O (95:2.5:2.5) were added, and the mixture was kept for 2.5 h. The volatile was evaporated to dryness and the crude peptide was dissolved in 20% AcOH in water (7 ml) and extracted with diisopropyl ether (4 ml) for three times. The aqueous layer was collected and evaporated to dryness, and the residue was purified by preparative reverse phase LC-MS.
After lyophilisation the products were obtained as white powders and analysed by ESI- MS analytical method as described above. The analytical data comprising purity after preparative HPLC and ESI-MS are shown in Table 1.
Examples 1-45, 52-63, 65-67, 70-71, 75-114, 129, 131-162 and 179-196 are shown in Table 1. The peptides were synthesized starting with the amino acid Pro which was grafted to the resin. Starting resin was Fmoc-Pro-2-chlorotrityl resin, which was prepared as described above. The linear peptides were synthesized on solid support according to the procedure described above in the following sequence: Resin-Pro- DPro-Pl 1-P10-P9- P8-P7-P6-P5-P4-P3-P2-P1. Ex. 1-6, 9-45, 52-63, 65-67, 70-71, 75-103 112-114, 129, 131, 133, 136-138, 140-141, 143-146, 148-153, 155, 157-162 and 179-196 were cleaved from the resin, subjected to the disulfide bridge formation, cyclized, deprotected and purified as indicated in procedure B. Ex. 82, 123, 149, 159, 161 and 178 were cleaved from the resin as indicated in procedure B. The disulfide bridges were formed using the following procedure: The crude product was solubilised in a ammonium acetate buffer 0.1 M (pH adjusted to 8) (concentration: lmg of crude product per ml). The mixture was stirred at room temperature in presence of air. The reaction was monitored by reverse phase LC-MS. After reaction completion, the solution was evaporated to dryness and the residue purified by preparative reverse phase LC-MS. The cyclisation of the backbone was perfomed as indicated in procedure A. The deprotection was performed using the following procedure:
To deprotect the peptide fully, 5 ml of cleavage cocktail TFA:H20:Pheπol:Thioanisol: Ethanedithiol (82.5:5:5:5:2.5) were added, and the mixture was kept for 5h at room temperature. The peptide was precipitated by addition of cold diethylether (10ml). After centrifugation, the supernatant phase was removed. The precipitate was washed three times with 5 ml of diethylether and was purified by preparative reverse phase LC-MS.
After lyophilisation the products were obtained as white powders and analysed by ESI- MS analytical method as described above.
Ex. 7, 8, 104-111, 132, 134, 135, 139, 142, 147, 154 and 156 were cleaved from the resin, cyclized, deprotected and purified as indicated in procedure A.
HPLC-retention times (minutes) were determined using the analytical method as described above:
Ex. 1 (15.37), Ex. 2 (11.54), Ex. 3 (7.82), Ex. 4 (8.62), Ex. 5 (16.51), Ex. 6 (13.67), Ex. 7 (3.61), Ex. 8 (4.1 1), Ex. 9 (5.82), Ex. 10 (7.98), Ex. 11 (8.38), Ex. 12 (6.80), Ex. 13 (7.41), Ex. 14 (6.20), Ex. 15 (8.68), Ex. 16 (9.82); Ex. 17 (5.59), Ex. 20 (7.32), Ex. 21 (8.66), Ex. 22 (8.68), Ex. 23 (12.66), Ex. 24 (8.67), Ex. 25 (7.53), Ex. 26 (9.02), Ex. 27 (8.06), Ex. 28 (9.62), Ex. 29 (8.78), Ex. 30 (10.49), Ex. 31 (5.50), Ex. 32 (7.45), Ex. 33 (8.39), Ex. 34 (10.16), Ex. 35 (9.04), Ex. 36 (10.98), Ex. 37 (7.56), Ex. 38 (9.29), Ex. 39 (8.32), Ex. 40 (10.11), Ex. 41 (7.23), Ex. 42 (8.83), Ex. 43 (7.92), Ex. 44 (9.87), Ex. 45 (8.26), Ex. 52 (6.20), Ex. 53 (8.68), Ex 54 (9.82), Ex. 55 (5.59), Ex. 56 (6.06), Ex. 57 (6.47), Ex. 58 (7.32), Ex. 59 (8.68), Ex. 60 (10.66), Ex. 61 (8.54), Ex. 62 (9.83), Ex. 63 (16.54), Ex. 65 (15.71), Ex. 66 (17.50), Ex. 67 (15.87), Ex. 70 (12.87), Ex. 71 (13.48), Ex. 75 (14.22), Ex. 76 (4.47), Ex. 77 (5.15), Ex. 78 (10.93), Ex. 79 (10.70), Ex. 80 (12.09), Ex. 81 (1 1.63), Ex. 82 (5.71), Ex. 83 (5.45), Ex. 84 (1 1.14), Ex. 85 (10.90), Ex. 86 (13.78), Ex. 87 (13.98), Ex. 88 (14.35), Ex. 89 (15.21), Ex. 90 (14.72), Ex. 91 (1 1.97), Ex. 92 (11.77), Ex. 93 (15.25), Ex. 94 (14.61), Ex. 95 (20.46), Ex. 96 (15.08), Ex. 97 (20.78), Ex. 98 (18.28), Ex. 99 (14.62), Ex. 100 (13.90), Ex. 101 (13.76), Ex. 102 (20.53), Ex. 103 (14.14), Ex. 104 (1 1.60), Ex. 105 (11.90), Ex. 106 (1 1.63), Ex. 107 (1 1.78), Ex. 108 (13.03), Ex. 109 (15.22), Ex. 110 (12.40), Ex. Ill (12.10), Ex. 112 (5.49), Ex. 113 (5.67), Ex. 114 (5.55), Ex. 129 (17.22), Ex. 131 (1 1.97), Ex. 132 (13.56), Ex. 133 (14.57), Ex. 134 (14.72), Ex. 135 (17.53), Ex. 136 (18.28), Ex. 137 (14.72), Ex. 138 (14.35), Ex.139 (15.40), Ex.140 (11.14), Ex.141 (5.71), Ex.142 (13.97), Ex.143 (13.94), Ex.144 (15.08), Ex.145 (20.87), Ex.146 (17.91), Ex.147 (17.11), Ex.148 (7.83), Ex.149 (16.22), Ex.150 (20.09), Ex.151 (20.72), Ex.152 (21.38), Ex.153 (17.97), Ex.154 (16.58), Ex.155 (19.46), Ex.156 (15.66), Ex.157 (22.04), Ex.158 (15.65), Ex.159 (17.89), Ex.160 (18.72), Ex.161 (19.91), Ex.162 (17.79), Ex.179 (4.25), Ex.180 (11.43), Ex.181 (12.30), Ex.182 (12.83), Ex.183 (10.51), Ex.184 (12.12), Ex.185 (10.14), Ex.186 (10.09), Ex.187 (10.14), Ex.188 (10.65), Ex.189 (10.73), Ex.190 (10.10), Ex.191 (10.17), Ex.192 (10.19), Ex.193 (11.02), Ex.194 (9.92), Ex.195 (10.74), Ex.196 (9.94).
Example 46 is shown in Table 1. The peptide was synthesized starting with the amino acid Pro which was grafted to the resin. Starting resin was Fmoc-Pro-2-chlorotrityl resin, which was prepared as described above. The linear peptide was synthesized on solid support according to procedure described above in the following sequence: Resin-Pro- DAsp(OtBu)-Pl 1-P10-P9-P8-P7-P6-P5-P4-P3-P2-P1. Thereafter the disulfide bridge was formed, and the peptide was cleaved from the resin, cyclized, deprotected and purified as indicated in procedure B.
HPLC-retention time (minutes) was determined using the analytical method described above: Ex. 46 (8.94).
Example 47 is shown in Table 1. The peptide was synthesized starting with the amino acid Asp which was grafted to the resin. Starting resin was Fmoc-Asp(OtBu)-2- chlorotrityl resin, which was prepared as described above. The linear peptide was synthesized on solid support according to procedure described above in the following sequence: Resin- Asp(OtBu)-DPro-Pl 1 -Pl 0-P9-P8-P7-P6-P5-P4-P3-P2-P1. Thereafter the disulfide bridge was formed, and the peptide was cleaved from the resin, cyclized, deprotected and purified as indicated in procedure B. HPLC-retention time (minutes) was determined using the analytical method as described above:
Ex. 47 (7.29).
Example 48 is shown in Table 1. The peptide was synthesized starting with the amino acid Pro(5RPhe) which was grafted to the resin. Starting resin was Fmoc-Pro(5RPhe)-2- chlorotrityl resin, which was prepared as described above. The linear peptide was synthesized on solid support according to procedure described above in the following sequence: Resin-Pro(5RPhe)-DPr o-Pl 1-P10-P9-P8-P7-P6-P5-P4-P3-P2-P1. Thereafter the disulfide bridge was formed, and the peptide was cleaved from the resin, cyclized, deprotected and purified as indicated in procedure B. HPLC-retention time (minutes) was determined using the analytical described above: Ex.48 (10.07).
Example 49 is shown in Table 1. The peptide was synthesized starting with the amino acid Pro which was grafted to the resin. Starting resin was Fmoc-Pro-2-chlorotrityl resin, which was prepared as described above. The linear peptide was synthesized on solid support according to procedure described above in the following sequence: Resin-Pro- 0AIa-Pl 1 -Pl 0-P9-P8-P7-P6-P5-P4-P3-P2-P1. Thereafter the disulfide bridge was formed, and the peptide was cleaved from the resin, cyclized, deprotected and purified as indicated in procedure B. HPLC-retention time (minutes) was determined using the analytical method described above: Ex.49 (8.09);
Example 50 is shown in Table 1. The peptide was synthesized starting with the amino acid Pro which was grafted to the resin. Starting resin was Fmoc-Pro-2-chlorotrityl resin, which was prepared as described above. The linear peptide was synthesized on solid support according to procedure described above in the following sequence: Resin- Pro- 0IIe-Pl 1-P10-P9-P8-P7-P6-P5-P4-P3-P2-P1. Thereafter the disulfide bridge was formed, and the peptide was cleaved from the resin, cyclized, deprotected and purified as indicated in procedure B.
HPLC-retention time (minutes) was determined using the analytical described above: Ex.50 (9.78).
Example 51 is shown in Table 1. The peptide was synthesized starting with the amino acid Leu which was grafted to the resin. Starting resin was Fmoc-Leu-2-chlorotrityl resin, which was prepared as described above. The linear peptide was synthesized on solid support according to procedure described above in the following sequence: Resin-Leu- DPro-Pl 1-P10-P9-P8-P7-P6-P5-P4-P3-P2-P1. Thereafter the disulfide bridge was formed, and the peptide was cleaved from the resin, cyclized, deprotected and purified as indicated in procedure B.
HPLC-retention time (minutes) was determined using the analytical method described above: Ex.51 (8.94);
Example 64 is shown in Table 1. The peptide was synthesized starting with the amino acid GIu which was grafted to the resin. Starting resin was Fmoc-Glu(OtBut)-2- chlorotrityl resin, which was prepared as described above. The linear peptide was synthesized on solid support according to procedure described above in the following sequence: Resin-Glu(OtBu)-DPro-Pl 1-P10-P9-P8-P7-P6-P5-P4-P3-P2-P1. Thereafter the disulfide bridge was formed, and the peptide was cleaved from the resin, cyclized, deprotected and purified as indicated in procedure B. HPLC-retention time (minutes) was determined using the analytical method described above:
Ex.64 (13.17).
Example 68 is shown in Table 1. The peptide was synthesized starting with the amino acid Asp which was grafted to the resin. Starting resin was Fmoc-Asp(OtBu)-2- chlorotrityl resin, which was prepared as described above. The linear peptide was synthesized on solid support according to procedure described above in the following sequence: Resin-Asp(OtBu)-D AIa-Pl 1-P10-P9-P8-P7-P6-P5-P4-P3-P2-P1. Thereafter the disulfide bridge was formed, and the peptide was cleaved from the resin, cyclized, deprotected and purified as indicated in procedure B. HPLC-retention time (minutes) was determined using the analytical method described above: Ex. 68 (12.44).
Example 69 is shown in Table 1. The peptide was synthesized starting with the amino acid Pro which was grafted to the resin. Starting resin was Fmoc-Pro-2 -chlorotrityl resin, which was prepared as described above. The linear peptide was synthesized on solid support according to procedure described above in the following sequence: Resin Pro- DAsn(Trt)-Pl l-P10-P9-P8-P7-P6-P5-P4-P3-P2-Pl . Thereafter the disulfide bridge was formed, and the peptide was cleaved from the resin, cyclized, deprotected and purified as indicated in procedure B. HPLC-retention time (minutes) was determined using the analytical method described above:
Ex. 69 (12.97).
Example 72 is shown in Table 1. The peptide was synthesized starting with the amino acid Pro which was grafted to the resin. Starting resin was Fmoc-Pro-2-chlorotrityl resin, which was prepared as described above. The linear peptide was synthesized on solid support according to procedure described above in the following sequence: Resin-Pro- DThr(OtBu)-Pl 1-P10-P9-P8-P7-P6-P5-P4-P3-P2-P1. Thereafter the disulfide bridge was formed, and the peptide was cleaved from the resin, cyclized, deprotected and purified as indicated in procedure B.
HPLC-retention time (minutes) was determined using the analytical method described above: Ex. 72 (13.34).
Example 73 is shown in Table 1. The peptide was synthesized starting with the amino acid Pro which was grafted to the resin. Starting resin was Fmoc-Pro-2-chlorotrityl resin, which was prepared as described above. The linear peptide was synthesized on solid support according to procedure described above in the following sequence: Resin-Pro- DIle-Pll-P10-P9-P8-P7-P6-P5-P4-P3-P2-Pl. Thereafter the disulfide bridge was formed, and the peptide was cleaved from the resin, cyclized, deprotected and purified as indicated in procedure B.
HPLC-retention time (minutes) was determined using the analytical method described above: Ex. 73 (9.78).
Example 74 is shown in Table 1. The peptide was synthesized starting with the amino acid Leu which was grafted to the resin. Starting resin was Fmoc-Leu-2-chlorotrityl resin, which was prepared as described above. The linear peptide was synthesized on solid support according to procedure described above in the following sequence: Resin-Leu-
DPro-Pl 1-P10-P9-P8-P7-P6-P5-P4-P3-P2-P1. Thereafter the disulfide bridge was formed, and the peptide was cleaved from the resin, cyclized, deprotected and purified as indicated in procedure B. HPLC-retention time (minutes) was determined using the analytical method described above: Ex. 74 (8.94).
Example 115 is shown in Table 1. The peptide was synthesized starting with the amino acid Pro which was grafted to the resin. Starting resin was Fmoc-Pro-2-chlorotrityl resin, which was prepared as described above. The linear peptide was synthesized on solid support according to procedure described above in the following sequence: Resin-Pro- DAsp(OtBu)-Pl l-P10-P9-P8-P7-P6-P5-P4-P3-P2-Pl. Thereafter the disulfide bridge was formed, and the peptide was cleaved from the resin, cyclized, deprotected and purified as indicated in procedure B. HPLC-retention time (minutes) was determined using the analytical method described above: Ex. 115 (4.82).
Example 116 is shown in Table 1. The peptide was synthesized starting with the amino acid Pro which was grafted to the resin. Starting resin was Fmoc-Pro-2-chlorotrityl resin, which was prepared as described above. The linear peptide was synthesized on solid support according to procedure described above in the following sequence: Resin-Pro- DPhe-Pl 1-P10-P9-P8-P7-P6-P5-P4-P3-P2-P1. Thereafter the disulfide bridge was formed, and the peptide was cleaved from the resin, cyclized, deprotected and purified as indicated in procedure B.
HPLC-retention time (minutes) was determined using the analytical method described above:
Ex. 116 (5.98).
Example 117 is shown in Table 1. The peptide was synthesized starting with the amino acid Pro which was grafted to the resin. Starting resin was Fmoc-Pro-2-chlorotrityl resin, which was prepared as described above. The linear peptide was synthesized on solid support according to procedure described above in the following sequence: Resin-Pro- DArg(Trt)-Pl 1-P10-P9-P8-P7-P6-P5-P4-P3-P2-P1. Thereafter the disulfide bridge was formed, and the peptide was cleaved from the resin, cyclized, deprotected and purified as indicated in procedure B.
HPLC-retention time (minutes) was determined using the analytical method described above: Ex. 117 (4.48). Example 118 is shown in Table 1. The peptide was synthesized starting with the amino acid Pro which was grafted to the resin. Starting resin was Fmoc-Pro-2-chlorotrityl resin, which was prepared as described above. The linear peptide was synthesized on solid support according to procedure described above in the following sequence: Resin-Pro- DSer(OtBu)-Pl 1-P10-P9-P8-P7-P6-P5-P4-P3-P2-P1. Thereafter the disulfide bridge was formed, and the peptide was cleaved from the resin, cyclized, deprotected and purified as indicated in procedure B.
HPLC-retention time (minutes) was determined using the analytical method described above: Ex. 118 (4.73).
Example 119 is shown in Table 1. The peptide was synthesized starting with the amino acid Pro which was grafted to the resin. Starting resin was Fmoc-Pro-2-chlorotrityl resin, which was prepared as described above. The linear peptide was synthesized on solid support according to procedure described above in the following sequence: Resin-Pro-
0VaI-Pl 1 -P10-P9-P8-P7-P6-P5-P4-P3-P2-P1. Thereafter the disulfide bridge was formed, and the peptide was cleaved from the resin, cyclized, deprotected and purified as indicated in procedure B. HPLC-retention time (minutes) was determined using the analytical method described above:
Ex. 119 (5.47).
Example 120 is shown in Table 1. The peptide was synthesized starting with the amino acid Pro which was grafted to the resin. Starting resin was Fmoc-Pro-2-chlorotrityl resin, which was prepared as described above. The linear peptide was synthesized on solid support according to procedure described above in the following sequence: Resin-Pro- 0PiC-Pl 1 -Pl 0-P9-P8-P7-P6-P5-P4-P3-P2-P1. Thereafter the disulfide bridge was formed, and the peptide was cleaved from the resin, cyclized, deprotected and purified as indicated in procedure B. HPLC-retention time (minutes) was determined using the gradient method 1 described above: Ex. 120 (5.48).
Example 121 is shown in Table 1. The peptide was synthesized starting with the amino acid Asp which was grafted to the resin. Starting resin was Fmoc-Asp(OtBu)-2- chlorotrityl resin, which was prepared as described above. The linear peptide was synthesized on solid support according to procedure described above in the following sequence: Resin-Asp(OtBu)-DPro-Pl 1-Pl 1-P10-P9-P8-P7-P6-P5-P4-P3-P2-P1. Thereafter the disulfide bridge was formed, and the peptide was cleaved from the resin, cyclized, deprotected and purified as indicated in procedure B.
HPLC-retention time (minutes) was determined using the analytical method described above:
Ex. 121 (4.56).
Examples 122 and 167 are shown in Table 1. The peptides were synthesized starting with the amino acid Phe which was grafted to the resin. Starting resin was Fmoc-Phe-2- chlorotrityl resin, which was prepared as described above. The linear peptides were synthesized on solid support according to procedure described above in the following sequence: Resin-Phe-DPro-Pl 1-P10-P9-P8-P7-P6-P5-P4-P3-P2-P1. Thereafter the disulfide bridge was formed, and the peptide was cleaved from the resin, cyclized, deprotected and purified as indicated in procedure B.
HPLC-retention time (minutes) was determined using the analytical method described above: Ex. 122 (5.75); 167 (5.75).
