WO2004080480A1 - Pharmaceutical preparations comprising acid-stabilised insulin - Google Patents

Pharmaceutical preparations comprising acid-stabilised insulin Download PDF

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Publication number
WO2004080480A1
WO2004080480A1 PCT/DK2004/000158 DK2004000158W WO2004080480A1 WO 2004080480 A1 WO2004080480 A1 WO 2004080480A1 DK 2004000158 W DK2004000158 W DK 2004000158W WO 2004080480 A1 WO2004080480 A1 WO 2004080480A1
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Prior art keywords
alkyl
aryl
independently selected
pharmaceutical composition
composition according
Prior art date
Application number
PCT/DK2004/000158
Other languages
French (fr)
Inventor
Søren Østergaard
Helle Birk Olsen
Niels C. Kaarsholm
Peter Madsen
Palle Jakobsen
Svend Ludvigsen
Gerd Schluckebier
Dorte Bjerre Steensgaard
Anders Klarskov Petersen
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Novo Nordisk A/S
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Publication date
Application filed by Novo Nordisk A/S filed Critical Novo Nordisk A/S
Priority to AU2004218808A priority Critical patent/AU2004218808A1/en
Priority to BRPI0408229-0A priority patent/BRPI0408229A/en
Priority to JP2006504320A priority patent/JP2007523842A/en
Priority to CA002522818A priority patent/CA2522818A1/en
Priority to MXPA05009565A priority patent/MXPA05009565A/en
Priority to EP04719368A priority patent/EP1610812A1/en
Publication of WO2004080480A1 publication Critical patent/WO2004080480A1/en
Priority to US11/227,760 priority patent/US20060069013A1/en
Priority to NO20054555A priority patent/NO20054555L/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K33/00—Medicinal preparations containing inorganic active ingredients
    • A61K33/24—Heavy metals; Compounds thereof
    • A61K33/30—Zinc; Compounds thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/22—Hormones
    • A61K38/28—Insulins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/34—Macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyesters, polyamino acids, polysiloxanes, polyphosphazines, copolymers of polyalkylene glycol or poloxamers
    • A—HUMAN NECESSITIES
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    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
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    • C07D317/48—Methylenedioxybenzenes or hydrogenated methylenedioxybenzenes, unsubstituted on the hetero ring
    • C07D317/50—Methylenedioxybenzenes or hydrogenated methylenedioxybenzenes, unsubstituted on the hetero ring with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to atoms of the carbocyclic ring
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    • C07D401/04—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings directly linked by a ring-member-to-ring-member bond
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    • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
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    • C07K14/62—Insulins

Definitions

  • the present invention discloses pharmaceutical preparations comprising ligands for the HisBlO Zn 2+ sites of the R-state insulin hexamer and acid-stabilsed insulin analogues.
  • the compositions release insulin slowly following subcutaneous injection.
  • the insulin hexamer is an allosteric protein that exhibits both positive and negative cooperativity and half-of-the-sites reactivity in ligand binding.
  • This allosteric behav- iour consists of two interrelated allosteric transitions designated L A 0 and- L B 0> three inter- converting allosteric conformation states (eq. 1),
  • T 6 , T 3 R 3 , and R 6 and two classes of allosteric ligand binding sites designated as the phenolic pockets and the His 810 anion sites. These allosteric sites are associated only with insulin subunits in the R conformation. Insulin Hexamer Structures and Ligand Binding.
  • the T- to R-transition of the insulin hexamer involves transformation of the first nine residues of the B chain from an extended conformation in the T-state to an ⁇ -helical conformation in the R-state. This coil-to-helix transition causes the N-terminal residue, Phe B1 , to undergo an ⁇ 30 A change in position.
  • This conformational change creates hydrophobic pockets (the phenolic pockets) at the sub- unit interfaces (three in T 3 R 3 , and six in Re), and the new B-chain helices form 3-helix bun- dies (one in T 3 R 3 and two in Re) with the bundle axis aligned along the hexamer three-fold symmetry axis.
  • the His 610 Zn 2+ in each R 3 unit is forced to change coordination geometry from octahedral to either tetrahedral (monodentate ligands) or pentahedral (bidentate ligands). Formation of the helix bundle creates a narrow hydrophobic tunnel in each R 3 unit that extends from the surface -12 A down to the His 810 metal ion. This tunnel and the His 610 Zn 2+ ion form the anion binding site.
  • Hexamer Ligand Binding and Stability of Insulin Formulations The in vivo role of the T to R transition is unknown. However, the addition of allosteric ligands (e.g. phenol and chloride ion) to insulin preparations is widely used. Hexamerization is driven by coordination of Zn 2+ at the His 610 sites to give T 6 , and the subsequent ligand-mediated transition of T 6 to T 3 R 3 and to Re is known to greatly enhance the physical and chemical stability of the resulting formulations.
  • allosteric ligands e.g. phenol and chloride ion
  • absorption rates vary between about 1 hour (for rapid-acting insulin analogues, such as Asp 828 human insulin) and about 4 hours (Co 3+ -hexamer).
  • Current Approaches Toward Slow Acting Insulins The inherent limitation of the absorption half-life to about 4 hours for a soluble human insulin hexamer necessitates further modifications to obtain the desired protraction.
  • this has been achieved by the use of preparations wherein the constituent insulin is in the form of a crystalline and/or amorphous precipitate. In this type of formulation, the dissolution of the precipitate in the subcutaneous depot becomes rate-limiting for the absorption.
  • NPH and Ultralente belong to this category of insulin preparations where crystallization/precipitation is effected by the addition of protamine and excessive zinc ion, respectively.
  • Another approach involves the use of insulin derivatives where the net charge is increased to shift the isoelectric point, and hence the pH of minimum solubility, from about 5.5 to the physiological range.
  • Such preparations may be injected as clear solutions at slightly acidic pH. The subsequent adjustment of the pH to neutral induces crystallization/precipitation in the subcutaneous depot and dissolution again becomes rate-limiting for the absorption.
  • Gly* 21 Arg 831 Arg 832 human insulin belongs to this category of insulin analogues.
  • soluble insulin derivatives with a hydrophobic moiety covalently attached to the side chain of Lys 829 have been synthesized. These derivatives may show prolonged action profile due to various mechanisms including albumin binding (e.g. B29-N ⁇ - myristoyl-des(B30) human insulin), extensive protein self-association and/or stickiness (e.g. B29-N ⁇ -(N-lithocholyl- ⁇ -glutamyl)-des(B30) human insulin) induced by the attached hydro- phobic group.
  • albumin binding e.g. B29-N ⁇ - myristoyl-des(B30) human insulin
  • extensive protein self-association and/or stickiness e.g. B29-N ⁇ -(N-lithocholyl- ⁇ -glutamyl)-des(B30) human insulin
  • the present invention provides a pharmaceutical preparation comprising ligands for the His 610 Zn 2+ sites of the R-state insulin hexamer, zinc ions and acid-stabilised insulin analogs.
  • the preparations form clear solutions at slightly acidic pH. When the pH is adjusted towards neutral upon subcutaneous injection, the ligands work to stabilize hexamers and modify solubility in the neutral pH range. As a result, the preparations release insulin slowly following subcutaneous injection.
  • the invention furthermore provides a method of preparing ligands for the His 810 Zn 2+ sites of the R-state insulin hexamer comprising the steps of
  • the invention also provides a method of prolonging the action of an acid-stabilised insulin preparation which comprises adding a zinc-binding ligand of the invention to the acid- stabilised insulin preparation.
  • the invention finally provides a method of treating type 1 or type 2 diabetes comprising administering to a patient in need thereof a theraputically effective amount of a pharmaceutical preparation of the invention.
  • Halogen designates an atom selected from the group consisting of F, Cl, Br and I.
  • CrCe-alkyl as used herein represents a saturated, branched or straight hydrocarbon group having from 1 to 6 carbon atoms. Representative examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, butyl, isobutyl, sec-butyl, tetf-butyl, n-pentyl, isopentyl, neopentyl, ferf-pentyl, n-hexyl, isohexyl and the like.
  • CrCe-alkylene represents a saturated, branched or straight bivalent hydrocarbon group having from 1 to 6 carbon atoms. Representative examples include, but are not limited to, methylene, 1 ,2-ethylene, 1 ,3-propylene, 1 ,2-propylene, 1 ,4-butylene, 1 ,5- pentylene, 1 ,6-hexylene, and the like.
  • C 2 -C 6 -alkenyl represents a branched or straight hydrocarbon group having from 2 to 6 carbon atoms and at least one double bond.
  • groups include, but are not limited to, vinyl, 1-propenyl, 2-propenyl, iso-propenyl, 1,3-buta- dienyl, 1-butenyl, 2-butenyl, 3-butenyl, 2-methyl-1-propenyl, 1-pentenyl, 2-pentenyl, 3- pentenyl, 4-pentenyl, 3-methyl-2-butenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 2,4-hexadienyl, 5- hexenyl and the like.
  • C 2 -C 6 -alkynyl represents a branched or straight hydrocarbon group having from 2 to 6 carbon atoms and at least one triple bond.
  • groups include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, 1-pentynyl, 2-pentynyl, 3-pentynyl, 4-pentynyl, 1-hexynyl, 2-hexynyl, 3-hexynyl, 4- hexynyl, 5-hexynyl, 2,4-hexadiynyl and the like.
  • d-Ce-alkoxy refers to the radical -O-d-C ⁇ -alkyl, wherein d-Ce-alkyl is as defined above. Representative examples are methoxy, ethoxy, n-propoxy, isopropoxy, butoxy, sec-butoxy, terf-butoxy, pentoxy, isopentoxy, hexoxy, isohexoxy and the like.
  • C 3 -C 8 -cycloalkyl as used herein represents a saturated, carbocyclic group having from 3 to 8 carbon atoms.
  • Representative examples are cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl and the like.
  • the term as used herein represents a non-aromatic, carbocyclic group having from 4 to 8 carbon atoms containing one or two double bonds.
  • Representative examples are 1-cyclopentenyl, 2-cyclopentenyl, 3-cyclopentenyl, 1-cyclohexenyl, 2-cyclohexenyl, 3- cyclohexenyl, 2-cycloheptenyl, 3-cycloheptenyl, 2-cyclooctenyl, 1 ,4-cyclooctadienyl and the like.
  • heterocyclyl represents a non-aromatic 3 to 10 membered ring containing one or more heteroatoms selected from nitrogen, oxygen and sulphur and optionally containing one or two double bonds. Representative examples are pyrrolidinyl, piperidyl, piperazinyl, morpholinyl, thiomorpholinyl, aziridinyl, tetrahydrofuranyl and the like.
  • aryl as used herein is intended to include carbocyclic, aromatic ring systems such as 6 membered monocyclic and 9 to 14 membered bi- and tricyclic, carbocyclic, aromatic ring systems.
  • Aryl is also intended to include the partially hydrogenated derivatives of the ring systems enumerated above. Non-limiting examples of such partially hydrogenated derivatives are 1,2,3,4-tetrahydronaphthyl, 1,4- dihydronaphthyl and the like.
  • arylene as used herein is intended to include divalent, carbocyclic, aromatic ring systems such as 6 membered monocyclic and 9 to 14 membered bi- and tricyclic, divalent, carbocyclic, aromatic ring systems.
  • Arylene is also intended to include the partially hydrogenated derivatives of the ring systems enumerated above.
  • Non-limiting examples of such partially hydrogenated derivatives are 1,2,3,4-tetrahydronaphthylene, 1 ,4-dihydronaphthylene and the like.
  • aryloxy as used herein denotes a group -O-aryl, wherein aryl is as defined above.
  • aroyl denotes a group -C(O)-aryl, wherein aryl is as defined above.
  • heteroaryl as used herein is intended to include aromatic, heterocyclic ring systems containing one or more heteroatoms selected from nitrogen, oxygen and sulphur such as 5 to 7 membered monocyclic and 8 to 14 membered bi- and tricyclic aromatic, heterocyclic ring systems containing one or more heteroatoms selected from nitrogen, oxygen and sulphur.
  • Representative examples are furyl, thienyl, pyrrolyl, pyrazolyl, 3-oxopyrazolyl, oxazolyl, thiazolyl, imidazolyl, isoxazoiyl, isothiazolyl, 1 ,2,3-triazolyl, 1,2,4-triazolyl, pyranyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, 1 ,2,3-triazinyl, 1 ,2,4-triazinyl, 1,3,5- triazinyl, 1,2,3- oxadiazolyl, 1 ,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1 ,3,4-oxadiazolyl, 1 ,2,3-thiadiazolyl, 1,2,4- thiadiazolyl, 1 ,2,5-thiadiazolyl, 1 ,3,4-thiadiazolyl, t
  • Heteroaryl is also intended to include the partially hydrogenated derivatives of the ring systems enumerated above.
  • Non-limiting examples of such partially hydrogenated derivatives are 2,3-dihydrobenzofuranyl, pyrrolinyl, pyrazolinyl, indolinyl, oxazolidinyl, oxazolinyl, oxazepinyl and the like.
  • heteroarylene as used herein is intended to include divalent, aromatic, heterocyclic ring systems containing one or more heteroatoms selected from nitrogen, oxygen and sulphur such as 5 to 7 membered monocyclic and 8 to 14 membered bi- and tricyclic aromatic, heterocyclic ring systems containing one or more heteroatoms selected from nitrogen, oxy- gen and sulphur.
  • Heteroaryl is also in- tended to include the partially hydrogenated derivatives of the ring systems enumerated above.
  • Non-limiting examples of such partially hydrogenated derivatives are 2,3-dihydro- benzofuranylene, pyrrolinylene, pyrazolinylene, indolinylene, oxazolidinylene, oxazolinylene, oxazepinylene and the like.
  • ArG1 as used herein is intended to include an aryl or arylene radical as applicable, where aryl or arylene are as defined above but limited to phenyl, biphenylyl, naphthyl, anthra- cenyl, phenanthrenyl, fluorenyl, indenyl, and azulenyl as well as the corrresponding divalent radicals.
  • ArG2 as used herein is intended to include an aryl or arylene radical as applicable, where aryl or arylene are as defined above but limited to phenyl, biphenylyl, naphthyl, fluo- renyl, and indenyl, as well as the corrresponding divalent radicals.
  • Het1 as used herein is intended to include a heteroaryl or heteroarylene radical as applicable, where heteroaryl or heteroarylene are as defined above but limited to furyl, thienyl, pyrrolyl, pyrazolyl, 3-oxopyrazolyl, oxazolyl, thiazolyl, imidazolyl, isoxazoiyl, isothiazolyl, 1 ,2,3-triazolyl, 1,2,4-triazolyl, pyranyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, 1 ,2,3- triazinyl, 1,2,4-triazinyl, 1,3,5- triazinyl, 1,2,3-oxadiazolyl, 1 ,2,4-oxadiazolyl, 1 ,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,3-thiadiazolyl, 1 ,
  • Het2 as used herein is intended to include a heteroaryl or heteroarylene radical as applicable, where heteroaryl or heteroarylene are as defined above but limited to furyl, thienyl, pyrrolyl, pyrazolyl, 3-oxopyrazolyl, oxazolyl, thiazolyl, imidazolyl, isoxazoiyl, isothiazolyl, 1 ,2,3-triazolyl, 1 ,2,4-triazolyl, pyranyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, 1,2,3- triazinyl, 1,2,4-triazinyl, 1,3,5- triazinyl, 1,2,3-oxadiazolyl, 1 ,2,4-oxadiazolyl, 1 ,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,3-thiadiazolyl, 1,2,4
  • Het3 is intended to include a heteroaryl or heteroarylene radical as applicable, where heteroaryl or heteroarylene are as defined above but limited to furyl, thienyl, pyrrolyl, pyrazolyl, 3-oxopyrazolyl, oxazolyl, thiazolyl, imidazolyl, isoxazoiyl, isothiazolyl, 1 ,2,3-triazolyl, 1,2,4-triazolyl, pyridyl, tetrazolyl, indolyl, isoindolyl, benzofuryl, benzothienyl, benzimidazolyl, benzthiazolyl, benzisothiazolyl, benzoxazolyl, benzisoxazolyl, quinolyl, isoqui- nolyl, quinoxalinyl, carbazolyl, thiazolidinyl, 2-thiooxothiazolidinyl,
  • Aryl-d-Ce-alkyl is intended to mean d-Ce- alkyl or C 2 -C 6 -alkenyl as defined above, substituted by an aryl or heteroaryl as defined above, for example:
  • treatment means the management and care of a patient for the purpose of combating a disease, disorder or condition.
  • the term is intended to include the delaying of the progression of the disease, disorder or condition, the alleviation or relief of symptoms and complications, and/or the cure or elimination of the disease, disorder or condition.
  • the patient to be treated is preferably a mammal, in particular a human being.
  • fragment as used herein is intended to mean a bivalent chemical group
  • Neutral amino acid as used herein is intended to mean any natural (codable) and non-natural amino acid, including ⁇ - or ⁇ -aminocarboxylic acids, including D-isomers of these (when applicable) without charges at physiologically relevant pH in the side chain, such as glycine, alanine, ⁇ -alanine, valine, leucine, isoleucine, phenylalanine, tyrosine, aspargine, glutamine, cysteine, methionine, 3-aminobenzoic acid, 4-aminobenzoic acid or the like.
  • positively charged group as used herein is intended to mean any pharmaceutically acceptable group that contains a positive charge at physiologically relevant pH, such as amino (primary, secondary and tertiary), ammonium and guanidino groups.
  • a amino acid as used herein is intended to mean mean any natural (codable) and non-natural ⁇ -aminocarboxylic acid, including D-isomers of these.
  • ⁇ amino acid as used herein is intended to mean any ⁇ -aminocarboxylic acid, such as ⁇ -alanine, isoserine or the like.
  • desB30 as used herein is intended to mean meant a natural insulin B chain or an analogue thereof lacking the B30 amino acid residue.
  • the amino acid residues are indicated in the three letter amino acid code or the one letter amino code.
  • B1 is intended to mean the amino acid residue in position 1 in the B chain of insulin or analogue thereof (counted from the N-terminal end) and the amino acid residue in position 1 in the A chain of insulin or analogue thereof (counted from the N-terminal end), respectively.
  • groups of compounds such as car- boxylates, dithiocarboxylates, phenolates, thiophenolates, alkylthiolates, sulfonamides, irni- dazoles, triazoles, 4-cyano-1 ,2,3-triazoles, benzimidazoles, benzotriazoles, purines, thia- zolidinediones, tetrazoles, 5-mercaptotetrazoles, rhodanines, N-hydroxyazoles, hydantoines, thiohydantoines, naphthoic acids and salicylic acids, these groups of compounds are intended to include also derivatives of the compounds from which the groups take their name.
  • acid-stabilised insulin refers to an insulin analog that does not deamidate or dimerize at pH values below 7. Specifically, the analog cannot have Asn or Asp as a C-terminal residue.
  • human insulin refers to naturally produced insulin or recombinantly produced insulin.
  • Recombinant human insulin may be produced in any suitable host cell, for example the host cells may be bacterial, fungal (including yeast), insect, animal or plant cells.
  • the expression "insulin derivative” as used herein (and related expressions) refers to human insulin or an analogue thereof in which at least one organic substituent is bound to one or more of the amino acids.
  • analogue of human insulin as used herein (and related expressions) is meant human insulin in which one or more amino acids have been deleted and/or replaced by other amino acids, including non-codeable amino acids, or human insulin comprising additional amino acids, i.e. more than 51 amino acids, such that the resulting analogue possesses insulin activity.
  • phenolic compound or similar expressions as used herein refers to a chemical compound in which a hydroxyl group is bound directly to a benzene or substituted benzene ring. Examples of such compounds include, but are not limited to, phenol, o-cresol, m-cresol and p-cresol.
  • physiologically relevant pH as used herein is intended to mean a pH of about 7.1 to 7.9.
  • Fig. 1 The effect of various concentrations of the ligand 4-[3-(2H-tetrazol-5-yl)carbazol-9- ylmethyl]benzoyl-Arg 1 -NH 2 on the pH-dependence of Gly* 21 , Asp 828 insulin solubility.
  • Fig. 2 The effect of a high concentrations of the ligand 4-[3-(2H-tetrazol-5-yl)carbazol-9- ylmethyl]benzoyl-Argi 2 -NH 2 on the pH-dependence of Gly* 21 insulin solubility.
  • Fig. 3 Disappearance from the subcutaneous depot (pig model) of insulin preparations.
  • Curves a)-c) are Gly* 21 , Asp 828 human insulin formulated with an excess concentration compared to Zn 2+ of a) 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]butyryl- Arg 3 -NH 2 , b) 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]butyryl-Arg 5 -NH 2 and c) 4-(2H-Tetrazol-5-yl)benzoyl-Abz-Gly 2 -Arg 5 -NH 2 .
  • Curve d) is B29-N ⁇ -myristoyl- des(B30) human insulin.
  • Curves e) and f) are Gly* 21 human insulin formulated with two dif- ferent excess concentrations compared to Zn 2+ of 4-[3-(2H-Tetrazol-5-yl)carbazol-9- ylmethy I] benzoy l-Arg 1 -N H 2.
  • Fig.4 4H3N-assay. UV/vis spectra resulting from a titration of hexameric insulin with the compound 3-hydroxy-2-naphthoic acid in the presence of 4-hydroxy-3-nitrobenzoic acid (4H3N). Inserted in the upper right corner is the absorbance at 444nm vs. the concentration of ligand
  • Fig. 5 TZD-assay. Fluorescence spectra resulting from a titration of hexameric insulin with 5-(3-methoxybenzylidene)thiazolidine-2,4-dione in the presence of 5-(4-dimethylamino- benzylidene)thiazolidine-2,4-dione (TZD). Inserted in the upper right corner is the fluorescence at 460 nm vs. the concentration of ligand
  • the present invention is based on the discovery that the His 810 Zn ++ ligand binding sites of the R-state insulin hexamer can be used to obtain an insulin preparation having prolonged action designed for flexible injection regimes including once-daily, based on acid-stabillised insulins.
  • the basic concept underlying the present invention involves reversible attachment of a ligand to the His 810 Zn 2+ site of the R-state hexamer.
  • a suitable ligand binds to the hexamer metal site with one end while other moieties are covalently attachment to the other end.
  • prolonged action via modification of preparation solubility may be obtained in a number of ways.
  • the anions currently used in insulin formulations as allosteric ligands for the R-state hexamers bind only weakly to the His 810 anion site.
  • the present invention which is based on the discovery of suitable higher affinity ligands for these anion sites, provides ligands which are extended to modify timing via changes in hexamer solubility as out- lined above.
  • the His 810 Zn + site consists of a tunnel or cavity with a triangular-shaped cross-section that extends -12 A from the surface of the hexamer down to the His 810 Zn 2+ ion.
  • the diameter of the tunnel varies along its length and, depending on the nature of the ligand occupying the site, the opening can be capped over by the Asn 83 and Phe 81 side chains.
  • the walls of the tunnel are made up of the side chains of the amino acid residues along one face each of the three ⁇ -helices.
  • the side chains from each helix that make up the lining of the tunnel are Phe B1 , Asn 83 , and Leu 86 . Therefore, except for the zinc ion, which is coordinated to three His 810 residues and is positioned at the bottom of the tunnel, the site is principally hydrophobic.
  • substituents on the ligand it may be possible for substituents on the ligand to make H-bonding interactions with Asn 83 and with the peptide linkage to Cys 87 .
  • the present invention originates from a search for compounds with suitable binding proper- ties by using UV-visible and fluorescence based competition assays described herein which are based on the displacement of chromophoric ligands from the R-state His 810 -Zn 2+ site by the incoming ligand in question. These compounds will be referred to as "starter compounds" in the following. These assays are easily transformed into a high-throughput format capable of handling libraries constructed around hits from the initial search of compound databases.
  • this chemical group can be attached (optionally using a spacer group using and synthesis procedures known to those skilled in the art) to a position on the starter compound remote from the Zn 2+ -binding functionality.