Examples 123, 164, 169, 170, 172, 173, 175, 177 and 178 are shown in Table 1. The peptides were synthesized starting with the amino acid GIn which was grafted to the resin. Starting resin was Fmoc-Gln(Trt)-2-chlorotrityl resin, which was prepared as described above. The linear peptides were synthesized on solid support according to procedure described above in the following sequence: Resin-Gln(Trt)-DPro-Pl 1 -P10-P9- P8-P7-P6-P5-P4-P3-P2-P1. Thereafter the disulfide bridge was formed, and the peptide was cleaved from the resin, cyclized, deprotected and purified as indicated in procedure B. HPLC-retention time (minutes) was determined using the analytical method described above:
Ex. 123 (4.35), 164 (13.20), 169 (16.81), 170 (14.57), 172 (16.78), 173 (13.57), 175 (15.94), 177 (16.78), 178 (17.45).
Example 124 is shown in Table 1. The peptide was synthesized starting with the amino acid Ser which was grafted to the resin. Starting resin was Fmoc-Ser(OtBu)-2-chlorotrityl resin, which was prepared as described above. The linear peptide was synthesized on solid support according to procedure described above in the following sequence: Resin- Ser(OtBu)-DPro-Pl l-P10-P9-P8-P7-P6-P5-P4-P3-P2-Pl. Thereafter the disulfide bridge was formed, and the peptide was cleaved from the resin, cyclized, deprotected and purified as indicated in procedure B.
HPLC-retention time (minutes) was determined using the analytical method described above:
Ex. 124 (4.46).
Example 125 is shown in Table 1. The peptide was synthesized starting with the amino acid VaI which was grafted to the resin. Starting resin was Fmoc-Val-2-chlorotrityl resin, which was prepared as described above. The linear peptide was synthesized on solid support according to procedure described above in the following sequence: Resin-Val- DPro-Pl l-Pl l-P10-P9-P8-P7-P6-P5-P4-P3-P2-Pl . Thereafter the disulfide bridge was formed, and the peptide was cleaved from the resin, cyclized, deprotected and purified as indicated in procedure B.
HPLC-retention time (minutes) was determined using the analytical method described above: Ex. 125 (18.42).
Example 126 is shown in Table 1. The peptide was synthesized starting with the amino acid Thr which was grafted to the resin. Starting resin was Fmoc-Thr(OtBu)-2- chlorotrityl resin, which was prepared as described above. The linear peptide was synthesized on solid support according to procedure described above in the following sequence: Resin-Thr(OtBu)-DThr(OtBu)-Pl 1-P10-P9-P8-P7-P6-P5-P4-P3-P2-P1.
Thereafter the disulfide bridge was formed, and the peptide was cleaved from the resin, cyclized, deprotected and purified as indicated in procedure B.
HPLC-retention time (minutes) was determined using the analytical method described above: Ex. 126 (4.35).
Examples 127, 163, 165 and 174 are shown in Table 1. The peptides were synthesized starting with the amino acid GIu which was grafted to the resin. Starting resin was Fmoc- Glu(OtBu)-2-chlorotrityl resin, which was prepared as described above. The linear peptides were synthesized on solid support according to procedure described above in the following sequence: Resin-Glu(OtBu)-DLys(Boc)-Pl 1-P10-P9-P8-P7-P6-P5-P4-P3-P2- Pl. Thereafter the disulfide bridge was formed, and the peptide was cleaved from the resin, cyclized, deprotected and purified as indicated in procedure B. HPLC-retention time (minutes) was determined using the analytical method described above:
Ex. 127 (4.11), 163 (14.93), 165 (14.40), 174 (12.73).
Example 128 is shown in Table 1. The peptide is synthesized starting with the amino acid Thr which was grafted to the resin. Starting resin was Fmoc-Thr(OtBu)-2- chlorotrityl resin, which was prepared as described above. The linear peptide was synthesized on solid support according to procedure described above in the following sequence: Resin-Thr(OtBu)-DPhe-Pl 1-Pl 1-P10-P9-P8-P7-P6-P5-P4-P3-P2-P1.
Thereafter the disulfide bridge was formed, and the peptide was cleaved from the resin, cyclized, deprotected and purified as indicated in procedure B. HPLC-retention time (minutes) was determined using the gradient method 1 described above:
Ex. 128 (5.26).
Example 130 is shown in Table 1. The peptide was synthesized starting with the amino acid Pro which was grafted to the resin. Starting resin was Fmoc-Pro-2-chlorotrityl resin, which was prepared as described above. The linear peptide was synthesized on solid support according to procedure described above in the following sequence: Resin-Pro- 0AIa-Pl 1-P10-P9-P8-P7-P6-P5-P4-P3-P2-P1. Thereafter the disulfide bridge was formed, and the peptide was cleaved from the resin, cyclized, deprotected and purified as indicated in procedure B.
HPLC-retention time (minutes) was determined using the analytical method described above:
Ex.130 (14.79).
Example 166 is shown in Table 1. The peptide was synthesized starting with the amino acid He which was grafted to the resin. Starting resin was Fmoc-Ile-2-chlorotrityl resin, which was prepared as described above. The linear peptide was synthesized on solid support according to procedure described above in the following sequence: Resin-Ile- DPhe-Pl 1-P10-P9-P8-P7-P6-P5-P4-P3-P2-P1. Thereafter the disulfide bridge was formed, and the peptide was cleaved from the resin, cyclized, deprotected and purified as indicated in procedure B.
HPLC-retention time (minutes) was determined using the analytical method described above: Ex. 166 (16.80).
Example 168 is shown in Table 1. The peptide was synthesized starting with the amino acid Asp which was grafted to the resin. Starting resin was Fmoc-Asp(OtBu)-2- chlorotrityl resin, which was prepared as described above. The linear peptide was synthesized on solid support according to procedure described above in the following sequence: Resin-Asp(OtBu)-DPro-Pl 1-P10-P9-P8-P7-P6-P5-P4-P3-P2-P1. Thereafter the disulfide bridge was formed, and the peptide was cleaved from the resin, cyclized, deprotected and purified as indicated in procedure B. HPLC-retention time (minutes) was determined using the analytical described above: Ex. 168 (4.56).
Examples 171 and 176 are shown in Table 1. The peptides were synthesized starting with the amino acid GIn which was grafted to the resin. Starting resin was Fmoc-Gln(Trt)-2- chlorotrityl resin, which was prepared as described above. The linear peptides were synthesized on solid support according to procedure described above in the following sequence: Resin-Gln(Trt)-DGln(Trt)-Pl 1-P10-P9-P8-P7-P6-P5-P4-P3-P2-P1. Thereafter the disulfide bridge was formed, and the peptide was cleaved from the resin, cyclized, deprotected and purified as indicated in procedure B. HPLC-retention time (minutes) was determined using the analytical method described above:
Ex. 171 (15.40), 176 (13.67).
Figure imgf000102_0001
Figure imgf000103_0001
Table 1 continued, Examples
Example Sequ.ID Pl P2 P3 P4 P5 P6 P7 P8 P9 PlO PI l Template Purity%a) [M+ H]
62 SEQ ID NO: 62 1-Nal Cys Thr Lys Ser Asp Pro Pro He Cys Ser DProLPro 95 1421.9
63 SEQ ID NO: 63 Chg Cys Thr Lys Ser Asp Pro Pro NIe Cys Tyr DProLPro 95 1439.
64 SEQ ID NO: 64 Phe Cys Thr Lys Ser Asp Pro Pro He Cys Ser DProLGlu 95 1403.8
65 SEQ ID NO: 65 Chg Cys Thr Lys Ser Asp Pro Pro Tyr Cys Tyr DProLPro 95 1489.5
66 SEQ ID NO: 66 Chg Cys Thr Lys Ser Asp Pro Pro Cha Cys Tyr DProLPro 95 1479.6
67 SEQ ID NO: 67 Chg Cys Thr Lys AlloThrAsp Pro Pro Tyr Cys Tyr DProLPro 95 1503.6
68 SEQ ID NO: 68 Phe Cys Thr Lys Ser Asp Pro Pro He Cys Ser DAlaLAsp 95 1363.6
69 SEQ ID NO: 69 Phe Cys Thr Lys Ser Asp Pro Pro He Cys Ser DAsnLPro 90 1388.8
70 SEQ ID NO: 70 4AmPheCys Thr Lys Ser Asp Pro Pro Cha Cys Asn DProLPro 92 1454.5
71 SEQ ID NO: 71 Chg Cys Thr Lys Ser Asp Pro Pro Cha Cys Arg 95 1472.6
72 SEQ ID NO: 72 Phe Cys Thr Lys Ser Asp Pro Pro He Cys Ser
Figure imgf000104_0001
95 1375.6
73 SEQ ID NO: 73 Phe Cys Thr Lys Ser Asp Pro Pro He Cys Ser DIleLPro 94 1387.9
74 SEQ ID NO: 74 Phe Cys Thr Lys Ser Asp Pro Pro He Cys Ser DProLeu 94 1387.9
75 SEQ ID NO: 75 Arg Cys Thr Lys Ser Asp Pro Pro He Cys Phe DProLPro 95 1440.5
76 SEQ ID NO: 76 He Cys Thr Ala Ser Leu Pro Pro GIn Cys Tyr DProLPro 95 1369.3
77 SEQ ID NO: 77 NIe Cys Thr Thr Ser He Pro Pro Tyr Cys Tyr DProLPro 95 1434.3
78 SEQ ID NO: 78 NIe Cys Thr Abu Ser He Pro Pro GIn Cys Tyr DProLPro 95 1383.6
79 SEQ ID NO: 79 NIe Cys Thr Ala Ser NIe Pro Pro GIn Cys Tyr DProLPro 95 1369.8
80 SEQ ID NO: 80 NIe Cys Thr Ala Ser Aoc Pro Pro GIn Cys Tyr DProLPro 95 1397.6
81 SEQ ID NO: 81 NIe Cys Thr Ala Ser OctG Pro Pro GIn Cys Tyr DProLPro 95 1425.6
82 SEQ ID NO: 82 NIe Cys Thr Ala Ser Cha Pro Pro GIn Cys Tyr DProLPro 95 1409.5
83 SEQ ID NO: 83 NIe Cys Thr Ala Ser hLeu Pro Pro GIn Cys Tyr DProLPro 95 1383.6
84 SEQ ID NO: 84 NIe Cys Thr Ala Ser Chg Pro Pro GIn Cys Tyr DProLPro 95 1395.7
85 SEQ ID NO: 85 NIe Cys Thr Ala Ser t-BuAla Pro Pro GIn Cys Tyr DProLPro 95 1383.6
86 SEQ ID NO: 86 NIe Cys Ala Ala Ser He Pro Pro GIn Cys Tyr DProLPro 95 1340.1
87 SEQ ID NO: 87 NIe Cys Abu Ala Ser He Pro Pro GIn Cys Tyr DProLPro 95 1354.0
88 SEQ ID NO: 88 NIe Cys Thr Ala Ser He Pro Pro(4NHCOPhe) GIn Cys Tyr DProLPro 95 1488.6
89 SEQ ID NO: 89 NIe Cys Thr Ala Ser He Pro Pro Phe Cys Tyr DProLPro 88 1388.7
90 SEQ ID NO: 90 NIe Cys Thr Ala Ser He Pro Pro GIn Cys Phe DProLPro 95 1353.6
91 SEQ ID NO: 91 NIe Cys Thr Ala Ser He Pro Pro GIn Cys GIn DProLPro 95 1334.5
92 SEQ ID NO: 92 NIe Cys Thr Ala Ser He Pro Pro GIn Cys Arg DProLPro 56 1362.6
93 SEQ ID NO: 93 NIe Cys Thr Ala Ser He Pro Pro GIn Cys Ser DProLPro 95 1293.7
Table 1 continued, Examples
Example Sequ.ID Pl P2 P3 P4 P5 P6 P7 P8 P9 PlO PI l Template Purity%a) [M+ H]
94 SEQ ID NO: 94 NIe Cys Thr Ala Ser He Pro Pro GIn Cys NIe DProLPro 95 1319.5
95 SEQ ID NO: 95 NIe Cys Thr Ala Ser He Pro Pro GIn Cys 2-Nal DProLPro 94 1404.0
96 SEQ ID NO: 96 NIe Cys Thr Ala Ser He Pro Pro GIn Cys 2Cl-PheDProLPro 94 1387.8
97 SEQ ID NO: 97 NIe Cys Thr Ala Ser He Pro Pro GIn Cys Cha DProLPro 95 1359.8
98 SEQ ID NO: 98 NIe Cys Thr Ala Ser He Pro Pro GIn Cys Phg DProLPro 95 1359.9
99 SEQ ID NO: 99 Aoc Cys Thr Ala Ser He Pro Pro Gh Cys Tyr DProLPro 93 1397.4
100 SEQ ID NO: 100 hLeu Cys Thr Ala Ser He Pro Pro GIn Cys Tyr DProLPro 95 1383.4
101 SEQ ID NO: 101 Chg Cys Thr Ala Ser He Pro Pro GIn Cys Tyr DProLPro 87 1395.6
102 SEQ ID NO: 102 OctG Cys Thr Ala Ser He Pro Pro Gb Cys Tyr DProLPro 95 1425.5
103 SEQ ID NO: 103 hPhe Cys Thr Ala Ser He Pro Pro GIn Cys Tyr DProLPro 95 1417.5
104 SEQ ID NO: 104 NIe GIu Thr Ala Ser He Pro Pro GIn Lys Tyr DProLPro 95 1422.8
105 SEQ ID NO: 105 NIe GIu Thr Ala Ser He Pro Pro GIn Arg Tyr DProLPro 95 1450.9
106 SEQ ID NO: 106 NIe Thr Thr Ala Ser He Pro Pro GIn Lys Tyr DProLPro 95 1394.7
107 SEQ ID NO: 107 NIe GIn Thr Ala Ser He Pro Pro GIn Arg Tyr DProLPro 90 1449.8
108 SEQ ID NO: 108 NIe Thr Thr Ala Ser He Pro Pro GIn Met Tyr DProLPro 96 1397.7
109 SEQ ID NO: 109 NIe GIn Thr Ala Ser He Pro Pro GIn Thr Tyr DProLPro 95 1394.7
110 SEQ ID NO: 110 NIe Thr Thr Ala Ser He Pro Pro GIn Gb Tyr DProLPro 81 1394.6
111 SEQ ID NO: 111 NIe GIn Thr Ala Ser He Pro Pro Gb Ser Tyr DProLPro 95 1380.7
112 SEQ ID NO: 112 NIe Cys Thr Ala Ser C5al Pro Pro GIn Cys Tyr DProLPro 85 1413.8
113 SEQ ID NO: 113 NIe Cys Thr Ala Ser Leu Pro Pro Tyr Cys Tyr DProLPro 95 1404.7
114 SEQ ID NO: 114 lie Cys Thr Ala Ser Leu Pro Pro Tyr Cys Tyr DProLPro 95 1404.7
115 SEQ ID NO: 115 NIe Cys Thr Ala Ser He Pro Pro GIn Cys Tyr DAspLPro 95 1387.8
116 SEQ ID NO: 116 NIe Cys Thr Ala Ser He Pro Pro GIn Cys Tyr DPheLPro 95 1419.9
117 SEQ ID NO: 117 NIe Cys Thr Ala Ser He Pro Pro Gb Cys Tyr DArgLPro 95 1428.6
118 SEQ ID NO: 118 NIe Cys Thr Ala Ser He Pro Pro Gb Cys Tyr DSerLPro 95 1359.9
119 SEQ ID NO: 119 NIe Cys Thr Ala Ser He Pro Pro Gb Cys Tyr DValLPro 95 1371.8
120 SEQ ID NO: 120 NIe Cys Thr Ala Ser He Pro Pro Gb Cys Tyr DPicLPro 95 1383.7
121 SEQ ID NO: 121 NIe Cys Thr Ala Ser He Pro Pro Gb Cys Tyr DProLAsp 95 1387.9
122 SEQ ID NO: 122 NIe Cys Thr Ala Ser He Pro Pro Gb Cys Tyr DProLPhe 95 1419.9
Table 1 continued, Examples
Example Sequ.ID Pl P2 P3 P4 P5 P6 P7 P8 P9 PlO PI l Template Purity%a) [M+ H]
123 SEQ ID NO: 123 NIe Cys Thr Ala Ser He Pro Pro GIn Cys Tyr DProLGb 95 1400.6
124 SEQ ID NO: 124 NIe Cys Thr Ala Ser He Pro Pro GIn Cys Tyr DProLSer 95 1359.5
125 SEQ ID NO: 125 NIe Cys Thr Ala Ser He Pro Pro GIn Cys Tyr DProLVal 95 1371.8
126 SEQ ID NO: 126 NIe Cys Thr Ala Ser He Pro Pro GIn Cys Tyr 1WThT 95 1377.4
127 SEQ ID NO: 127 NIe Cys Thr Ala Ser He Pro Pro GIn Cys Tyr DLysLGlu 95 1433.5
128 SEQ ID NO: 128 NIe Cys Thr Ala Ser He Pro Pro GIn Cys Tyr DPheLThr 95 1423.5
129 SEQ ID NO: 129 NIe Cys Thr Ala Ser OctG Pro Pro GIn Cys Gb DProLPro 91 1390.4
130 SEQ ID NO: 130 NIe Cys Thr Ala Ser He Pro Pro GIn Cys Tyr DAlaLPro 95 1343.5
131 SEQ ID NO: 131 NIe Cys Thr Ala Ser He Pro Pro GIn Cys Gb DProLPro 95 1334.5
132 SEQ ID NO: 132 hPhe GIu Thr Ala Ser He Pro Pro GIn Lys Tyr DProLPro 95 1470.6
133 SEQ ID NO: 133 NIe Cys Thr Ala Ser Cha Pro Pro GIn Cys Gb DProLPro 95 1440.5
134 SEQ ID NO: 134 hPhe Thr Thr Ala Ser He Pro Pro GIn GIn Tyr DProLPro 95 1442.5
135 SEQ ID NO: 135 NIe Thr Thr Ala Ser OctG Pro Pro GIn GIn Tyr DProLPro 88 1450.7
136 SEQ ID NO: 136 NIe Cys Thr Ala Ser He Pro Pro GIn Cys Phg DProLPro 95 1339.9
137 SEQ ID NO: 137 NIe Cys Thr Ala Ser He Pro Pro GIn Cys Phe DProLPro 95 1353.6
138 SEQ ID NO: 138 NIe Cys Thr Ala Ser He Pro Pro(4NHCOPhe) GIn Cys Tyr DProLPro 95 1488.6
139 SEQ ID NO: 139 NIe Thr Thr Ala Ser Cha Pro Pro GIn GIn Tyr DProLPro 95 1434.8
140 SEQ ID NO: 140 NIe Cys Thr Ala Ser Chg Pro Pro GIn Cys Tyr DProLPro 95 1395.7
141 SEQ ID NO: 141 NIe Cys Thr Ala Ser Cha Pro Pro GIn Cys Tyr DProLPro 95 1409.5
142 SEQ ID NO: 142 hPhe GIn Thr Ala Ser He Pro Pro GIn Thr Tyr DProLPro 91 1406.5
143 SEQ ID NO: 143 hPhe Cys Thr Ala Ser He Pro Pro GIn Cys Gb DProLPro 94 1383.5
144 SEQ ID NO: 144 NIe Cys Thr Ala Ser He Pro Pro GIn Cys 2Cl-Phe DProLPro 94 1387.8
145 SEQ ID NO: 145 OctG Cys Thr Ala Ser He Pro Pro GIn Cys Phe DProLPro 95 1409.4
146 SEQ ID NO: 146 hPhe Cys Thr Ala Ser He Pro Pro GIn Cys Phe DProLPro 95 1401.5
147 SEQ ID NO: 147 OctG Thr Thr Ala Ser He Pro Pro Gb GIn Tyr DProLPro 95 1450.9
148 SEQ ID NO: 148 OctG Cys Thr Ala Ser OctG Pro Pro GIn Cys Gb DProLPro 95 1446.6
149 SEQ ID NO: 149 OctG Cys Thr Ala Ser He Pro Pro GIn Cys Gb DProLPro 95 1390.4
150 SEQ ID NO: 150 OctG Cys Thr Ala Ser Cha Pro Pro GIn Cys Tyr DProLPro 95 1465.6
151 SEQ ID NO: 151 OctG Cys Thr Ala Ser OctG Pro Pro(4NHCOPhe) GIn Cys Gb DProLPro 94 1565.7
152 SEQ ID NO: 152 hPhe Cys Thr Ala Ser OctG Pro Pro GIn Cys Phe DProLPro 95 1457.6
153 SEQ ID NO: 153 hPhe Cys Thr Ala Ser OctG Pro Pro GIn Cys Gb DProLPro 95 1438.5
154 SEQ ID NO: 154 OctG GIn Thr Ala Ser He Pro Pro GIn Thr Tyr DProLPro 93 1450.9
155 SEQ ID NO: 155 hPhe Cys Thr Ala Ser Cha Pro Pro GIn Cys Phe DProLPro 90 1441.5
Table 1 continued, Examples
Example Sequ.ID Pl P2 P3 P4 P5 P6 P7 P8 P9 PlO PIl Template Purityo/o° [M+ H]
156 SEQIDNO: 156 OctG GIu Thr Ala Ser He Pro Pro GIn Lys Tyr DProLPro 95 1478.7
157 SEQIDNO: 157 OctG Cys Thr Ala Ser Cha Pro Pro GIn Cys Phe DProLPro 95 1449.8
158 SEQIDNO: 158 hPhe Cys Thr Ala Ser Cha Pro Pro GIn Cys GIn DProLPro 94 1422.7
159 SEQIDNO: 159 OctG Cys Thr Ala Ser Cha Pro Pro GIn Cys GIn DProLPro 93 1430.0
160 SEQIDNO: 160 OctG Cys Thr Ala Ser Cha Pro Pro(4NHCOPhe) GIn Cys GIn DProLPro 95 1549.6
161 SEQIDNO: 161 hPhe Cys Thr Ala Ser OctG Pro Pro GIn Cys Tyr DProLPro 95 1473.4
162 SEQIDNO: 162 hPhe Cys Thr Ala Ser Cha Pro Pro GIn Cys Tyr DProLPro 95 1457.3
163 SEQIDNO: 163 NIe Cys Thr Ala Ser OctG Pro Pro GIn Cys Tyr DLysLGlu 95 1374.4
164 SEQIDNO: 164 NIe Cys Thr Ala Ser Cha Pro Pro GIn Cys GIn DProLGln 95 1405.5
165 SEQIDNO: 165 OctG Cys Thr Ala Ser He Pro Pro GIn Cys Tyr DLysLGlu 95 1488.0
166 SEQIDNO: 166 NIe Cys Thr Ala Ser He Pro Pro GIn Cys Tyr DProLIle 95 1385.6
167 SEQIDNO: 167 NIe Cys Thr Ala Ser He Pro Pro GIn Cys Tyr DProLPhe 95 1419.9
168 SEQIDNO: 168 NIe Cys Thr Ala Ser He Pro Pro GIn Cys Tyr DProLAsp 95 1387.9
169 SEQIDNO: 169 NIe Cys Thr Ala Ser OctG Pro Pro GIn Cys Tyr DProLGln 95 1456.5 O
ON
170 SEQIDNO: 170 NIe Cys Thr Ala Ser Cha Pro Pro GIn Cys Tyr DProLGln 95 1440.5
171 SEQIDNO: 171 NIe Cys Thr Ala Ser Cha Pro Pro GIn Cys Cha DGlnLGln 95 1461.0
172 SEQIDNO: 172 NIe Cys Thr Ala Ser Cha Pro Pro GIn Cys Cha DProLGln 95 1430.6
173 SEQIDNO: 173 hPhe Cys Thr Ala Ser He Pro Pro GIn Cys Tyr DProLGln 95 1448.6
174 SEQIDNO: 174 hPhe Cys Thr Ala Ser He Pro Pro GIn Cys Tyr DLysLGlu 95 1480.0
175 SEQIDNO: 175 NIe Cys Thr Ala Ser Cha Pro Pro GIn Cys 2Cl-PheDProLGln 95 1458.5
176 SEQIDNO: 176 NIe Cys Thr Ala Ser Cha Pro Pro(4NHCOPhe) GIn Cys GIn DGlnLGln 95 1555.5
177 SEQIDNO: 177 OctG Cys Thr Ala Ser He Pro Pro GIn Cys Tyr DProLGln 95 1430.6
178 SEQIDNO: 178 OctG Cys Thr Ala Ser OctG Pro Pro GIn Cys GIn DProLGln 95 1477.6
179 SEQIDNO: 179 NIe Cys Thr Ala Ser He Pro Pro GIn Cys Tyr DProLPro 90 1369.7
180 SEQIDNO: 180 He Cys Thr Lys Ser Leu Pro Pro He Cys Arg DProLPro 94 1404.8
181 SEQIDNO: 181 He Cys Thr Lys Ser hPhe Pro Pro He Cys Arg DProLPro 92 1452.6
182 SEQIDNO: 182 lie Cys Thr Lys Ser Cha Pro Pro He Cys Arg DProLPro 95 1444.6
183 SEQIDNO: 183 He Cys Thr Lys Ser Tyr Pro Pro He Cys Arg DProLPro 91 1454.5
184 SEQIDNO: 184 Phe Cys Thr Lys Ser Leu Pro Pro He Cys Arg DProLPro 95 1438.6
Table 1 continued, Examples
Example Sequ.ID Pl P2 P3 P4 P5 P6 P7 P8 P9 PlO PI l Template Puπty%d; [M+ H]
185 SEQ ID NO: 185 lie Cys Thr Lys Ser Leu Pro Pro Arg Cys Arg DProLPro 95 1447.5
186 SEQ ID NO: 186 He Cys Thr Lys Ser Leu Pro Pro Lys Cys Arg DProLPro 95 1419.9
187 SEQ ID NO: 187 He Cys Thr Lys Ser Leu Pro Pro His Cys Arg DProLPro 95 1428.6
188 SEQ ID NO: 188 He Cys Thr Lys Ser Leu Pro Pro GIn Cys Arg DProLPro 95 1419.8
189 SEQ ID NO: 189 He Cys Thr Lys Ser Leu Pro Pro Thr Cys Arg DProLPro 95 1392.4
190 SEQ ID NO: 190 He Cys Thr Lys Ser Leu Pro Pro Arg Cys Lys DProLPro 95 1420.1
191 SEQ ID NO: 191 Leu Cys Thr Lys Ser Leu Pro Pro Lys Cys Arg DProLPro 95 1420.0
192 SEQ ID NO: 192 NIe Cys Thr Lys Ser Leu Pro Pro Lys Cys Arg DProLPro 95 1420.0
193 SEQ ID NO: 193 Cha Cys Thr Lys Ser Leu Pro Pro Lys Cys Arg DProLPro 95 1459.7
194 SEQ ID NO: 194 Tyr Cys Thr Lys Ser Leu Pro Pro Lys Cys Arg DProLPro 95 1469.6
195 SEQ ID NO: 195 Tip Cys Thr Lys Ser Leu Pro Pro Lys Cys Arg DProLPro 92 1492.6
196 SEQ ID NO: 196 Arg Cys Thr Lys Ser Leu Pro Pro Lys Cys Tyr DProLPro 95 1469.6 a) %-purity of compounds after prep. HPLC
Cys in pos. 2 and 10 in Ex. 1-6, 9-103, 112-131, 133, 136-138, 140-141, 143-146, 148-153, 155, 157-196 form a disulfide bridge
2. Biological methods
2.1. Preparation of the peptide samples.
Lyophilized peptides were weighed on a Microbalance (Mettler MT5) and dissolved in sterile water to a final concentration of 1 niM unless stated otherwise. Stock solutions were kept at +4°C, light protected.
2.2. Enzymatic assays
Enzyme and substrate conditions were as indicated Table 2.
Kinetic measurements were made in a total reaction volume of 100 μl in 96 well flat bottomed plates (Greiner) on a Genios plate reader (Tecan). The enzyme was combined with the peptides (inhibitors) in a buffer containing 10OmM HEPES (pH 7.5), 5OmM CaCl2, 0.025% Tween-20, 5% DMSO, and 1 mM of the substrate. The rate of substrate hydrolysis was measured by monitoring the change in absorbance at 405 nm over 30 minutes to verify linearity of the reaction curve. The average rate from minute 1 through minute 10 was used for all calculations. Initial calculations of background subtraction, average rate, duplicate averaging and % inhibition were made using the Magellan software (version 5) from Tecan. IC50% calculations were made using Grafit (version 5.0.10) from Erithacus Software by fitting inhibition data from 6 different inhibitor concentrations to a 4-parameter equation:
100%
Figure imgf000109_0001
In this equation 5 is the slope factor, x is the inhibitor concentration and y is % inhibition at a given concentration of the inhibitor.
Km/Kj determination
The K1n for the serine protease substrate was determined from a Lineweaver-Burke plot (Grafit v5). The Kj values for inhibitors were calculated using the formula K1 = IC50%/(l+([substrate]/Km)).
Increasing concentrations of substrate were reacted with the enzyme and the rate of each reaction (ABS/mSec) was plotted vs. substrate concentration. The reciprocal plot (Lineweaver-Burke) was also plotted to give Km and Vmax (inset) (see ref. 1 below). Table 2
Figure imgf000110_0001
Figure imgf000111_0001
2.3. Cytotoxicity assay
The cytotoxicity of the peptides to HELA cells (Acc57) and COS-7 cells (CRL-1651 ) was determined using the MTT reduction assay [see ref. 2 and 3, below]. Briefly the method was as follows: HELA cells and COS-7 cells were seeded at 7.OxIO3 and, respectively, 4.5xlO3 cells per well and grown in 96-well microtiter plates for 24 hours at 370C at 5% CO2. At this point, time zero (Tz) was determined by MTT reduction (see below).The supernatant of the remaining wells was discarded, and fresh medium and the peptides in serial dilutions of 12.5, 25 and 50 μM were pipetted into the wells. Each peptide concentration was assayed in triplicate. Incubation of the cells was continued for 48 hours at 370C at 5% CO2. Wells were then washed once with phosphate buffered saline (PBS) and subsequently 100 μl MTT reagent (0.5 mg/ml in medium RPMIl 640 and, respectively, DMEM) were added to the wells. This was incubated at 370C for 2 hours and subsequently the medium was aspirated and 100 μl isopropanol were added to each well. The absorbance at 595 nm of the solubilized product was measured (OD595peptide). For each concentration averages were calculated from triplicates. The percentage of growth was calculated as follows: (OD595peptide-OD595Tz-OD595Empty well) / (OD595TZ- OD595Empty well) x 100% and was plotted for each peptide concentration. The LC 50 values (Lethal Concentration, defined as the concentration that kills 50% of the cells) were determined for each peptide by using the trend line function of EXCEL (Microsoft Office 2000) for the concentrations (50, 25, 12.5 and 0 μM), the corresponding growth percentages and the value -50, (=TREND(C50:C0,%50:%0,-50)). The GI 50 (Growth Inhibition) concentrations were calculated for each peptide by using a trend line function for the concentrations (50, 25, 12.5 and 0 μg/ml), the corresponding percentages and the value 50, (=TREND (C5o:Co,%5o:%o,50). 2.4. Hemolysis
The peptides were tested for their hemolytic activity against human red blood cells (hRBC). Fresh hRBC were washed three times with phosphate buffered saline (PBS) by centrifugation for 10 min at 2000 x g. Peptides at a concentration of 100 μM were incubated with 20% v/v hRBC for 1 hour at 370C. The final erythrocyte concentration was approximately 0.9x109 cells per ml. A value of 0% and, respectively,. 100% cell lysis was determined by incubation of the hRBC in the presence of PBS alone and, respectively, 0.1% Triton X-100 in H2O. The samples were centrifuged, the supernatant was 20-fold diluted in PBS buffer and the optical density (OD) of the sample at 540 nM was measured. The 100% lysis value (OD540H20) gave an OD540 of approximately 1.3-1.8. Percent hemolysis was calculated as follows: (OD54opeptide/OD54oH20) xlOO%.
2.5 Plasma stability
405 μl of plasma/albumin solution were placed in a polypropylene (PP) tube and spiked with 45 μl of compound from a 100 mM solution B, derived from 135 μl of PBS and 15 μl of 1 mM peptide in PBS, pH 7.4. 150 μl aliquots were transferred into individual wells of the 10 kDa filter plate (Millipore MAPPB 1010 Biomax membrane). For "0 minutes controls": 270 μl of PBS were placed in a PP tube and 30 μl of stock solution B was added and vortexed. 150 μl of control solution were placed into one well of the filter plate and served as "filtered control".
Further 150 μl of control solution were placed directly into a receiver well (reserved for filtrate) and served as "not-filtered control". The entire plate including evaporation lid was incubated for 60 min at 37°C. Plasma samples (rat plasma: Harlan Sera lab UK, human plasma: Blutspendezentrum Zurich) were centrifuged at least for 2 h at 4300 rpm (3500 g) and 150C in order to yield 100 μl filtrate. For "serum albumin"-samples (freshly prepared human albumin: Sigma A-4327, rat albumin: Sigma A-6272, all at 40 mg/ml concentration in PBS) approximately 1 hour of centrifugation was sufficient. The filtrates in the receiver PP plate were analysed by LC/MS as followes: Column: Jupiter Cl 8 (Phenomenex), mobile phases: (A) 0.1% formic acid in water and (B) acetonitrile, gradient: 5%-100% (B) in 2 minutes, electrospray ionization, MRM detection (triple quadrupole). The peak areas were determined and triplicate values were averaged. The binding was expressed in percent of the (filtered and not-filtered time point 0 min) control 1 and 2 by: 100-(100 x T6o/To). The average from these values was then calculated. 2.6. Pharmacokinetic study (PK)
Pharmacokinetic study after single oral (gavage) and intravenous administration in rats Pharmacokinetic study after single intravenous (i.v.) and oral (p.o., gavage) administration was performed for the compound of Example 75 ("Ex. 75"). 332 g (± 10 g) male Wistar mice obtained from RCC Ltd, Laboratory animal Services, CH-4414 Fϋllinsdorf, Switzerland were used in the study. The vehicle, physiological saline, was added to give a final concentration of 2.5 mg/ml of the compound. The volume was 2 ml/kg i.v. and 10 ml/kg p.o. and the peptide Ex. 75 was injected to give a final intravenous dose of 5 mg/kg and an oral dose of 50 mg/kg. Blood samples (approx. 0.24 ml) were taken following the schedule below at different time points into heparinized tubes by automated blood sampling using the DiLab AccuSampler. When a problem occured during automated blood sampling, blood was sampled by retro-orbital bleeding under light isoflurane anesthesia. Samples where taken at the following time points:
0, 5 min (only i.v.), 15, 30 min and 1, 2, 4, 8, 16, 24 and 36 (only p.o.) hours and added to heparinized tubes. Plasma was removed from pelleted cells upon centrifugation and frozen at -800C prior to HPLC-MS analysis.
Preparation of the plasma calibration samples
"Blank" rat plasma from untreated animals was used. Aliquots of plasma of 0.1 ml each were spiked with 50 ng of propranolol (Internal Standard, IS), (sample preparation by solid phase extraction on OASIS® HLB cartridges (Waters)) and with known amounts of Ex. 75 in order to obtain 9 plasma calibration samples in the range 5 - 2000 ng/ml. The OASIS® HLB cartridges were conditioned with 1 ml of methanol and then with 1 ml of 1% NH3 in water. Samples were then diluted with 400 μl of 1% NH3 in water and loaded. The plate was washed with 1 ml of methanol/1% NH3 in water 5/95. Elution was performed using 1 ml of 0.1% TFA in methanol. The plate containing eluates was introduced into the concentrator system and taken to dryness. The residues were dissolved in 100 μl of formic acid 0.1%/acetonitrile, 95/5 (v/v) and analysed in the HPLC/MS on a reverse phase analytical column (Jupiter Cl 8, 50 x 2.0 mm, 5 μm, Phenomenex), using gradient elution (mobile phases A: 0.1% formic acid in water, B: Acetonitrile; from 5%B to 100%B in 2 min.).
Preparation of plasma samples From each sample 100 μl of plasma were taken for the extraction. If the volume was less than 100 μl the appropriate amount of "blank" mouse plasma was added in order to keep the matrix identical to the calibration curve. Samples were then spiked with IS and processed as described for the calibration curve.
Pharmacokinetic evaluation
PK analysis was performed on pooled data (generally n=2 or 3) using the software PK solutions 2.0™ (Summit Research Service, Montrose, CO 81401 USA). The area under the curve AUC was calculated by the linear trapezoidal rule. AUC^-00) was estimated as Ct/b (b: elimination rate constant). AUC(K0) is the sum of AUC(0.t) and AUC^j.
Elimination half-life was calculated by the linear regression on at least three data points during the elimination phase. The time intervals selected for the half-life determinations were evaluated by the correlation coefficient (r2), which should be at least above 0.85 and most optimally above 0.96. In case of i.v. administration the initial concentration at tzero was determined by extrapolation of the curve through the first two time points. Finally bioavailability after i.p. administration was calculated from the normalised AUQo-∞) ratio after i.p. versus i.v. administration.
3.0 Results The results of the experiments described under 2.2 - 2.5, above, are indicated in Table 3 herein below.
Table 3
Figure imgf000115_0001
Table 3 continued
Figure imgf000116_0001
Table 3 continued
Figure imgf000117_0001
Table 3 continued
Figure imgf000118_0001
Table 3 continued
OO
Figure imgf000119_0001
Table 3 continued
Figure imgf000120_0001
Table 3 continued
Figure imgf000121_0001
Table 3 continued
Figure imgf000122_0001
Table 3 continued
Figure imgf000123_0001
Table 3 continued
Figure imgf000124_0001
Nd: not determined
The results of the experiment described in 2.5 above are indicated in Table 4 herein below.
Table 4
Figure imgf000125_0001
The results of the experiment described in 2.6 (PK), above, are indicated in Table 5 herein below.
Table 5
Figure imgf000125_0002
The large inter-individual variation in plasma concentration of Ex. 75 was most pronounced after single oral administration (lor i.v.: %C.V = 6 - 68%, except for one value at the lowest measurable concentration 173%; for p.o. %C.Y.: 113 - 173%).
Intravenous administration
After intravenous administration of Ex. 75 at a dose level of 5 mg/kg body weight, Ex. 75 followed intravenous kinetic characteristics. After PK analysis, Ex 75 showed an extrapolated CinMaI of 14069 ng/ml and a Cmax observed of 10762 ng/ml at 5 min (0.083 hour). Plasma levels rapidly decreased to 5774 and 3455 ng/ml at 15 min and 30 min, respectively. From 1 to 2 hours plasma levels decreased with a terminal tiβ of 0.46 hours to 18 ng/ml at 4 hours. The AUC0.t and AUCo-infinite amounted to 6044 and 6047 ng x h/ml, respectively; the initial distribution volume amounted to 355 ml/kg. The apparent distribution volume was 547 ml/kg.