  • the invention thus provides pharmaceutical preparation comprising
  • CGr is a chemical group which reversibly binds to a His 810 Zn 2+ site of an insulin hexamer;
  • Lnk is a linker selected from
  • Frg1 is a fragment consisting of 0 to 5 neutral ⁇ - or ⁇ -amino acids
  • Frg2 is a fragment comprising 1 to 20 positively charged groups independently selected from amino or guanidino groups.
  • X is -OH, -NH 2 or a diamino group
  • any optical isomer or mixture of optical isomers including a racemic mixture, or any tautomeric forms.
  • CGr is a chemical structure selected from the group consisting of car- boxylates, dithiocarboxylates, phenolates, thiophenolates, alkylthiolates, sulfonamides, imi- dazoles, triazoles, 4-cyano-1 ,2,3-triazoles, benzimidazoles, benzotriazoles, purines, thia- zolidinediones, tetrazoles, 5-mercaptotetrazoles, rhodanines, N-hydroxyazoles, hydantoines, thiohydantoines, barbiturates, naphthoic acids and salicylic acids.
  • CGr is a chemical structure selected from the group consisting of benzotriazoles, 3-hydroxy 2-napthoic acids, salicylic acids, tetrazoles, thiazolidinediones, 5- mercaptotetrazoles, or 4-cyano-1 ,2,3-triazoles. In another embodiment CGr is
  • R 1 and R 4 are independently selected from hydrogen or CrCe-alkyl
  • R 2 is hydrogen or d-C 6 -alkyl or aryl, R 1 and R 2 may optionally be combined to form a double bond,
  • R 3 and R 5 are independently selected from hydrogen, halogen, aryl, d-C ⁇ -alkyl, or -C(O)NR 11 R 12 ,
  • a and B are independently selected from d-C 6 -alkylene, arylene, aryl-d-C 6 -alkyl-, aryl-C - Ce-alkenyl- or heteroarylene, wherein the alkylene or alkenylene is optionally substituted with one or more substituents independently selected from R 6 and the arylene or heteroarylene is optionally substituted with up to four substituents R 7 , R 8 , R 9 , and R 10 ,
  • a and R 3 may be connected through one or two valence bonds
  • B and R 5 may be connected through one or two valence bonds
  • R 6 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , aryl, -COOH and -NH 2 ,
  • R 7 , R 8 , R 9 and R 10 are independently selected from
  • each cyclic moiety may optionally be substituted with one or more substituents independently selected from R 14 ,
  • R 11 and R 12 are independently selected from hydrogen, OH, C 1 -C 20 -alkyl, aryl-d-C 6 -alkyl or aryl, wherein the alkyl groups may optionally be substituted with one or more substituents independently selected from R 15 , and the aryl groups may optionally be substituted one or more substituents independently selected from R 16 ; R 11 and R 12 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
  • R 13 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -OR 11 , -C(O)OR 11 , -NR 1 R 12 , and -C(O)NR 11 R 12 ,
  • R 14 is independently selected from halogen, -C(O)OR 11 , -CH 2 C(O)OR 11 , -CH 2 OR 11 , -CN, - CF 3 , -OCF 3 , -NO 2 , -OR 11 , -NR 11 R 12 , S(O) 2 R 11 , aryl and C C 6 -alkyl,
  • R 15 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -Od-C 6 -alkyl, -C(O)OC Ce- alkyl, -COOH and -NH 2 ,
  • Crg is arylene optionally substituted with up to four substituents, R 7 , R 8 , R 9 , and R 10 which may be the same or different.
  • A is selected from ArG1 optionally substituted with up to four substituents, R 7 , R 8 , R 9 , and R 10 which may be the same or different.
  • A is phenylene or naphtylene optionally substituted with up to four substituents, R 7 , R 8 , R 9 , and R 10 which may be the same or different.
  • R 7 , R 8 , R 9 , and R 10 which may be the same or different.
  • A is phenylene
  • A is heteroarylene optionally substituted with up to four substituents
  • R 7 , R 8 , R 9 , and R 0 which may be the same or different.
  • A is selected from Het1 optionally substituted with up to four substituents, R 7 , R 8 , R 9 , and R 10 which may be the same or different.
  • A is selected from Het2 optionally substituted with up to four substituents, R 7 , R 8 , R 9 , and R 10 which may be the same or different.
  • A is selected from Het3 optionally substituted with up to four sub- stituents, R 7 , R 8 , R 9 , and R 10 which may be the same or different.
  • A is selected from the group consisting of indolylene, benzofu- ranylidene, quinolylene, furylene, thienylene, or pyrrolylene, wherein each heteroaryl may optionally substituted with up to four substituents, R 7 , R 8 , R 9 , and R 10 which may be the same or different.
  • A is benzofuranylene optionally substituted with up to four substituents R 7 , R 8 , R 9 , and R 10 which may be the same or different.
  • A is
  • A is carbazolylidene optionally substituted with up to four substituents R 7 , R 8 , R 9 , and R 10 which may be the same or different.
  • R 7 , R 8 , R 9 , and R 10 which may be the same or different.
  • A is quinolylidene optionally substituted with up to four substituents R 7 , R 8 , R 9 , and R 10 which may be the same or different. In another embodiment A is
  • A is indolylene optionally substituted with up to four substituents R 7 , R , R , and R -10 which may be the same or different.
  • R 7 , R , R , and R -10 which may be the same or different.
  • R 1 is hydrogen
  • R 2 is hydrogen
  • R 1 and R 2 are combined to form a double bond.
  • R 3 is d-Ce-alkyl, halogen, or C(O)NR 16 R 17 .
  • R 3 is d-C 6 -alkyl or C(O)NR 16 R 17 .
  • R 3 is methyl
  • B is phenylene optionally substituted with up to four substituents, R 7 ,
  • R 8 , R 9 , and R 10 which may be the same or different.
  • R 4 is hydrogen
  • R 5 is hydrogen
  • R 6 is aryl. In another embodiment R 6 is phenyl.
  • R 7 , R 8 , R 9 and R 10 are independently selected from
  • R 7 , R 8 , R 9 and R 10 are independently selected from
  • each of the cyclic moieties optionally may be substituted with one or more substituents independently selected from R 14
  • each of the cyclic moieties optionally may be substituted with one or more substituents independently selected from R 14 .
  • R 7 , R 8 , R 9 and R 10 are independently selected from
  • each of the cyclic moieties optionally may be substituted with one or more substituents independently selected from R 4 .
  • R 7 , R 8 , R 9 and R 10 are independently selected from • hydrogen, halogen, -OR 11 , -OCrC 6 -alkyl-C(O)OR 11 , or -C(O)OR 11 ,
  • R 11 and R 12 are independently selected from hydrogen, CrC 20 -alkyl, aryl or aryl-d-Ce-alkyl, wherein the alkyl groups may optionally be substituted with one or more substituents independently selected from R 15 , and the aryl groups may optionally be substituted one or more substituents independently selected from R 16 ; R 11 and R 12 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds.
  • R 1 and R 12 are independently selected from hydrogen, CrC 20 -alkyl, aryl or aryl-d-Ce-alkyl, wherein the alkyl groups may optionally be substituted with one or more substituents independently selected from R 15 , and the aryl groups may optionally be substituted one or more substituents independently selected from R 16 .
  • R 11 and R 12 are independently selected from phenyl or phenyl-CrC 6 - alkyl.
  • R 11 and R 12 are methyl.
  • R 13 is independently selected from halogen, CF 3 , OR 11 or NR 11 R 12 . In another embodiment R 13 is independently selected from halogen or OR 11 .
  • R 13 is OR 11 .
  • R 14 is independently selected from halogen, -C(O)OR 11 , -CN, -CF 3 , -
  • R 14 is independently selected from halogen, -C(O)OR 11 , or -OR 11 .
  • R 15 is independently selected from halogen, -CN, -CF 3 , -C(O)Od-C 6 - alkyl.and -COOH.
  • R 15 is independently selected from halogen or -C(O)OCrC 6 -alkyl.
  • R 16 is independently selected from halogen, -C(O)OCrC 6 -alkyl,
  • R 1 ⁇ is independently selected from halogen, -C(O)OCrC 6 -alkyl,
  • R 19 is hydrogen or d-C ⁇ -alkyl
  • R 20 is hydrogen or CrCe-alkyl
  • D and F are a valence bond or d-C 6 -alkylene optionally substituted with one or more substituents independently selected from R 72 ,
  • R 72 is independently selected from hydroxy, d-C ⁇ -alkyl, or aryl,
  • E is CrCe-alkylene, arylene or heteroarylene, wherein the arylene or heteroarylene is optionally substituted with up to three substituents R 21 , R 22 and R 23 ,
  • G is CrC 6 -alkylene, arylene or heteroarylene, wherein the arylene or heteroarylene is op- tionally substituted with up to three substituents R 24 , R 25 and R 26 ,
  • R 17 , R 18 , R 21 , R 22 , R 23 , R 24 , R 25 and R 26 are independently selected from
  • R 27 and R 28 are independently selected from hydrogen, d-Ce-alkyl, aryl-d-C 6 -alkyl or aryl, or R 27 and R 28 when attached to the same nitrogen atom together with the said nitrogen atom may form a 3 to 8 membered heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
  • R is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -OR 27 , and -NR 27 R 28 ,
  • R 30 is independently selected from halogen, -C(O)OR 27 , -CN, -CF 3 , -OCF 3 , -NO 2 , -OR 27 , -NR 27 R 28 and d-Ce-alkyl, or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base.
  • D is a valence bond
  • D is d-Ce-alkylene optionally substituted with one or more hydroxy, d-Ce-alkyl, or aryl.
  • E is arylene or heteroarylene, wherein the arylene or heteroarylene is optionally substituted with up to three substituents independently selected from R 21 , R 22 and
  • E is arylene optionally substituted with up to three substituents independently selected from R 21 , R 22 and R 23 .
  • E is selected from ArG1 and optionally substituted with up to three substituents independently selected from R 21 , R 22 and R 23 .
  • E is phenylene optionally substituted with up to three substituents independently selected from R 21 , R 22 and R 23 .
  • R , R and R are independently selected from
  • R 2 , R 22 and R 23 are independently selected from
  • the cyclic moieties optionally may be substituted with one or more substituents selected from R 30 .
  • R 21 , R 22 and R 23 are independently selected from
  • R 21 , R 22 and R 23 are independently selected from
  • R 19 is hydrogen or methyl.
  • R 19 is hydrogen.
  • R 27 is Hydrogen, d-C 6 -alkyl or aryl.
  • R 27 is hydrogen or d-C ⁇ -alkyl.
  • R 28 is hydrogen or d-C ⁇ -alkyl.
  • F is a valence bond.
  • F is d-C 6 -alkylene optionally substituted with one or more hydroxy, d-C ⁇ -alkyl, or aryl.
  • G is d-C e -alkylene or arylene, wherein the arylene is optionally substituted with up to three substituents R 24 , R 25 and R 26 .
  • G is CrCe-alkylene or ArG1 , wherein the arylene is optionally substituted with up to three substituents R 24 , R 25 and R 26 .
  • G is d-C ⁇ -alkylene.
  • G is phenylene optionally substituted with up to three substituents R 24 , R 25 and R 26 .
  • R 24 , R 25 and R 26 are independently selected from
  • R 29 • aryl, aryloxy, aryloxycarbonyl, aroyl, aryl-d-C ⁇ -alkoxy, aryl-CrC 6 -alkyl, a ⁇
  • the cyclic moieties optionally may be substituted with one or more substituents selected from R 30 .
  • R 24 , R 25 and R 26 are independently selected from
  • R 24 , R 25 and R 26 are independently selected from
  • the cyclic moieties optionally may be substituted with one or more substituents selected from R 30 .
  • R 24 , R 25 and R 26 are independently selected from
  • R 24 , R 25 and R 26 are independently selected from
  • R 24 , R 25 and R 26 are independently selected from
  • R 20 is hydrogen or methyl. In another embodiment R 20 is hydrogen.
  • R 27 is hydrogen, CrC 6 -alkyl or aryl.
  • R 27 is hydrogen or d-Ce-alkyl or ArG1.
  • R 27 is hydrogen or d-Ce-alkyl.
  • R 28 is hydrogen or d-C 6 -alkyl.
  • R 17 and R 18 are independently selected from
  • R 17 and R 18 are independently selected from • hydrogen, halogen, -CN, -CF 3 , -N ⁇ 2 , -OR 27 , -NR 27 R 28 , or -C(O)OR 27 ,
  • R 17 and R 18 are independently selected from
  • R 17 and R 18 are independently selected from
  • R 17 and R 18 are independently selected from
  • R 27 is hydrogen or d-C ⁇ -alkyl.
  • R 27 is hydrogen, methyl or ethyl.
  • R 28 is hydrogen or d-C 6 -alkyl.
  • R 28 is hydrogen, methyl or ethyl.
  • R 72 is -OH or phenyl.
  • CGr is
  • I is selected from »a valence bond
  • Z 1 is S(O) 2 or CH 2
  • Z 2 is -NH-, -O-or -S-
  • n is 1 or 2
  • R 31 is independently selected from hydrogen, halogen, -CN, -CH 2 CN, -CHF 2 , -CF 3 , -OCF 3 , -OCHF 2 , -OCH 2 CF 3 , -OCF 2 CHF 2 , -S(O) 2 CF 3
  • R 32 and R 33 are independently selected from hydrogen, d-C 6 -alkyl or d-Ce-alkanoyl,
  • R ⁇ is independently selected from halogen, -CN, -CF 3 , -OCF 3l -OR 35 , and -NR 35 R 36 ,
  • R 35 and R 36 are independently selected from hydrogen, d-Ce-alkyl, aryl-d-C 6 -alkyl or aryl, or R 35 and R 36 when attached to the same nitrogen atom together with the said nitrogen atom may form a 3 to 8 membered heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
  • R 37 is independently selected from halogen, -C(O)OR 35 , -C(O)H, -CN, -CF 3 , -OCF 3l -NO 2 , - OR 35 , -NR 35 R 36 , d-Ce-alkyl or d-Ce-alkanoyl,
  • I is a valence bond, -CH 2 N(R 32 )-, or -SO 2 N(R 33 )-. In another embodiment I is a valence bond. In another embodiment J is
  • R 32 and R 33 are independently selected from hydrogen or d-C 6 -alkyl.
  • R 34 is hydrogen, halogen, -CN, -CF 3 , -OCF 3 , -SCF 3 , -NO 2 , -OR 35 ,
  • R 34 is hydrogen, halogen, -CF 3 , -NO 2 , -OR 35 , -NR ⁇ R 36 , -SR 35 ,
  • R 34 is hydrogen, halogen, -CF 3 , -NO 2 , -OR 35 , -NR ⁇ R 36 , or
  • R 34 is hydrogen, halogen, or -OR 35 .
  • R 35 and R 36 are independently selected from hydrogen, d-C ⁇ -alkyl, or aryl.
  • R 35 and R 36 are independently selected from hydrogen or d-C ⁇ alkyl.
  • R 37 is halogen, -C(O)OR 35 , -CN, -CF 3 , -OR 35 , -NR 35 R 36 , C C ⁇ -alkyl or CrC ⁇ -alkanoyl.
  • R 37 is halogen, -C(O)OR 35 , -OR 35 , -NR 35 R 38 , d-C 6 -alkyl or d-Ce- alkanoyl.
  • R 37 is halogen, -C(O)OR 35 or -OR 35 .
  • CGr is
  • U is a valence bond, d-C 6 -alkenylene, -d-C 6 -alkyl-O- or d-C 6 -alkylene wherein any d- Ce-alkyl moiety is optionally substituted with d-Ce-alkyl,
  • R 38 is CrCe-alkyl, aryl, wherein the alkyl or aryl moieties are optionally substituted with one or more substituents independently selected from R 39 ,
  • R 39 is independently selected from halogen, cyano, nitro, amino,
  • M is a valence bond, arylene or heteroarylene, wherein the aryl or heteroaryl moieties are optionally substituted with one or more substituents independently selected from R 40 ,
  • R 40 is selected from
  • R 41 and R 42 are independently selected from hydrogen, -OH, d-Ce-alkyl, d-Ce-alkenyl, aryl- d-C ⁇ -alkyl or aryl, wherein the alkyl moieties may optionally be substituted with one or more substituents independently selected from R 45 , and the aryl moieties may optionally be substituted with one or more substituents independently selected from R 46 ; R 41 and R 42 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
  • R 43 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -OR 41 , and -NR 41 R 42
  • R 44 is independently selected from halogen, -C(O)OR 41 , -CH 2 C(O)OR 41 , -CH 2 OR 41 , -CN, -
  • R 45 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -O-d-C ⁇ -alkyl, -C(O)-O-d-
  • R 49 is independently selected from halogen and -COOH
  • any alkylene, alkenylene , alkynylene, arylene and heteroarylene moiety is optionally substituted with one or more substituents independently selected from R 50 ,
  • R 51 and R 52 are independently selected from hydrogen and d-C ⁇ -alkyl
  • R 53 is independently selected from C C ⁇ -alkyl, d-C 6 -alkoxy, -CrC 6 -alkyl-COOH, -C 2 - C 6 -alkenyl-COOH, -OR 51 , -NO 2 , halogen, -COOH, -CF 3 , -CN, or -N(R 51 R 52 ),
  • CrCe-alkyl moiety is optionally substituted with R 38 .
  • K is a valence bond or d-Ce-alkylene, wherein any d-C 6 -alkyl moiety is optionally substituted with R 38 .
  • K is a valence bond
  • U is a valence bond or -d-C ⁇ -alkyl-O-.
  • M is arylene or heteroarylene, wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from
  • M is ArG1 or Het1 , wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from R 40 .
  • M is ArG1 or Het2, wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from R 40 .
  • M is ArG1 or Het3, wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from R 40 .
  • M is phenylene optionally substituted with one or more substituents independently selected from R 40 .
  • M is indolylene optionally substituted with one or more substituents independently selected from R 40 .
  • M is carbazolylene optionally substituted with one or more substitu- ents independently selected from R 40 .
  • M is
  • R 40 is selected from
  • R 40 is selected from
  • R 40 is selected from
  • R 41 and R 42 are independently selected from hydrogen, d-Ce-alkyl, or aryl, wherein the aryl moieties may optionally be substituted with halogen or -COOH.
  • R 41 and R 42 are independently selected from hydrogen, methyl, ethyl, or phenyl, wherein the phenyl moieties may optionally be substituted with halogen or - COOH.
  • R 47 and R 48 are independently selected from hydrogen, methyl and phenyl.
  • T is
  • R 50 is d-C ⁇ -alkyl, d-Ce-alkoxy, aryl, aryloxy, aryl-d-C 6 -alkoxy , -OR 51 , -N0 2 , halogen, -COOH, -CF 3 , wherein any aryl moiety is optionally substituted with one or more R 53 .
  • R 50 is d-Ce-alkyl, aryloxy, aryl-d-C 6 -alkoxy , -OR 51 , halogen, -COOH, -CF 3 , wherein any aryl moiety is optionally substituted with one or more R 53 .
  • R 50 is d-Ce-alkyl, ArG1-O-, ArG1-d-C 6 -alkoxy , -OR 51 , halogen, -COOH, -CF 3 , wherein any aryl moiety is optionally substituted with one or more R 53 .
  • R 50 is phenyl, methyl or ethyl.
  • R 50 is methyl or ethyl.
  • R 51 is methyl.
  • R 53 is d-Ce-alkyl, d-C 6 -alkoxy, -OR 51 , halogen.or -CF 3 .
  • V is Ci-C 6 -alkylene, arylene, heteroarylene, arylene-d-e-alkylene or arylene-C 2 - 6 - alkenylene, wherein the alkylene or alkenylene is optionally substituted with one or more substituents independently selected from R 54 , and the arylene or heteroarylene is optionally substituted with one or more substituents independently selected from R 55 ,
  • R 54 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , aryl, -COOH and -NH 2
  • R 55 is independently selected from
  • R 58 which may optionally be substituted with one or more substituents selected from R 58 , • aryl, aryloxy, aryloxycarbonyl, aroyl, arylsulfanyl, aryl-d-C ⁇ -alkoxy, aryl-d-C ⁇ -alkyl, aryl-C 2 -C 6 -alkenyl, aroyl-C 2 -C 6 -alkenyl, aryl-C 2 -C 6 -alkynyl, heteroaryl, heteroaryl-d- C ⁇ -alkyl, heteroaryl-C 2 -C 6 -alkenyl or heteroaryl-C 2 -C 6 -alkynyl,
  • R 58 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -OR 56 , and -NR 56 R 57 ,
  • R 59 is independently selected from halogen, -C(0)OR 56 , -CH 2 C(0)OR 56 , -CH 2 OR 56 , -CN, - CF 3 , -OCF 3 , -N0 2 , -OR 56 , -NR 56 R 57 and d-C ⁇ -alkyl,
  • R 62 is d-C ⁇ -alkyl, aryl optionally substituted with one or more substituents independently selected from halogen, or heteroaryl optionally substituted with one or more d-C 6 -alkyl jnde- pendently, or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base.
  • V is arylene, heteroarylene, or arylene-d.C ⁇ -alkylene, wherein the alkylene is optionally substituted with one or more substituents independently selected R 54 , and the arylene or heteroarylene is optionally substituted with one or more substituents independently selected from R 55 .
  • V is arylene, Het1, or arylene-d-C ⁇ -alkylene, wherein the alkylene is optionally substituted with one or more substituents independently selected from R 54 , and the arylene or heteroarylene moiety is optionally substituted with one or more substituents independently selected from R 55 .
  • V is arylene, Het2, or arylene-d.C 6 -alkylene, wherein the alkylene is optionally substituted with one or more substituents independently selected from R 54 , and the arylene or heteroarylene moiety is optionally substituted with one or more substituents inde- pendently selected from R 55 .
  • V is arylene, Het3, or arylene-d.C 6 -alkylene, wherein the alkylene is optionally substituted with one or more substituents independently selected from R 54 , and the arylene or heteroarylene moiety is optionally substituted with one or more substituents independently selected from R 55 .
  • V is arylene optionally substituted with one or more substituents independently selected from R 55 .
  • V is ArG1 optionally substituted with one or more substituents independently selected from R 55 .
  • V is phenylene, naphthylene or anthranylene optionally substituted with one or more substituents independently selected from R 55 .
  • V is phenylene optionally substituted with one or more substituents independently selected from R 55 .
  • R 55 is independently selected from
  • R 55 is independently selected from
  • R 55 is independently selected from halogen, -OR 56 , -NR 56 R 57 ,
  • R 55 is independently selected from halogen, -OR 56 , -NR 56 R 57 ,
  • R 56 and R 57 are independently selected from hydrogen or d-Ci 2 -alkyl, R 56 and R 57 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom.
  • R 56 and R 57 are independently selected from hydrogen or methyl, ethyl, propyl butyl, R 56 and R 57 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom.
  • AA is d-C ⁇ -alkylene, arylene, heteroarylene, arylene-Ci-C ⁇ -alkylene or arylene-C 2 .
  • C ⁇ -alkenylene wherein the alkylene or alkenylene is optionally substituted with one or more substituents independently selected from R 63 , and the arylene or heteroarylene is optionally substituted with one or more substituents independently selected from R 64 ,
  • R 63 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , aryl, -COOH and -NH 2 ,
  • R 64 is independently selected from
  • R 67 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -OR 65 , and -NR 65 R 66 ,
  • R 68 is independently selected from halogen, -C(0)OR 65 , -CH 2 C(0)OR 65 , -CH 2 OR 65 , -CN, - CF 3 , -OCF 3 , -N0 2 , -OR 65 , -NR 65 R 66 and CrC 6 -alkyl,
  • R 69 is independently selected from d-Ce-alkyl, aryl optionally substituted with one or more halogen, or heteroaryl optionally substituted with one or more d-C ⁇ -alkyl
  • R 70 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -Od-Ce-alkyl, -C(0)OCrC 6 - alkyl, -COOH and -NH 2 ,
  • AA is arylene, heteroarylene or arylene-d-Ce-alkylene, wherein the alkylene is optionally substituted with one or more R 63 , and the arylene or heteroarylene is optionally substituted with one or more substituents independently selected from R 64 .