Oral administration Alter oral administration of Ex 75 at a dose level of 50 mg /kg body weight, plasma levels of Ex. 75 followed oral kinetic characteristics. After PK analysis, Ex. 75 showed an observed Cmax of 464 ng/ml at 0.25 hour (15 min). From 0.25 hours, plasma levels decreased with a terminal tm of 0.87 hours to 24 ng/ml at 4 hours. The AUC0.t and AUC0- iπfimte amounted to 782 and 813 ng x h/ml. respectively. Taking into account the absorption of 1.3%, the apparent distribution volume was 1008 ml/kg.
Oral versus intravenous administration
Due to the different dose levels between the oral group versus the i.v. group, values were compared after dose normalisation. Compared to the normalized AUCo-infinιte value after i.v. administration of Ex. 75 (100%: 6047 ng-h/ml), the percentage of Ex. 75 absorbed (F) after oral administration amounted to 1.3% (81 ng x h/ml) at an about 234 times lower normalised Cmax value after oral administration (46 versus 10762 ng/ml; Table 3). The apparent distribution volume after oral administration was about 1.8 fold higher than after i.v. administration (1008 versus 547 ml/kg).
References
1. Barrtt, AJ. Methods in Enzymology 1981, 80, 561-565; Leatherbarrow, R. J. 1992, GraFit, Erithacus Software Ltd., Staines, U.K.
2. Mossman T. J. Immunol. Meth 1983, (55:55-63
3. Berridge MV, Tan AS. Arch.Biochem.Biophys. 1993, 303:474-482

Claims

1. Compounds of the general formula
Figure imgf000127_0001
(I) wherein
Figure imgf000127_0002
is a group of one of the formulae
Figure imgf000127_0003
(a3)
Figure imgf000127_0004
Figure imgf000128_0001
Figure imgf000129_0001
(k) (I) (m)
Figure imgf000129_0002
(n) (o) (P)
wherein
Figure imgf000129_0003
is GIy or 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;
s* is a group of one of the formulae
Figure imgf000129_0004
A1 A2 A3 A4
Figure imgf000130_0001
A5 A6 A7 A8 A9
Figure imgf000130_0002
A10 A11 A12 A13 A14
Figure imgf000130_0003
A15 A16 A17 A18 A1Θ
Figure imgf000130_0004
A20 A21 A22 A23 A24
Figure imgf000130_0005
A25 A26 A27 A28
Figure imgf000130_0006
A29 A30 A31 A32 A33
Figure imgf000131_0001
A34 A35 A36 A37
Figure imgf000131_0002
A48 A50 A51 A52
Figure imgf000131_0003
Figure imgf000132_0001
A63 A64 A65 A66
Figure imgf000132_0002
A67 A68 A69
Figure imgf000132_0003
A70 A71 A72 A73 A74
Figure imgf000132_0004
A75 A76 A77 A78 A79
Figure imgf000132_0005
A80 A81 A82 A83 A84
Figure imgf000132_0006
A85 A86
Figure imgf000133_0001
R is H; lower alkyl; or aryl-lower alkyl;
R2 is H; alkyl; alkenyl; -(CH2)m(CHR61)sOR55; -(CH2)m(CHR61)sSR56;
-(CH2)m(CHR61)sNR33R34; -(CH2)m(CHR61)sOCONR33R75;
-(CH2)m(CHR61)sNR20CONR33R82; -(CH2)o(CHR61)sCOOR57;
-(CH2)o(CHR61)sCONR58R59; -(CH2)o(CHR61)sPO(OR60)2;
-(CH2)o(CHR61)s SO2R62; or -(CH2)o(CHR61)sC6H4R8; R3 is H; alkyl; alkenyl; -(CH2)m(CHR61)sOR55; -(CH2)m(CHR61)sSR56;
-(CH2)m(CHR61)sNR33R34; -(CH2)m(CHR61)sOCONR33R75;
-(CH2)m(CHR61)sNR20CONR33R82; -(CH2)o(CHR61)sCOOR57 ;
-(CH2)o(CHR61)sCONR58R59 ; -(CH2)o(CHR61)sPO(OR60)2;
-(CH2)o(CHR61)s SO2R62; or -(CH2)O(CHR01XC6H4R
R4 is H; alkyl; alkenyl; -(CH2)ra(CHR61)sOR55; -(CH2)m(CHR61)sSR5
(CH2)m(CHR°')sNR33R34; -(CH2)m(CHR61)sOCONR33R75; -(CH2)m(CHR61)sNR20CONR33R82; -(CH2)P(CHR61)sCOOR57; -(CH2)P(CHR61)sCONR58R59; - (CH2)p(CHR61)sPO(OR60)2;
-(CH2)p(CHR61)s SO2R62; or -(CH2)o(CHR61)sC6H4R8; R5 is alkyl; alkenyl; -(CH2)O(CHR61)SOR55; -(CH2)O(CHR61)SSR56; -
(CH2)O(CHR61)SNR33R34;
-(CH2)o(CHR61)sOCONR33R75; -(CH2)o(CHR61)sNR20CONR33R82; -(CH2)o(CHR61)sCOOR57; -(CH2)0(CHR61)SCONR58R59; - (CH2)o(CHR61)sPO(OR60)2;
-(CH2)o(CHR61)s SO2R62; or -(CH2)O(CHR61^C6H4R8; R6 is H; alkyl; alkenyl; -(CH2)0(CHR61)SOR55; -(CH2)O(CHR61)SSR56; - (CH2)O(CHR61)SNR33R34;
-(CH2)o(CHR61)sOCONR33R75; -(CH2)o(CHR61)sNR20CONR33R82; -(CH2)o(CHR61)sCOOR57; -(CH2)o(CHR61)sCONR58R59; -
(CH2)o(CHR61)sPO(OR60)2;
-(CH2)O(CHR61)S SO2R62; or -(CH2)O(CHR61)SC6H4R8; R7 is alkyl; alkenyl; -(CH2)q(CHR61)sOR55; -(CH2)q(CHR61)sNR33R34;
-(CH2)q(CHR61)sOCONR33R75; -(CH2)q(CHR61)sNR20CONR33R82; -(CH2)r(CHR61)sCOOR57; -(CH2)r(CHR61)sCONR58R59; -
(CH2)r(CHR61)sPO(OR60)2;
-(CH2)r(CHR61)sSO2R62; or -(CH2)r(CHR61)s C6H4R8;
R8 is H; Cl; F; CF3; NO2; lower alkyl; lower alkenyl; aryl; aryl-lower alkyl; -(CH2)O(CHR61)SOR55; -(CH2)O(CHR61)SSR56; -(CH2)O(CHR6I)NR33R34 ; -(CH2)0(CHR61)SOCONR33R75; -(CH2)o(CHR61)sNR20CONR33R82;
-(CH2)o(CHR61)sCOOR57; -(CH2)0(CHR6I)SCONR58R59; - (CH2)o(CHR61)sPO(OR60)2;
-(CH2)O(CHR61)SSO2R62; or -(CH2)o(CHRδl)sCOR64; R9 is alkyl; alkenyl; -(CH2)0(CHR61)SOR55; -(CH2)O(CHR61)SSR56; - (CH2)O(CHR61)SNR33R34;
-(CH2)o(CHR61)sOCONR33R75; -(CH2)o(CHR61)sNR20CONR33R82; -(CH2)O(CHR61)SCOOR57; -(CH2)0(CHR61)SCONR58R59; - (CH2)0(CHR61)SPO(OR60)2;
-(CH2)O(CHR61)S SO2R62; or -(CH2)O(CHR61)SC6H4R8; R10 is alkyl; alkenyl; -(CH2)O(CHR61 )SOR55; -(CH2)O(CHR61)SSR56; - (CH2)O(CHR61)SNR33R34;
-(CH2)o(CHR61)sOCONR33R75; -(CH2)o(CHR61)sNR20CONR33R82; -(CH2)O(CHR6!)SCOOR57; -(CH2)0(CHR61)SCONR58R59; - (CH2)o(CHR6I)sPO(OR60)2; -(CH2)O(CHR61)S SO2R62; or -(CH2)O(CHR61)SC6H4R8;
R1 1 is H; alkyl; alkenyl; -(CH2)m(CHR61)sOR55; -(CH2)m(CHR61)sNR33R34; -(CH2)m(CHR61)sOCONR33R75; -(CH2)ra(CHR61)sNR20CONR33R82;
-(CH2)0(CHR61)SCOOR57; -(CH2)O(CHR61)SCONR58R59; - (CH2)o(CHR61)sPO(OR60)2;
-(CH2)o(CHR61)sSO2R62; or -(CH2)O(CHR61)S C6H4R8; R12 is H; alkyl; alkenyl; -(CH2)m(CHR6I)sOR55; -(CH2)m(CHR61)sSR56; -(CH2)m(CHR61)sNR33R34; -(CH2)m(CHR61)sOCONR33R75; -(CH2)m(CHR61)sNR20CONR33R82; -(CH2)r(CHR61)sCOOR57; - (CH2)r(CHR61)sCONR58R59; -(CH2)r(CHR61)sPO(OR60)2; -(CH2)r(CHR61)s SO2R62; or -(CH2)r(CHR61)sC6H4R8; R13 is alkyl; alkenyl; -(CH2)q(CHR61)sOR55; -(CH2)q(CHR61)sSR56; - (CH2)q(CHR61)sNR33R34;
-(CH2)q(CHR61)sOCONR33R75; -(CH2)q(CHR61)sNR20CONR33R82; -(CH2)q(CHRδl)sCOOR57; -(CH2)q(CHR61)sCONR58R59; - (CH2)q(CHR6!)sPO(OR60)2; -(CH2)q(CHR61)s SO2R62; or -(CH2)q(CHR61)sC6H4R8;
R14 is H; alkyl; alkenyl; -(CH2)m(CHR61)sOR55; -(CH2)m(CHR61)sNR33R34; -(CH2)m(CHR61)sOCONR33R75; -(CH2)m(CHR61)sNR20CONR33R82; -(CH2)q(CHR61)sCOOR57; -(CH2)q(CHR61)sCONR58R59; - (CH2)q(CHR61)sPO(OR60)2;. -(CH2)q(CHR61)sSOR62; or -(CH2)q(CHR61)s C6H4R8;
R15 is alkyl; alkenyl; -(CH2)O(CHR61)SOR55; -(CH2)O(CHR61)SSR56; - (CH2)O(CHR61)SNR33R34;
-(CH2)o(CHR61)sOCONR33R75; -(CH2)o(CHR61)sNR20CONR33R82; -(CH2)0(CHR61)SCOOR57; -(CH2)O(CHR61)SCONR58R59; - (CH2)o(CHR61)sPO(OR60)2;
-(CH2)O(CHR61)S SO2R62; or -(CH2)O(CHR61^C6H4R8; R16 is alkyl; alkenyl; -(CH2)0(CHR61)SOR55; -(CH2)O(CHR61)SSR56; - (CH2)O(CHR61)SNR33R34;
-(CH2)o(CHR61)sOCONR33R75; -(CH2)o(CHR6i)sNR20CONR33R82; -(CH2)0(CHR61)SCOOR57; -(CH2)O(CHR61)SCONR58R59; -
(CH2)0(CHR61)SPO(OR60)2; -(CH2)O(CHR6I)S SO2R62; or -(CH2)O(CHR61)SC6H4R8 ;
R17 is alkyl; alkenyl; -(CH2)q(CHR61)sOR55; -(CH2)q(CHR61)sSR56; -
(CH2)q(CHR61)sNR33R34; -(CH2)q(CHR61)sOCONR33R75; -(CH2)q(CHR61)sNR20CONR33R82; -(CH2)q(CHR61)sCOOR57; -(CH2)q(CHR61^CONR58R59; -
(CH2)q(CHR61)sPO(OR60)2;
-(CH2)q(CHR61)s SO2R62; or -(CH2)q(CHR61)sC6H4R8; R18 is alkyl; alkenyl; -(CH2)P(CHR61)sOR55; -(CH2)P(CHR61 )s SR56; - (CH2)P(CHR61)sNR33R34;
-(CH2)p(CHR61)sOCONR33R75; -(CH2)p(CHR61)sNR20CONR33R82; -(CH2)P(CHR61)sCOOR57; -(CH2)P(CHR61)sCONR58R59; - (CH2)P(CHR61)sPO(OR60)2;
-(CH2)p(CHR61)s SO2R62; or -(CH2)o(CHR61)sC6H4R8; R19 is lower alkyl; -(CH2)p(CHR61)sOR55; -(CH2)P(CHR61)sSR56 ; - (CH2)P(CHR61)sNR33R34;
-(CH2)p(CHR61)sOCONR33R75; -(CH2)p(CHR61)sNR20CONR33R82; -(CH2)p(CHR61)sCOOR57; -(CH2)P(CHR61)sCONR58R59; - (CH2)P(CHR61)sPO(OR60)2; -(CH2)p(CHR61)s SO2R62; or -(CH2)o(CHR61)sC6H4R8; or
R18 and R19 taken together can form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-;
R20 is H; alkyl; alkenyl; or aryl-lower alkyl;
R21 is H; alkyl; alkenyl; -(CH2)o(CHR61)sOR55; -(CH2)o(CHR61)sSR56; - (CH2)o(CHR61)sNR33R34;
-(CH2)o(CHR61)sOCONR33R75; -(CH2)o(CHR61)sNR20CONR33R82; -(CH2)o(CHR61)sCOOR57; -(CH2)o(CHR61^CONR58R59; - (CH2)o(CHR61)sPO(OR60)2;
-(CH2)o(CHR61)s SO2R62; or -(CH2)o(CHR61)sC6H4R8; R22 is H; alkyl; alkenyl; -(CH2)o(CHR61)sOR55; -(CH2)o(CHR61)sSR56; - (CH2)o(CHR61)sNR33R34;
-(CH2)o(CHR61)sOCONR33R75; -(CH2)o(CHR61)sNR20CONR33R82; -(CH2)o(CHR61)sCOOR57; -(CH2)o(CHR61)sCONR58R59; - (CH2)o(CHR61)sPO(OR60)2; -(CH2)o(CHR61)s SO2R62; or -(CH2)o(CHR61)sC6H4R8;
R23 is alkyl; alkenyl; -(CH2)o(CHR61)sOR55; -(CH2)o(CHR61)sSR56; - (CH2)o(CHR61)sNR33R34;
-(CH2)o(CHR61)sOCONR33R75; -(CH2)o(CHR61)sNR20CONR33R82; -(CH2)o(CHR61)sCOOR57; -(CH2)o(CHR61)sCONR58R59; - (CH2)o(CHR61)sPO(OR60)2;
-(CH2)o(CHR61)s SO2R62; or -(CH2)o(CHR61^C6H4R8; R24 is alkyl; alkenyl; -(CH2)o(CHR61)sOR55; -(CH2)o(CHR61)sSR56; -
(CH2)o(CHR61)sNR33R34;
-(CH2)o(CHR61)sOCONR33R75; -(CH2)o(CHR61)sNR20CONR33R82;
-(CH2)o(CHR61)sCOOR57; -(CH2)o(CHR61)sCONR58R59; - (CH2)o(CHR61)sPO(OR60)2;
-(CH2)o(CHR61)s SO2R62; or -(CH2)o(CHR61)sC6H4R8; R25 is H; alkyl; alkenyl; -(CH2)m(CHR61)sOR55; -(CH2)m(CHR61)sSR56;
-(CH2)m(CHR61)sNR33R34; -(CH2)m(CHR61)sOCONR33R75;
-(CH2)m(CHR61)sNR20CONR33R82; -(CH2)o(CHR61)sCOOR57; -(CH2)o(CHR61)sCONR58R59; -(CH2)o(CHRδl)sPO(OR60)2;
-(CH2)o(CHR61)sSO2R62; or -(CH2)o(CHR61)sC6H4R8; R26 is H; alkyl; alkenyl; -(CH2)m(CHR61)sOR55; -(CH2)m(CHR61)sSR56;
-(CH2)m(CHR61)sNR33R34; -(CH2)m(CHR61)sOCONR33R75;
-(CH2)m(CHR61)sNR20CONR33R82; -(CH2)o(CHR61)sCOOR57; - (CH2)o(CHR61)sCONR58R59; -(CH2)o(CHR61)sPO(OR60)2;
-(CH2)o(CHR61)s SO2R62; or -(CH2)o(CHR61)sC6H4R8; or R25 and R26 taken together can form: -(CH2)2-6-; -(CH2)rO(CH2)r-; -(CH2)rS(CH2)r-; or
-(CH2)rNR57(CH2)r-;
R27 is H; alkyl; alkenyl; -(CH2)o(CHR61)sOR55; -(CH2)o(CHR61)sSR56; - (CH2)o(CHR61)sNR33R34;
-(CH2)o(CHR61)sCOOR57; -(CH2)o(CHR61)sCONR58R59; -
(CH2)o(CHR61)sOCONR33R75;
-(CH2)o(CHR61)sNR20CONR33R82; -(CH2)o(CHR61)sPO(OR60)2;
-(CH2)o(CHR61)s SO2R62; or -(CH2)o(CHR61)sC6H4R8; R28 is alkyl; alkenyl; -(CH2)o(CHR61 )s-OR55; -(CH2)o(CHR61)s SR56; -(CH2)o(CHR6 I)s
NR33R34;
-(CH2)o(CHR61)sOCONR33R75; -(CH2)o(CHR61)sNR20CONR33R82;
-(CH2)o(CHR61)s COOR57; -(CH2)o(CHR61)s CONR58R59; -(CH2)o(CHR61)s
PO(OR60)2; -(CH2)o(CHR61)s SO2R62; or -(CH2)o(CHR61)s C6H4R8;
R29 is alkyl; alkenyl; -(CH2)o(CHR61)sOR55; -(CH2)o(CHR61)sSR56; -
(CH2)o(CHR61)sNR33R34;
-(CH2)o(CHR61)sOCONR33R75; -(CH2)o(CHR61)sNR20CONR33R82;
-(CH2)o(CHR61)sCOOR57; -(CH2)o(CHR61)sCONR58R59; - (CH2)o(CHR61)sPO(OR60)2;
-(CH2)o(CHR61)s SO2R62; or -(CH2)o(CHR61)sC6H4R8; R30 is H; alkyl; alkenyl; or aryl-lower alkyl;
R31 is H; alkyl; alkenyl; -(CH2)P(CHR61)sOR55; -(CH2)P(CHR61)sNR33R34; -(CH2)p(CHR61)sOCONR33R75; -(CH2)p(CHR61)sNR20CONR33R82; -(CH2)o(CHR61)sCOOR57; -(CH2)o(CHR61)sCONR58R59; - (CH2)o(CHR61)sPO(OR60)2;
-(CH2)o(CHR61)sSO2R62; or -(CH2)o(CHR61)s C6H4R8; R32 is H; lower alkyl; or aryl-lower alkyl;
R33 is H; alkyl, alkenyl; -(CH2)m(CHR61)sOR55; -(CH2)m(CHR61)sNR34R63; -(CH2)m(CHR61)sOCONR75R82; -(CH2)m(CHR61)sNR20CONR78R82; -(CH2)o(CHR61)sCOR64; -(CH2)o(CHR61)s-CONR58R59, -
(CH2)o(CHR61)sPO(OR60)2;
-(CH2)o(CHR61)s SO2R62; or -(CH2)o(CHR61)sC6H4R8; R34 is H; lower alkyl; aryl, or aryl-lower alkyl;
R33 and R34 taken together can form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-;
R35 is H; alkyl; alkenyl; -(CH2)m(CHR61)sOR55; -(CH2)m(CHR61)sNR33R34; -(CH2)m(CHR61)sOCONR33R75; -(CH2)m(CHR61)sNR20CONR33R82; -(CH2)p(CHR61)sCOOR57; -(CH2)P(CHR61)sCONR58R59; - (CH2)p(CHR61)sPO(OR60)2; -(CH2)P(CHR61)sSO2R62; or -(CH2)P(CHR61)s C6H4R8;
R36 is H, alkyl; alkenyl; -(CH2)o(CHR61)sOR55; -(CH2)P(CHR61)sNR33R34; -(CH2)P(CHR61)sOCONR33R75; -(CH2)P(CHR61)sNR20CONR33R82; -(CH2)p(CHR61)sCOOR57; -(CH2)P(CHR61)sCONR58R59; - (CH2)P(CHR61)sPO(OR60)2; -(CH2)P(CHR61)sSO2R62; or -(CH2)o(CHR61)s C6H4R8;
R37 is H; F; Br; Cl; NO2; CF3; lower alkyl; -(CH2)P(CHR61)sOR55; - (CH2)p(CHR61)sNR33R34;
-(CH2)p(CHR61)sOCONR33R75; -(CH2)p(CHR61)sNR20CONR33R82; -(CH2)o(CHR61)sCOOR57; -(CH2)o(CHR61)sCONR58R59; - (CH2)o(CHR61)sPO(OR60)2;
-(CH2)o(CHR61)sSO2R62; or -(CH2)o(CHR61)s C6H4R8; R38 is H; F; Br; Cl; NO2; CF3; alkyl; alkenyl; -(CH2)P(CHR61)sOR55; - (CH2)p(CHR61)sNR33R34;
-(CH2)p(CHR61)sOCONR33R75; -(CH2)p(CHR61)sNR20CONR33R82; -(CH2)o(CHR61)sCOOR57; -(CH2)o(CHR61)sCONR58R59; -
(CH2)o(CHR61)sPO(OR60)2; -(CH2)o(CHR61)sSO2R62; or -(CH2)o(CHR61)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; NO2; CF3; alkyl; alkenyl; -(CH2)P(CHR61)sOR55; - (CH2)P(CHR61)sNR33R34;
-(CH2)p(CHR61)sOCONR33R75; -(CH2)p(CHR61)sNR20CONR33R82; -(CH2)o(CHR61)sCOOR57; -(CH2)o(CHR61)sCONR58R59; - (CH2)o(CHR61)sPO(OR60)2;
-(CH2)o(CHR61)sSO2R62; or -(CH2)o(CHR61)s C6H4R8; R42 is H; F; Br; Cl; NO2; CF3; alkyl; alkenyl; -(CH2)P(CHR61)sOR55; - (CH2)P(CHR61)sNR33R34;
-(CH2)P(CHR61)sOCONR33R75; -(CH2)P(CHR61)sNR20CONR33R82; -(CH2)o(CHR6l)sCOOR57; -(CH2)o(CHR61)sCONR58R59; - (CH2)o(CHR61)sPO(OR60)2; -(CH2)o(CHR61)sSO2R62; or -(CH2)o(CHR61)s C6H4R8;
R43 is H; alkyl; alkenyl; -(CH2)m(CHR61)sOR55; -(CH2)m(CHR61)sNR33R34; -(CH2)m(CHR61)sOCONR33R75; -(CH2)m(CHR61)sNR20CONR33R82; -(CH2)o(CHR61)sCOOR57; -(CH2)o(CHR61)sCONR58R59; - (CH2)o(CHR61)sPO(OR60)2; -(CH2)o(CHR61)sSO2R62; or -(CH2)o(CHR61)s C6H4R8;
R44 is alkyl; alkenyl; -(CH2)r(CHR61)sOR55; -(CH2)r(CHR61)sSR56; - (CH2)r(CHR61)sNR33R34;
-(CH2)r(CHR61)sOCONR33R75; -(CH2)r(CHR61)sNR20CONR33R82; -(CH2)r(CHR61)sCOOR57; -(CH2)r(CHR61)sCONR58R59; - (CH2)r(CHR61)sPO(OR60)2;
-(CH2)r(CHR61)s SO2R62; or -(CH2)r(CHR61^C6H4R8; R45 is H; alkyl; alkenyl; -(CH2)o(CHR61)sOR55; -(CH2)o(CHR61)sSR56; - (CH2)o(CHR61)sNR33R34;
-(CH2)o(CHR61)sOCONR33R75; -(CH2)o(CHR61)sNR20CONR33R82; -(CH2)o(CHR61)sCOOR57; -(CH2)s(CHR61)sCONR58R59; -
(CH2)s(CHR61)sPO(OR60)2;
-(CH2)s(CHR61)s SO2R62; or -(CH2)s(CHR61)sC6H4R8; R46 is H; alkyl; alkenyl; or -(CH2)o(CHR61^C6H4R8; R47 is H; alkyl; alkenyl; or -(CH2)o(CHR61)sOR55; R48 is H; lower alkyl; lower alkenyl; or aryl-lower alkyl;
R49 is H; alkyl; alkenyl; -(CHR61)sCOOR57; (CHR61)sCONR58R59; (CHR61)sPO(OR60)2; -(CHR61)sSOR62; or -(CHR61)sC6H4R8;
R50 is H; lower alkyl; or aryl-lower alkyl;
R51 is H; alkyl; alkenyl; -(CH2)ra(CHR61)sOR55; -(CH2)m(CHR61)sSR56;
-(CH2)m(CHR61)sNR33R34; -(CH2)m(CHR61)sOCONR33R75; -(CH2)m(CHR61)sNR20CONR33R82; -(CH2)o(CHR61)sCOOR57;
-(CH2)o(CHR61)sCONR58R59; -(CH2)o(CHR61)pPO(OR60)2; -(CH2)P(CHR61)s SO2R62; or -(CH2)P(CHR61)sC6H4R8; R52 is H; alkyl; alkenyl; -(CH2)m(CHR61)sOR55; -(CH2)m(CHR61)sSR56;
-(CH2)m(CHR61)sNR33R34; -(CH2)m(CHR61)sOCONR33R75; -(CH2)m(CHR61)sNR20CONR33R82; -(CH2)o(CHR61)sCOOR57;
-(CH2)o(CHR61)sCONR58R59; -(CH2)o(CHR6i)pPO(OR60)2; -(CH2)P(CHR61)s SO2R62; or -(CH2)P(CHR61)sC6H4R8; R53 is H; alkyl; alkenyl; -(CH2)m(CHR61)sOR55; -(CH2)m(CHR61)sSR56; -
(CH2)m(CHR61)sNR33R34; -(CH2)ra(CHR61)sOCONR33R75; -(CH2)m(CHR61)sNR20CONR33R82; -(CH2)o(CHR61)sCOOR57;
-(CH2)o(CHR61)sCONR58R59; -(CH2)o(CHR61)pPO(OR60)2; -(CH2)P(CHR61)s SO2R62; or -(CH2)P(CHR61^C6H4R8; R54 is H; alkyl; alkenyl; -(CH2)m(CHR61)sOR55; -(CH2)m(CHR61)sNR33R34; -(CH2)m(CHR61)sOCONR33R75; -(CH2)m(CHR61)sNR20CONR33R82; -(CH2)o(CHR61)COOR57; -(CH2)o(CHR61)sCONR58R59; or -(CH2)o(CHR61)s
C6H4R8;
R55 is H; lower alkyl; lower alkenyl; aryl-lower alkyl; -(CH2)ra(CHR61)sOR57; -(CH2)m(CHR61)sNR34R63; -(CH2)m(CHR61)sOCONR75R82; -(CH2)m(CHR61)sNR20CONR78R82; -(CH2)o(CHR6')s-COR64; - (CH2)o(CHR61)COOR57; or
-(CH2)o(CHR61)sCONR58R59;
R56 is H; lower alkyl; lower alkenyl; aryl-lower alkyl; -(CH2)m(CHR6l)sOR57; -(CH2)m(CHR61)sNR34R63; -(CH2)m(CHR61)sOCONR75R82; -(CH2)m(CHR61)sNR20CONR78R82; -(CH2)o(CHR61)s-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)MS -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-;
R60 is H; lower alkyl; lower alkenyl; aryl; or aryl-lower alkyl; R61 is alkyl; alkenyl; aryl; heteroaryl; aryl-lower alkyl; heteroaryl-lower alkyl; - (CH2)mOR55; -(CH2)mNR33R34; -(CH2)mOCONR75R82; -(CH2)raNR20CONR78R82; -
(CH2)oCOOR37;
-(CH2)oNR58R59; or -(CH2)oPO(COR60)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; -SO2R62; or -PO(OR60)2; R34and R63 taken together can form: -(CH2)M-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-;
R64 is H; lower alkyl; lower alkenyl; aryl; heteroaryl; aryl-lower alkyl; heteroaryl-lower alkyl;
-(CH2)P(CHR61)sOR65; -(CH2)P(CHR61)sSR66; or -(CH2)P(CHR61)sNR34R63; -(CH2)P(CHR61)sOCONR75R82; -(CH2)P(CHR61)sNR20CONR78R82; R65 is H; lower alkyl; lower alkenyl; aryl, aryl-lower alkyl; heteroaryl-lower alkyl; -
COR57; -COOR57; or -CONR58R59;
R66 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 O or 1 ;
Z is a chain of 11 α-amino acid residues, 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, GIy, Pro, Pro(4NHCOPhe), or of formula -A-CO-, or of formula -B-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-;
-NR20-CH(CO-)-(-(CH2)pNR20CO(CH2)p-CH(CO-)-NR20-; and -NR20-CH(CO-)-(-(CH2)pNR20CONR20(CH2)p-CH(CO-)-NR20-;
R71 is lower alkyl; lower alkenyl; -(CH2)P(CHR61)sOR75; -(CH2)p(CHRβl)sSR75;
-(CH2)P(CHR61)sNR33R34; -(CH2)P(CHR61)sOCONR33R75; -
(CH2)P(CHR61)sNR20CONR33R82; -(CH2)o(CHR61)sCOOR75; -(CH2)pCONR58R59; -(CH2)PPO(OR62)2; -(CH2)PSO2R62; or
-(CH2)o-C6R67R68R69R70R76;
R72 is H, lower alkyl; lower alkenyl; -(CH2)P(CHR61)sORS5; or -(CH2)P(CHR61)sSR85; R73 is -(CR86R87)OR77; -(CH2)rO(CH2)oR77; -(CH2)rS(CH2)oR77; or -(CH2)rNR20(CH2)oR77; R74 is -(CH2)pNR78R79; -(CH2)PNR77R80; -(CH2)pC(=NR80)NR78R79; - (CH2)pC(=NOR50)NR78R79;
-(CH2)pC(=NNR78R79)NR78R79; -(CH2)pNR80C(=NR80)NR78R79;
-(CH2)pN=C(NR78R80)NR79R80;-(CH2)pC6H4NR78R79; -(CH2)pC6H4NR77R80;
-(CH2)pC6H4C(=NR80)NR78R79; -(CH2)pC6H4C(=NOR50)NR7SR79;
-(CH2)pC6H4C(=NNR78R79)NR78R79; -(CH2)pC6H4NR80C(=NR80)NR78R79; -(CH2)pC6H4N=C(NR78R80)NR79R80; -(CH2)rO(CH2)mNR78R79; -
(CH2)rO(CH2)mNR77R80;
-(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)mN=C(NR78R80)NR79R80; -(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)mNR77R80;-(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)mN=C(NR78RS0)NR79R80;
-(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)pNR80COR64; - (CH2)pNR80COR77;
-(CH2)PNRSOCONR78R79; -(CH2)PC6H4NR80CONR78R79; or
-(CH2)pNR20CO-[(CH2)u-X]rCH3 where X is -O-; -NR20-, or -S-; u is 1 -3, and t is 1-
6;
R75 is lower alkyl; lower alkenyl; or aryl-lower alkyl; R33 and R75 taken together can form: -(CH2)2-6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; R75 and R82 taken together can form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-; R76 is H; lower alkyl; lower alkenyl; aryl-lower alkyl; -(CH2)oOR72; -(CH2)oSR72;
,33r,34
-(CH2)oNR33R34; -(CH2)oOCONR33R75; -(CH2)oNR20CONR33R82; -(CH2)oCOOR75; -(CH2)oCONR58R59; -(CH2)oPO(OR60)2; -(CH2)pSO2R62; or -(CH2)oCOR64; R77 is -C6R67R68R69R70R76; or a heteroaryl group of one of the formulae
Figure imgf000143_0001
Figure imgf000144_0001
R78 is H; lower alkyl; aryl; or aryl-lower alkyl;
R78 and R82 taken together can form: -(CH2)2-6-; -(CH2)2θ(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-;
R79 is H; lower alkyl; aryl; or aryl-lower alkyl; or
R78 and R79, taken together, can be -(CH2)2-7-; -(CH2)2O(CH2)2-; or -(CH2)2NR57(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;
R33 and R82 taken together can form: -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-;
R83 is H; lower alkyl; aryl; or -NR78R79; R84 is -(CH2)m(CHR61)sOH; -(CR86R87)pORS0;-(CR86R87)pCOOR80; -(CH2)m(CHR61)sSH; -
(CR86R87)pSR80;-(CH2)pCONR78R79; -(CH2)PNR80CONR78R79; -(CH2)PC6H4CONR78R79; or -
(CH2)PC6H4NR80CONR78R79; -(CR86R87)oPO(OR60)2; -(CR86R87)PSO2R60; -(CR86R87)PSOR60; (CH2)m(CHR61)s OPO(OR60)2,; or -(CH2)m(CHR61)s OSO2R60,;
R85 is lower alkyl; or lower alkenyl; R86 is H; lower alkyl, where H is maybe substituted by halogen; or halogen;
R87 is H; lower alkyl, where H is maybe substituted by halogen; or halogen;
with the proviso that in said chain of 11 α-amino acid residues Z the amino acid residues in positions 1 to 11 are:
Pl : of type C, or of type D, or of type E, or of type F;
P2: of type C, or of Type D, or of type E, or of type F;
P3: or of type C, of type F, or the residue is GIy;
P4: of type C, or of type D, or of type F, or of type E5 or the residue is GIy or Pro;
P5: of type E, or of type C, or of type F, or the residue is GIy or Pro;
P6: of type E, or of type F, or of type E, or of type C, or the residue is
GIy or Pro;
P7: of type C, or of type E, or of type, F or of formula -A-CO-, or the residue is GIy or Pro;
P8: of type D, or of type C, or of type F, or of formula -A-CO, or the residue is GIy or Pro or Pro(4NHCOPhe);
P9: of type C, or of type D, or of type E, or of type F;
PlO: of type D, or of type C, or of type F, or of type E; and - PI l : of type C, or of type D, or of type E, or of type F; or P2 and PlO, taken together, can form a group of type H; and with the further proviso that if the template is DProLPro the amino acid residues in positions Pl to Pl 1 are other than
Pl : Arg
P2: Cys, linked with Cys in position PlO by a disulfide bridge
P3: Thr
P4 Lys
P5 Ser
P6 He
P7 Pro
P8 Pro
P9 He
PlO Cys, linked with Cys in position PlO by a disulfide bridge; and
PI l Phe;
and pharmaceutically acceptable salts thereof.
2. Compounds according to claim 1 or 2 wherein
Figure imgf000146_0001
is a group of formula (al) or (a2) or (a3).
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)mSR56 (where R56 is lower alkyl; or lower alkenyl); -(CH2)mNR33R34
(where R33 is lower alkyl; or lower alkenyl; R34 is H; or lower alkyl; or R33 and R34 taken together are -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -
(CH2)mOCONR33R75 (where R33 is H; lower alkyl; or lower alkenyl; R75 is lower alkyl; or
R33 and R75 taken together are
57/
-(CH2)M-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NRD /(CH2)2-; where R" is H; or lower alkyl);
-(CH2)mNR20CONR33R82 (where R20 is H; or lower alkyl; R33 is H; or lower alkyl; or lower alkenyl; R82 is H; or lower alkyl; or R33 and R82 taken together are -(CH2)2.6-; -
(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -
(CH2)oN(R20)COR64(where: R20 is H; or lower alkyl; R64 is lower alkyl; or lower alkenyl);
(CH2)oCOOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where
R58 is lower alkyl; or lower alkenyl; and R59 is H; or lower alkyl; or R58 and R59 taken together are -(CH2W; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)oPO(OR60)2 (where R60 is lower alkyl; or lower alkenyl); -(CH2)oSO2R62 (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)raSR56 (where R56 is lower alkyl; or lower alkenyl); -(CH2)mNR33R34 (where R33 is lower alkyl; or lower alkenyl; R34 is H; or lower alkyl; or R33 and R34 taken together are -
(CH2)M-;
-(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl);
-(CH2)mOCONR33R75 (where R33 is H; or lower alkyl; or lower alkenyl; R75 is lower alkyl; or R33 and R75 taken together are -(CH2W; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -
(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)mNR20CONR33R82 (where R20 is H; or lower alkyl; R33 is H; or lower alkyl; or lower alkenyl; R82 is H; or lower alkyl; or R33 and R82 taken together are -(CH2W;
-(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl);
-(CH2)oN(R20)COR64 (where: R20 is H; or lower alkyl; R64 is lower alkyl; or lower alkenyl); -(CH2)oCOOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where
R58 is lower alkyl; or lower alkenyl; and R59 is H; lower alkyl; or R38 and R59 taken together are -(CH2W;
-(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)oPO(OR60)2 (where R60 is lower alkyl; or lower alkenyl); -(CH2)oSO2R62 (where R62 is lower alkyl; or lower alkenyl); or -(CH2)C1C6H4R8 (where R8 is H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
R4 is H; lower alkyl; lower alkenyl; -(CH2)mOR55 (where R55 is lower alkyl; or lower alkenyl); -(CH2)mSR56 (where R56 is lower alkyl; or lower alkenyl); -(CH2)mNR33R34 (where R33 is lower alkyl; or lower alkenyl; R34 is H; or lower alkyl; or R33 and R34 taken together are -
(CHz)2-6-;
-(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)mOCONR33R75 (where R33 is H; or lower alkyl; or lower alkenyl; R75 is lower alkyl; or R33 and R75 taken together are -(CH2W; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -
(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)JSIR20CONR33R82 (where R20 is H; or lower alkyl; R33 is H; or lower alkyl; or lower alkenyl; R82 is H; or lower alkyl; or
R33 and R82 taken together are -(CH2W; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl);
-(CH2)mN(R20)COR64(where: R20 is H; or lower alkyl; R64 is lower alkyl; or lower alkenyl);
-(CH2)oCOOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where R58 is lower alkyl; or lower alkenyl; and R59 is H; or lower alkyl; or R58 and R59 taken together are -(CH2)2.6-;
-(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: isH; or lower alkyl);
-(CH2)oPO(OR60)2 (where R60 is lower alkyl; or lower alkenyl); -(CH2)oSO2R62 (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).