  • AA is arylene or heteroarylene, wherein the arylene or heteroarylene is optionally substituted with one or more substituents independently selected from R 64 .
  • AA is ArG1 or Het1 optionally substituted with one or more substitu- ents independently selected from R 64 .
  • AA is ArG1 or Het2 optionally substituted with one or more substituents independently selected from R 64 .
  • AA is ArG1 or Het3 optionally substituted with one or more substituents independently selected from R 64 .
  • AA is phenylene, naphtylene, anthrylene, carbazolylene, thienylene, pyridylene, or benzodioxylene optionally substituted with one or more substituents independently selected from R 64 .
  • AA is phenylene or naphtylene optionally substituted with one or more substituents independently selected from R 64 .
  • R 64 is independently selected from hydrogen, halogen, -CF 3 , -OCF 3 , -OR 65 , -NR 65 R 66 , d-C 6 -alkyl, -OC(0)R 65 , -Od-C 6 -alkyl-C(0)OR 65 , aryl-C 2 -C 6 -alkenyl, aryloxy or aryl, wherein d-C 6 -alkyl is optionally substituted with one or more substituents independently selected from R 67 , and the cyclic moieties optionally are substituted with one or more substituents independently selected from R 68 .
  • R 64 is independently selected from halogen, -CF 3 , -OCF 3 , -OR 65 , -NR 65 R 66 , methyl, ethyl, propyl, -OC(O)R 65 , -OCH 2 -C(0)OR 65 , -OCH 2 -CH 2 -C(0)OR 65 , phenoxy optionally substituted with one or more substituents independently selected from R 68 .
  • R 65 and R 66 are independently selected from hydrogen, CF 3 , d-C 12 -alkyl, aryl, or heteroaryl optionally substituted with one or more substituents independently selected from R 71 .
  • R 85 and R 66 are independently hydrogen, d-C 12 -alkyl, aryl, or het- eroaryl optionally substituted with one or more substituents independently selected from R 71 .
  • R 65 and R 66 are independently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, ArG1 or Het1 optionally substituted with one or more substituents independently selected from R 71 .
  • R 65 and R 66 are independently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, ArG1 or Het2 optionally substituted with one or more substituents independently selected from R 71 .
  • R 65 and R 66 are independently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, ArG1 or Het3 optionally substituted with one or more substituents independently selected from R 71 .
  • R 65 and R 66 are independently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, phenyl, naphtyl, thiadiazolyl optionally substituted with one or more R 71 independently; or isoxazoiyl optionally substituted with one or more substituents independently selected from R 71 .
  • R 71 is halogen or d-C 6 -alkyl.
  • R 71 is halogen or methyl.
  • Frg1 consists of 0 to 5 neutral amino acids independently selected from the group consisting of Gly, Ala, Thr, and Ser. In another embodiment Frg1 consists of 0 to 5 Gly. In another embodiment Frg1 consists of 0 Gly. In another embodiment Frg1 consists of 1 Gly. In another embodiment Frg1 consists of 2 Gly. In another embodiment Frg1 consists of 3 Gly. In another embodiment Frg 1 consists of 4 Gly. In another embodiment Frg1 consists of 5 Gly. In another embodiment G B is of the formula B 1 -B 2 -C(0)-, B 1 -B 2 -S0 2 - or B 1 -B 2 -CH 2 -, wherein B 1 and B 2 are as defined in claim 1.
  • G 8 is of the formula B 1 -B 2 -C(0)-, B 1 -B 2 -S0 2 - or B 1 -B 2 -NH-, wherein B 1 and B 2 are as defined in claim 1.
  • G B is of the formula B 1 -B 2 -C(0)-, B 1 -B 2 -CH 2 - or B 1 -B 2 -NH-, wherein B 1 and B 2 are as defined in claim 1.
  • G 8 is of the formula B 1 -B 2 -CH 2 -, B 1 -B 2 -S ⁇ 2 - or B 1 -B 2 -NH-, wherein B 1 and B 2 are as defined in claim 1.
  • G 8 is of the formula B 1 -B 2 -C(0)- or B 1 -B 2 -S0 2 -, wherein B 1 and B 2 are as defined in claim 1.
  • G 8 is of the formula B 1 -B 2 -C(0)- or B 1 -B 2 -CH 2 -, wherein B 1 and B 2 are as defined in claim 1.
  • G B is of the formula B 1 -B 2 -C(0)- or B 1 -B 2 -NH-, wherein B 1 and B 2 are as defined in claim 1.
  • G 8 is of the formula B 1 -B 2 -CH 2 - or B 1 -B 2 -S0 2 - , wherein B 1 and B 2 are as defined in claim 1.
  • G 8 is of the formula B 1 -B 2 -NH- or B 1 -B 2 -S0 2 - , wherein B 1 and B 2 are as defined in claim 1.
  • G 8 is of the formula B 1 -B 2 -CH 2 - or B -B 2 -NH- , wherein B 1 and B 2 are as defined in claim 1.
  • G 8 is of the formula B 1 -B 2 -C(0)-.
  • G 8 is of the formula B 1 -B 2 -CH 2 -.
  • G 8 is of the formula B 1 -B 2 -S0 2 -.
  • G B is of the formula B 1 -B -NH-.
  • B 1 is a valence bond, -0-, or -S-. In another embodiment B 1 is a valence bond, -0-, or -N(R 6 )-. In another embodiment B 1 is a valence bond, -S-, or -N(R 6 )-. In another embodiment B 1 is -O-, -S- or -N(R 6 )-. In another embodiment B 1 is a valence bond or -O-. In another embodiment B 1 is a valence bond or -S-. In another embodiment B 1 is a valence bond or -N(R 6 )-. In another embodiment B 1 is -O-or -S-.
  • B 1 is -0-or -N(R 6 )-. In another embodiment B 1 is -S-or -N(R 6 )-. In another embodiment B 1 is a valence bond. In another embodiment B 1 is -0-. In another embodiment B 1 is -S-. In another embodiment B 1 is -N(R 6 )-.
  • B z is a valence bond, d-C 18 -alkylene, arylene, heteroarylene, -C
  • B 2 is a valence bond, d-da-alkylene, arylene, heteroarylene, -d- C 18 -alkyl-aryl-, and the alkylene and arylene moieties are optionally substituted as defined in claim 1.
  • B 2 is a valence bond, C C 18 -alkylene, arylene, -CrC ⁇ 8 -alkyl-aryl-, and the alkylene and arylene moieties are optionally substituted as defined in claim 1.
  • B 2 is a valence bond or -d-C 18 -alkylene, and the alkylene moieties are optionally substituted as defined in claim 1.
  • Frg2 comprises 1 to 16 positively charged groups.
  • Frg2 comprises 1 to 12 positively charged groups.
  • Frg2 comprises 1 to 10 positively charged groups.
  • Frg2 is a fragment containing basic amino acids independently selected from the group consisting of Lys and Arg and D-isomers of these.
  • the basic amino acid is Arg.
  • X is -OH or -NH 2 .
  • X is -NH 2 .
  • the pharmaceutical preparation further comprises at least 3 phenolic molecules.
  • the acid-stabilised insulin is selected from the group consisting of A21G
  • A21G, B28K, B29P A21G, B28D A21G, B28E A21G, B3K, B29E A21G, desB27
  • zinc ions are present in an amount corresponding to 10 to 40 ⁇ g
  • zinc ions are present in an amount corresponding to 10 to 26 ⁇ g
  • the ratio between insulin and the zinc-binding ligand of the invention is in the range from 99:1 to 1 :99.
  • the ratio between insulin and the zinc-binding ligand of the invention is in the range from 95:5 to 5:95
  • the ratio between between insulin and the zinc-binding ligand of the invention is in the range from 80:20 to 20:80 In another embodiment the ratio between between insulin and the zinc-binding ligand of the invention is in the range from 70:30 to 30:70
  • the invention relates to a method of preparing a zinc-binding ligand of the invention comprising the steps of
  • the invention in another aspect relates to a method of prolonging the action of an acid- stabilised insulin preparation which comprises adding a zinc-binding ligand of the invention to the acid-stabilised insulin preparation.
  • the invention in another aspect relates to a method of treating type 1 or type 2 diabetes comprising administering to a patient in need thereof a theraputically effective amount of a pharmaceutical preparation comprising 1. Acid-stabilised insulin 2. Zinc ions
  • an embodiment 1 which is a pharmaceutical prepa- ration comprising 1.
  • CGr is a chemical group which reversibly binds to a His 810 Zn 2+ site of an insulin hexamer
  • Lnk is a linker selected from
  • Frg1 is a fragment consisting of 0 to 5 neutral ⁇ - or ⁇ -amino acids
  • Frg2 is a fragment comprising 1 to 20 positively charged groups independently selected from amino or guanidino groups.
  • X is -OH, -NH 2 or a diamino group
  • Embodiment 2 A pharmaceutical preparation according to embodiment 1 wherein CGr is a chemical structure selected from the group consisting of carboxylates, dithiocarboxylates, .
  • Embodiment 3 A pharmaceutical preparation according to embodiment 2 wherein CGr is a chemical structure selected from the group consisting of benzotriazoles, 3-hydroxy 2- napthoic acids, salicylic acids, tetrazoles, thiazolidinediones, 5-mercaptotetrazoles, or 4- cyano-1 ,2,3-triazoles.
  • Embodiment 4 A pharmaceutical composition according to any one of the embodiments 1 to 3 wherein CGr is
  • R 1 , R 1 * and R 4 are independently selected from hydrogen or d-Ce-alkyl, R 2 and R 2 * are hydrogen or d-C 6 -alkyl or aryl, R 1 and R 2 may optionally be combined to form a double bond, R 1 * and R 2 * may optionally be combined to form a double bond,
  • R 3 , R 3 * and R 5 are independently selected from hydrogen, halogen, aryl optionally substituted with one or more substituents independently selected from R 16 , d-C 6 -alkyl, or -C(0)NR 11 R 12 ,
  • A, A 1 and B are independently selected from d-Ce-alkyl, aryl, aryl-d-Ce-alkyl, -NR 11 -aryl, aryl-C 2 -C 6 -alkenyl or heteroaryl, wherein the alkyl or alkenyl is optionally substituted with one or more substituents independently selected from R 6 and the aryl or heteroaryl is optionally substituted with up to four substituents R 7 , R 8 , R 9 , and R 10 , A and R 3 may be connected through one or two valence bonds, B and R 5 may be connected through one or two valence bonds,
  • R 6 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , aryl, -COOH and -NH 2
  • R 7 , R 8 , R 9 and R 10 are independently selected from
  • each cyclic moiety may optionally be substituted with one or more substituents independently selected from R 14 ,
  • R 11 and R 12 are independently selected from hydrogen, OH, CrC 20 -alkyl, aryl-d-Ce-alkyl or aryl, wherein the alkyl groups may optionally be substituted with one or more substituents independently selected from R 15 , and the aryl groups may optionally be substituted one or more substituents independently selected from R 16 ; R 11 and R 12 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
  • R 13 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -OR 11 , -C(0)OR 11 , -NR 11 R 12 , and -C(0)NR 11 R 12 ,
  • Embodiment 8. A pharmaceutical composition according to any one of the embodiment s 4 to 7 wherein Y is -O- or -S-.
  • Embodiment 9. A pharmaceutical composition according to embodiment 8 wherein Y is -O-.
  • Embodiment 10. A pharmaceutical composition according to embodiment 8 wherein Y is -NH-.
  • Embodiment 11 A pharmaceutical composition according to embodiment 8 wherein Y is -S-.
  • Embodiment 12 A pharmaceutical composition according to any one of the embodiment s 4 to 11 wherein A is aryl optionally substituted with up to four substituents, R 7 , R 8 , R 9 , and R 10 which may be the same or different.
  • Embodiment 13 A pharmaceutical composition according to embodiment 12 wherein A is selected from ArG1 optionally substituted with up to four substituents, R 7 , R 8 , R 9 , and R 10 which may be the same or different.
  • Embodiment 14 A pharmaceutical composition according to embodiment 13 wherein A is phenyl or naphtyl optionally substituted with up to four substituents, R 7 , R 8 , R 9 , and R 10 which may be the same or different.
  • Embodiment 15 A pharmaceutical composition according to embodiment 14 wherein A is
  • Embodiment 17 A pharmaceutical composition according to any one of the embodiment s 4 to 11 wherein A is heteroaryl optionally substituted with up to four substituents, R 7 , R a , R 9 , and R 10 which may be the same or different.
  • Embodiment 18 A pharmaceutical composition according to embodiment 17 wherein A is selected from Het1 optionally substituted with up to four substituents, R 7 , R 8 , R 9 , and R 10 which may be the same or different.
  • Embodiment 19 A pharmaceutical composition according to embodiment 18 wherein A is selected from Het2 optionally substituted with up to four substituents, R 7 , R 8 , R 9 , and R 10 which may be the same or different.
  • Embodiment 20 A pharmaceutical composition according to embodiment 19 wherein A is selected from Het3 optionally substituted with up to four substituents, R 7 , R 8 , R 9 , and R 10 which may be the same or different.
  • Embodiment 21 A pharmaceutical composition according to embodiment 20 wherein A is selected from the group consisting of indolyl, benzofuranyl, quinolyl, furyl, thienyl, or pyrrolyl, wherein each heteroaryl may optionally substituted with up to four substituents, R 7 , R 8 , R 9 , and R 10 which may be the same or different.
  • Embodiment 22 A pharmaceutical composition according to embodiment 20 wherein A is benzofuranyl optionally substituted with up to four substituents R 7 , R 8 , R 9 , and R 10 which may be the same or different.
  • Embodiment 23 A pharmaceutical composition according to embodiment 22 wherein A is
  • Embodiment 26 A pharmaceutical composition according to embodiment 20 wherein A is quinolyl optionally substituted with up to four substituents R 7 , R 8 , R 9 , and R 10 which may be the same or different.
  • Embodiment 27 A pharmaceutical composition according to embodiment 26 wherein A is
  • Embodiment 28 A pharmaceutical composition according to embodiment 20 wherein A is indolyl optionally substituted with up to four substituents R 7 , R 8 , R 9 , and R 10 which may be the same or different.
  • Embodiment 29 A pharmaceutical composition according to embodiment 28 wherein A is
  • Embodiment 30 A pharmaceutical composition according to any one of the embodiment s 4 to 29 wherein R 1 is hydrogen.
  • Embodiment 31 A pharmaceutical composition according to any one of the embodiment s 4 to 30 wherein R 2 is hydrogen.
  • Embodiment 32. A pharmaceutical composition according to any one of the embodiment s 4 to 29 wherein R 1 and R 2 are combined to form a double bond.
  • Embodiment 33 A pharmaceutical composition according to any one of the embodiment s 4 to 32 wherein R 3 is d-Ce-alkyl, halogen, or C(0)NR 16 R 17 .
  • Embodiment 34 A pharmaceutical composition according to embodiment 33 wherein R 3 is d-Ce-alkyl or C(0)NR 16 R 17 .
  • Embodiment 35 A pharmaceutical composition according to embodiment 34 wherein R 3 is methyl.
  • Embodiment 36 A pharmaceutical composition according to any one of the embodiment s 4 to 11 wherein B is phenyl optionally substituted with up to four substituents, R 7 , R 8 , R 9 , and R 10 which may be the same or different.
  • Embodiment 37 A pharmaceutical composition according to any one of the embodiment s 4 to 11 or 36 wherein R 4 is hydrogen.
  • Embodiment 38. A pharmaceutical composition according to any one of the embodiment s 4 to 11 or 36 to 37 wherein R 5 is hydrogen.
  • Embodiment 39 A pharmaceutical composition according to any one of the embodiment s 4 to 38 wherein R 6 is aryl.
  • Embodiment 40. A pharmaceutical composition according to embodiment 39 wherein R 6 is phenyl.
  • Embodiment 41 A pharmaceutical composition according to any one of the embodiment s 4 to 40 wherein R 7 , R 8 , R 9 and R 10 are independently selected from
  • Embodiment 43 A pharmaceutical composition according to embodiment 42 wherein R ? , R 8 , R 9 and R 10 are independently selected from
  • Embodiment 44 A pharmaceutical composition according to embodiment 43 wherein R 7 , R 8 , R 9 and R 10 are independently selected from
  • Embodiment 45 A pharmaceutical composition according to embodiment 44 wherein R 7 , R 8 , R 9 and R 10 are independently selected from • hydrogen, halogen, -OR 11 , -Od-Ce-alkyl-C(0)OR 11 , or -C(0)OR 11 ,
  • Embodiment 46 A pharmaceutical composition according to embodiment 45 wherein R 7 , R 8 , R 9 and R 10 are independently selected from
  • Embodiment 47 A pharmaceutical composition according to any one of the embodiment s 4 to 46 wherein R 11 and R 12 are independently selected from hydrogen, CrC 20 -alkyl, aryl or aryl-d-Ce-alkyl, wherein the alkyl groups may optionally be substituted with one or more substituents independently selected from R 15 , and the aryl groups may optionally be substituted one or more substituents independently selected from R 16 ; R 11 and R 12 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitro- gen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds.
  • Embodiment 48 A pharmaceutical composition according to embodiment 47 wherein R 11 and R 12 are independently selected from hydrogen, CrC 20 -alkyl, aryl or aryl-d-Ce-alkyl, wherein the alkyl groups may optionally be substituted with one or more substituents independently selected from R 15 , and the aryl groups may optionally be substituted one or more substituents independently selected from R 16 .
  • Embodiment 49 A pharmaceutical composition according to embodiment 48 wherein R 11 and R 12 are independently selected from phenyl or phenyl-d-C e -alkyl.
  • Embodiment 50 A pharmaceutical composition according to embodiment 48 wherein one or both of R 11 and R 12 are methyl.
  • Embodiment 51 A pharmaceutical composition according to any one of the embodiment s 4 to 50 wherein R 13 is independently selected from halogen, CF 3 , OR 11 or NR 1 R 12 .
  • Embodiment 52 A pharmaceutical composition according to embodiment 51 wherein R 13 is independently selected from halogen or OR 11 .
  • Embodiment 53 A pharmaceutical composition according to embodiment 52 wherein R 13 is
  • Embodiment 54 A pharmaceutical composition according to any one of the embodiment s 4 to 53 wherein R 14 is independently selected from halogen, -C(0)OR 11 , -CN, -CF 3 , -OR 11 ,
  • Embodiment 55 A pharmaceutical composition according to embodiment 54 wherein R 14 is independently selected from halogen, -C(0)OR 11 , or -OR 11 .
  • Embodiment 56 A pharmaceutical composition according to any one of the embodiment s 4 to 55 wherein R 15 is independently selected from halogen, -CN, -CF 3 , -C(0)Od-C 6 -alkyl,and
  • Embodiment 57 A pharmaceutical composition according to embodiment 56 wherein R 15 is independently selected from halogen or -C(0)OCrC 6 -alkyl.
  • Embodiment 58 A pharmaceutical composition according to any one of the embodiment s 4 to 57 wherein R 16 is independently selected from halogen, -C(0)OCrC 6 -alkyl, -COOH, -N0 2 ,
  • Embodiment 59 A pharmaceutical composition according to embodiment 58 wherein R 16 is independently selected from halogen, -C(0)OC Ce-alkyl, -COOH, -N0 2 , or d-C 6 -alkyl.
  • Embodiment 60 A pharmaceutical composition according to any one of the embodiment s 1 to 3 wherein CGr is
  • R 19 is hydrogen or d-C 6 -alkyl
  • R 20 is hydrogen or d-Ce-alkyl
  • D, D 1 and F are a valence bond, d-C 6 -alkylene or d-C ⁇ -alkenylene optionally substituted with one or more substituents independently selected from R 72 ,
  • R is independently selected from hydroxy, d-C 6 -alkyl, or aryl
  • E is CrC 6 -alkyl, aryl or heteroaryl, wherein the aryl or heteroaryl is optionally substituted with up to three substituents R 21 , R 22 and R 23 ,
  • G and G 1 are d-C ⁇ -alkyl, aryl or heteroaryl, wherein the aryl or heteroaryl is optionally sub- stituted with up to three substituents R 24 , R 25 and R 26 ,
  • R 17 , R 18 , R 21 , R 22 , R 23 , R 24 , R 25 and R 26 are independently selected from
  • R 27 and R 28 are independently selected from hydrogen, d-C ⁇ -alkyl, aryl-d-C ⁇ -alkyl or aryl, or R 27 and R 28 when attached to the same nitrogen atom together with the said nitrogen atom may form a 3 to 8 membered heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds, R 29 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -OR 27 , and -NR 27 R 28 ,
  • R 30 is independently selected from halogen, -C(0)OR 27 , -CN, -CF 3 , -OCF 3 , -N0 2 , -OR 27 , -NR 27 R 28 and d-Ce-alkyl, or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base.
  • Embodiment 61 A pharmaceutical composition according to embodiment 60 wherein D is a valence bond.
  • Embodiment 62 A pharmaceutical composition according to embodiment 60 wherein D is CrC 6 -alkylene optionally substituted with one or more hydroxy, d-Ce-alkyl, or aryl.
  • Embodiment 63 A pharmaceutical composition according to any one of the embodiment s 60 to 62 wherein E is aryl or heteroaryl, wherein the aryl or heteroaryl is optionally substituted with up to three substituents independently selected from R 21 , R 22 and R 23 .
  • Embodiment 64 A pharmaceutical composition according to embodiment 63 wherein E is aryl optionally substituted with up to three substituents independently selected from R 21 , R 22 and R 23 .
  • Embodiment 65 A pharmaceutical composition according to embodiment 64 wherein E is selected from ArG1 and optionally substituted with up to three substituents independently selected from R 21 , R 22 and R 23 .
  • Embodiment 66 A pharmaceutical composition according to embodiment 65 wherein E is phenyl optionally substituted with up to three substituents independently selected from R 21 ,
  • Embodiment 67 A pharmaceutical composition according to embodiment 66 wherein CGr is
  • Embodiment 68 A pharmaceutical composition according to any one of the embodiment s 60 to 67 wherein R 21 , R 22 and R 23 are independently selected from
  • Embodiment 69 A pharmaceutical composition according to embodiment 68 wherein R 21 , R 22 and R 23 are independently selected from
  • Embodiment 71 A pharmaceutical composition according to embodiment 70 wherein R 21 ,
  • R 22 and R 23 are independently selected from
  • Embodiment 72 A pharmaceutical composition according to embodiment 71 wherein R 21 , R 22 and R 23 are independently selected from
  • Embodiment 73 A pharmaceutical composition according to any one of the embodiment s 60 to 72 wherein R 9 is hydrogen or methyl.
  • Embodiment 74 A pharmaceutical composition according to embodiment 73 wherein R 19 is hydrogen.
  • Embodiment 75 A pharmaceutical composition according to any one of the embodiment s 60 to 74 wherein R 27 is Hydrogen, d-C 6 -alkyl or aryl.
  • Embodiment 76 A pharmaceutical composition according to embodiment 75 wherein R 27 is hydrogen or d-C 6 -alkyl.
  • Embodiment 77 A pharmaceutical composition according to any one of the embodiment s 60 to 76 wherein R 28 is hydrogen or d-C 6 -alkyl.
  • Embodiment 78 A pharmaceutical composition according to embodiment 60 wherein F is a valence bond.
  • Embodiment 79 A pharmaceutical composition according to embodiment 60 wherein F is d- C ⁇ -alkylene optionally substituted with one or more hydroxy, CrC 6 -alkyl, or aryl.
  • Embodiment 80 A pharmaceutical composition according to any one of the embodiment s 60 or 78 to 79 wherein G is d-C 6 -alkyl or aryl, wherein the aryl is optionally substituted with up to three substituents R 24 , R 25 and R 26 .
  • Embodiment 81 A pharmaceutical composition according to any one of the embodiment s 60 or 78 to 79 wherein G is d-Ce-alkyl or ArG1 , wherein the aryl is optionally substituted with up to three substituents R 24 , R 25 and R 26 .
  • Embodiment 82 A pharmaceutical composition according to embodiment 80 wherein G is d-Ce-alkyl.