R5 is lower alkyl; lower alkenyl; -(CH2)oOR55 (where R55 is lower alkyl; or lower alkenyl);
-(CH2)oSR56 (where R56 is lower alkyl; or lower alkenyl); (CH2)oNR33R34 (where R33 is lower alkyl; or lower alkenyl; R34 is H; or lower alkyl; or R33 and R34 taken together are -
(CH2W; -(CH2)2O(CH2)2-;
-(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -
(CH2)oOCONR33R75 (where R33 is H; or lower alkyl; or lower alkenyl; R75 is lower alkyl; or R33 and R75 taken together are -(CH2)M-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)oNR20CONR33R82 (where R20 is H; or lower lower alkyl; R33 is H; or lower alkyl; or lower alkenyl; R82 is H; or lower alkyl; or R33 and R82 taken together are -
(CH2)M-; -(CH2)2O(CH2)r;
-(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); - (CH2)oN(R20)COR64 (where: R20 is H; or lower alkyl; R64 is alkyl; alkenyl; aryl; aryl- lower alkyl; or heteroaryl-lower alkyl);
Figure imgf000149_0001
(where R57 is lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where R58 is lower alkyl; or lower alkenyl; and R59 is
H; or lower alkyl; or R58 and R59 taken together are -(CH2)2.6-; -(CH2)2O(CH2)2-; -
(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl);
-(CH2)oPO(OR60)2 (where R60 is lower alkyl; or lower alkenyl); -(CH2)oSO2R62 (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):
R6 is H; lower alkyl; lower alkenyl; -(CH2)oOR55 (where R55 is lower alkyl; or lower alkenyl);
-(CH2)oSR56 (where R56 is lower alkyl; or lower alkenyl); -(CH2)oNR33R34 (where R33 is lower alkyl; or lower alkenyl; R34 is H; or lower alkyl; or R33 and R34 taken together are -
(CH2)MS -(CH2)2O(CH2)2-;
-(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -
(CH2)oOCONR33R75 (where R33 is H; or lower alkyl; or lower alkenyl; R75 is lower alkyl; or R33 and R75 taken together are -(CH2)MS -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)oNR20CONR33R82 (where R20 is H; or lower lower alkyl; R33 is H; or lower alkyl; or lower alkenyl; R82 is H; or lower alkyl; or R33 and R82 taken together are -
(CH2)MS -(CH2)2O(CH2)2-;
-(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); - (CH2)oN(R20)COR64 (where R20 is H; or lower alkyl; R64 is lower alkyl; or lower alkenyl);
-(CH2)oCOOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where
R58 is lower alkyl; or lower alkenyl; and R59 is H; or lower alkyl; or R58 and R59 taken together are -(CH2)M-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)oPO(OR60)2 (where R60 is lower alkyl; or lower alkenyl); -(CH2)oSO2R62 (where R62 is lower alkyl; or lower alkenyl); or -(CH2)C1C6H4R8 (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 lower alkyl; or lower alkenyl); -(CH2)qNR33R34 (where R33 is lower alkyl; or lower alkenyl; R34 is H; or lower alkyl; or R33 and R34 taken together are - (CHz)2-6-; -(CH2)2O(CH2)2-;
-(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); - (CH2)qOCONR33R75 (where R33 is H; or lower alkyl; or lower alkenyl; R75 is lower alkyl; or R33 and R75 taken together are -(CH2)M-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)qNR20CONR33R82 (where R20 is H; or lower alkyl; R33 is H; or lower alkyl; or lower alkenyl; R82 is H; or lower alkyl; or R33 and R82 taken together are -(CH2)2_ 6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)qN(R20)COR64(where: R20 is H; or lower alkyl; R64 is lower alkyi; or lower alkenyl); -(CH2)rCOOR57 (where R57 is lower alkyl; or lower alkenyl); -
(CH2)qCONR58R59 (where R5S is lower alkyl; or lower alkenyl; and R59 is H; or lower alkyl; or R58 and R59 taken together are -(CH2)M-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 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 -(CHz)11C6H4R8 (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)oOR55 (where R55 is lower alkyl; or lower alkenyl); -(CH2)oSR56 (where R56 is lower alkyl; or lower alkenyl); -(CH2)oNR33R34
(where R33 is lower alkyl; or lower alkenyl; R34 is H; or lower alkyl; or R33 and R34 taken together are -(CH2)2-6-;
-(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl);
-(CH2)oOCONR33R75 (where R33 is H; or lower alkyl; or lower alkenyl; R75 is lower alkyl; . or R33 and R75 taken together are -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or - (CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)oNR20CONR33R82 (where R20 is H; or lower alkyl; R33 is H; or lower alkyl; or lower alkenyl; R82 is H; or lower alkyl; or
R33 and R82 taken together are -(CH2)2-6-;
-(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)oN(R20)COR64 (where R20 is H; or lower alkyl; R64 is lower alkyl; or lower alkenyl);
-(CH2)oCOOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where
R58 is lower alkyl; or lower alkenyl; and R59 is H; or lower alkyl; or R58 and R59 taken together are -(CH2)2_6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)oPO(OR60)2 (where R60 is lower alkyl; or lower alkenyl); -(CH2)oSO2R62 (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);
R9 is lower alkyl; lower alkenyl; -(CH2)oOR55 (where R55 is lower alkyl; or lower alkenyl);
-(CH2)oSR56 (where R56 is lower alkyl; or lower alkenyl); -(CH2)oNR33R34 (where R33 is lower alkyl; or lower alkenyl; R34 is H; or lower alkyl; or R33 and R34 taken together are -
(CH2)2.6-; -(CH2)2O(CH2)2-;
-(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); - (CH2)oOCONR33R75 (where R33 is H; or lower alkyl; or lower alkenyl; R75 is lower alkyl; or R33 and R75 taken together are
-(CH2)2-6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)mNR20CONR33R82 (where R20 is H; or lower lower alkyl; R33 is H; or lower alkyl; or lower alkenyl; R82 is H; or lower alkyl; or R33 and R82 taken together are - (CH2)2.6-; -(CH2)2O(CH2)2-;
-(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -
(CH2)oN(R20)COR64(where R20 is H; or lower alkyl; R64 is lower alkyl; or lower alkenyl);
-(CH2)oCOOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where
R58 is lower alkyl; or lower alkenyl; and R59 is H; or lower alkyl; or R58 and R59 taken together are -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)oPO(OR60)2 (where R60 is lower alkyl; or lower alkenyl); -(CH2)oSO2R62 (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); R10 is lower alkyl; lower alkenyl; -(CH2)oOR55 (where R55 is lower alkyl; or lower alkenyl);
-(CH2)oSR56 (where R56 is lower alkyl; or lower alkenyl); -(CH2)oNR33R34 (where R33 is lower alkyl; or lower alkenyl; R34 is H; or lower alkyl; or R33 and R34 taken together are -
(CH2)M-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -
(CH2)oOCONR33R75 (where R33 is H; or lower alkyl; or lower alkenyl; R75 is lower alkyl; or R33 and R75 taken together are
-(CH2W; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H is or lower alkyl); -(CH2)oNR20CONR33R82 (where R20 is H; or lower lower alkyl; R33 is H; or lower alkyl; or lower alkenyl; R82 is H; or lower alkyl; or R33 and R82 taken together are - (CH2)MS -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -
(CH2)oN(R20)COR64(where R20 is H; or lower alkyl; R64 is lower alkyl; or lower alkenyl);
-(CH2)oCOOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where
R58 is lower alkyl; or lower alkenyl; and R59 is H; lower alkyl; or R58 and R59 taken together are -(CH2)2-6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)r; or
-(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)oPO(OR60)2 (where R60 is lower alkyl; or lower alkenyl); -(CH2)oSO2R62 (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); R11 is H; lower alkyl; lower alkenyl; -(CH2)mOR55 (where R55 is lower alkyl; or lower alkenyl);
-(CH2)mSR56 (where R56 is lower alkyl; or lower alkenyl); -(CH2)mNR33R34 (where R33 is lower alkyl; or lower alkenyl; R34 is H; or lower alkyl; or R33 and R34 taken together are -
(CH2)2_6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl);
-(CH2)mOCONR33R75 (where R33 is H; or lower alkyl; or lower alkenyl; R75 is lower alkyl; or R33 and R75 taken together ar -(CH2)2-6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -
(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)mNR20CONR33R82 (where R20 is H; or lower alkyl; R33 is H; or lower alkyl; or lower alkenyl; R82 is H; or lower alkyl; or
R33 and R82 taken together are -(CH2)2.6-;
-(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl);
-(CH2)mN(R20)COR64 (where R20 is H; or lower alkyl; R64 is lower alkyl; or lower alkenyl);
-(CH2)oCOOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where
R58 is lower alkyl; or lower alkenyl; and R59 is H; lower alkyl; or R58 and R59 taken together are -(CH2)2.6-;
-(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl);
-(CH2)oPO(OR60)2 (where R60 is lower alkyl; or lower alkenyl); -(CH2)oSO2R62 (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);
R12 is H; lower alkyl; lower alkenyl; -(CH2)mOR55 (where R55 is lower alkyl; or lower alkenyl);
-(CH2)mSR56 (where R56 is lower alkyl; or lower alkenyl); -(CH2)mNR33R34 (where R33 is lower alkyl; or lower alkenyl; R34 is H; or lower alkyl; or R33 and R34 taken together are -
(CH2)MS
-(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)mOCONR33R75 (where R33is H; or lower alkyl; or lower alkenyl; R75 is lower alkyl; or R33 and R75 taken together are -(CH2)2.6s -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -
(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)mNR20CONR33R82 (where R20 is H; or lower lower alkyl; R33 is H; or lower alkyl; or lower alkenyl; R82 is H; or lower alkyl; or R33 and R82 taken together are -(CH2)2.6s -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl);
-(CH2)mN(R20)COR64 (where: R20 is H; or lower alkyl; 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; or R58 and R59 taken together are -(CH2)2.6-;
-(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl);
-(CH2)rPO(OR60)2 (where R60 is lower alkyl; or lower alkenyl); -(CH2)oSO2R62 (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)qOR55 (where R55 isis lower alkyl; or lower alkenyl);
-(CH2)qSR56 (where R56 is lower alkyl; or lower alkenyl); -(CH2)qNR33R34 (where R33 is lower alkyl; or lower alkenyl; R34 is H; or lower alkyl; or R33 and R34 taken together are -
(CH2)MS -(CH2)2O(CH2)2-;
-(CH2)2S(CH2)2s or -(CH2)2NR57(CH2)r; where R57 is H; or lower alkyl); -
(CH2) AqTOCONR33R75 (where R33 is H; or lower alkyl; or lower alkenyl; R75 is lower alkyl; or R ,33 and j R r>75 t . aken together are -(CHj)2-6S -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2S where R57 is H; or lower alkyl); -(CH2)qNR20CONR33R82 (where R20 is H; or lower lower alkyl; R33 is H; or lower alkyl; or lower alkenyl; R82 is H; or lower alkyl; or R33 and R82 taken together are -
(CH2)MS -(CH2)2O(CH2)2-;
-(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); - (CH2)qN(R20)COR64 (where: R20 is H; or lower alkyl; 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; or R58 and R59 taken together are -(CH2)2-6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-; where R57 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);
R14 is H; lower alkyl; lower alkenyl; -(CH2)mOR55 (where R55 is lower alkyl; or lower alkenyl);
-(CH2)mSR56 (where R56 is lower alkyl; or lower alkenyl); -(CH2)mNR33R34 (where R33 is lower alkyl; or lower alkenyl; R34 is H; or lower alkyl; or R33 and R34 taken together are -
(CH2)2.6-;
-(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl);
-(CH2)mOCONR33R75 (where R33 is H; or lower alkyl; or lower alkenyl; R75 is lower alkyl; or R33 and R75 taken together are -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -
(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)mNR20CONR33R82 (where R20 is H; or lower lower alkyl; R33 is H; or lower alkyl; or lower alkenyl is R82: H; or lower alkyl; or R33 and R82 taken together are -(CH2)2-6-;
-(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl);
-(CH2)mN(R20)COR64 (where: R20 is H; lower alkyl; R64 is lower alkyl; or lower alkenyl);
-(CH2)oCOOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where
R58 is lower alkyl; or lower alkenyl; and R59 is H; or lower alkyl; or R58 and R59 taken together are -(CH2)2-6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl);
-(CH2)oPO(OR60)2 (where R60 is lower alkyl; or lower alkenyl); -(CH2)oSO2R62 (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)oOR55 (where R55 is lower alkyl; or lower alkenyl);
-(CH2)oSR56 (where R56 is lower alkyl; or lower alkenyl); -(CH2)oNR33R34 (where R33 is lower alkyl; or lower alkenyl; R34 is H; or lower alkyl; or R33 and R34 taken together are -
(CH2)M-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -
(CH2)oOCONR33R75 (where R33 is H; or lower alkyl; or lower alkenyl; R75 is lower alkyl; or R33 and R75 taken together are
-(CH2)2.6-; -(CH2)2θ(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)oNR20CONR33R82 (where R20 is H; or lower lower alkyl; R33 is H; or lower alkyl; or lower alkenyl; R82 is H; or lower alkyl; or R33 and R82 taken together are - (CHo)2-5-; -(CH2)2O(CH2)2-;
-(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); - (CH2)oN(R20)COR64 (where R20 is H; or lower alkyl; R64 is lower alkyl; or lower alkenyl); -NR20COlower alkyl (R20=H; or lower alkyl); being particularly favoured; - (CH2)oCOOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where R58 is lower alkyl, or lower alkenyl; and R59 is H; lower alkyl; or R58 and R59 taken together are -(CH2)2-6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)oPO(OR60)2 (where R60 is lower alkyl; or lower alkenyl); -(CH2)oSO2R62 (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); R16 is lower alkyl; lower alkenyl; -(CH2)oOR55 (where R55 is lower alkyl; or lower alkenyl);
-(CH2)oSR56 (where R56 is lower alkyl; or lower alkenyl); -(CH2)oNR33R34 (where R33 is lower alkyl; or lower alkenyl; R34 is H; or lower alkyl; or R33 and R34 taken together are - (CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -
(CH2)oOCONR33R75 (where R33 is H; or lower alkyl; or lower alkenyl; R75 is lower alkyl; or R33 and R75 taken together are
-(CH2)2-6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)oNR20CONR33R82 (where R20 is H; or lower lower alkyl; R33 is H; or lower alkyl; or lower alkenyl; R82 is H; or lower alkyl; or R33 and R82 taken together are - (CHz)2-6-; -(CH2)2O(CH2)2-;
-(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); - (CH2)oN(R20)COR64 (where R20 is H; or lower alkyl; R64 is lower alkyl; or lower alkenyl); -(CH2)oCOOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where R58 is lower alkyl; or lower alkenyl; and R59 is H; or lower alkyl; or R58 and R59 taken together are -(CH2)2-6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)oPO(OR60)2 (where R60 is lower alkyl; or lower alkenyl); -(CH2)oSO2R62 (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 lower alkyl; or lower alkenyl); -(CH2)qNR33R34 (where R33 is lower alkyl; or lower alkenyl; R34 is H; or lower alkyl; or R33 and R34 taken together are - (CH2)2.6-; -(CH2)2O(CH2)2-;
-(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); - (CH2)C1OCONR33R75 (where R33 is H; or lower alkyl; or lower alkenyl; R75 is lower alkyl; or R33 and R75 taken together are -(CH2)M-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)qNR20CONR33R82 (where R20 is H; or lower alkyl; R33 is H; or lower alkyl; or lower alkenyl; R82 is H; or lower alkyl; or R33 and R82 taken together are -(CH2)2. 6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)qN(R20)COR64(where: R20 is H; or lower alkyl; R64 is lower alkyl; or lower alkenyl); -(CH2)rCOOR57 (where R57 is lower alkyl; or lower alkenyl); - (CH2)C1CONR58R59 (where R58 is lower alkyl; or lower alkenyl; and R59 is H; lower alkyl; or R58 and R59 taken together are -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or - (CH2)2NR57(CH2)2-; where R57 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)C1C6H4R8 (where R8 is H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy).
4. Compounds according to claim 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
Figure imgf000156_0001
A8' wherein R20 is H or lower alkyl; and R64 is alkyl; alkenyl; -[(CH2)u-X]rCH3, wherein X is -O-, -NR20- or -S-, u is 1-3 and t is 1-6; aryl; aryl-lower alkyl; or heteroaryl-lower alkyl.
6. Compounds according to claim 5 wherein R64 is n-hexyl; n-heptyl; 4-
(phenyl)benzyl; diphenylmethyl, 3 -amino-propyl; 5-amino-pentyl; methyl; ethyl; isopropyl; isobutyl; n-propyl; cyclohexyl; cyclohexylmethyl; n-butyl; phenyl; benzyl; (3- indolyl)methyl; 2-(3-indolyl)ethyl; (4-phenyl)phenyl; n-nonyl; CH3-OCH2CH2-OCH2- or CH3-(OCH2CH2)2-OCH2-.
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)PSR56 (where R56 is lower alkyl; or lower alkenyl); -(CH2)PNR33R34 (where R33 is lower alkyl; or lower alkenyl; R34 is H; or lower alkyl; or R33 and R34 taken together are - " (CH2)W-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)r; where R57 is H; or lower alkyl); - (CH2)POCONR33R75 (where R33 is H; or lower alkyl; or lower alkenyl; R75 is lower alkyl; or R33 and R75 taken together are
-(CH2)2-6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or " lower alkyl); -(CH2)PNR20CONR33R82 (where R20 is H; or lower lower alkyl; R33 is H; or lower alkyl; or lower alkenyl; R82 is H; or lower alkyl; or R33 and R82 taken together are - (CH2W; -(CH2)2O(CH2)2-;
-(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); - (CH2)pN(R20)COR64 (where: R20 is H; or lower alkyl; R64 is lower alkyl; or lower alkenyl); (CH2)PCOOR57 (where R57: lower alkyl; or lower alkenyl); (CH2)PCONR58R59 (where R58 is lower alkyl; or lower alkenyl; and R59 is H; or lower alkyl; or R5S and R59 taken together are -(CH2W; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 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)oC6H4R8 (where R8 is H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy); R21 is H; lower alkyl; lower alkenyl; -(CH2)oOR55 (where R55 is lower alkyl; or lower alkenyl);
-(CH2)oSR56 (where R56 is lower alkyl; or lower alkenyl); -(CH2)oNR33R34 (where R33 is lower alkyl; or lower alkenyl; R34 is H; or lower alkyl; or R33 and R34 taken together are - (CH2)M-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)r; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl) ; - (CH2)oOCONR33R75 (where R33 is H; or lower alkyl; or lower alkenyl; R75 is lower alkyl; . or R33 and R75 taken together are -(CH2)M-; -(CH2)2O(CH2)r; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl);
-(CH2)oNR20CONR33R82 (where R20 is H; or lower lower alkyl; R33 is H; or lower alkyl; or lower alkenyl; R82 is H; or lower alkyl; or R33 and R82 taken together are -(CH2)2.6-; - (CH2)2O(CH2)2-;
-(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); - (CH2)oN(R20)COR64 (where: R20 is H; or lower alkyl; R64 is lower alkyl; or lower alkenyl); -(CH2)oCOOR57 (where R57 is lower alkyl; or lower alkenyl); - (CH2)oCONR58R59 (where R58 is lower alkyl, or lower alkenyl; and R59 is H; lower alkyl; or R58 and R59 taken together are -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)oPO(OR60)2 (where R60 is lower alkyl; or lower alkenyl); -(CH2)oSO2R62 (where R62 is lower alkyl; or lower alkenyl); or -(CH2^C6H4R8 (where R8 is H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy); R22 is lower alkyl; lower alkenyl; -(CH2)oOR55 (where R55 is lower alkyl; or lower alkenyl);
-(CH2)oSR56 (where R56 is lower alkyl; or lower alkenyl); -(CH2)oNR33R34 (where R33 is lower alkyl; or lower alkenyl; R34 is H; or lower alkyl; or R33 and R34 taken together are -
(CH2)2.6-; -(CH2)2O(CH2)2-; -(CHz)2S(CHz)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -
(CH2)oOCONR33R75 (where R33 is H; or lower alkyl; or lower alkenyl; R75 is lower alkyl; or R33 and R75 taken together are
-(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)oNR20CONR33R82 (where R20 is H; or lower alkyl; R33 is H; or lower alkyl; or lower alkenyl; R82 is H; or lower alkyl; or R33 and R82 taken together are -(CH2)2.