  • Embodiment 83 A pharmaceutical composition according to embodiment 82 wherein G is phenyl optionally substituted with up to three substituents R 24 , R 25 and R 26 .
  • Embodiment 84 A pharmaceutical composition according to any one of the embodiment s 60 to 83 wherein R 24 , R 25 and R 26 are independently selected from
  • the cyclic moieties optionally may be substituted with one or more substituents selected from R 30 .
  • Embodiment 85 A pharmaceutical composition according to embodiment 84 wherein R 24 ,
  • R 25 and R 26 are independently selected from
  • Embodiment 86 A pharmaceutical composition according to embodiment 85 wherein R 24 ,
  • R 25 and R 26 are independently selected from
  • Embodiment 87 A pharmaceutical composition according to embodiment 86 wherein R 21 , R 22 and R 23 are independently selected from
  • Embodiment 89 A pharmaceutical composition according to embodiment 88 wherein R 21 , R 22 and R 23 are independently selected from
  • Embodiment 90 A pharmaceutical composition according to any one of the embodiment s 60 or 78 to 89 wherein R 20 is hydrogen or methyl.
  • Embodiment 91 A pharmaceutical composition according to embodiment 90 wherein R 20 is hydrogen.
  • Embodiment 92 A pharmaceutical composition according to any one of the embodiment s 60 or 78 to 91 wherein R 27 is hydrogen, d-d-alkyl or aryl.
  • Embodiment 93 A pharmaceutical composition according to embodiment 92 wherein R 27 is hydrogen or d-Ce-alkyl or ArG1.
  • Embodiment 94 A pharmaceutical composition according to embodiment 93 wherein R 27 is hydrogen or d-C 6 -alkyl.
  • Embodiment 95 A pharmaceutical composition according to any one of the embodiment s 60 or 78 to 93 wherein R 28 is hydrogen or d-C 6 -alkyl.
  • Embodiment 96 A pharmaceutical composition according to embodiment 60 wherein R l 7 and R 18 are independently selected from
  • Embodiment 97 A pharmaceutical composition according to embodiment 96 wherein R 17 and R 18 are independently selected from
  • Embodiment 98 A pharmaceutical composition according to embodiment 97 wherein R 17 and R 18 are independently selected from • hydrogen, halogen, -CN, -CF 3 , -N0 2 , -OR 27 , -NR 27 R 28 , or -C(0)OR 27 • methyl, ethyl propyl optionally substituted with one or more substituents independently selected from R 29
  • Embodiment 99 A pharmaceutical composition according to embodiment 98 wherein R 1 r and R 18 are independently selected from
  • Embodiment 100 A pharmaceutical composition according to embodiment 99 wherein R 17 and R 18 are independently selected from
  • Embodiment 101 A pharmaceutical composition according to any one of the embodiment s 60 to 100 wherein R 27 is hydrogen or d-Ce-alkyl.
  • Embodiment 102 A pharmaceutical composition according to embodiment 101 wherein R 27 is hydrogen, methyl or ethyl.
  • Embodiment 103 A pharmaceutical composition according to any one of the embodiment s
  • Embodiment 104 A pharmaceutical composition according to embodiment 103 wherein R 28 is hydrogen, methyl or ethyl.
  • Embodiment 105 A pharmaceutical composition according to any one of the embodiment s
  • R 72 is -OH or phenyl.
  • Embodiment 106 A pharmaceutical composition according to embodiment 60 wherein CGr is
  • Embodiment 107 A pharmaceutical composition according to any one of the embodiment s 1 to 3 wherein CGr is of the form H-l-J-
  • n 1 or 2
  • R 31 is independently selected from hydrogen, halogen, -CN, -CH 2 CN, -CHF 2 , -CF 3 , -OCF 3
  • R 32 and R 33 are independently selected from hydrogen, d-C 6 -alkyl or d-Ce-alkanoyl,
  • R 34 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -OR 35 , and -NR R 36 ,
  • R 35 and R 36 are independently selected from hydrogen, d-Ce-alkyl, aryl-d-Ce-alkyl or aryl, or R 35 and R 36 when attached to the same nitrogen atom together with the said nitrogen atom may form a 3 to 8 membered heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
  • R 37 is independently selected from halogen, -C(0)OR 35 , -C(0)H, -CN, -CF 3 , -OCF 3 , -NO 2 , - OR 35 , -NR 35 R 36 , Ci-C 6 -alkyl or C C 6 -alkanoyl,
  • Embodiment 108 A pharmaceutical composition according to embodiment 107 wherein CGr is of the form H-l-J, wherein H is
  • phenyl, naphthalene or benzocarbazole rings are optionally substituted with one or more substituents independently selected from R 31 ,
  • R 31 is independently selected from hydrogen, halogen, -CN, -CH 2 CN, -CHF 2 , -CF 3 , -OCF 3 , -OCHF 2 , -OCH 2 CF 3 , -OCF 2 CHF 2 , -S(0) 2 CF 3 , -SCF 3 , -N0 2 , -OR 35 , -C(0)R 35 , -NR 35 R 3e , -SR 35 , -NR 35 S(0) 2 R 36 , -S(0) 2 NR 35 R 36 , -S(0)NR 35 R 36 , -S(0)R 35 , -S(0) 2 R 35 , -C(0)NR 35 R 36 , -OC(0)NR 35 R 36 , -NR 35 C(0)R 36 , -CH 2 C(0)NR 35 R 36 , -OCH 2 C(0)NR 35 R 36 , -CH 2 OR 35 , -CH 2 NR 35 R 36 , -OC(0)R 35
  • R 32 and R 33 are independently selected from hydrogen, CrCe-alkyl or d-Ce-alkanoyl,
  • R 34 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -OR 35 , and -NR 35 R 36 ,
  • R 35 and R 36 are independently selected from hydrogen, d-Ce-alkyl, aryl-d-C 6 -alkyl or aryl, or R 35 and R 36 when attached to the same nitrogen atom together with the said nitrogen atom may form a 3 to 8 membered heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
  • R 37 is independently selected from halogen, -C(0)OR 35 , -C(0)H, -CN, -CF 3 , -OCF 3 , -N0 2 , - OR 35 , -NR 35 R 36 , d-C e -alkyl or C,-C 6 -alkanoyl, or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base, With the proviso that R 31 and J cannot both be hydrogen.
  • Embodiment 109 A pharmaceutical composition according to any one of the embodiment s 107 or 108 wherein H is
  • Embodiment 110 A pharmaceutical composition according to embodiment 109 wherein H is
  • Embodiment 111 A pharmaceutical composition according to embodiment 109 wherein H is
  • Embodiment 112 A pharmaceutical composition according to any one of the embodiment s 107 to 111wherein I is a valence bond, -CH 2 N(R 32 )-, or -SO 2 N(R 33 )-.
  • Embodiment 113 A pharmaceutical composition according to embodiment 112 wherein I is a valence bond.
  • Embodiment 114 A pharmaceutical composition according to any one of the embodiment s 107 to 113 wherein J is
  • Embodiment 115 A pharmaceutical composition according to embodiment 114 wherein J is
  • Embodiment 116 A pharmaceutical composition according to embodiment 114 wherein J is
  • Embodiment 117 A pharmaceutical composition according to embodiment 116 wherein J is
  • Embodiment 118 A pharmaceutical composition according to embodiment 117 wherein J is hydrogen.
  • Embodiment 119 A pharmaceutical composition according to any one of the embodiment s
  • Embodiment 120 A pharmaceutical composition according to any one of the embodiment s
  • R 34 is hydrogen, halogen, -CN, -CF 3 , -OCF 3 , -SCF 3 , -N0 2 , -OR 35 ,
  • Embodiment 121 A pharmaceutical composition according to embodiment 120 wherein R 34 is hydrogen, halogen, -CF 3 , -N0 2 , -OR 35 , -NR 35 R 36 , -SR 35 , -NR 35 C(0)R 36 , or -C(0)OR 35 .
  • Embodiment 122 A pharmaceutical composition according to embodiment 121 wherein R 34 is hydrogen, halogen, -CF 3 , -N0 2 , -OR 35 , -NR 35 R 36 , or -NR 35 C(0)R 36 .
  • Embodiment 123 A pharmaceutical composition according to embodiment 122 wherein R 34 is hydrogen, halogen, or -OR 35 .
  • Embodiment 124. A pharmaceutical composition according to any one of the embodiment s
  • R 35 and R 36 are independently selected from hydrogen, d-C ⁇ -alkyl, or aryl.
  • Embodiment 125 A pharmaceutical composition according to embodiment 124 wherein R 35 and R 36 are independently selected from hydrogen or d-Ce-alkyl.
  • Embodiment 126. A pharmaceutical composition according to any one of the embodiment s
  • R 37 is halogen, -C(0)OR 35 , -CN, -CF 3 , -OR 35 , -NR 35 R 36 , d-C 6 -alkyl or d-
  • Embodiment 127 A pharmaceutical composition according to embodiment 126 wherein R 37 is halogen, -C(0)OR 35 , -OR 35 , -NR 35 R 36 , d-C 6 -alkyl or d-C 6 -alkanoyl.
  • Embodiment 128 A pharmaceutical composition according to embodiment 127 wherein R 37 is halogen, -C(0)OR 35 or -OR 35 .
  • Embodiment 129 A pharmaceutical composition according to any one of the embodiment s 1 to 3 wherein CGr is
  • U is a valence bond, d-Ce-alkenylene, -d-Ce-alkyl-O- or d-C 6 -alkylene wherein any d- Ce-alkyl moiety is optionally substituted with d-C 6 -alkyl,
  • R 38 is CrCe-alkyl, aryl, wherein the alkyl or aryl moieties are optionally substituted with one or more substituents independently selected from R 39 ,
  • R is independently selected from halogen, cyano, nitro, amino,
  • M is a valence bond, arylene or heteroarylene, wherein the aryl or heteroaryl moieties are optionally substituted with one or more substituents independently selected from R 40 ,
  • R 41 and R 42 are independently selected from hydrogen, -OH, d-Ce-alkyl, d-Ce-alkenyl, aryl- d-Ce-alkyl or aryl, wherein the alkyl moieties may optionally be substituted with one or more substituents independently selected from R 45 , and the aryl moieties may optionally be substituted with one or more substituents independently selected from R 46 ; R 41 and R 42 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
  • R 43 is independently selected from halogen, -CN, -CF 3l -OCF 3 , -OR 41 , and -NR 41 R 42
  • R 44 is independently selected from halogen, -C(0)OR 41 , -CH 2 C(0)OR 41 , -CH 2 OR 41 , -CN, - CF 3 , -OCF 3 , -N0 2 , -OR 41 , -NR 41 R 42 and d-Ce-alkyl,
  • R 45 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -0-d-C 6 -alkyl, -C(0)-0-C Ce-alkyl, -COOH and -NH 2 ,
  • R 47 and R 48 are independently selected from hydrogen, d-Ce-alkyl, aryl optionally substituted with one or more R 49 ,
  • R is independently selected from halogen and -COOH
  • T is
  • any alkyl, alkenyl , alkynyl, aryl and heteroaryl moiety is optionally substituted with one or more substituents independently selected from R 50 ,
  • Embodiment R 50A and R 508 are independently selected from -C(0)OCrC 6 -alkyl, -COOH, -d- C 6 -alkyl-C(0)OCrC 6 -alkyl, -Ci-Ce-alkyl-COOH, or d-C 6 -alkyl
  • R 51 and R 52 are independently selected from hydrogen and d-Ce-alkyl
  • R 53 is independently selected from CrC 6 -alkyl, d-Ce-alkoxy, -d-Ce-alkyl-COOH, -C 2 - Ce-alkenyl-COOH, -OR 51 , -N0 2 , halogen, -COOH, -CF 3 , -CN, or -N(R 51 R 52 ),
  • Embodiment 133 A pharmaceutical composition according to embodiment 132 wherein K is a valence bond or C C ⁇ -alkylene, wherein any d-C e -alkyl moiety is optionally substituted with R 38 .
  • Embodiment 135. A pharmaceutical composition according to embodiment 133 wherein K is a valence bond.
  • Embodiment 136 A pharmaceutical composition according to any one of the embodiment s
  • U is a valence bond or -CrC 6 -alkyl-0-.
  • Embodiment 137 A pharmaceutical composition according to embodiment 136 wherein U is a valence bond.
  • Embodiment 138 A pharmaceutical composition according to any one of the embodiment s
  • M is arylene or heteroarylene, wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from
  • Embodiment 139 A pharmaceutical composition according to embodiment 138 wherein M is ArG1 or Het1 , wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from R 40 .
  • Embodiment 140 A pharmaceutical composition according to embodiment 139 wherein M is
  • Embodiment 141 A pharmaceutical composition according to embodiment 140 wherein M is
  • Embodiment 142 A pharmaceutical composition according to embodiment 141 wherein M is phenylene optionally substituted with one or more substituents independently selected from R 40 .
  • Embodiment 143 A pharmaceutical composition according to embodiment 141 wherein M is indolylene optionally substituted with one or more substituents independently selected from
  • Embodiment 144 A pharmaceutical composition according to embodiment 143 wherein M is
  • Embodiment 146. A pharmaceutical composition according to embodiment 145 wherein is
  • Embodiment 147 A pharmaceutical composition according to any one of the embodiment s 129 to 146 wherein R 40 is selected from
  • Embodiment 148 A pharmaceutical composition according to embodiment 147 wherein R 40 is selected from
  • Embodiment 150 A pharmaceutical composition according to embodiment 149 wherein R 40 is hydrogen.
  • Embodiment 151 A pharmaceutical composition according to embodiment 149 wherein R 40 is selected from .halogen, -N0 2 , -OR 41 , -NR 41 R 42 , -C(0)OR 41 , or -NR 41 C(0)R 42 ,
  • Embodiment 152 A pharmaceutical composition according to any one of the embodiment s 129 to 151 wherein R 41 and R 42 are independently selected from hydrogen, d-Ce-alkyl, or aryl, wherein the aryl moieties may optionally be substituted with halogen or -COOH.
  • Embodiment 153 A pharmaceutical composition according to embodiment 152 wherein R 41 and R 42 are independently selected from hydrogen, methyl, ethyl, or phenyl, wherein the phenyl moieties may optionally be substituted with halogen or -COOH.
  • Embodiment 154 A pharmaceutical composition according to embodiment 152 wherein R 41 and R 42 are independently selected from hydrogen, methyl, ethyl, or phenyl, wherein the phenyl moieties may optionally be substituted with halogen or -COOH.
  • Embodiment 155 Embodiment 155.
  • Q is a valence bond.
  • Embodiment 156 Embodiment 156.
  • Embodiment 157. A pharmaceutical composition according to any one of the embodiment s 129 to 156 wherein T is
  • Embodiment 158 A pharmaceutical composition according to embodiment 157 wherein T is
  • Embodiment 159 A pharmaceutical composition according to embodiment 158 wherein T is •hydrogen,
  • Embodiment 160 A pharmaceutical composition according to embodiment 159 wherein T is phenyl substituted with R 50 .
  • Embodiment 165 Embodiment 165.
  • Embodiment 166 A pharmaceutical composition according to embodiment 164 wherein R 50 is phenyl, methyl or ethyl.
  • Embodiment 167 A pharmaceutical composition according to embodiment 166 wherein R 50 is methyl or ethyl.
  • Embodiment 168 A pharmaceutical composition according to any one of the embodiment s
  • Embodiment 169 A pharmaceutical composition according to any one of the embodiment s 129 to 168 wherein R 51 is methyl.
  • Embodiment 170 A pharmaceutical composition according to any one of the embodiment s
  • R 53 is d-C ⁇ -alkyl, d-C 6 -alkoxy, -OR 51 , halogen.or -CF 3 .
  • Embodiment 171 A pharmaceutical composition according to any one of the embodiment s
  • R 50 * is -C(0)OCH 3 , -C(0)OCH 2 CH 3 -COOH, -CH 2 C(O)OCH 3 , - CH 2 C(0)OCH 2 CH 3 , -CH 2 CH 2 C(0)OCH 3 , -CH 2 CH 2 C(0)OCH 2 CH 3 , -CH 2 COOH, methyl, or ethyl.
  • Embodiment 172 A pharmaceutical composition according to any one of the embodiment s
  • R 508 is -C(0)OCH 3 , -C(0)OCH 2 CH 3 -COOH, -CH 2 C(O)OCH 3 , -
  • Embodiment 173 A pharmaceutical composition according to any one of the embodiment s 1 to 3 wherein CGr is
  • V is d-Ce-alkyl, aryl, heteroaryl, aryl-d-e-alkyl- or aryl-C 2 -e-alkenyl-, wherein the alkyl or alkenyl is optionally substituted with one or more substituents independently selected from R 54 , and the aryl or heteroaryl is optionally substituted with one or more substituents independently selected from R 55 ,
  • R 54 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , aryl, -COOH and -NH 2
  • R 55 is independently selected from • hydrogen, halogen, -CN, -CH 2 CN, -CHF 2 , -CF 3 , -OCF 3> -OCHF 2 , -OCH;>CF 3 , -OCF 2 CHF 2 , -S(0) 2 CF 3 , -OS(0) 2 CF 3 , -SCF 3 , -N0 2 .
  • R 58 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -OR 56 , and -NR 56 R 57 ,
  • R 59 is independently selected from halogen, -C(0)OR 56 , -CH 2 C(0)OR 56 , -CH 2 OR 56 , -CN, - CF 3 , -OCF 3 , -N0 2 , -OR 56 , -NR 56 R 57 and d-Ce-alkyl
  • R 62 is CrCe-alkyl, aryl optionally substituted with one or more substituents independently selected from halogen, or heteroaryl optionally substituted with one or more d-C 6 -alkyl independently, or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base.
  • Embodiment 174 A pharmaceutical composition according to embodiment 173 wherein V is aryl, heteroaryl, or aryl-d-e-alkyl-, wherein the alkyl is optionally substituted with one or more substituents independently selected R 54 , and the aryl or heteroaryl is optionally substituted with one or more substituents independently selected from R 55 .
  • Embodiment 175. A pharmaceutical composition according to embodiment 174 wherein V is aryl, Het1 , or aryl-d-e-alkyl-, wherein the alkyl is optionally substituted with one or more substituents independently selected from R 54 , and the aryl or heteroaryl moiety is optionally substituted with one or more substituents independently selected from R 55 .
  • Embodiment 176. A pharmaceutical composition according to embodiment 175 wherein V is aryl, Het2, or aryl-d-e-alkyl-, wherein the alkyl is optionally substituted with one or more substituents independently selected from R 54 , and the aryl or heteroaryl moiety is optionally substituted with one or more substituents independently selected from R 55 .
  • Embodiment 177 A pharmaceutical composition according to embodiment 176 wherein V is aryl, Het3, or aryl-d-e-alkyl-, wherein the alkyl is optionally substituted with one or more substituents independently selected from R 54 , and the aryl or heteroaryl moiety is optionally substituted with one or more substituents independently selected from R 55 .
  • Embodiment 178. A pharmaceutical composition according to embodiment 177 wherein V is aryl optionally substituted with one or more substituents independently selected from R 55 .
  • Embodiment 179. A pharmaceutical composition according to embodiment 178 wherein V is ArG1 optionally substituted with one or more substituents independently selected from R 55 .
  • Embodiment 180 A pharmaceutical composition according to embodiment 176 wherein V is aryl, Het3, or aryl-d-e-alkyl-, wherein the alkyl is optionally substituted with one or more substituents independently selected from R 54 , and the aryl or heteroaryl moiety
  • Embodiment 182. A pharmaceutical composition according to any one of the embodiment s 173 to 181 wherein R 55 is independently selected from •halogen, d-Ce-alkyl, -CN, -OCF 3 ,-CF 3 , -N0 2 , -OR 56 , -NR 56 R 57 , -NR 56 C(O)R 57
  • Embodiment 184 A pharmaceutical composition according to embodiment 183 wherein R 55 is independently selected from halogen, -OR 56 , -NR ⁇ R 57 , -C(0)OR 56 , -Od-C 8 - alkyl-C(0)OR 56 , -NR 56 C(0)R 57 or d-C 6 -alkyl.
  • Embodiment 185 A pharmaceutical composition according to embodiment 184 wherein R 55 is independently selected from halogen, -OR 56 , -NR 56 R 57 , -C(0)OR 56 , -Od-C 8 - alkyl-C(0)OR 56 , -NR 56 C(0)R 57 , methyl or ethyl.
  • Embodiment 187 A pharmaceutical composition according to embodiment 186 wherein R 56 and R 57 are independently selected from hydrogen or d-C ⁇ 2 -alkyl, R 56 and R 57 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom.
  • a pharmaceutical composition according to embodiment 187 wherein R 56 and R 57 are independently selected from hydrogen or methyl, ethyl, propyl butyl, R 56 and R 57 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom.
  • Embodiment 189 A pharmaceutical composition according to any one of the embodiment s 1 to 3 wherein CGr is
  • AA is d-Ce-alkyl, aryl, heteroaryl, aryl-d-e-alkyl- or aryl-C 2 - 6 -alkenyl-, wherein the alkyl or alkenyl is optionally substituted with one or more substituents independently selected from R 63 , and the aryl or heteroaryl is optionally substituted with one or more substituents independently selected from R 64 ,
  • R 63 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , aryl, -COOH and -NH 2 ,
  • R 64 is independently selected from
  • R 67 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -OR 65 , and -NR 65 R 66 ,
  • R 68 is independently selected from halogen, -C(0)OR 65 , -CH 2 C(0)OR 65 , -CH 2 OR 65 , -CN, - CF 3 , -OCF 3 , -N0 2 , -OR 65 , -NR 65 R 66 and d-C ⁇ -alkyl,
  • R 69 is independently selected from d-Ce-alkyl, aryl optionally substituted with one or more halogen, or heteroaryl optionally substituted with one or more d-Ce-alkyl,
  • R 70 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -Od-Ce-alkyl, -C(0)OCrC 6 - alkyl, -COOH and -NH 2 ,
  • Embodiment 190 A pharmaceutical composition according to embodiment 189 wherein AA is aryl, heteroaryl or aryl-d-e-alkyl-, wherein the alkyl is optionally substituted with one or more R 63 , and the aryl or heteroaryl is optionally substituted with one or more substituents independently selected from R 64 .
  • Embodiment 191 A pharmaceutical composition according to embodiment 190 wherein AA is aryl or heteroaryl optionally substituted with one or more substituents independently se- lected from R 64 .
  • Embodiment 192 A pharmaceutical composition according to embodiment 191 wherein AA is ArG1 or Het1 optionally substituted with one or more substituents independently selected from R 64 .
  • Embodiment 193. A pharmaceutical composition according to embodiment 192 wherein AA is ArG1 or Het2 optionally substituted with one or more substituents independently selected from R 64 .
  • Embodiment 194 A pharmaceutical composition according to embodiment 193 wherein AA is ArG1 or Het3 optionally substituted with one or more substituents independently selected from R 64 .
  • Embodiment 195 A pharmaceutical composition according to embodiment 194 wherein AA is phenyl, naphtyl, anthryl, carbazolyl, thienyl, pyridyl, or benzodioxyl optionally substituted with one or more substituents independently selected from R 64 .
  • Embodiment 196 A pharmaceutical composition according to embodiment 195 wherein AA is phenyl or naphtyl optionally substituted with one or more substituents independently se- lected from R 64 .
  • Embodiment 197 A pharmaceutical composition according to any one of the embodiment s 189 to 196 wherein R 64 is independently selected from hydrogen, halogen, -CF 3 , -OCF 3 , -OR 65 , -NR 65 R 66 , d-Ce-alkyl , -OC(0)R 65 , -OCrC 6 -alkyl-C(0)OR 65 , aryl-C 2 -C 6 -alkenyl, aryloxy or aryl, wherein d-d-alkyl is optionally substituted with one or more substituents inde- pendently selected from R 67 , and the cyclic moieties optionally are substituted with one or more substituents independently selected from R 68 .