6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)oN(R20)COR64(where R20 is H; or lower alkyl; R64 is lower alkyl; or lower alkenyl); -(CH2)oCOOR57 (where R57 is lower alkyl; or lower alkenyl); -
(CH2)oCONR58R59 (where R58 is lower alkyl, or lower alkenyl; and R59 is H; lower alkyl; or R58 and R59 taken together are -(CH2)2-6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -
(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)oPO(OR60)2 (where R60 is lower alkyl; or lower alkenyl);
-(CH2)oSO2R62 (where R62 is lower alkyl; or lower alkenyl); or -(CH2)qC6H4R8 (where R8 is H; F; Cl; CF; lower alkyl; lower alkenyl; or lower alkoxy); R23 is H; lower alkyl; lower alkenyl; -(CH2)oOR55 (where R55 is lower alkyl; or lower alkenyl); -(CH2)oSR56 (where R56 is lower alkyl; or lower alkenyl); -(CH2)oNR33R34 (where R33 is lower alkyl; or lower alkenyl; R34 is H; or lower alkyl; or R33 and R34 taken together are - (CH2W; -(CH2)2O(CH2)2-;
-(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); - (CH2)oOCONR33R75 (where R33 is H; or lower alkyl; or lower alkenyl; R75 is lower alkyl; or R33 and R75 taken together are
-(CH2)M-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)oNR20CONR33R82 (where R20 is H; or lower alkyl; R33 is H; or lower alkyl; or lower alkenyl; R82 is H; or lower alkyl; or R33 and R82 taken together are -(CH2)2. 6-; -(CH2)2O(CH2)2-;
-(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); - (CH2)oN(R20)COR64 (where: R20 is H; or lower alkyl; R64 is lower alkyl; or lower alkenyl); -NR20COlower alkyl (R20=H; or lower alkyl) being particularly favoured; - (CH2)oCOOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where R58 is lower alkyl, or lower alkenyl; and R59 is H; lower alkyl; or R58 and R59 taken together are -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where
R57 is H; or lower alkyl); -(CH2)oPO(OR60)2 (where R60 is lower alkyl; or lower alkenyl);
-(CH2)oSO2R62 (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); R24 is lower alkyl; lower alkenyl; -(CH2)oOR55 (where R55 is lower alkyl; or lower alkenyl);
-(CH2)oSR56 (where R56 is lower alkyl; or lower alkenyl); -(CH2)oNR33R34 (where R33 is lower alkyl; or lower alkenyl; R34 is H; or lower alkyl; or R33 and R34 taken together are -
(CH2W; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -
(CH2)oOCONR33R75 (where R33 is H; or lower alkyl; or lower alkenyl; R75 is lower alkyl; or R33 and R75 taken together are
-(CH2W; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)oNR20CONR33R82 (where R20 is H; or lower lower alkyl; R33 is H; or lower alkyl; or lower alkenyl; R82 is H; or lower alkyl; or R33 and R82 taken together are -
(CH2W; -(CH2)2O(CH2)2-;
-(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -
(CH2)oN(R20)COR64 (where: R20 is H; or lower alkyl; R64 is lower alkyl; or lower alkenyl); -NR20COlower alkyl (R20=H ; or lower alkyl) being particularly favoured; - (CH2)oCOOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where
R58 is lower alkyl, or lower alkenyl; and R59 is H; lower alkyl; or R58 and R59 taken together are -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where
R57 is H; or lower alkyl); -(CH2)oPO(OR60)2 (where R60 is lower alkyl; or lower alkenyl); • -(CH2)oSO2R62 (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; R34 is H; or lower alkyl; or R33 and R34 taken together are -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or - (CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)mOCONR33R75 (where R33 is H; or lower alkyl; or lower alkenyl; R75 is lower alkyl; or R33 and R75 taken together are - (CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)mNR20CONR33R82 (where R20 is H; or lower alkyl; R33 is H; or lower alkyl; or lower alkenyl; R82 is H; or lower alkyl; or R33 and R82 taken together are -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or - (CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)mN(R20)COR64 (where: R20 is H; or lower alkyl; R64 is lower alkyl; or lower alkenyl); -(CH2)oCOOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)oCONR5SR59 (where R58 is lower alkyl; or lower alkenyl; and R59 is H; lower alkyl; or R58 and R59 taken together are -(CH2W; - (CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl);
-(CH2)oPO(OR60)2 (where R60 is lower alkyl; or lower alkenyl); -(CH2)oSO2R62 (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); 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; R34 is H; or lower alkyl; or R33 and R34 taken together are -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or - (CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)mOCONR33R75 (where R33 is H; or lower alkyl; or lower alkenyl; R75 is lower alkyl; or R33 and R75 taken together are - (CH2W; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)r; where R57 is H; or lower alkyl); -(CH2)mNR20CONR33R82 (where R20 is H; or lower alkyl; R33 is H; or lower alkyl; or lower alkenyl; R82 is H; or lower alkyl; or R33 and R82 taken together are -(CH2W; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or - (CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)mN(R20)COR64 (where: R20 is H; or lower alkyl; R64 is lower alkyl; or lower alkenyl); -(CH2)oCOOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where R58 is lower alkyl; or lower alkenyl; and R59 is H; lower alkyl; or R58 and R59 taken together are -(CH2)2-6-; -
(CH2)2O(CH2)r; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl);
-(CH2)oPO(OR60)2 (where R60 is lower alkyl; or lower alkenyl); -(CH2)oSO2R62 (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, alternatively, R25 and R26 taken together are -
(CH2)2.6-; -(CH2)2O(CH2)2-;
-(CH2)2S(CH2)2-; or -(CH2)2NR34(CH2)2-;
R27 is H; lower alkyl; lower alkenyl; -(CH2)oOR55 (where R55 is lower alkyl; or lower alkenyl);
-(CH2)oSR56 (where R56 is lower alkyl; or lower alkenyl); -(CH2)oNR33R34 (where R33 is lower alkyl; or lower alkenyl; R34 is H; or lower alkyl; or R33 and R34 taken together are -
(CH2)2-6-; -(CH2)2O(CH2)2-;
-(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); - (CH2)oOCONR33R75 (where R33 is H; or lower alkyl; or lower alkenyl; R75 is lower alkyl; or R33 and R75 taken together are
-(CH2)2.6-; -(CH2)2O(CH2)r; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)oNR20CONR33R82 (where R20 is H; or lower lower alkyl; R33 is H; or lower alkyl; or lower alkenyl; R82 is H; or lower alkyl; or R33 and R82 taken together are - (CH2)2-6-; -(CH2)2O(CH2)2-;
-(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -
(CH2)oN(R20)COR64 (where R20 is H; or lower alkyl; R64 is lower alkyl; or lower alkenyl);
-(CH2)oCOOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where
R58 is lower alkyl, or lower alkenyl; and R59 is H; lower alkyl; or R58 and R59 taken together are -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)r; where R57 is H; or lower alkyl); -(CH2)oPO(OR60)2 (where R60 is lower alkyl; or lower alkenyl); -(CH2)oSO2R62 (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)oOR55 (where R55 is lower alkyl; or lower alkenyl);
-(CH2)oSR56 (where R56 is lower alkyl; or lower alkenyl); -(CH2)oNR33R34 (where R33 is lower alkyl; or lower alkenyl; R34 is H; or lower alkyl; or R33 and R34 taken together are -
(CH2)W-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -
(CH2)oOCONR33R75 (where R33 is H; or lower alkyl; or lower alkenyl; R75 is lower alkyl; or R33 and R75 taken together are
-(CH2W; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR37(CH2)2-; where R57 is H; or lower alkyl); -(CH2)oNR20CONR33R82 (where R20 is H; or lower alkyl; R33 is H; or lower alkyl; or lower alkenyl; R82 is H; or lower alkyl; or R33 and R82 taken together are -(CH2)2. 6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower • alkyl); -(CH2)oN(R20)COR64(where: R20 is H; or lower alkyl; R64 is lower alkyl; or lower alkenyl); -(CH2)oCOOR57 (where R57 is lower alkyl; or lower alkenyl); - (CH2)oCONR58R59 (where R58 is lower alkyl, or lower alkenyl; and R59 is H; lower alkyl; or R58 and R59 taken together are -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or - (CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)oPO(OR60)2 (where R60 is lower alkyl; or lower alkenyl);
-(CH2)oSO2R62 (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 R29 is lower alkyl; lower alkenyl; -(CH2)oOR55 (where R55 is lower alkyl; or lower alkenyl);
-(CH2)oSR56 (where R56 is lower alkyl; or lower alkenyl); -(CH2)oNR33R34 (where R33 is lower alkyl; or lower alkenyl; R34 is H; or lower alkyl; or R33 and R34 taken together are - (CH2W; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); - (CH2)oOCONR33R75 (where R33 is H; or lower alkyl; or lower alkenyl; R75 is lower alkyl; or R33 and R75 taken together are
-(CHz)2-6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)oNR20CONR33R82 (where R20 is H; or lower alkyl; R33 is H; or lower alkyl; or lower alkenyl; R82 is H; or lower alkyl; or R33 and R82 taken together are -(CH2)2. 6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)r; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)oN(R20)COR64(where: R20 is H; or lower alkyl; R64 is lower alkyl; or lower alkenyl); -NR20COlower-alkyl (R20=H; or lower alkyl) being particularly favoured; - (CH2)oCOOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)oCONR5SR59 (where R58 is lower alkyl, or lower alkenyl; and R59 is H; lower alkyl; or R38 and R59 taken together are -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or - (CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)oPO(OR60)2 (where R60 is lower alkyl; or lower alkenyl);
-(CH2)oSO2R62 (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).
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; GIy; His; He; Leu; Lys; Met; Phe; Pro; Pro(5RPhe), Ser; Thr; Trp; Tyr; VaI; 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; C4al; C5al; NIe; 2-Nal; 1 -NaI; 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(BzI); Bip; S(BzI); T(BzI); hCha; hCys; hSer, hArg; hPhe; Bpa; Pip; OctG; MePhe; MeNIe; MeAIa; MeIIe; MeVaI; or MeLeu.
12. Compounds according to claim 10 wherein B is a group, having (L)- configuration, of formula
Figure imgf000163_0001
A8"
wherein R20 is H; or lower alkyl; and R64 is alkyl; alkenyl; -[(CH2)u-X]rCH3, wherein X is -O-, -NR20- or -S-, u is 1-3 and t is 1-6; aryl; aryl-lower alkyl; or heteroaryl-lower alkyl.
13. Compounds according to claim 12 wherein R64 is n-hexyl; n-heptyl; 4- (phenyl)benzyl; diphenylmethyl, 3-amino-propyl; 5-amino-pentyl; methyl; ethyl; isopropyl; isobutyl; n-propyl; cyclohexyl; cyclohexylmethyl; n-butyl; phenyl; benzyl; (3- indolyl)methyl; 2-(3-indolyl)ethyl; (4-phenyl)phenyl; n-nonyl; CH3-OCH2CH2-OCH2- or CH3-(OCH2CH2)O-OCH2-.
14. Compounds according to claim 1 wherein
Figure imgf000164_0001
is a group of formula (bl) or (1); 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; R34 is H; or lower alkyl; or .
R33 and R34 taken together are -(CH2W; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -
(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)POCONR33R75 (where R33 is
H; or lower alkyl; or lower alkenyl; R75 is lower alkyl; or R33 and R75 taken together are -
(CH2W; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)PNR20CONR33R82 (where R20 is H; or lower alkyl; R33 is H; or lower alkyl; or lower alkenyl; R82 is H; or lower alkyl; or
R33 and R82 taken together are -(CH2)M-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -
(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)PN(R20)COR64 (where: R20 is
H; or lower alkyl; R64 is lower alkyl; or lower alkenyl); -(CH2)oCOOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where R58 is lower alkyl, or lower alkenyl; and R59 is H; lower alkyl; or R58 and R59 taken together are -(CH2)2.6-; -
(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl);
-(CH2)oPO(OR60)2 (where R60 is lower alkyl; or lower alkenyl); -(CH2)oSO2R62 (where R62 is lower alkyl; or lower alkenyl); or -(CH2)AH4R8 (where R8 is H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy); most preferably -CH2CONR58R59 (where R58 is H; or lower alkyl; and R59 is lower alkyl; or lower alkenyl);
R32 is H; or methyl;
R33 is lower alkyl; lower alkenyl; -(CH2)mOR55 (where R55 is lower alkyl; or lower alkenyl);
-(CH2)mNR34R63 (where R34 is lower alkyl; or lower alkenyl; R63 is H; or lower alkyl; or
R34 and R63 taken together are -(CH2W; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or - (CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)mOCONR75R82 (where R75 is lower alkyl; or lower alkenyl; R82 is H; or lower alkyl; or R75 and R82 taken together are - (CH2)M-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or
-(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)mNR20CONR78R82 (where R20 is H; or lower alkyl; R78 is H; or lower alkyl; or lower alkenyl; R82 is H; or lower alkyl; or R78 and R82 taken together are -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or - (CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)mN(R20)COR64 (where: R20 is H; or lower alkyl; R64 is lower alkyl; or lower alkenyl); -(CH2)oCOOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where R58 is lower alkyl; or lower alkenyl; and R59 is H; lower alkyl; or R58 and R59 taken together are -(CH2)2.6-; -
(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57: H; or lower alkyl);
R34 is H; or lower alkyl;
R35: isH; lower alkyl; lower alkenyl; (CH2)mOR55 (where R55: lower alkyl; or lower alkenyl); -(CH2)mNR33R34 (where R33 is lower alkyl; or lower alkenyl; R34 is H; or lower alkyl; or R33 and R34 taken together are -(CH2W; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or - (CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)mOCONR33R75 (where R33 is H; or lower alkyl; or lower alkenyl; R75 is lower alkyl; or R33 and R75 taken together are - (CH2W; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)mNR20CONR33R82 (where R20 is H; or lower alkyl; R33 is H; or lower alkyl; or lower alkenyl; R82 is H; or lower alkyl; or R33 and R82 taken together are -(CH2W; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or - (CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)mN(R20)COR64 (where: R20 is H; or lower alkyl; R64 is lower alkyl; or lower alkenyl); -(CH2)oCOOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where R58 is lower alkyl; or lower alkenyl; and R59 is H; lower alkyl; or R58 and R59 taken together are -(CH2)2.6-; - (CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); R36: 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; R34 is H; or lower alkyl; or R33 and R34 taken together are -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or - (CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl);- (CH2)POCONR33R75 (where R33 is H; or lower alkyl; or lower alkenyl; R75 is lower alkyl; or R33 and R75 taken together are - (CH2)W-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)PNR20CONR33R82 (where R20 is H; or lower alkyl; R33 is H; or lower alkyl; or lower alkenyl; R82 is H; or lower alkyl; or R33 and R82 taken together are -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or - (CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)PN(R20)COR64 (where: R20 is H; or lower alkyl; R64 is lower alkyl; or lower alkenyl); -(CH2)oCOOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where R58 is lower alkyl, or lower alkenyl; and R59 is H; lower alkyl; or R58 and R59 taken together are -(CH2)2-6-; - (CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)oPO(OR60)2 (where R60 is lower alkyl; or lower alkenyl); -(CH2)oSO2R62 (where R62 is lower alky; or lower alkenyl); or -(CH2)C1C6H4R8 (where R8 is H; F; Cl; CF3; lower alkyl; lower alkenyl; or lower alkoxy); and
R3S 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; R34 is H; or lower alkyl; or R33 and R34 taken together sre -(CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or - (CH2)2NR57(CH2)r; where R57 is H; or lower alkyl); -(CH2)POCONR33R75 (where R33 is H; or lower alkyl; or lower alkenyl; R75 is lower alkyl; or R33 and R75 taken together are - " (CH2)2.6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)PNR20CONR33R82 (where R20 is H; or lower alkyl; R33 is H; or lower alkyl; or lower alkenyl; R82 is H; or lower alkyl; or R33 and R82 taken together are -(CH2)2-6-; -(CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or - (CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)PN(R20)COR64 (where: R20 is H; or lower alkyl; R64 is lower alkyl; or lower alkenyl); -(CH2)oCOOR57 (where R57 is lower alkyl; or lower alkenyl); -(CH2)oCONR58R59 (where R58 is lower alkyl, or lower alkenyl; and R59 is H; lower alkyl; or R58 and R59 taken together are -(CH2)2.6-; - (CH2)2O(CH2)2-; -(CH2)2S(CH2)2-; or -(CH2)2NR57(CH2)2-; where R57 is H; or lower alkyl); -(CH2)oPO(OR60)2 (where R60 is lower alkyl; or lower alkenyl); -(CH2)oSO2R62 (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 the α-amino acid residues in positions 1 to 11 in the chain Z are:
Pl : of type C, or of type D, or of type E, or of type F;
P2: of type E, or of type F, or of type C; - P3: or of type C, of type F or the residue is GIy;
P4: of type C, or of type E, or of type F, or the residue is GIy or Pro;
P5: of type E, or of type F, or the residue is GIy or Pro;
P6: of type C, or of type D, or of type F, or the residue is GIy or Pro;
P7: of type F or of formula -A-CO-or the residue is GIy or Pro; - P8: of type D, or of type C, or of formula -A-CO or the residue is GIy or Pro or Pro(4NHCOPhe);
P9: of type C, or of type D, or of type E, or of type F;
PlO: of type F, or of type C, or type E;
Pl 1 : of type E, or of type F, or of type C or of type D; or - P2 and PlO, taken together, form a group of type H; with the proviso that if template is DPro-LPro the amino acid residues in positions Pl to PI l are other than
Pl : Arg
P2: Cys, linked with Cys in position PlO by a disulfide bridge - P3: Thr
P4 Lys
P5 Ser
P6 He
P7 Pro - P8 Pro
P9 He
PlO Cys, linked with Cys in position PlO by a disulfide bridge; and
Pl 1 Phe.
17. Compounds according to claim 16 wherein the α-amino acid residues in positions 1 to 11 of the chain Z are:
Pl : NIe, He, Aoc, hLeu, Chg, OctG, hPhe, 4AmPhe, Cha, Phe, Ty r,
2C1-Phe, Trp, 1-Nal, Leu, Cha or Arg;
P2: Cys, GIu, NIe, Thr, or GIn ; - P3: Thr, Ala or Abu;
P4: Lys, NIe, Ala, Abu, or Thr; P5: Ser, AlloThr or Dpr;
P6: He, c5al, Leu, NIe, Aoc, OctG, Cha, hLeu, hPhe, Chg, t-BuA, GIu, or Asp; P7: Pro; - P8: Pro, Ala, or Pro(4NHCOPhe);
P9: Tyr, Phe, He, NIe, Cha, GIn, Arg, Lys, His, Thr, or Ala; PlO: Cys, Arg, NIe, GIn, Lys, Met, Thr, or Ser; Pl 1 : Tyr, GIn, Arg, Ser, NIe, 2-NaI, 2C1-Phe, Cha, Phg, Tyr, Phe, Asp Asn, or Thr; and - Cys, if present at P2 and PlO, may form a disulfide bridge.