  • R 64 is independently selected from hydrogen, halogen, -CF 3 , -OCF 3 , -OR 65 , -NR 65 R 66 , d-Ce-alkyl , -OC(0)R 65 ,
  • Embodiment 198 A pharmaceutical composition according to embodiment 197 wherein R 64 is independently selected from halogen, -CF 3 , -OCF 3 , -OR 65 , -NR 65 R 66 , methyl, ethyl, propyl, -OC(0)R 65 , -OCH 2 -C(0)OR 65 , -OCH 2 -CH 2 -C(0)OR 65 , phenoxy optionally substituted with one or more substituents independently selected from R 68 .
  • R 64 is independently selected from halogen, -CF 3 , -OCF 3 , -OR 65 , -NR 65 R 66 , methyl, ethyl, propyl, -OC(0)R 65 , -OCH 2 -C(0)OR 65 , -OCH 2 -CH 2 -C(0)OR 65 , phenoxy optionally substituted with one or more substituents independently selected from R 68 .
  • Embodiment 199 A pharmaceutical composition according to any one of the embodiment s 189 to 198 wherein R 65 and R 66 are independently selected from hydrogen, CF 3 , d-C ⁇ 2 -alkyl, aryl, or heteroaryl optionally substituted with one or more substituents independently selected from R 71 .
  • Embodiment 200 A pharmaceutical composition according to embodiment 199 wherein R 65 and R 66 are independently hydrogen, d-C 12 -alkyl, aryl, or heteroaryl optionally substituted with one or more substituents independently selected from R 71 .
  • Embodiment 201 A pharmaceutical composition according to embodiment 200 wherein R 65 and R 66 are independently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, ArG1 or Het1 optionally substituted with one or more substituents independently selected from R 71 .
  • Embodiment 202 A pharmaceutical composition according to embodiment 201 wherein R 65 and R 66 are independently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, ArG1 or Het2 optionally substituted with one or more substituents independently selected from R 71 .
  • Embodiment 203 A pharmaceutical composition according to embodiment 202 wherein R 65 and R 66 are independently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, ArG1 or Het3 optionally substituted with one or more substituents independently selected from R 71 .
  • Embodiment 204 A pharmaceutical composition according to embodiment 203 wherein R 65 and R 66 are independently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, phenyl, naphtyl, thiadiazolyl optionally substituted with one or more R 71 independently; or isoxazoiyl optionally substituted with one or more substituents independently selected from R 7 .
  • Embodiment 205 A pharmaceutical composition according to any one of the embodiment s
  • R 71 is halogen or d-d-alkyl.
  • Embodiment 206 A pharmaceutical composition according to embodiment 205 wherein R 71 is halogen or methyl.
  • Embodiment 207 A pharmaceutical preparation according to any one of the embodiment s 1 to 205 wherein Frg1 consists of 0 to 5 neutral amino acids independently selected from the group consisting of Gly, Ala, Thr, and Ser.
  • Embodiment 208 A pharmaceutical preparation according to embodiment 207 wherein Frg1 consists of 0 to 5 Gly.
  • Embodiment 209 A pharmaceutical preparation according to embodiment 208 wherein Frg1 consists of 0 Gly.
  • Embodiment 210 A pharmaceutical preparation according to embodiment 208 wherein Frg1 consists of 1 Gly.
  • Embodiment 211. A pharmaceutical preparation according to embodiment 208 wherein Frg1 consists of 2 Gly.
  • Embodiment 212 A pharmaceutical preparation according to embodiment 208 wherein Frg1 consists of 3 Gly.
  • Embodiment 213. A pharmaceutical preparation according to embodiment 208 wherein Frg1 consists of 4 Gly.
  • Embodiment 214. A pharmaceutical preparation according to embodiment 208 wherein Frg1 consists of 5 Gly.
  • Embodiment 215. A pharmaceutical preparation according to any one of the embodiment s 1 to 214 wherein G B is of the formula B 1 -B 2 -C(0)-, B -B 2 -S0 2 - or B 1 -B 2 -CH 2 -, wherein B 1 and B 2 are as defined in embodiment 1.
  • Embodiment 216 A pharmaceutical preparation according to any one of the embodiment s 1 to 214 wherein G B is of the formula B 1 -B 2 -C(0)-, B 1 -B 2 -S0 2 - or B 1 -B 2 -NH-, wherein B 1 and B 2 are as defined in embodiment 1.
  • Embodiment 217 A pharmaceutical preparation according to any one of the embodiment s 1 to 214 wherein G B is of the formula B -B -C(0)-, B 1 -B 2 -CH 2 - or B 1 -B 2 -NH-, wherein B 1 and B 2 are as defined in embodiment 1.
  • Embodiment 218 A pharmaceutical preparation according to any one of the embodiment s 1 to 214 wherein G B is of the formula B 1 -B 2 -CH 2 -, B 1 -B 2 -S0 2 - or B 1 -B 2 -NH-, wherein B 1 and B 2 are as defined in embodiment 1.
  • Embodiment 219. A pharmaceutical preparation according to any one of the embodiment s
  • G B is of the formula B 1 -B 2 -C(0)- or B 1 -B 2 -S0 2 -, wherein B 1 and B 2 are as defined in embodiment 1.
  • Embodiment 220 A pharmaceutical preparation according to any one of the embodiment s
  • G B is of the formula B 1 -B 2 -C(0)- or B 1 -B 2 -CH 2 -, wherein B 1 and B 2 are as defined in embodiment 1.
  • Embodiment 22 A pharmaceutical preparation according to any one of the embodiment s
  • G B is of the formula B 1 -B 2 -C(0)- or B 1 -B 2 -NH-, wherein B 1 and B 2 are as defined in embodiment 1.
  • Embodiment 222 A pharmaceutical preparation according to any one of the embodiment s 215 or 218 wherein G 8 is of the formula B 1 -B -CH 2 - or B 1 -B 2 -S0 2 - , wherein B 1 and B 2 are as defined in embodiment 1.
  • Embodiment 223. A pharmaceutical preparation according to any one of the embodiment s
  • G B is of the formula B 1 -B 2 -NH- or B 1 -B 2 -S0 2 - , wherein B 1 and B 2 are as defined in embodiment 1.
  • Embodiment 224 A pharmaceutical preparation according to any one of the embodiment s
  • G 8 is of the formula B 1 -B 2 -CH 2 - or B 1 -B 2 -NH- , wherein B 1 and B 2 are as defined in embodiment 1.
  • Embodiment 225 A pharmaceutical preparation according to any one of the embodiment s 219, 220, or 221 wherein G B is of the formula B 1 -B 2 -C(0)-.
  • Embodiment 226 A pharmaceutical preparation according to any one of the embodiment s
  • G B is of the formula B 1 -B 2 -CH 2 -.
  • Embodiment 227 A pharmaceutical preparation according to any one of the embodiment s
  • G B is of the formula B 1 -B 2 -NH-.
  • Embodiment 229. A pharmaceutical preparation according to any one of the embodiment s 1 to 228 wherein B 1 is a valence bond, -0-, or -S-.
  • Embodiment 230 A pharmaceutical preparation according to any one of the embodiment s 1 to 228 wherein B 1 is a valence bond, -0-, or -N(R 68 )-.
  • Embodiment 23 A pharmaceutical preparation according to any one of the embodiment s 1 to 228 wherein B 1 is a valence bond, -S-, or -N(R 68 )-.
  • Embodiment 232 A pharmaceutical preparation according to any one of the embodiment s 1 to 228 wherein B 1 is -0-, -S- or -N(R 68 )-.
  • Embodiment 233 A pharmaceutical preparation according to any one of the embodiment s
  • B 1 is a valence bond or -0-.
  • Embodiment 23 A pharmaceutical preparation according to any one of the embodiment s
  • B 1 is a valence bond or -S-.
  • Embodiment 235 A pharmaceutical preparation according to any one of the embodiment s 230 or 231 wherein B 1 is a valence bond or -N(R 68 )-.
  • Embodiment 236 A pharmaceutical preparation according to any one of the embodiment s
  • B 1 is -O-or -S-.
  • Embodiment 237 A pharmaceutical preparation according to any one of the embodiment s
  • B 1 is -S-or -N(R 68 )-.
  • Embodiment 239. A pharmaceutical preparation according to any one of the embodiment s
  • Embodiment 240 A pharmaceutical preparation according to any one of the embodiment s 233, 236 or 237 wherein B 1 is -0-.
  • Embodiment 241 A pharmaceutical preparation according to any one of the embodiment s
  • Embodiment 242 A pharmaceutical preparation according to any one of the embodiment s
  • a pharmaceutical preparation according to embodiment 246 wherein B 2 is a valence bond, d-C 18 -alkylene, arylene, heteroarylene, -CrC 18 -alkyl-aryl-, and the alkylene and arylene moieties are optionally substituted as defined in embodiment 1.
  • Embodiment 248. A pharmaceutical preparation according to embodiment 247 wherein B 2 is a valence bond, C C 18 -alkylene, arylene, -d-C 18 -alkyl-aryl-, and the alkylene and arylene moieties are optionally substituted as defined in embodiment 1.
  • Embodiment 249. A pharmaceutical preparation according to embodiment 248 wherein B 2 is a valence bond or -CrC 18 -alkylene, and the alkylene moieties are optionally substituted as defined in embodiment 1.
  • Embodiment 250. A pharmaceutical preparation according to any one of the embodiment s 1 to 249 whereiin Frg2 comprises 1 to 16 positively charged groups.
  • Embodiment 251 A pharmaceutical preparation according to embodiment 250 wherein Frg2 comprises 1 to 12 positively charged groups.
  • Embodiment 252 A pharmaceutical preparation according to embodiment 251 wherein Frg2 comprises 1 to 10 positively charged groups.
  • Embodiment 253. A pharmaceutical preparation according to any one of the embodiment s 1 to 249 wherein Frg2 comprises 10 to 20 positively charged groups.
  • Embodiment 254. A pharmaceutical preparation according to embodiment 253 wherein Frg2 comprises 12 to 20 positively charged groups.
  • Embodiment 255. A pharmaceutical preparation according to embodiment 254 wherein Frg2 comprises 16 to 20 positively charged groups.
  • Embodiment 256 A pharmaceutical preparation according to any one of the embodiment s 250 to 255 wherein the positively charged groups of Frg2 are basic amino acids independently selected from the group consisting of Lys and Arg and D-isomers of these.
  • Embodiment 257 A pharmaceutical preparation according to embodiment 256 wherein the basic amino acids are all Arg.
  • Embodiment 258 A pharmaceutical preparation according to any one of the embodiment s 250 to 257, wherein Frg2 comprises one or more neutral amino acids independently selected from the group consisting of Gly, Ala, Thr, and Ser.
  • Embodiment 259. A pharmaceutical preparation according to embodiment 258, wherein Frg2 comprises one or more Gly.
  • Embodiment 260 A pharmaceutical preparation according to any one of the embodiment s 1 to 259 wherein X is -OH or -NH 2 .
  • Embodiment 261. A pharmaceutical preparation according to embodiment 260 wherein X is - NH 2 .
  • Embodiment 262 A pharmaceutical preparation according to any one of the embodiment s 1 to 261 which further comprises at least 3 phenolic molecules per putative insulin hexamer.
  • Embodiment 263. A pharmaceutical preparation according to any one of the embodiment s 1 to 262 wherein the acid-stabilised insulin is an analogue of human insulin wherein A21 is Ala, Gin, Glu, Gly, His, He, Leu, Met, Phe, Ser, Thr, Trp, Tyr, Val, and hSer.
  • Embodiment 264 A pharmaceutical preparation according to embodiment 263 wherein the acid-stabilised insulin is an analogue of human insulin wherein A21 is Ala, Gly, He, Leu, Phe, Ser, Thr, Val, and hSer.
  • Embodiment 265. A pharmaceutical preparation according to embodiment 264 wherein the acid-stabilised insulin is an analogue of human insulin wherein A21 is Ala or Gly.
  • Embodiment 266 A pharmaceutical preparation according to embodiment 265 wherein the acid-stabilised insulin is an analogue of human insulin wherein A21 is Gly.
  • Embodiment 267 A pharmaceutical preparation according to any one of the embodiment s 263 to 266 wherein the acid-stabilised insulin is an analogue of human insulin further modi- fied by exchange or deletion of one or more amino acid residues according to the following: B3 is selected from Thr, Ser, Lys or Ala
  • B28 is Lys, Asp or Glu
  • B29 is Pro or Glu B9 is Glu or Asp
  • Embodiment 268 A pharmaceutical preparation according to embodiment 267 wherein the acid-stabilised insulin is an analogue of human insulin further modified by exchange of B28 to Lys or Asp.
  • Embodiment 269. A pharmaceutical preparation according to embodiment 268 wherein the acid-stabilised insulin is an analogue of human insulin further modified by exchange of B28 to Asp.
  • Embodiment 270. A pharmaceutical preparation according to embodiment 268 wherein the acid-stabilised insulin is an analogue of human insulin further modified by exchange of B28 to Lys.
  • Embodiment 271 A pharmaceutical preparation according any one of the embodiment s 263 to 270 wherein the acid-stabilised insulin is an analogue of human insulin further modified by exchange of B29 to Pro.
  • Embodiment 272 A pharmaceutical preparation according any one of the embodiment s 263 to 271 wherein the acid-stabilised insulin is an analogue of human insulin further modified by exchange of B3 to Lys or Ala.
  • Embodiment 273 A pharmaceutical preparation according any one of the embodiment s 263 to 272 wherein the acid-stabilised insulin is an analogue of human insulin further modified by exchange of A18 to Gin.
  • Embodiment 274 A pharmaceutical preparation according any one of the embodiment s 263 to 273 wherein the acid-stabilised insulin is an analogue of human insulin further modified by deletion of B25.
  • Embodiment 275 A pharmaceutical preparation according any one of the embodiment s 263 to 274 wherein the acid-stabilised insulin is an analogue of human insulin further modified by deletion of B30.
  • Embodiment 276 A pharmaceutical preparation according to embodiment 263 wherein the acid-stabilised insulin is selected from the group A21G A21G, B28K, B29P
  • A21G, B3K, B29E, desB30 A21G, desB27, desB30
  • Embodiment 277 A pharmaceutical preparation according to any one of the embodiment s 1 to 276 wherein zinc ions are present in an amount corresponding to 10 to 40 ⁇ g Zn/100 U insulin.
  • Embodiment 278 A pharmaceutical preparation according to embodiment 277 wherein zinc ions are present in an amount corresponding to 10 to 26 ⁇ g Zn/100 U insulin.
  • Embodiment 280 A pharmaceutical preparation according to embodiment 279 wherein the ratio between insulin and the zinc-binding ligand according to any one of the embodiment s 1 to 261 is in the range from 95:5 to 5:95.
  • Embodiment 281. A pharmaceutical preparation according to embodiment 280 wherein the ratio between between insulin and the zinc-binding ligand according to any one of the embodiment s 1 to 261 is in the range from 80:20 to 20:80. Embodiment 282. A pharmaceutical preparation according to embodiment 281 wherein the ratio between between insulin and the zinc-binding ligand according to any one of the embodiment s 1 to 261 is in the range from 70:30 to 30:70.
  • Embodiment 283 A pharmaceutical preparation according to any one of the embodiment s 1 to 282 wherein the concentration of insulin is 60 to 3000 nmol/ml.
  • Embodiment 284. A pharmaceutical preparation according to embodiment 283 wherein the concentration of insulin is 240 to 1200 nmol/ml.
  • Embodiment 285. A pharmaceutical preparation according to embodiment 284 wherein the concentration of insulin is about 600 nmol/ml.
  • Embodiment 286. A method of preparing a zinc-binding ligand according to embodiment 1 comprising the steps of
  • Embodiment 287 Method of prolonging the action of an acid-stabilised insulin preparation which comprises adding a zinc-binding ligand according to any of embodiment s 1 to 261 to the acid-stabilised insulin preparation.
  • Embodiment 288 A method of treating type 1 or type 2 diabetes comprising administering to a patient in need thereof a theraputically effective amount of a pharmaceutical preparation according to any one of the embodiment s 1 to 282.
  • Embodiment 28 Use of a preparation according to any one of the embodiment s 1 to 282 for the preparation of a medicament for treatment of type 1 or type 2 diabetes.
  • the present invention also relates to a pharmaceutical preparation for the treatment of diabetes in a patient in need of such a treatment comprising an R-state hexamer of insulin according to the invention together with a pharmaceutically acceptable carrier.
  • the insulin preparation comprises 60 to 3000 nmol/ml of insulin.
  • the insulin preparation comprises 240 to 1200 nmol/ml of insulin. In another embodiment of the invention the insulin preparation comprises about 600 nmol/ml of insulin.
  • Zinc ions may be present in an amount corresponding to 10 to 40 ⁇ g Zn/100 U insulin, more preferably 10 to 26 ⁇ g Zn/100 U insulin.
  • Insulin formulations of the invention are usually administered from multi-dose containers where a preservative effect is desired. Since phenolic preservatives also stabilize the R-state hexamer the formulations may contain up to 50 mM of phenolic molecules.
  • the phenolic molecules in the insulin formulation may be selected from the group consisting of phenol, m- cresol, chloro-cresol, thymol, 7-hydroxyindole or any mixture thereof. In one embodiment of the invention 0.5 to 4.0 mg/ml of phenolic compound may be employed.
  • 0.6 to 4.0 mg/ml of m-cresol may be employed.
  • 0.5 to 4.0 mg/ml of phenol may be employed.
  • 0.5 to 4.0 mg/ml of a mixture of m-cresol or phenol may be employed.
  • the pharmaceutical preparation may further comprises a buffer substance, such as a TRIS, phosphate, glycine or glycylglycine (or another zwitterionic substance) buffer, an isotonicity agent, such as NaCl, glycerol, mannitol and/or lactose.
  • a buffer substance such as a TRIS, phosphate, glycine or glycylglycine (or another zwitterionic substance) buffer
  • an isotonicity agent such as NaCl, glycerol, mannitol and/or lactose.
  • Chloride would be used at moderate concentrations (e.g. up to 50 mM) to avoid competition with the zinc-site ligands of the present invention.
  • the action of insulin may further be slowed down in vivo by the addition of physiologically acceptable agents that increase the viscosity of the pharmaceutical preparation.
  • the pharmaceutical preparation according to the invention may furthermore comprise an agent which increases the viscosity, such as polyethylene glycol, polypropylene glycol, copolymers thereof, dextrans and/or polylactides.
  • the insulin preparation of the invention comprises between 0.001
  • a non-ionic surfactant for example tween 20 or Polox 188.
  • a nonionic detergent can be added to stabilise insulin against fibrillation during storage and handling.
  • the insulin preparation of the present invention may have a pH value in the range of 2.5 to
  • the preparations of the invention are used in connection with insulin pumps.
  • the insulin pumps may be prefilled and disposable, or the insulin preparations may be supplied from a reservoir which is removable. Insulin pumps may be skin-mounted or carried, and the path of the insulin preparation from the storage compartment of the pump to the patient may be more or less tortuous.
  • Non-limiting examples of insulin pumps are disclosed in US 5,957,895, US 5,858,001 , US 4,468,221, US 4,468,221 , US 5,957,895, US 5,858,001 , US 6,074,369, US 5,858,001 , US 5,527,288, and US 6,074,369.
  • the preparations of the invention are used in connection with pen-like injection devices, which may be prefilled and disposable, or the insulin preparations may be supplied from a reservoir which is removable.
  • pen-like injection devices are FlexPen ® , InnoLet ® , InDuoTM, Innovo ® .
  • preparations of the invention are used in connection with devices for pulmonary administration of aqueous insulin preparations, a non-limiting example of which is the AerX ® device.
  • the invention furthermore relates to treatment of a patient in which the pharmaceutical preparation of the invention, i.e. comprising zinc ions, acid-stabilised insulin analogue and a ligand for the R-state His 810 Zn 2+ site, is combined with another form of treatment.
  • the pharmaceutical preparation of the invention i.e. comprising zinc ions, acid-stabilised insulin analogue and a ligand for the R-state His 810 Zn 2+ site
  • treatment of a patient with the pharmaceutical preparation of the invention is combined with diet and/or exercise.
  • the pharmaceutical preparation of the invention is administered in combination with one or more further active substances in any suitable ratios.
  • Such further active substances may e.g. be selected from antiobesity agents, antidiabetics, antihypertensive agents, agents for the treatment of complications resulting from or associated with diabetes and agents for the treatment of complications and disorders resulting from or associated with obesity.
  • the pharmaceutical preparation of the invention may be administered in combination with one or more antiobesity agents or appetite regulating agents.
  • agents may be selected from the group consisting of CART (cocaine amphetamine regulated transcript) agonists, NPY (neuropeptide Y) antagonists, MC4 (melano- cortin 4) agonists, MC3 (melanocortin 3) agonists, orexin antagonists, TNF (tumor necrosis factor) agonists, CRF (corticotropin releasing factor) agonists, CRF BP (corticotropin releas- ing factor binding protein) antagonists, urocortin agonists, ⁇ 3 adrenergic agonists such as CL-316243, AJ-9677, GW-0604, LY362884, LY377267 or AZ-40140, MSH (melanocyte- stimulating hormone) agonists, MCH (melanocyte-concentrating hormone
  • the antiobesity agent is leptin.
  • the antiobesity agent is dexamphetamine or amphetamine. In another embodiment the antiobesity agent is fenfluramine or dexfenfluramine.
  • the antiobesity agent is sibutramine.
  • the antiobesity agent is orlistat.
  • the antiobesity agent is mazindol or phentermine.
  • the antiobesity agent is phendimetrazine, diethylpropion, fluoxetine, bupropion, topiramate or ecopipam.
  • the orally active hypoglycemic agents comprise imidazolines, sulphonylureas, biguanides, meglitinides, oxadiazolidinediones, thiazolidinediones, insulin sensitizers, insulin secretagogues such as glimepride, ⁇ -glucosidase inhibitors, agents acting on the ATP- dependent potassium channel of the ⁇ -cells eg potassium channel openers such as those disclosed in WO 97/26265, WO 99/03861 and WO 00/37474 (Novo Nordisk A/S) which are incorporated herein by reference, or mitiglinide, or a potassium channel blocker, such as BTS-67582, nateglinide, glucagon antagonists such as those disclosed in WO 99/01423 and WO 00/39088 (Novo Nordisk A/S and Agouron Pharmaceuticals, Inc.), which are incorpo- rated herein by reference, GLP-1 agonists such as those
  • the pharmaceutical preparation of the invention is administered in combination with a sulphonylurea e.g. tolbutamide, chlorpropa- mide, tolazamide, glibenclamide, glipizide, glimepiride, glicazide or glyburide.
  • a sulphonylurea e.g. tolbutamide, chlorpropa- mide, tolazamide, glibenclamide, glipizide, glimepiride, glicazide or glyburide.
  • the pharmaceutical preparation of the invention is administered in combination with a biguanide, e.g. metformin.
  • a biguanide e.g. metformin.
  • the pharmaceutical preparation of the invention is administered in combination with a meglitinide eg repaglinide or nateglinide.
  • the pharmaceutical preparation of the invention is administered in combination with a thiazolidinedione insulin sensitizer, e.g. trogli- tazone, ciglitazone, pioglitazone, rosiglitazone, isaglitazone, darglitazone, englitazone, CS- 011/CI-1037 or T 174 or the compounds disclosed in WO 97/41097, WO 97/41119, WO 97/41120, WO 00/41121 and WO 98/45292 (Dr. Reddy's Research Foundation), which are incorporated herein by reference.
  • the pharmaceutical preparation of the invention may be administered in combination with an insulin sensitizer, e.g. such as GI 262570, YM-440, MCC-555, JTT-501, AR-H039242, KRP-297, GW-409544, CRE-16336, AR-H049020, LY510929, MBX-102, CLX-0940, GW-501516 or the compounds disclosed in WO 99/19313, WO 00/50414, WO 00/63191 , WO 00/63192, WO 00/63193 (Dr.
  • an insulin sensitizer e.g. such as GI 262570, YM-440, MCC-555, JTT-501, AR-H039242, KRP-297, GW-409544, CRE-16336, AR-H049020, LY510929, MBX-102, CLX-0940, GW-501516 or the compounds disclosed in WO 99/19313, WO 00/50414,
  • the pharmaceutical preparation of the in- vention is administered in combination with an ⁇ -glucosidase inhibitor, e.g. voglibose, emigli- tate, miglitol or acarbose.
  • an ⁇ -glucosidase inhibitor e.g. voglibose, emigli- tate, miglitol or acarbose.
  • the pharmaceutical preparation of the invention is administered in combination with an agent acting on the ATP-dependent potassium channel of the ⁇ -cells, e.g. tolbutamide, glibenclamide, glipizide, glicazide, BTS-67582 or re- paglinide.
  • an agent acting on the ATP-dependent potassium channel of the ⁇ -cells e.g. tolbutamide, glibenclamide, glipizide, glicazide, BTS-67582 or re- paglinide.
  • the pharmaceutical preparation of the invention may be administered in combination with nateglinide.
  • the pharmaceutical preparation of the invention is administered in combination with an antilipidemic agent, e.g. cholestyramine, colestipol, clofibrate, gemfibrozil, lovastatin, pravastatin, simvastatin, probucol or dextrothyroxine.
  • an antilipidemic agent e.g. cholestyramine, colestipol, clofibrate, gemfibrozil, lovastatin, pravastatin, simvastatin, probucol or dextrothyroxine.
  • the pharmaceutical preparation of the invention is administered in combination with more than one of the above-mentioned compounds, e.g. in combination with metformin and a sulphonylurea such as glyburide; a sulphonylurea and acarbose; nateglinide and metformin; acarbose and metformin; a sulphonylurea, metformin and troglitazone; metformin and a sulphonylurea; etc.
  • metformin and a sulphonylurea such as glyburide
  • a sulphonylurea and acarbose such as glyburide
  • a sulphonylurea and acarbose such as glyburide
  • a sulphonylurea and acarbose such as glyburide
  • the pharmaceutical preparation of the invention may be administered in combination with one or more antihypertensive agents.
  • antihypertensive agents are ⁇ -blockers such as alprenolol, atenolol, timolol, pindolol, propranolol and metoprolol, ACE (angiotensin converting enzyme) inhibitors such as benazepril, captopril, enalapril, fosinopril, lisinopril, quinapril and ramipril, calcium channel blockers such as nifedipine, felodipine, nicardipine, isradipine, nimodipine, diltiazem and verapamil, and ⁇ - blockers such as doxazosin, urapidil, prazosin and terazosin.
  • ⁇ -blockers such as alprenolol, atenolol, timolo
  • the pharmaceutical preparation of the invention may also be combined with NEP inhibitors such as candoxatril. Further reference can be made to Remington: The Science and Practice of Pharmacy, 19 th Edition, Gennaro, Ed., Mack Publishing Co., Easton, PA, 1995.

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Abstract

Novel ligands for the HisB10 Zn2+ sites of the R-state insulin hexamer that are capable of prolonging the action of insulin preparations are disclosed.

Description

PHARMACEUTICAL PREPARATIONS COMPRISING ACID-STABILISED INSULIN.
FIELD OF THE INVENTION
The present invention discloses pharmaceutical preparations comprising ligands for the HisBlO Zn2+ sites of the R-state insulin hexamer and acid-stabilsed insulin analogues. The compositions release insulin slowly following subcutaneous injection.
BACKGROUND OF THE INVENTION
Insulin Allostery. The insulin hexamer is an allosteric protein that exhibits both positive and negative cooperativity and half-of-the-sites reactivity in ligand binding. This allosteric behav- iour consists of two interrelated allosteric transitions designated LA 0 and- LB 0> three inter- converting allosteric conformation states (eq. 1),
I A I B
L o L o
Figure imgf000002_0001
designated T6, T3R3, and R6 and two classes of allosteric ligand binding sites designated as the phenolic pockets and the His810 anion sites. These allosteric sites are associated only with insulin subunits in the R conformation. Insulin Hexamer Structures and Ligand Binding. The T- to R-transition of the insulin hexamer involves transformation of the first nine residues of the B chain from an extended conformation in the T-state to an α-helical conformation in the R-state. This coil-to-helix transition causes the N-terminal residue, PheB1, to undergo an ~ 30 A change in position. This conformational change creates hydrophobic pockets (the phenolic pockets) at the sub- unit interfaces (three in T3R3, and six in Re), and the new B-chain helices form 3-helix bun- dies (one in T3R3 and two in Re) with the bundle axis aligned along the hexamer three-fold symmetry axis. The His610 Zn2+ in each R3 unit is forced to change coordination geometry from octahedral to either tetrahedral (monodentate ligands) or pentahedral (bidentate ligands). Formation of the helix bundle creates a narrow hydrophobic tunnel in each R3 unit that extends from the surface -12 A down to the His810 metal ion. This tunnel and the His610 Zn2+ ion form the anion binding site.
Hexamer Ligand Binding and Stability of Insulin Formulations. The in vivo role of the T to R transition is unknown. However, the addition of allosteric ligands (e.g. phenol and chloride ion) to insulin preparations is widely used. Hexamerization is driven by coordination of Zn2+ at the His610 sites to give T6, and the subsequent ligand-mediated transition of T6 to T3R3 and to Re is known to greatly enhance the physical and chemical stability of the resulting formulations.
Ligand Binding and Long Acting Insulin Formulations. Although the conversion of T6 to T3R3 and Rβ improves the stability of the preparation, the rate of absorption following subcutaneous injection of a soluble hexameric preparation is not much affected by the addition of phenol and cloride.
Putative events following injection of a soluble hexameric preparation. The small molecule ligands initially diffuse away from the protein. The affinity of the ligands for insulin may help to slow this process. On the other hand, the affinity of Zn2+ for e.g. albumin and the large effective space available for diffusion of the lipophilic phenol will tend to speed up the separation. In about 10-15 minutes after injection, the distribution of insulin species in the subcutaneous tissue will roughly correspond to that of a zinc-free insulin preparation at the same dilution. Then, the equilibrium distribution of species at this point will determine the observed absorption rate. In this regimen, absorption rates vary between about 1 hour (for rapid-acting insulin analogues, such as Asp828 human insulin) and about 4 hours (Co3+-hexamer). Current Approaches Toward Slow Acting Insulins. The inherent limitation of the absorption half-life to about 4 hours for a soluble human insulin hexamer necessitates further modifications to obtain the desired protraction. Traditionally, this has been achieved by the use of preparations wherein the constituent insulin is in the form of a crystalline and/or amorphous precipitate. In this type of formulation, the dissolution of the precipitate in the subcutaneous depot becomes rate-limiting for the absorption. NPH and Ultralente belong to this category of insulin preparations where crystallization/precipitation is effected by the addition of protamine and excessive zinc ion, respectively. Another approach involves the use of insulin derivatives where the net charge is increased to shift the isoelectric point, and hence the pH of minimum solubility, from about 5.5 to the physiological range. Such preparations may be injected as clear solutions at slightly acidic pH. The subsequent adjustment of the pH to neutral induces crystallization/precipitation in the subcutaneous depot and dissolution again becomes rate-limiting for the absorption. Gly*21 Arg831 Arg832 human insulin belongs to this category of insulin analogues.
Most recently, a series of soluble insulin derivatives with a hydrophobic moiety covalently attached to the side chain of Lys829 have been synthesized. These derivatives may show prolonged action profile due to various mechanisms including albumin binding (e.g. B29-Nε- myristoyl-des(B30) human insulin), extensive protein self-association and/or stickiness (e.g. B29-Nε-(N-lithocholyl-γ-glutamyl)-des(B30) human insulin) induced by the attached hydro- phobic group.
SUMMARY OF THE INVENTION
The present invention provides a pharmaceutical preparation comprising ligands for the His610 Zn2+ sites of the R-state insulin hexamer, zinc ions and acid-stabilised insulin analogs. The preparations form clear solutions at slightly acidic pH. When the pH is adjusted towards neutral upon subcutaneous injection, the ligands work to stabilize hexamers and modify solubility in the neutral pH range. As a result, the preparations release insulin slowly following subcutaneous injection.
The invention furthermore provides a method of preparing ligands for the His810 Zn2+ sites of the R-state insulin hexamer comprising the steps of
• Identifying a starter compound that binds to the R-state HisB10-Zn2+ site
• optionally attaching a fragment consisting of 0 to 5 neutral α- or β-amino acids •attaching a fragment comprising 1 to 20 positively charged groups independently selected from amino or guanidino groups. The invention also provides a method of prolonging the action of an acid-stabilised insulin preparation which comprises adding a zinc-binding ligand of the invention to the acid- stabilised insulin preparation. The invention finally provides a method of treating type 1 or type 2 diabetes comprising administering to a patient in need thereof a theraputically effective amount of a pharmaceutical preparation of the invention.
DEFINITIONS
The following is a detailed definition of the terms used to describe the invention: "Halogen" designates an atom selected from the group consisting of F, Cl, Br and I. The term "CrCe-alkyl" as used herein represents a saturated, branched or straight hydrocarbon group having from 1 to 6 carbon atoms. Representative examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, butyl, isobutyl, sec-butyl, tetf-butyl, n-pentyl, isopentyl, neopentyl, ferf-pentyl, n-hexyl, isohexyl and the like.
The term "CrCe-alkylene" as used herein represents a saturated, branched or straight bivalent hydrocarbon group having from 1 to 6 carbon atoms. Representative examples include, but are not limited to, methylene, 1 ,2-ethylene, 1 ,3-propylene, 1 ,2-propylene, 1 ,4-butylene, 1 ,5- pentylene, 1 ,6-hexylene, and the like.
The term "C2-C6-alkenyl" as used herein represents a branched or straight hydrocarbon group having from 2 to 6 carbon atoms and at least one double bond. Examples of such groups include, but are not limited to, vinyl, 1-propenyl, 2-propenyl, iso-propenyl, 1,3-buta- dienyl, 1-butenyl, 2-butenyl, 3-butenyl, 2-methyl-1-propenyl, 1-pentenyl, 2-pentenyl, 3- pentenyl, 4-pentenyl, 3-methyl-2-butenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 2,4-hexadienyl, 5- hexenyl and the like. The term "C2-C6-alkynyl" as used herein represents a branched or straight hydrocarbon group having from 2 to 6 carbon atoms and at least one triple bond. Examples of such groups include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, 1-pentynyl, 2-pentynyl, 3-pentynyl, 4-pentynyl, 1-hexynyl, 2-hexynyl, 3-hexynyl, 4- hexynyl, 5-hexynyl, 2,4-hexadiynyl and the like. The term "d-Ce-alkoxy" as used herein refers to the radical -O-d-Cβ-alkyl, wherein d-Ce-alkyl is as defined above. Representative examples are methoxy, ethoxy, n-propoxy, isopropoxy, butoxy, sec-butoxy, terf-butoxy, pentoxy, isopentoxy, hexoxy, isohexoxy and the like. The term "C3-C8-cycloalkyl" as used herein represents a saturated, carbocyclic group having from 3 to 8 carbon atoms. Representative examples are cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl and the like. The term
Figure imgf000005_0001
as used herein represents a non-aromatic, carbocyclic group having from 4 to 8 carbon atoms containing one or two double bonds. Representative examples are 1-cyclopentenyl, 2-cyclopentenyl, 3-cyclopentenyl, 1-cyclohexenyl, 2-cyclohexenyl, 3- cyclohexenyl, 2-cycloheptenyl, 3-cycloheptenyl, 2-cyclooctenyl, 1 ,4-cyclooctadienyl and the like. The term "heterocyclyl" as used herein represents a non-aromatic 3 to 10 membered ring containing one or more heteroatoms selected from nitrogen, oxygen and sulphur and optionally containing one or two double bonds. Representative examples are pyrrolidinyl, piperidyl, piperazinyl, morpholinyl, thiomorpholinyl, aziridinyl, tetrahydrofuranyl and the like. The term "aryl" as used herein is intended to include carbocyclic, aromatic ring systems such as 6 membered monocyclic and 9 to 14 membered bi- and tricyclic, carbocyclic, aromatic ring systems. Representative examples are phenyl, biphenylyl, naphthyl, anthracenyl, phe- nanthrenyl, fluorenyl, indenyl, azulenyl and the like. Aryl is also intended to include the partially hydrogenated derivatives of the ring systems enumerated above. Non-limiting examples of such partially hydrogenated derivatives are 1,2,3,4-tetrahydronaphthyl, 1,4- dihydronaphthyl and the like. The term "arylene" as used herein is intended to include divalent, carbocyclic, aromatic ring systems such as 6 membered monocyclic and 9 to 14 membered bi- and tricyclic, divalent, carbocyclic, aromatic ring systems. Representative examples are phenylene, biphenylylene, naphthylene, anthracenylene, phenanthrenylene, fluorenylene, indenylene, azulenylene and the like. Arylene is also intended to include the partially hydrogenated derivatives of the ring systems enumerated above. Non-limiting examples of such partially hydrogenated derivatives are 1,2,3,4-tetrahydronaphthylene, 1 ,4-dihydronaphthylene and the like. The term "aryloxy" as used herein denotes a group -O-aryl, wherein aryl is as defined above. The term "aroyl" as used herein denotes a group -C(O)-aryl, wherein aryl is as defined above. The term "heteroaryl" as used herein is intended to include aromatic, heterocyclic ring systems containing one or more heteroatoms selected from nitrogen, oxygen and sulphur such as 5 to 7 membered monocyclic and 8 to 14 membered bi- and tricyclic aromatic, heterocyclic ring systems containing one or more heteroatoms selected from nitrogen, oxygen and sulphur. Representative examples are furyl, thienyl, pyrrolyl, pyrazolyl, 3-oxopyrazolyl, oxazolyl, thiazolyl, imidazolyl, isoxazoiyl, isothiazolyl, 1 ,2,3-triazolyl, 1,2,4-triazolyl, pyranyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, 1 ,2,3-triazinyl, 1 ,2,4-triazinyl, 1,3,5- triazinyl, 1,2,3- oxadiazolyl, 1 ,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1 ,3,4-oxadiazolyl, 1 ,2,3-thiadiazolyl, 1,2,4- thiadiazolyl, 1 ,2,5-thiadiazolyl, 1 ,3,4-thiadiazolyl, tetrazolyl, thiadiazinyl, indolyl, isoindolyl, benzofuryl, benzothienyl, indazolyl, benzimidazolyl, benzthiazolyl, benzisothiazolyl, benzoxazolyl, benzisoxazolyl, purinyl, quinazolinyl, quinolizinyl, quinolinyl, isoquinolinyl, quinoxalinyl, naphthyridinyl, pteridinyl, carbazolyl, azepinyl, diazepinyl, acridinyl, thiazolidinyl, 2- thiooxothiazolidinyl and the like. Heteroaryl is also intended to include the partially hydrogenated derivatives of the ring systems enumerated above. Non-limiting examples of such partially hydrogenated derivatives are 2,3-dihydrobenzofuranyl, pyrrolinyl, pyrazolinyl, indolinyl, oxazolidinyl, oxazolinyl, oxazepinyl and the like.
The term "heteroarylene" as used herein is intended to include divalent, aromatic, heterocyclic ring systems containing one or more heteroatoms selected from nitrogen, oxygen and sulphur such as 5 to 7 membered monocyclic and 8 to 14 membered bi- and tricyclic aromatic, heterocyclic ring systems containing one or more heteroatoms selected from nitrogen, oxy- gen and sulphur. Representative examples are furylene, thienylene, pyrrolylene, oxa- zolylene, thiazolylene, imidazolylene, isoxazolylene, isothiazolylene, 1 ,2,3-triazolylene, 1 ,2,4- triazolylene, pyranylene, pyridylene, pyridazinylene, pyrimidinylene, pyrazinylene, 1 ,2,3- triazinylene, 1 ,2,4-triazinylene, 1 ,3,5- triazinylene, 1 ,2,3-oxadiazolylene, 1 ,2,4-oxadiazolylene, 1 ,2,5-oxadiazolylene, 1 ,3,4-oxadiazolylene, 1 ,2,3-thiadiazolylene, 1 ,2,4-thiadiazolylene, 1,2,5- thiadiazolylene, 1 ,3,4-thiadiazolylene, tetrazolylene, thiadiazinylene, indolylene, isoindolylene, benzofurylene, benzothienylene, indazolylene, benzimidazolylene, benzthiazolylene, ben- zisothiazolylene, benzoxazolylene, benzisoxazolylene, purinylene, quinazolinylene, quinoliz- inylene, quinolinylene, isoquinolinylene, quinoxalinylene, naphthyridinylene, pteridinylene, carbazolylene, azepinylene, diazepinylene, acridinylene and the like. Heteroaryl is also in- tended to include the partially hydrogenated derivatives of the ring systems enumerated above. Non-limiting examples of such partially hydrogenated derivatives are 2,3-dihydro- benzofuranylene, pyrrolinylene, pyrazolinylene, indolinylene, oxazolidinylene, oxazolinylene, oxazepinylene and the like. The term "ArG1" as used herein is intended to include an aryl or arylene radical as applicable, where aryl or arylene are as defined above but limited to phenyl, biphenylyl, naphthyl, anthra- cenyl, phenanthrenyl, fluorenyl, indenyl, and azulenyl as well as the corrresponding divalent radicals.
The term "ArG2" as used herein is intended to include an aryl or arylene radical as applicable, where aryl or arylene are as defined above but limited to phenyl, biphenylyl, naphthyl, fluo- renyl, and indenyl, as well as the corrresponding divalent radicals.
The term "Het1" as used herein is intended to include a heteroaryl or heteroarylene radical as applicable, where heteroaryl or heteroarylene are as defined above but limited to furyl, thienyl, pyrrolyl, pyrazolyl, 3-oxopyrazolyl, oxazolyl, thiazolyl, imidazolyl, isoxazoiyl, isothiazolyl, 1 ,2,3-triazolyl, 1,2,4-triazolyl, pyranyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, 1 ,2,3- triazinyl, 1,2,4-triazinyl, 1,3,5- triazinyl, 1,2,3-oxadiazolyl, 1 ,2,4-oxadiazolyl, 1 ,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,3-thiadiazolyl, 1 ,2,4-thiadiazolyl, 1 ,2,5-thiadiazolyl, 1 ,3,4-thiadiazolyl, tetrazolyl, thiadiazinyl, indolyl, isoindolyl, benzofuryl, benzothienyl, indazolyl, benzimidazolyl, benzthiazolyl, benzisothiazolyl, benzoxazolyl, benzisoxazolyl, purinyl, quinazolinyl, quinolizinyl, quinolinyl, isoquinolinyl, quinoxalinyl, naphthyridinyl, pteridinyl, carbazolyl, azepinyl, di- azepinyl, acridinyl, thiazolidinyl, 2-thiooxothiazolidinyl, as well as the corrresponding divalent radicals.
The term "Het2" as used herein is intended to include a heteroaryl or heteroarylene radical as applicable, where heteroaryl or heteroarylene are as defined above but limited to furyl, thienyl, pyrrolyl, pyrazolyl, 3-oxopyrazolyl, oxazolyl, thiazolyl, imidazolyl, isoxazoiyl, isothiazolyl, 1 ,2,3-triazolyl, 1 ,2,4-triazolyl, pyranyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, 1,2,3- triazinyl, 1,2,4-triazinyl, 1,3,5- triazinyl, 1,2,3-oxadiazolyl, 1 ,2,4-oxadiazolyl, 1 ,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1 ,2,5-thiadiazolyl, 1 ,3,4-thiadiazolyl, tetrazolyl, thiadiazinyl, indolyl, isoindolyl, benzofuryl, benzothienyl, benzimidazolyl, benzthiazolyl, benzisothiazolyl, benzoxazolyl, benzisoxazolyl, quinolinyl, isoquinolinyl, quinoxalinyl, carbazolyl, thiazolidinyl, 2-thiooxothiazolidinyl, as well as the corrresponding divalent radicals.
The term "Het3" as used herein is intended to include a heteroaryl or heteroarylene radical as applicable, where heteroaryl or heteroarylene are as defined above but limited to furyl, thienyl, pyrrolyl, pyrazolyl, 3-oxopyrazolyl, oxazolyl, thiazolyl, imidazolyl, isoxazoiyl, isothiazolyl, 1 ,2,3-triazolyl, 1,2,4-triazolyl, pyridyl, tetrazolyl, indolyl, isoindolyl, benzofuryl, benzothienyl, benzimidazolyl, benzthiazolyl, benzisothiazolyl, benzoxazolyl, benzisoxazolyl, quinolyl, isoqui- nolyl, quinoxalinyl, carbazolyl, thiazolidinyl, 2-thiooxothiazolidinyl, as well as the corrresponding divalent radicals. "Aryl-d-Ce-alkyl", "heteroaryl-d-C6-alkyl", "aryl-C2-C6-alkenyr etc. is intended to mean d-Ce- alkyl or C2-C6-alkenyl as defined above, substituted by an aryl or heteroaryl as defined above, for example:
Figure imgf000008_0001
The term "optionally substituted" as used herein means that the groups in question are either unsubstituted or substituted with one or more of the substituents specified. When the groups in question are substituted with more than one substituent the substituents may be the same or different.
Certain of the above defined terms may occur more than once in the structural formulae, and upon such occurrence each term shall be defined independently of the other. Furthermore, when using the terms "independently are" and "independently selected from" it should be understood that the groups in question may be the same or different. The terms "treatment" and "treating" as used herein means the management and care of a patient for the purpose of combating a disease, disorder or condition. The term is intended to include the delaying of the progression of the disease, disorder or condition, the alleviation or relief of symptoms and complications, and/or the cure or elimination of the disease, disorder or condition. The patient to be treated is preferably a mammal, in particular a human being. The term "fragment" as used herein is intended to mean a bivalent chemical group The term "Neutral amino acid" as used herein is intended to mean any natural (codable) and non-natural amino acid, including α- or β-aminocarboxylic acids, including D-isomers of these (when applicable) without charges at physiologically relevant pH in the side chain, such as glycine, alanine, β-alanine, valine, leucine, isoleucine, phenylalanine, tyrosine, aspargine, glutamine, cysteine, methionine, 3-aminobenzoic acid, 4-aminobenzoic acid or the like. The term "positively charged group" as used herein is intended to mean any pharmaceutically acceptable group that contains a positive charge at physiologically relevant pH, such as amino (primary, secondary and tertiary), ammonium and guanidino groups. The term "a amino acid" as used herein is intended to mean mean any natural (codable) and non-natural α-aminocarboxylic acid, including D-isomers of these.
The term "β amino acid" as used herein is intended to mean any β-aminocarboxylic acid, such as β-alanine, isoserine or the like.
The term "desB30" as used herein is intended to mean meant a natural insulin B chain or an analogue thereof lacking the B30 amino acid residue. The amino acid residues are indicated in the three letter amino acid code or the one letter amino code.
The terms "B1", "A1" and the like as used herein is intended to mean the amino acid residue in position 1 in the B chain of insulin or analogue thereof (counted from the N-terminal end) and the amino acid residue in position 1 in the A chain of insulin or analogue thereof (counted from the N-terminal end), respectively.
When in the specification or claims mention is made of groups of compounds such as car- boxylates, dithiocarboxylates, phenolates, thiophenolates, alkylthiolates, sulfonamides, irni- dazoles, triazoles, 4-cyano-1 ,2,3-triazoles, benzimidazoles, benzotriazoles, purines, thia- zolidinediones, tetrazoles, 5-mercaptotetrazoles, rhodanines, N-hydroxyazoles, hydantoines, thiohydantoines, naphthoic acids and salicylic acids, these groups of compounds are intended to include also derivatives of the compounds from which the groups take their name.
The term acid-stabilised insulin as used herein refers to an insulin analog that does not deamidate or dimerize at pH values below 7. Specifically, the analog cannot have Asn or Asp as a C-terminal residue.
The term human insulin as used herein refers to naturally produced insulin or recombinantly produced insulin. Recombinant human insulin may be produced in any suitable host cell, for example the host cells may be bacterial, fungal (including yeast), insect, animal or plant cells. The expression "insulin derivative" as used herein (and related expressions) refers to human insulin or an analogue thereof in which at least one organic substituent is bound to one or more of the amino acids. By "analogue of human insulin" as used herein (and related expressions) is meant human insulin in which one or more amino acids have been deleted and/or replaced by other amino acids, including non-codeable amino acids, or human insulin comprising additional amino acids, i.e. more than 51 amino acids, such that the resulting analogue possesses insulin activity.