18. Compounds according to any one of claims 1 to 15 wherein the α-amino acid residues in positions 1 to 11 in the chain Z are:
- Pl : of type C, or of type D, or of type E;
P2: of type F, or of type C;
P3: of type F;
P4: of type C, or of type E;
P5: of type E, or of type F; - P6: of type F;
P7: of type F, or of formula -A-CO-, or the residue is GIy or Pro;
P8: of type C, or of formula -A-CO-, or the residue is GIy or Pro or
Pro(4NHCOPhe);
P9: of type C, or of type D, or of type F; - PlO: of type F, or of type C, or type E;
Pl 1 : of type E, or of type D, or of type F; or
P2 and PlO, taken together, form a group of type H;
19. Compounds according to claim 18 wherein the α-amino acid residues in positions 1 to 11 are:
Pl : Phe, hPhe, 4AmPhe, NIe, Chg, He, Tyr, Arg, Trp, 2C1-Phe, Arg, 1- NaI, or Cha; P2: Cys, GIu, or NIe; - P3: Thr;
P4: Lys, or NIe; P5: Ser, AlloThr, or Dpr;
P6: Asp, or GIu;
P7: Pro;
P8: Pro;
P9: lie, NIe, Cha, GIn, Tyr, or Ala;
PlO: Cys, Arg, or NIe;
PI l : Thr, Asp, Ser, Tyr, Phe, Asn, or Arg; and
Cys, if present at P2 and PlO, may form a disulfide bridge
20. Compounds according to any one of claims 1 to 15 wherein the α-amino acid residues in positions 1 to 11 in the chain Z are:
Pl : of type C, or of type D;
P2: of type F;
P3: oftype F or oftype C; - P4: oftype C or oftype F;
P5: oftype F;
P6: oftype C;
P7: of formula A-CO-, or the residue is GIy or Pro;
P8: of formula -A-CO-, or the residue is GIy or Pro or Pro(4NHCOPhe);
P9: of type D, or of type F or of type C;
PlO: of type F, or of type C, or type E;
PI l : of type E, or of type F, or of type D; or
P2 and PlO, taken together, form a group of type H.
21. Compounds according to claim 20 wherein the α-amino acid residues in positions 1 to 11 are:
Pl : He, NIe, Aoc, hLeu, Chg, OctG, or hPhe;
P2: Cys, GIu, Thr, or GIn; - P3: Thr, Ala, or Abu;
P4: Ala, Thr, or Abu;
P5: Ser;
P6: OctG, He, Cha, Leu, c5al, NIe, Aoc, Chg, tBuA, or hLeu;
P7: Pro; - P8: Pro, or Pro(4NHCOPhe);
P9: GIn, Tyr, ILe, or Phe; PlO: Cys, Lys, GIn, Thr, Met, or Arg;
PI l : Tyr, Ser, Arg, GIn, NIe, 2-Nal, 2C1-Phe, Phe, Cha, or Phg; and Cys, if present at P2 and PlO, may form a disulfide bridge.
22. Compounds according to any one of claims 1 to 15 wherein the α-amino acid residues in positions 1 to 11 in the chain Z are:
Pl : of type C, or of type D, or of type E;
P2: of type F;
P3: oftype F; - P4: oftype E;
P5: oftype F;
P6: of type C or of type D;
P7: of type F or of formula -A-CO-, or the residue is GIy or Pro;
P8: of type C, or of formula -A-CO-, or the residue is GIy or Pro; - P9: of type C, or of type E, or of type F;
PlO: of type F;
PI l : of type E, or of type D; or
P2 and PlO, taken together, form a group of type H; with the proviso that if the template is DPro-LPro, the amino acid residues in positions Pl to Pl 1 are other than
Pl: Arg
P2: Cys, linked with Cys in position PlO by a disulfide bridge
P3: Thr
P4 Lys - P5 Ser
P6 He
P7 Pro
P8 Pro
P9 He - PlO Cys, linked with Cys in position Pl 0 by a disulfide bridge; and
PI l Phe.
23. Compounds according to claim 22 wherein the α-amino acid residues in positions 1 to 11 of the chain Z are: - Pl: Cha, Tyr , or Trp
P2: Cys P3: Thr
P4: Lys
P5: Ser
P6: Leu
P7: Pro
P8: Pro
P9: Lys
PlO: Cys
PI l : Arg; and the Cys residues present at P2 and PlO may form a disulfide bridge.
24. A compound of formula I according to claim 1 wherein the template is Pro- Pro and the amino acid residues in position 1 - 11 are:
Pl : Phe;
P2: Cys;
P3: Thr;
P4: Lys;
P5: Ser;
P6: Asp;
P7: Pro;
P8: Pro;
P9: He;
PlO: Cys;
PI l : Tyr;
Cys at P2 and PlO forming a disulfide bridge.
25. A compound of formula I according to claim 1 wherein the template is DPro-LPro and the amino acid residues in position 1 - 11 are: Pl : He;
P2: Cys; P3: Thr; P4: Lys; P5: Ser; - P6: Asp;
P7: Pro; P8: Pro;
P9: NIe;
PlO: Cys;
PI l : Are;
Cys at P2 and PlO forming a disulfide bridge.
26. A compound of formula I according to claim 1 wherein the template is Pro- Pro and the amino acid residues in position 1 - 11 are:
Pl : He;
P2: Cys;
P3: Thr;
P4: Lys;
P5: Ser;
P6: Asp;
P7: Pro;
P8: Pro;
P9: Cha;
PlO: Cys;
PI l: Arg; Cys at P2 and PlO forming a disulfide bridge.
27. A compound of formula I according to claim 1 wherein the template is Pro- Pro and the amino acid residues in position 1 - 1 1 are:
Pl : He;
P2: Cys;
P3: Thr;
P4: NIe;
P5: Ser;
P6: Asp;
P7: Pro;
P8: Pro;
P9: He;
PlO: Cys;
PI l : Arg Cys at P2 and PlO forming a disulfide bridge.
28. A compound of formula I according to claim 1 wherein the template is DPro-LPro and the amino acid residues in position 1 — 11 are:
Pl : Phe;
P2: Cys; - P3: Thr;
P4: NIe;
P5: Ser;
P6: Asp;
P7: Pro; - P8: Pro;
P9: He;
PlO: Cys;
PI l : Tyr;
Cys at P2 and PlO forming a disulfide bridge.
29. A compound of formula I according to claim 1 wherein the template is DPro-LPro and the amino acid residues in position 1 - 11 are:
Pl : Chg;
P2: Cys; - P3: Thr;
P4: Lys;
P5: Ser;
P6: Asp;
P7: Pro; - P8: Pro;
P9: He;
PlO: Cys;
PI l : Tyr;
Cys at P2 and PlO forming a disulfide bridge.
30. A compound of formula I according to claim 1 wherein the template is DPro-LPro and the amino acid residues in position 1 - 11 are:
Pl : Arg;
P2: Cys; P3: Thr;
P4: Lys; P5: Ser;
P6: Asp;
P7: Pro;
P8: Pro;
P9: He;
PlO: Cys;
PI l : Phe;
Cys at P2 and PlO forming a disulfide bridge.
31. A compound of formula I according to claim 1 wherein the template is DPro-LPro and the amino acid residues in position 1 - 11 are:
Pl : NIe;
P2: Cys;
P3: Thr;
P4: Ala;
P5: Ser;
P6: He;
P7: Pro;
P8: Pro;
P9: GIn;
PlO: Cys;
PI l : GIn;
Cys at P2 and PlO forming a disulfide bridge.
32. A compound of formula I according to claim 1 wherein the template is DPro-LPro and the amino acid residues in position 1 - 11 are: Pl : NIe;
P2: Cys;
P3: Thr;
P4: Ala;
P5: Ser;
P6: He;
P7: Pro;
P8: Pro;
P9: GIn;
PlO: Cys; PIl: Tyr;
Cys at P2 and PlO forming a disulfide bridge.
33. A compound of formula I according to claim 1 wherein the template is Pro- Pro and the amino acid residues in position 1-11 are:
Pl: hPhe;
P2: Cys;
P3: Thr;
P4: Ala;
P5: Ser;
P6: OctG;
P7: Pro;
P8: Pro;
P9: GIn;
PlO: Cys;
PIl: GIn
Cys at P2 and PlO forming a disulfide bridge.
34. A compound of formula I according to claim 1 wherein the template is Pro- Pro and the amino acid residues in position 1-11 are:
Pl: OctG;
P2: GIn;
P3: Thr;
P4: Ala;
P5: Ser;
P6: He;
P7: Pro;
P8: Pro;
P9: GIn;
PlO: Thr;
PIl: Tyr.
35. A compound of formula I according to claim 1 wherein the template is DPro-LPro and the amino acid residues in position 1-11 are: - Pl: hPhe;
P2: Cys; P3: Thr;
P4: Ala;
P5: Ser;
P6: Cha;
P7: Pro;
P8: Pro;
P9: GIn;
PlO: Cys;
PI l : Phe;
Cys at P2 and PlO forming a disulfide bridge.
36. A compound of formula I according to claim 1 wherein the template is DPro-LPro and the amino acid residues in position 1 - 11 are:
Pl : OctG;
P2: GIu;
P3: Thr;
P4: Ala;
P5: Ser;
P6: lie;
P7: Pro;
P8: Pro;
P9: GIn;
PlO: Lys;
PI l : Tyr.
37. A compound of formula I according to claim 1 wherein the template is DPro-LPro and the amino acid residues in position 1 - 11 are:
Pl : OctG;
P2: Cys;
P3: Thr;
P4: Ala;
P5: Ser;
P6: Cha;
P7: Pro;
P8: Pro;
P9: GIn; PlO: Cys;
PI l : Phe; Cys at P2 and PlO forming a disulfide bridge.
38. A compound of formula I according to claim 1 wherein the template is DPro-LPro and the amino acid residues in position 1 - 1 1 are:
Pl : hPhe;
P2: Cys;
P3: Thr;
P4: Ala;
P5: Ser;
P6: Cha;
P7: Pro;
P8: Pro;
P9: GIn;
PlO: Cys;
PI l : GIn;
Cys at P2 and PlO forming a disulfide bridge.
39. A compound of formula I according to claim 1 wherein the template is Pro- Pro and the amino acid residues in position 1 - 11 are:
Pl : OctG;
P2: Cys;
P3: Thr;
P4: Ala;
P5: Ser;
P6: Cha;
P7: Pro;
P8: Pro(4NHCOPhe);
P9: GIn;
PlO: Cys;
PI l : GIn;
Cys at P2 and PlO forming a disulfide bridge.
40. A compound of formula I according to claim 1 wherein the template is DPro-LPro and the amino acid residues in position 1 - 11 are: - Pl : hPhe;
- P2: Cys;
- P3: Thr;
- P4: Ala;
- P5: Ser;
- P6: OctG;
- P7: Pro;
- P8: Pro;
- P9: GIn;
- PlO: Cys;
- PI l : Tyr;
Cys at P2 and PlO forming a disulfide bridge.
41. A compound of formula I according to claim 1 wherein the template is DPro-LPro and the amino acid residues in position 1 - 11 are:
- Pl : Chg;
- P2: Cys;
- P3: Thr;
- P4: Lys;
- P5: Ser;
- P6: Asp;
- P7: Pro;
- P8: Pro;
- P9: He;
- PlO: Cys;
PI l : Thr;
Cys at P2 and PlO forming a disulfide bridge.
42. A compound of formula I according to claim 1 wherein the template is DPro-LPro and the amino acid residues in position 1 - 11 are:
Pl: Chg;
P2: Cys;
P3: Thr;
P4: Lys; - P5: Ser;
P6: Asp; - P7: Pro;
- P8: Pro;
- P9: He;
- PlO: Cys;
- PI l : Asp;
Cys at P2 and PlO forming a disulfide bridge.
43 , A compound of formula I according to claim 1 wherein the template is DPro-LPro and the amino acid residues in position 1 - 11 are:
- Pl : OctG;
- P2: Cys;
- P3: Thr;
- P4: Ala;
- P5: Ser;
- P6: Cha;
- P7: Pro;
- P8: Pro;
- P9: GIn;
- PlO: Cys;
I PI l : GIn;
Cys at P2 and PlO forming a disulfide bridge.
44. A compound of formula I according to claim 1 wherein the template is Pro- Pro and the amino acid residues in position 1 - 1 1 are:
Pl : OctG;
P2: Cys;
P3: Thr;
P4: Ala;
P5: Ser;
P6: He;
P7: Pro;
P8: Pro;
P9: GIn;
PlO: Cys;
PI l : Tyr;
Cys at P2 and PlO forming a disulfide bridge.
45. A compound of formula I according to claim 1 wherein the template is DPro-LPro and the amino acid residues in position 1 — 11 are:
Pl: OctG;
P2: Cys;
P3: Thr;
P4: Ala;
P5: Ser;
P6: OctG;
P7: Pro;
P8: Pro;
P9: GIn;
PlO: Cys;
PI l : GIn; Cys at P2 and PlO forming a disulfide bridge.
46. A compound of formula I according to claim 1 wherein the template is DPro- Pro and the amino acid residues in position 1 - 11 are:
Pl: Cha;
P2: Cys;
P3: Thr;
P4: Lys;
P5: Ser;
P6: Leu;
P7: Pro;
P8: Pro;
P9: Lys;
PlO: Cys;
PI l : Arg; Cys at P2 and PlO forming a disulfide bridge.
47. A compound of formula I according to claim 1 wherein the template is DPro-LPro and the amino acid residues in position 1 - 11 are:
Pl: Tyr; - P2: Cys;
P3: Thr; P4: Lys;
P5: Ser;
P6: Leu;
P7: Pro;
P8: Pro;
P9: Lys;
PlO: Cys;
PI l : Arg;
Cys at P2 and PlO forming a disulfide bridge.
48. A compound of formula I according to claim 1 wherein the template is DPro-LPro and the amino acid residues in position 1 — 11 are:
Pl : Trp;
P2: Cys;
P3: Thr;
P4: Lys;
P5: Ser;
P6: Leu;
P7: Pro;
P8: Pro;
P9: Lys;
PlO: Cys;
PI l : Arg;
Cys at P2 and PlO forming a disulfide bridge.
49. Enantiomers of the compounds of formula I as defined in claim 1.
50. Compounds according to any one of claims 1 to 49 for use as therapeutically active substances.
51. Compounds according to claim 50 having selective protease inhibitory activity in particular against Cathepsin G or Elastase or Tryptase and/or anticancer activity and/or anti inflammatory activity and/or anti infective activity and/or anticardiovascular activity and/or antiimmunological activity and/or antineurodegenerative activity.
52. A pharmaceutical composition containing a compound according to any one of claims 1 to 49 and a pharmaceutically inert carrier.
53. Compositions according to claim 52 in a form suitable for oral, topical, transdermal, buccal, transmucosal or pulmonary administration or for administration by injection or inhalation.
54. Compositions according to claim 52 or 53 in form of tablets, dragees, capsules, solutions, liquids, gels, plaster, creams, ointments, syrup, slurries, suspensions, spray, nebuliser or suppositories.
55. The use of compounds according to any one of claims 1 to 49 for the manufacture of a medicament for use as an inhibitor of protease enzymes.
56. The use according to claim 55 wherein said protease inhibiting medicament is intended to be used for preventing infections in healthy individuals or for slowing infections in infected patients; or where cancer is mediated or resulting from; or where an immunological diseases are mediated or resulting from protease activity or where inflammation is mediated or resulting from protease activity; or where immunological reaction is mediated or resulting from protease activity.
57. The use of compounds according to any one of claims 18, 19 and 24-30 for the manufacture of a medicament for use as an inhibitor of Cathepsin G.
58. The use of compounds according to any one of claims 20, 21 and 31-42 for the manufacture of a medicament for use as an inhibitor of Elastase.
59. The use of compounds according to any one of claims 22, 23 and 43-45 for the manufacture of a medicament for use as an inhibitor of Tryptase.
60. A process for the manufacture of compounds according to any one of claims 1 -48 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 5, 6 or 7, 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 steps (c) and (d) until the N-terminal amino acid residue has been introduced;
(f) coupling the product thus obtained with a compound of the general formula
Figure imgf000183_0001
π wherein
Figure imgf000183_0002
is as defined in claim 1 and X is an N-protecting group or, alternatively, if
Figure imgf000183_0003
is to be group (al ) or (a2) as defined in claim 1 ,
(fa) coupling the product obtained in step (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 in claim 1 , 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, III, any functional group which may be present in said N-protected amino acid derivative being likewise appropriately protected; and, respectively, if
Figure imgf000184_0001
is to be group (a3) as defined in claim 1 ,
(fa1) coupling the product obtained in step (e) with an appropriately N- protected derivative of an amino acid of the above general formula III, any functional group which may be present in said N-protected amino acid derivative being likewise appropriately protected;
(fbτ) removing the N-protecting group from the product thus obtained; and (fc1) coupling the product thus obtained with an appropriately N-protected derivative of an amino acid of the above general formula III, 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) or
(fc1);
(h) coupling the product thus obtained with an appropriately N-protected derivative of that amino acid which in the desired end-product is in position 1 1, 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 with an appropriately N-protected derivative of that amino acid which in the desired end-product is one position farther away from position 1 1, 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 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; (n) if desired, forming an interstrand linkage between side-chains of appropriate amino acid residues at positions 2 and 10;
(o) detaching the product thus obtained from the solid support;
(p) cyclizing the product cleaved from the solid support;
(q) 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.
61. A process for the manufacture of compounds according to any one of claims 1-48 which process comprises
(a1) coupling an appropriately functionalized solid support with a compound of the general formula
Figure imgf000185_0001
II wherein
Figure imgf000185_0002
is as defined in claim 1 and X is an N-protecting group or, alternatively, if
Figure imgf000185_0003
is to be group (al) or (a2) as defined in claim 1 , (a'a) coupling said appropriately functionalized solid support 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 in claim 1 , any functional group which may be present in said N-protected amino acid derivative being likewise appropriately protected;
(a'b) removing the N-protecting group from the product thus obtained; and (a'c) coupling the product thus obtained with an appropriately N-protected derivative of an amino acid of the above general formula IV and, respectively, III, any functional group which may be present in said N-protected amino acid derivative being likewise appropriately protected; and, respectively, if
Figure imgf000186_0001
is to be group (a3) as defined in claim 1 , (a'a') coupling said appropriately functionalized solid support with an appropriately N-protected derivative of an amino acid of the above general formula III, any functional group which may be present in said N-protected amino acid derivative being likewise appropriately protected; (a'b1) removing the N-protecting group from the product thus obtained; and (a'c1) coupling the product thus obtained with an appropriately N-protected derivative of an amino acid of the above general formula III, any functional group which may be present in said N-protected amino acid derivative being likewise appropriately protected;
(b1) removing the N-protecting group from the product obtained in step (a1), (a'c) or (a'c1);
(c1) coupling the product thus obtained with an appropriately N-protected derivative of that amino acid which in the desired end-product is in position 1 1 , any functional group which may be present in said N-protected amino acid derivative being likewise appropriately protected; (d1) removing the N-protecting group from the product thus obtained;
(e1) coupling the product thus obtained with an appropriately N-protected derivative of that amino acid which in the desired end-product is one position farther away from position, any functional group which may be present in said N-protected amino acid derivative being likewise appropriately protected;
(f ) removing the N-protecting group from the product thus obtained;
(g1) repeating steps (e1) and (f ) until all amino acid residues have been introduced; (h1) if desired, selectively deprotecting one or several protected functional group(s) present in the molecule and appropriately substituting the reactive group(s) thus liberated;
(i1) if desired forming an interstrand linkage between side-chains of appropriate amino acid residues at positions 2 and 10;
(j1) detaching the product thus obtained from the solid support; (k1) cyclizing the product cleaved from the solid support;
(V) 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
(m1) 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.
62. A modification of the processes according to claim 60 or 61 for the manufacture of compounds according to claim 49 in which enantiomers of all chiral starting materials are used.
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