The term "phenolic compound" or similar expressions as used herein refers to a chemical compound in which a hydroxyl group is bound directly to a benzene or substituted benzene ring. Examples of such compounds include, but are not limited to, phenol, o-cresol, m-cresol and p-cresol.
The term "physiologically relevant pH" as used herein is intended to mean a pH of about 7.1 to 7.9.
Abbreviations:
4H3N 4-hydroxy-3-nitrobeπzoic acid
AcOH acetic acid BT Benzotriazol-5-oyl
DMF N,N-Dimethylformamide
DMSO Dimethylsulfoxide
DIC Diisopropylcarbodiimide
EDAC 1-ethyl-3-(3'-dimethylamino-propyl)carbodiimide, hydrochloride Fmoc 9/-/-Fluorene-9-ylmethoxycarbonyl
G. GIy Glycine
HOAt 1 -hydroxy-7-azabenzotriazole
HOBT 1 -Hydroxybenzotriazole . Lys Lysine NMP N-methyl-2-pyrrolidone
Pbf 2,2,4,6,7-pentamethyldihydrobenzofuran-5-sulfonyl
Pmc 2,2,5,7,8-pentamethylchroman-6-sulfonyl
R. Arg Arginine
TFA Trifluoroacetic acid
Abbreviations for non-natural amino acid residues:
Figure imgf000011_0001
BRIEF DESCRIPTION OF DRAWINGS
Fig. 1 : The effect of various concentrations of the ligand 4-[3-(2H-tetrazol-5-yl)carbazol-9- ylmethyl]benzoyl-Arg1 -NH2 on the pH-dependence of Gly*21, Asp828 insulin solubility. Fig. 2: The effect of a high concentrations of the ligand 4-[3-(2H-tetrazol-5-yl)carbazol-9- ylmethyl]benzoyl-Argi2-NH2on the pH-dependence of Gly*21 insulin solubility. Fig. 3: Disappearance from the subcutaneous depot (pig model) of insulin preparations. Curves a)-c) are Gly*21, Asp828 human insulin formulated with an excess concentration compared to Zn2+ of a) 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]butyryl- Arg3-NH2, b) 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]butyryl-Arg5-NH2 and c) 4-(2H-Tetrazol-5-yl)benzoyl-Abz-Gly2-Arg5-NH2. Curve d) is B29-Nε-myristoyl- des(B30) human insulin. Curves e) and f) are Gly*21 human insulin formulated with two dif- ferent excess concentrations compared to Zn2+ of 4-[3-(2H-Tetrazol-5-yl)carbazol-9- ylmethy I] benzoy l-Arg 1 -N H2.
Fig.4: 4H3N-assay. UV/vis spectra resulting from a titration of hexameric insulin with the compound 3-hydroxy-2-naphthoic acid in the presence of 4-hydroxy-3-nitrobenzoic acid (4H3N). Inserted in the upper right corner is the absorbance at 444nm vs. the concentration of ligand
Fig. 5: TZD-assay. Fluorescence spectra resulting from a titration of hexameric insulin with 5-(3-methoxybenzylidene)thiazolidine-2,4-dione in the presence of 5-(4-dimethylamino- benzylidene)thiazolidine-2,4-dione (TZD). Inserted in the upper right corner is the fluorescence at 460 nm vs. the concentration of ligand
DESCRIPTION OF THE INVENTION
The present invention is based on the discovery that the His810 Zn++ ligand binding sites of the R-state insulin hexamer can be used to obtain an insulin preparation having prolonged action designed for flexible injection regimes including once-daily, based on acid-stabillised insulins. The basic concept underlying the present invention involves reversible attachment of a ligand to the His810 Zn2+ site of the R-state hexamer. A suitable ligand binds to the hexamer metal site with one end while other moieties are covalently attachment to the other end. On this basis, prolonged action via modification of preparation solubility may be obtained in a number of ways. However, all cases involve the same point of protein-ligand attachment and the delivery of human insulin (or analogues or derivatives thereof) as the active species. Use of a acid-stabilized analog allows a stable, clear solution with ligand to be formulated at slightly acidic pH. Following subcutaneous injection, the pH is gradually adjusted towards neutral. As a result the ligand binds to and precipitates insulin in the subcutaneous tissue. The release of insulin analog into the blood stream is then limited by the rate of redissolution of the precipitate. Of particular advantage is the possibility of adjusting the amount of added ligand as well as the charge and affinity of the ligand. Variation of these parameters allows adjustment of the rate of dissolution following precipitation in the subcutis and hence the proportion of slow and fast acting analog in the formulationon. Hence formulations covering a wide range of re- lease rates may be prepared by this principle.
The anions currently used in insulin formulations as allosteric ligands for the R-state hexamers (notably chloride ion) bind only weakly to the His810 anion site. The present invention, which is based on the discovery of suitable higher affinity ligands for these anion sites, provides ligands which are extended to modify timing via changes in hexamer solubility as out- lined above.
Most ligand binding sites in proteins are highly asymmetric. Because the His810 Zn2+ sites reside on the three-fold symmetry axis, these sites posses a symmetry that is unusual, but not unique. Several other proteins have highly symmetric ligand binding sites. The His810 Zn + site consists of a tunnel or cavity with a triangular-shaped cross-section that extends -12 A from the surface of the hexamer down to the His810 Zn2+ ion. The diameter of the tunnel varies along its length and, depending on the nature of the ligand occupying the site, the opening can be capped over by the Asn83 and Phe81 side chains. The walls of the tunnel are made up of the side chains of the amino acid residues along one face each of the three α-helices. The side chains from each helix that make up the lining of the tunnel are PheB1, Asn83, and Leu86. Therefore, except for the zinc ion, which is coordinated to three His810 residues and is positioned at the bottom of the tunnel, the site is principally hydrophobic. Depending on the ligand structure, it may be possible for substituents on the ligand to make H-bonding interactions with Asn83 and with the peptide linkage to Cys87. The present invention originates from a search for compounds with suitable binding proper- ties by using UV-visible and fluorescence based competition assays described herein which are based on the displacement of chromophoric ligands from the R-state His810-Zn2+ site by the incoming ligand in question. These compounds will be referred to as "starter compounds" in the following. These assays are easily transformed into a high-throughput format capable of handling libraries constructed around hits from the initial search of compound databases.
These starter compounds provide the starting point for the task of constructing a chemical handle that allows for attachment of the positively charged fragment Frg2 (see below).
Thus, from the structure-activity relationship (SAR) information obtained from the binding as- say(s) it will be apparent for those skilled in the art to modify the starter compounds in question by introduction of a chemical group that will allow for coupling to a peptide containing e.g. one or more arginine or lysine residues. These chemical groups include carboxylic acid (amide bond formation with the peptide), carbaldehyde (reductive alkylation of the peptide), sulfonyl chloride (sulphonamide formation with the peptide) or the like.
The decision where and how to introduce this chemical group can be made in various ways. For example: From the SAR of a series of closely related starter compounds, a suitable position in the starter compound can be identified and the chemical group can be attached to this position, optionally using a spacer group, using synthesis procedures known to those skilled in the art.
Alternatively, this chemical group can be attached (optionally using a spacer group using and synthesis procedures known to those skilled in the art) to a position on the starter compound remote from the Zn2+-binding functionality.
The invention thus provides pharmaceutical preparation comprising
1. Acid-stabilised insulin
2. Zinc ions
3. A zinc-binding ligand of the following general formula (I)
CGr-Lnk-Frg1-Frg2-X (I)
wherein:
CGr is a chemical group which reversibly binds to a His810 Zn2+ site of an insulin hexamer; Lnk is a linker selected from
• a valence bond
• a chemical group G8 of the formula -B1-B2-C(O)-, -B1-B2-SO2-, -B1-B2-CH2-, or -B1- B2-NH-; wherein B1 is a valence bond, -O-, -S-, or -NR6-,
B2 is a valence bond, d-C18-alkylene, C2-C18-alkenylene, C2-C18-alkynylene, arylene, heteroarylene, -d-C18-alkyl-aryl-, -C2-C18-alkenyl-aryl-, -C2-C18-alkynyl-aryl-, -C(=O)- d-C18-alkyl-C(=O)-, -C(=O)-d-C18-alkenyl-C(=O)-,
Figure imgf000014_0001
alkyl-C(=O)-, -C(=O)- C1-C18-alkyl-S-Cι-C18-alkyl-C(=O)-, -C(=O)-C1-C18-alkyl-NRβ-C1- C18-alkyl-C(=O)-, -C(=O)-aryl-C(=O)-, -C(=O)-heteroaryl-C(=O)-; wherein the alkylene, alkenylene, and alkynylene moieties are optionally substituted by -CN, -CF3, -OCF3, -OR6, or -NR6R7 and the arylene and heteroarylene moieties are optionally substituted by halogen, -C(O)OR6, -C(O)H, OCOR6, -SO2, -CN, -CF3, - OCF3, -NO2, -OR6, -NR6R7, d-dβ-alkyl, or d-C18-alkanoyl; R6and R7 are independently H, Cι-C4-alkyl;
Frg1 is a fragment consisting of 0 to 5 neutral α- or β-amino acids
Frg2 is a fragment comprising 1 to 20 positively charged groups independently selected from amino or guanidino groups; and
X is -OH, -NH2 or a diamino group,
or a salt thereof with a pharmaceutically acceptable acid or base, or any optical isomer or mixture of optical isomers, including a racemic mixture, or any tautomeric forms.
In one embodiment CGr is a chemical structure selected from the group consisting of car- boxylates, dithiocarboxylates, phenolates, thiophenolates, alkylthiolates, sulfonamides, imi- dazoles, triazoles, 4-cyano-1 ,2,3-triazoles, benzimidazoles, benzotriazoles, purines, thia- zolidinediones, tetrazoles, 5-mercaptotetrazoles, rhodanines, N-hydroxyazoles, hydantoines, thiohydantoines, barbiturates, naphthoic acids and salicylic acids.
In another embodiment CGr is a chemical structure selected from the group consisting of benzotriazoles, 3-hydroxy 2-napthoic acids, salicylic acids, tetrazoles, thiazolidinediones, 5- mercaptotetrazoles, or 4-cyano-1 ,2,3-triazoles. In another embodiment CGr is
Figure imgf000015_0001
wherein X is =O, =S or =NH Y is -S-, -O- or -NH-
R1 and R4 are independently selected from hydrogen or CrCe-alkyl,
R2 is hydrogen or d-C6-alkyl or aryl, R1 and R2 may optionally be combined to form a double bond,
R3 and R5 are independently selected from hydrogen, halogen, aryl, d-Cβ-alkyl, or -C(O)NR11R12,
A and B are independently selected from d-C6-alkylene, arylene, aryl-d-C6-alkyl-, aryl-C - Ce-alkenyl- or heteroarylene, wherein the alkylene or alkenylene is optionally substituted with one or more substituents independently selected from R6 and the arylene or heteroarylene is optionally substituted with up to four substituents R7, R8, R9, and R10,
A and R3 may be connected through one or two valence bonds, B and R5 may be connected through one or two valence bonds, R6 is independently selected from halogen, -CN, -CF3, -OCF3, aryl, -COOH and -NH2,
R7, R8, R9 and R10 are independently selected from
• hydrogen, halogen, -CN, -CH2CN, -CHF2, -CF3, -OCF3, -OCHF2, -OCH2CF3, -OCF2CHF2, -S(O)2CF3, -OS(O)2CF3, -SCF3, -NO2, -OR11, -NR11R12, -SR11, -NR11S(O)2R12, -S(O)2NR R12, -S(O)NR11R12, -S(O)R11, -S(O)2R11, -OS(O)2 R11,
-C(O)NR11R12, -OC(O)NR11R12, -NR11C(O)R12, -CH2C(O)NR11R12,
-Od-C6-alkyl-C(O)NR11R12, -CH2OR11, -CH2OC(O)R11, -CH2NR11R12, -OC(0)R11, -Od-C15-alkyl-C(O)OR11, -Od-Cβ-alkyl-OR11, -Sd-C6-alkyl-C(O)OR11 -C2-C6-alkenyl-C(=O)OR11 , -NR11-C(=O)-d-C6-alkyl-C(=O)OR11 , -NR11-C(=O)-d-C6-alkenyl-C(=O)OR11 , -C(O)OR11, C(O)R11, or -C2-Cβ-alkenyl-
C(=O)R11, =O, or -C2-C6-alkenyl-C(=O)-NR11R12, • d-Ce-alkyl, C2-C6-alkenyl or C2-C6-alkynyl, each of which may optionally be substituted with one or more substituents independently selected from R13,
• aryl, aryloxy, aryloxycarbonyl, aroyl, arylsulfanyl, aryl-d-C6-alkoxy, aryl-d-C6-alkyl, aryl-C2-C6-alkenyl, aroyl-C2-C6-alkenyl, aryl-C2-C6-alkynyl, heteroaryl, heteroaryl-d- C6-alkyl, heteroaryl-C2-C6-alkenyl, heteroaryl-C2-C6-alkynyl, or C3-C6 cycloalkyl,
of which each cyclic moiety may optionally be substituted with one or more substituents independently selected from R14,
R11 and R12 are independently selected from hydrogen, OH, C1-C20-alkyl, aryl-d-C6-alkyl or aryl, wherein the alkyl groups may optionally be substituted with one or more substituents independently selected from R15, and the aryl groups may optionally be substituted one or more substituents independently selected from R16; R11 and R12 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
R13 is independently selected from halogen, -CN, -CF3, -OCF3, -OR11, -C(O)OR11 , -NR1 R12, and -C(O)NR11R12,
R14 is independently selected from halogen, -C(O)OR11, -CH2C(O)OR11, -CH2OR11, -CN, - CF3, -OCF3, -NO2, -OR11, -NR11R12, S(O)2R11, aryl and C C6-alkyl,
R15 is independently selected from halogen, -CN, -CF3, -OCF3, -Od-C6-alkyl, -C(O)OC Ce- alkyl, -COOH and -NH2,
R16 is independently selected from halogen, -C(O)OC Ce-alkyl, -COOH, -CN, -CF3, -OCF3, - NO2, -OH, -OCι-C6-alkyl, -NH2, C(=O) or CrCe-alkyl, or any enantiomer, diastereomer, in- eluding a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base.
In another embodiment X is =O or =S. In another embodiment X is =O. In another embodiment X is =S. In another embodiment Y is -O- or -S-. In another embodiment Y is -O-. In another embodiment wherein Y is -S-.
In another embodiment Crg is arylene optionally substituted with up to four substituents, R7, R8, R9, and R10 which may be the same or different.
In another embodiment A is selected from ArG1 optionally substituted with up to four substituents, R7, R8, R9, and R10 which may be the same or different.
In another embodiment A is phenylene or naphtylene optionally substituted with up to four substituents, R7, R8, R9, and R10 which may be the same or different. In another embodiment A is
Figure imgf000017_0001
In another embodiment A is phenylene.
In another embodiment A is heteroarylene optionally substituted with up to four substituents,
R7, R8, R9, and R 0 which may be the same or different. In another embodiment A is selected from Het1 optionally substituted with up to four substituents, R7, R8, R9, and R10 which may be the same or different.
In another embodiment A is selected from Het2 optionally substituted with up to four substituents, R7, R8, R9, and R10 which may be the same or different. In another embodiment A is selected from Het3 optionally substituted with up to four sub- stituents, R7, R8, R9, and R10 which may be the same or different.
In another embodiment A is selected from the group consisting of indolylene, benzofu- ranylidene, quinolylene, furylene, thienylene, or pyrrolylene, wherein each heteroaryl may optionally substituted with up to four substituents, R7, R8, R9, and R10 which may be the same or different. In another embodiment A is benzofuranylene optionally substituted with up to four substituents R7, R8, R9, and R10 which may be the same or different. In another embodiment A is
Figure imgf000017_0002
In another embodiment A is carbazolylidene optionally substituted with up to four substituents R7, R8, R9, and R10 which may be the same or different. In another embodiment A is
Figure imgf000018_0001
In another embodiment A is quinolylidene optionally substituted with up to four substituents R7, R8, R9, and R10 which may be the same or different. In another embodiment A is
Figure imgf000018_0002
In another embodiment A is indolylene optionally substituted with up to four substituents R7, R , R , and R -10 which may be the same or different. In another embodiment A is
Figure imgf000018_0003
In another embodiment R1 is hydrogen.
In another embodiment R2 is hydrogen.
In another embodiment R1 and R2 are combined to form a double bond.
In another embodiment R3 is d-Ce-alkyl, halogen, or C(O)NR16R17.
In another embodiment R3 is d-C6-alkyl or C(O)NR16R17.
In another embodiment R3 is methyl.
In another embodiment B is phenylene optionally substituted with up to four substituents, R7,
R8, R9, and R10 which may be the same or different.
In another embodiment R4 is hydrogen.
In another embodiment R5 is hydrogen.
In another embodiment R6 is aryl. In another embodiment R6 is phenyl.
In another embodiment R7, R8, R9 and R10 are independently selected from
• hydrogen, halogen, -NO2, -OR11, -NR11R12, -SR11, -NR11S(O)2R12, -S(O)2NR11R12, -S(O)NR11R12, -S(O)R11, -S(O)2R11, -OS(O)2 R11, -NR11C(O)R12, -CH2OR11, -
CH2OC(O)R11, -CH2NR11R12, -OC(O)R11, -Od-C6-alkyl-C(O)OR11, -OCrC6- alkyl-C(O)NR11R12, -Od-Ce-alkyl-OR11, -Sd-C6-alkyl-C(O)OR11, -C2-C6-alkenyl- C(=O)OR11, -C(O)OR11, or -C2-C6-alkenyl-C(=O)R11,
• CrCe-alkyl, C2-C6-alkenyl or C2-C6-alkynyl, which may each optionally be substituted with one or more substituents independently selected from R13
• aryl, aryloxy, aroyl, arylsulfanyl, aryl-d-Ce-alkoxy, aryl-d-Ce-alkyl, aryl-C2- C6-alkenyl, aroyl-C2-C6-alkenyl, aryl-C2-C6-alkynyl, heteroaryl, heteroaryl-CrC6-alkyl, wherein each of the cyclic moieties optionally may be substituted with one or more substituents independently selected from R14
In another embodiment R7, R8, R9 and R10 are independently selected from
- hydrogen, halogen, -NO2, -OR11, -NR11R12, -SR11, -S(O)2R11, -OS(O)2 R11, -
CH2OC(O)R11, -OC(O)R11, -Od-C6-alkyl-C(O)OR11, -Od-Ce-alkyl-OR11, -Sd-C6- alkyl-C(O)OR11, -C(O)OR11, or -C2-C6-alkenyl-C(=O)R11,
• CrCe-alkyl or CrC6-alkenyl which may each optionally be substituted with one or more substituents independently selected from R13
• aryl, aryloxy, aroyl, aryl-d-Ce-alkoxy, aryl-d-Ce-alkyl, heteroaryl,
of which each of the cyclic moieties optionally may be substituted with one or more substituents independently selected from R14
In another embodiment R7, R8, R9 and R 0 are independently selected from • hydrogen, halogen, -NO2, -OR11, -NR11R12, -SR11, -S(O)2R11, -0S(O)2 R1Λ - CH2OC(O)R11, -OC(O)R11, -OCrC6-alkyl-C(O)OR11, -Od-Ce-alkyl-OR11, -SC4-C6- alkyl-C(O)OR11, -C(O)OR11, or -C2-C6-alkenyl-C(=O)R11,
-CrCe-alkyl or C C6- which may each optionally be substituted with one or more substituents independently selected from R13
• aryl, aryloxy, aroyl, aryl-d-Ce-alkoxy, aryl-d-Ce-alkyl, heteroaryl,
of which each of the cyclic moieties optionally may be substituted with one or more substituents independently selected from R14.
In another embodiment R7, R8, R9 and R10 are independently selected from
• hydrogen, halogen, -OR11, -Od-C6-alkyl-C(O)OR11, or -C(O)OR11,
• CrCe-alkyl which may each optionally be substituted with one or more substituents independently selected from R13
• aryl, aryloxy, aryl-d-Ce-alkoxy,
of which each of the cyclic moieties optionally may be substituted with one or more substituents independently selected from R 4.
In another embodiment R7, R8, R9 and R10 are independently selected from • hydrogen, halogen, -OR11, -OCrC6-alkyl-C(O)OR11, or -C(O)OR11,
• d-Ce-alkyl which may optionally be substituted with one or more substituents independently selected from R 3
• phenyl, phenyloxy, phenyl-CrC6-alkoxy, wherein each of the cyclic moieties optionally may be substituted with one or more substituents independently selected from R14.
In another embodiment R11 and R12 are independently selected from hydrogen, CrC20-alkyl, aryl or aryl-d-Ce-alkyl, wherein the alkyl groups may optionally be substituted with one or more substituents independently selected from R15, and the aryl groups may optionally be substituted one or more substituents independently selected from R16; R11 and R12 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds.
In another embodiment R1 and R12 are independently selected from hydrogen, CrC20-alkyl, aryl or aryl-d-Ce-alkyl, wherein the alkyl groups may optionally be substituted with one or more substituents independently selected from R15, and the aryl groups may optionally be substituted one or more substituents independently selected from R16.
In another embodiment R11 and R12 are independently selected from phenyl or phenyl-CrC6- alkyl.
In another embodiment R11 and R12 are methyl.
In another embodiment R13 is independently selected from halogen, CF3, OR11 or NR11R12. In another embodiment R13 is independently selected from halogen or OR11.
In another embodiment R13 is OR11.
In another embodiment R14 is independently selected from halogen, -C(O)OR11, -CN, -CF3, -
OR11, S(O)2R11, and d-C6-alkyl.
In another embodiment R14 is independently selected from halogen, -C(O)OR11, or -OR11. In another embodiment R15 is independently selected from halogen, -CN, -CF3, -C(O)Od-C6- alkyl.and -COOH.
In another embodiment R15 is independently selected from halogen or -C(O)OCrC6-alkyl.
In another embodiment R16 is independently selected from halogen, -C(O)OCrC6-alkyl,
-COOH, -NO2, -Od-Ce-alkyl, -NH2, C(=O) or d-Cβ-alkyl. In another embodiment R1β is independently selected from halogen, -C(O)OCrC6-alkyl,
-COOH, -NO2, or d-Cβ-alkyl.
In another embodiment CGr is
Figure imgf000021_0001
wherein
R19 is hydrogen or d-Cβ-alkyl, R20 is hydrogen or CrCe-alkyl, D and F are a valence bond or d-C6-alkylene optionally substituted with one or more substituents independently selected from R72,
R72 is independently selected from hydroxy, d-Cβ-alkyl, or aryl,
E is CrCe-alkylene, arylene or heteroarylene, wherein the arylene or heteroarylene is optionally substituted with up to three substituents R21, R22 and R23,
G is CrC6-alkylene, arylene or heteroarylene, wherein the arylene or heteroarylene is op- tionally substituted with up to three substituents R24, R25 and R26,
R17, R18, R21, R22, R23, R24, R25 and R26 are independently selected from
• hydrogen, halogen, -CN, -CH2CN, -CHF2, -CF3, -OCF3, -OCHF2, -OCH2CF3, -OCF2CHF2, -S(O)2CF3, -SCF3, -NO2, -OR27, -NR27R28, -SR27, -NR27S(O)2R28,
-S(O)2NR27R28, -S(O)NR27R28, -S(O)R27, -S(O)2R27, -C(O)NR27R28, -OC(O)NR27R28, -NR27C(O)R28, -NR27C(O)OR28, -CH2C(O)NR27R28, -OCH2C(O)NR27R28, -CH2OR27, -CH2NR27R28, -OC(O)R27, -OCrC6-alkyl-C(O)OR27, -SCrC6-alkyl-C(0)OR27, -C2-C6- alkenyl-C(=O)OR27, -NR27-C(=O)-d-Ce-alkyl-C(=O)OR27, -NR27-C(=O)-CrC6- alkenyl-C(=O)OR27, -C(=O)NR 7-CrC6-alkyl-C(=O)OR27, -d-C6-alkyl-C(=O)OR27,or
-C(O)OR27,
• CrCe-alkyl, C2-Cβ-alkenyl or C2-C6-alkynyl,
which may optionally be substituted with one or more substituents independently selected from R29,
• aryl, aryloxy, aryloxycarbonyl, aroyl, aryl-d-C6-alkoxy, aryl-d-Cβ-alkyl, aryl-C2- Cβ-alkenyl, aryl-C2-C6-alkynyl, heteroaryl, heteroaryl-d-Ce-alkyl, heteroaryl-C2-C6- alkenyl or heteroaryl-C2-C6-alkynyl,
of which the cyclic moieties optionally may be substituted with one or more substituents selected from R30, R27 and R28 are independently selected from hydrogen, d-Ce-alkyl, aryl-d-C6-alkyl or aryl, or R27 and R28 when attached to the same nitrogen atom together with the said nitrogen atom may form a 3 to 8 membered heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
R is independently selected from halogen, -CN, -CF3, -OCF3, -OR27, and -NR27R28,
R30 is independently selected from halogen, -C(O)OR27, -CN, -CF3, -OCF3, -NO2, -OR27, -NR27R28 and d-Ce-alkyl, or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base.
In another embodiment D is a valence bond.
In another embodiment D is d-Ce-alkylene optionally substituted with one or more hydroxy, d-Ce-alkyl, or aryl. In another embodiment E is arylene or heteroarylene, wherein the arylene or heteroarylene is optionally substituted with up to three substituents independently selected from R21, R22 and
R23.
In another embodiment E is arylene optionally substituted with up to three substituents independently selected from R21, R22 and R23. In another embodiment E is selected from ArG1 and optionally substituted with up to three substituents independently selected from R21, R22 and R23.
In another embodiment E is phenylene optionally substituted with up to three substituents independently selected from R21, R22 and R23.
In another embodiment CGr is
Figure imgf000023_0001
In another embodiment R , R and R are independently selected from
• hydrogen, halogen, -CHF2, -CF3, -OCF3, -OCHF2, -OCH2CF3, -OCF2CHF2, -SCF3, - NO2, -OR27, -NR 7R28, -SR27, -C(O)NR27R28, -OC(O)NR27R28, -NR27C(O)R28,
-NR27C(O)OR28, -CH2C(O)NR27R28, -OCH2C(O)NR27R28, -CH2OR27, -CH2NR27R28,
-OC(O)R27, -Od-Ce-alkyl-C(O)OR27, -Sd-C6-alkyl-C(O)OR27, -C2-C6-alkenyl- C(=O)OR27, -NR27-C(=O)-d-Cβ-alkyl-C(=O)OR27, -NR27-C(=O)-C C6- alkenyl-C(=O)OR27-, -C(=O)NR27-C C6-alkyl-C(=O)OR27, -CrC6-alkyl-C(=O)OR27, or -C(O)OR27,
•d-Ce-alkyl, C2-C6-alkenyl or C2-Cβ-alkynyl,
which may optionally be substituted with one or more substituents independently selected from R29
'aryl, aryloxy, aryloxycarbonyl, aroyl, aryl-d-Cβ-alkoxy, aryl-CrC6-alkyl, aryl-C2-
Cβ-alkenyl, aryl-C2-C6-alkynyl, heteroaryl, heteroaryl-d-Ce-alkyl, heteroaryl-C2-Cβ- alkenyl or heteroaryl-C2-C6-alkynyl,
of which the cyclic moieties optionally may be substituted with one or more substitu- ents selected from R30.
In another embodiment R2 , R22 and R23 are independently selected from
• hydrogen, halogen, -OCF3, -OR27, -NR27R28, -SR27, -NR27C(O)R28, -NR 7C(O)OR28, -OC(O)R27, -OCrC6-alkyl-C(O)OR27, -SCrC6-alkyl-C(O)OR27, -C2-C6-alkenyl-
C(=O)OR27, -C(=O)NR27-CrC6-alkyl-C(=O)OR27, -d-C6-alkyl-C(=O)OR27, or -C(O)OR27,
• d-Ce-alkyl optionally substituted with one or more substituents independently se- lected from R29
• aryl, aryloxy, aroyl, aryl-d-Cβ-alkoxy, aryl-CrC6-alkyl, heteroaryl, heteroaryl-d-Cβ- alkyl,
of which the cyclic moieties optionally may be substituted with one or more substituents selected from R30.
In another embodiment R21, R22 and R23 are independently selected from • hydrogen, halogen, -OCF3, -OR27, -NR27R28, -SR27, -NR27C(O)R28, -NR27C(0) R28, -OC(O)R27, -Od-Ce-alkyl-C(0)OR27, -SCrC6-alkyl-C(O)OR27, -C2-C6-all enyl- C(=O)OR27, -C(=O)NR27-d-C6-alkyl-C(=O)OR27, -CrC6-alkyl-C(=0)OR27, or -C(O)OR27,
• methyl, ethyl propyl optionally substituted with one or more substituents independently selected from R29
• aryl, aryloxy, aroyl, aryl-d-Ce-alkoxy, aryl-d-Ce-alkyl, heteroaryl, heteroaryl-d-Cβ- alkyl of which the cyclic moieties optionally may be substituted with one or more sub stitu- ents selected from R30. In another embodiment R21, R22 and R23 are independently selected from
•hydrogen, halogen, -OCF3, -OR27, -NR27R28, -SR27, -NR27C(O)R28, -NR27C(O)OR28,
-OC(O)R27, -OCrC6-alkyl-C(O)OR27, -SCrC6-alkyl-C(O)OR27, -C2-C6-alkenyl- C(=O)OR27, -C(=O)NR27-d-Ce-alkyl-C(=O)OR27, -d-C6-alkyl-C(=O)OR27, or -C(O)OR27,
• methyl, ethyl propyl optionally substituted with one or more substituents independently selected from R29
• ArG1 , ArG1-O-, ArG1-C(O)-, ArG1-CrC6-alkoxy, ArG1-d-C6-alkyl, Het3, Het3-d- Cβ-alkyl of which the cyclic moieties optionally may be substituted with one or more substituents selected from R30. In another embodiment R21, R22 and R23 are independently selected from
• hydrogen, halogen, -OCF3, -OR27, -NR27R28, -SR27, -NR27C(O)R28, -NR27C(O)OR28, -OC(O)R27, -Od-C6-alkyl-C(O)OR27, -Sd-C6-alkyl-C(O)OR27, -C2-C6-alkenyl-
C(=O)OR27, -C(=O)NR27-CrC6-alkyl-C(=O)OR27, -d-C6-alkyl-C(=O)OR27, or -C(O)OR27,
• CrCe-alkyl optionally substituted with one or more substituents independently se- lected from R29 • phenyl, phenyloxy, phenyl-d-Cβ-alkoxy, phenyl-CrC6-alkyl, of which the cyclic moieties optionally may be substituted with one or more substituents selected from R30. In another embodiment R19 is hydrogen or methyl. In another embodiment R19 is hydrogen. In another embodiment R27 is Hydrogen, d-C6-alkyl or aryl. In another embodiment R27 is hydrogen or d-Cβ-alkyl. In another embodiment R28 is hydrogen or d-Cβ-alkyl. In another embodiment F is a valence bond.
In another embodiment F is d-C6-alkylene optionally substituted with one or more hydroxy, d-Cβ-alkyl, or aryl.
In another embodiment G is d-Ce-alkylene or arylene, wherein the arylene is optionally substituted with up to three substituents R24, R25 and R26. In another embodiment G is CrCe-alkylene or ArG1 , wherein the arylene is optionally substituted with up to three substituents R24, R25 and R26. In another embodiment G is d-Cβ-alkylene.
In another embodiment G is phenylene optionally substituted with up to three substituents R24, R25 and R26. In another embodiment R24, R25 and R26 are independently selected from
• hydrogen, halogen, -CHF2, -CF3, -OCF3, -OCHF2, -OCH2CF3, -OCF2CHF2, -SCF3, - NO2, -OR27, -NR27R28, -SR27, -C(O)NR27R28, -OC(O)NR27R28, -NR27C(O)R28, -NR27C(O)OR28, -CH2C(O)NR27R28, -OCH2C(O)NR27R28, -CH2OR27, -CH2NR27R28, -OC(O)R27, -Od-C6-alkyl-C(O)OR27, -Sd-C6-alkyl-C(O)OR27, -C2-Ce-alkenyl-
C(=O)OR27, -NR27-C(=O)-CrC6-alkyl-C(=O)OR27, -NR27-C(=O)-d-C6- alkenyl-C(=O)OR27-, -C(=O)NR27-CrC6-alkyl-C(=O)OR27, -d-C6-alkyl-C(=O)OR27, or -C(O)OR27,
• d-Cβ-alkyl, C2-Cβ-alkenyl or C2-C6-alkynyl,
which may optionally be substituted with one or more substituents independently selected from R29 • aryl, aryloxy, aryloxycarbonyl, aroyl, aryl-d-Cβ-alkoxy, aryl-CrC6-alkyl, aι |-C2- C6-alkenyl, aryl-C2-C6-alkynyl, heteroaryl, heteroaryl-CrC6-alkyl, heteroaryl-C -C6- alkenyl or heteroaryl-C2-Cβ-alkynyl,
of which the cyclic moieties optionally may be substituted with one or more substituents selected from R30.
In another embodiment R24, R25 and R26 are independently selected from
• hydrogen, halogen, -OCF3, -OR27, -NR27R28, -SR27, -NR27C(O)R28, -NR27C(O)OR28,
-OC(0)R27, -OCrC6-alkyl-C(O)OR27, -SCrC6-alkyl-C(O)OR27, -C2-C6-alkenyl- C(=O)OR27, -C(=O)NR27-CrC6-alkyl-C(=O)OR27, -CrC6-alkyl-C(=O)OR27, or -C(O)OR27,
• d-Cβ-alkyl, C2-C6-alkenyl or C2-C6-alkynyl,
which may optionally be substituted with one or more substituents independently selected from R29
»aryl, aryloxy, aryloxycarbonyl, aroyl, aryl-d-Cβ-alkoxy, aryl-d-Cβ-alkyl, aryl-C2-
C6-alkenyl, aryl-C2-C6-alkynyl, heteroaryl, heteroaryl-d-Ce-alkyl, heteroaryl-C2-C6- alkenyl or heteroaryl-C -C6-alkynyl,
of which the cyclic moieties optionally may be substituted with one or more substitu- ents selected from R30.
In another embodiment R24, R25 and R26 are independently selected from
• hydrogen, halogen, -OCF3, -OR27, -NR27R28, -SR27, -NR27C(O)R28, -NR27C(O)OR28, -OC(O)R27, -Od-Cβ-alkyl-C(O)OR27, -Sd-C6-alkyl-C(O)OR27, -C2-Cβ-alkenyl-
C(=O)OR27, -C(=O)NR27-d-Ce-alkyl-C(=O)OR27, -CrC6-alkyl-C(=O)OR27, or -C(O)OR27,
• d-Cβ-alkyl optionally substituted with one or more substituents independently se- lected from R29 • aryl, aryloxy, aroyl, aryl-d-Ce-alkoxy, aryl-d-Ce-alkyl, heteroaryl, heteroaryl-CrC6- alkyl,
of which the cyclic moieties optionally may be substituted with one or more substituents selected from R30.
In another embodiment R24, R25 and R26 are independently selected from
•hydrogen, halogen, -OCF3, -OR27, -NR27R28, -SR27, -NR27C(O)R28, -NR27C(O)OR28,
-OC(O)R27, -OCrC6-alkyl-C(O)OR27, -Sd-C6-alkyl-C(O)OR27, -C2-Cβ-alkenyl- C(=O)OR27, -C(=O)NR27-CrC6-alkyl-C(=O)OR27, -CrC6-alkyl-C(=O)OR27, or -C(O)OR27,
• methyl, ethyl propyl optionally substituted with one or more substituents independently selected from R29
• ArG1, ArG1-O-, ArG1-C(O)-, ArG1-d-C6-alkoxy, ArG1-d-C6-alkyl, Het3, Het3-d- Ce-alkyl of which the cyclic moieties optionally may be substituted with one or more substituents selected from R30. In another embodiment R24, R25 and R26 are independently selected from
• hydrogen, halogen, -OCF3, -OR27, -NR27R28, -SR27, -NR27C(O)R28, -NR27C(O)OR28, -OC(O)R27, -Od-C6-alkyl-C(O)OR27, -SCrC6-alkyl-C(O)OR27, -C2-C6-alkenyl-
C(=O)OR27, -C(=O)NR27-CrC6-alkyl-C(=O)OR27, -d-C6-alkyl-C(=O)OR27, or -C(O)OR27,
• methyl, ethyl propyl optionally substituted with one or more substituents independ- ently selected from R29
• ArG1 , ArG1-O-, ArG1-C(O)-, ArG1-CrC6-alkoxy, ArG1-CrC6-alkyl, Het3, Het3-d- Cβ-alkyl of which the cyclic moieties optionally may be substituted with one or more substitu- ents selected from R30. In another embodiment R24, R25 and R26 are independently selected from
• hydrogen, halogen, -OCF3, -OR27, -NR27R28, -SR27, -NR27C(O)R28, -NR27C(0)OR28, -OC(O)R27, -Od-C6-alkyl-C(O)OR27, -SCrC6-alkyl-C(O)OR27, -C2-C6-allcenyl- C(=O)OR27, -C(=O)NR27-CrC6-alkyl-C(=O)OR27, -CrC6-alkyl-C(=O)OR27, or
-C(O)OR27,
• methyl, ethyl propyl optionally substituted with one or more substituents independently selected from R29
»ArG1, ArG1-O-, ArG1-Cι-C6-alkoxy, ArG1-d-C6-alkyl, of which the cyclic moieties optionally may be substituted with one or more substituents selected from R30.
In another embodiment R20 is hydrogen or methyl. In another embodiment R20 is hydrogen.
In another embodiment R27 is hydrogen, CrC6-alkyl or aryl.
In another embodiment R27 is hydrogen or d-Ce-alkyl or ArG1.
In another embodiment R27 is hydrogen or d-Ce-alkyl.
In another embodiment R28 is hydrogen or d-C6-alkyl. In another embodiment R17 and R18 are independently selected from
• hydrogen, halogen, -CN, -CF3, -OCF3, -NO2, -OR27, -NR27R28, -SR27, -S(O)R27, -S(O)2R27, -C(O)NR27R28, -CH2OR27, -OC(O)R27, -OCrC6-alkyl-C(O)OR27, -SCrC6- alkyl-C(O)OR27, or -C(O)OR27,
• CrCe-alkyl, C2-C6-alkenyl or C2-C6-alkynyl, optionally substituted with one or more substituents independently selected from R29
• aryl, aryloxy, aroyl, aryl-d-Cβ-alkoxy, aryl-d-Ce-alkyl, heteroaryl, heteroaryl-d-C6- alkyl,
of which the cyclic moieties optionally may be substituted with one or more substituents selected from R30. In another embodiment R17and R18are independently selected from • hydrogen, halogen, -CN, -CF3, -Nθ2, -OR27, -NR27R28, or -C(O)OR27,
• CrCe-alkyl optionally substituted with one or more substituents independently selected from R29
• aryl, aryloxy, aroyl, aryl-d-Cβ-alkoxy, aryl-d-Ce-alkyl, heteroaryl, heteroaryl-Cι-C6- alkyl,
of which the cyclic moieties optionally may be substituted with one or more substitu- ents selected from R30.
In another embodiment R17and R18are independently selected from
• hydrogen, halogen, -CN, -CF3, -NO2, -OR27, -NR27R28, or -C(O)OR27
• methyl, ethyl propyl optionally substituted with one or more substituents independ- ently selected from R29
• aryl, aryloxy, aroyl, aryl-d-Cβ-alkoxy, aryl-d-C6-alkyl, heteroaryl, heteroaryl-d-Ce- alkyl of which the cyclic moieties optionally may be substituted with one or more substituents selected from R30. In another embodiment R17 and R18 are independently selected from
• hydrogen, halogen, -CN, -CF3, -NO2, -OR27, -NR27R28, or -C(O)OR27
• methyl, ethyl propyl optionally substituted with one or more substituents independently selected from R29
• ArG1 , ArG1-O-, ArG1-C(O)-, ArG1-C C6-alkoxy, ArG1-d-C6-alkyl, Het3, Het3-d- Ce-alkyl of which the cyclic moieties optionally may be substituted with one or more substituents selected from R30. In another embodiment R17and R18are independently selected from
• hydrogen, halogen, -CN, -CF3, -NO2, -OR27, -NR27R28, or -C(O)OR27 "CrCe-alkyl optionally substituted with one or more substituents independently selected from R29
• phenyl, phenyloxy, phenyl-d-Cβ-alkoxy, phenyl-d-Ce-alkyl, of which the cyclic moieties optionally may be substituted with one or more substituents selected from R30. In another embodiment R27 is hydrogen or d-Cβ-alkyl. In another embodiment R27 is hydrogen, methyl or ethyl. In another embodiment R28 is hydrogen or d-C6-alkyl. In another embodiment R28 is hydrogen, methyl or ethyl. In another embodiment R72 is -OH or phenyl. In another embodiment CGr is
Figure imgf000031_0001
In another embodiment CGr is of the form H-l-J-
Figure imgf000031_0002
wherein the phenyl, naphthalene or benzocarbazole rings are optionally substituted with one or more substituents independently selected from R31
I is selected from »a valence bond,
• -CH2N(R32)- or -SO2N(R33)-,
Figure imgf000031_0003
wherein Z1 is S(O)2 or CH2, Z2 is -NH-, -O-or -S-, and n is 1 or 2,
J is
• CrCe-alkylene, C2-C6-alkenylene or C2-C6-alkynylene, which may each optionally be substituted with one or more substituents selected from R34,
• Arylene, -aryloxy-, arylene-oxycarbonyl-, -aroyl, arylene-CrC6-alkoxy-, ary- lene-d-Cβ-alkylene, arylene-C2-C6-alkenylene, arylene-C2-C6-alkynylene, heteroarylene, heteroarylene-d-Cβ-alkylene-, heteroarylene-C2-C6-alkenylene or heteroary- lene-C2-C6-alkynylene, wherein the cyclic moieties are optionally substituted with one or more substituents selected from R37, R31 is independently selected from hydrogen, halogen, -CN, -CH2CN, -CHF2, -CF3, -OCF3, -OCHF2, -OCH2CF3, -OCF2CHF2, -S(O)2CF3, -SCF3, -NO2, -OR35, -C(O)R35, -NR35R36, -SR35, -NR35S(O)2R36, -S(O)2NR35R36, -S(O)NR35R36, -S(O)R35, -S(O)2R35, -C(O)NR35R36, -OC(O)NR35R36, -NR35C(O)R36, -CH2C(O)NR35R36, -OCH2C(O)NR35R36, -CH2OR35, -CH2NR35R36, -OC(O)R35, -OCrCβ-alkyl-C(O)OR35, -SCrC6-alkyl-C(O)OR35 -C2-C6-alkenyl- C(=O)OR35, -NR35-C(=O)-C1-C6-alkyl-C(=O)OR35, -NR35-C(=O)-C1-C6-alkenyl-C(=O)OR35-, d-Ce-alkyl, d-C6-alkanoyl or -C(O)OR35,
R32 and R33 are independently selected from hydrogen, d-C6-alkyl or d-Ce-alkanoyl,
R^ is independently selected from halogen, -CN, -CF3, -OCF3l -OR35, and -NR35R36,
R35 and R36 are independently selected from hydrogen, d-Ce-alkyl, aryl-d-C6-alkyl or aryl, or R35 and R36 when attached to the same nitrogen atom together with the said nitrogen atom may form a 3 to 8 membered heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
R37 is independently selected from halogen, -C(O)OR35, -C(O)H, -CN, -CF3, -OCF3l -NO2, - OR35, -NR35R36, d-Ce-alkyl or d-Ce-alkanoyl,
or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base.
In another embodiment H is
Figure imgf000032_0001
In another embodiment H is
Figure imgf000033_0001
In another embodiment H is
Figure imgf000033_0002
In another embodiment I is a valence bond, -CH2N(R32)-, or -SO2N(R33)-. In another embodiment I is a valence bond. In another embodiment J is
• d-Cβ-alkylene, C2-C6-alkenylene or C2-C6-alkynylene, which may optionally be substituted with one or more substituents selected from halogen, -CN, -CF3, -OCF3, -OR35, and -NR35R36, • arylene, or heteroarylene, wherein the cyclic moieties are optionally substituted with one or more substituents independently selected from R37. In another embodiment J is
• arylene or heteroarylene, wherein the cyclic moieties are optionally substituted with one or more substituents independently selected from R37. In another embodiment J is
• ArG1 or Het3, wherein the cyclic moieties are optionally substituted with one or more substituents independently selected from R37.
In another embodiment J is
• phenylene or naphthylene optionally substituted with one or more substitu- ents independently selected from R37.
In another embodiment R32 and R33 are independently selected from hydrogen or d-C6-alkyl.
In another embodiment R34 is hydrogen, halogen, -CN, -CF3, -OCF3, -SCF3, -NO2, -OR35,
-C(O)R35, -NR35R36, -SR35, -C(O)NR35R36, -OC(O)NR35R36, -NR35C(O)R36, -OC(0)R35, -Od-
C6-alkyl-C(O)OR35, -Sd-C6-alkyl-C(O)OR35 or -C(O)OR35. In another embodiment R34 is hydrogen, halogen, -CF3, -NO2, -OR35, -NR∞R36, -SR35,
-NR35C(O)R36, or -C(0)OR35.
In another embodiment R34 is hydrogen, halogen, -CF3, -NO2, -OR35, -NR^R36, or
-NR35C(O)R36. In another embodiment R34 is hydrogen, halogen, or -OR35.
In another embodiment R35 and R36 are independently selected from hydrogen, d-Cβ-alkyl, or aryl.
In another embodiment R35 and R36 are independently selected from hydrogen or d-Cβ~alkyl. In another embodiment R37 is halogen, -C(O)OR35, -CN, -CF3, -OR35, -NR35R36, C Cβ-alkyl or CrCβ-alkanoyl.
In another embodiment R37 is halogen, -C(O)OR35, -OR35, -NR35R38, d-C6-alkyl or d-Ce- alkanoyl.
In another embodiment R37 is halogen, -C(O)OR35 or -OR35. In another embodiment CGr is
Figure imgf000034_0001
wherein K is a valence bond, d-Ce-alkylene, -NH-C(=O)-U-, -CrC6-alkyl-S-, -d-Ce-alkyl-O-, -C(=O)-, or -C(=O)-NH-, wherein any d-Ce-alkyl moiety is optionally substituted with R38,
U is a valence bond, d-C6-alkenylene, -d-C6-alkyl-O- or d-C6-alkylene wherein any d- Ce-alkyl moiety is optionally substituted with d-Ce-alkyl,
R38 is CrCe-alkyl, aryl, wherein the alkyl or aryl moieties are optionally substituted with one or more substituents independently selected from R39,
R39 is independently selected from halogen, cyano, nitro, amino,
M is a valence bond, arylene or heteroarylene, wherein the aryl or heteroaryl moieties are optionally substituted with one or more substituents independently selected from R40,
R40 is selected from
• hydrogen, halogen, -CN, -CH2CN, -CHF2, -CF3, -OCF3, -OCHF2, -OCH2CF3, -OCF2CHF2, -S(O)2CF3, -OS(O)2CF3, -SCF3, -NO2, -OR41, -NR41R42, -SR41, -NR41S(O)2R42, -S(O)2NR 1R42, -S(O)NR 1R42, -S(O)R41, -S(O)2R41, -OS(0)2 R41,
-C(O)NR41R42, -OC(O)NR41R42, -NR41C(O)R42, -CH2C(O)NR4 R42, -OCrdr alkyl-C(O)NR41R42, -CH2OR41, -CH2OC(O)R41 , -CH2NR41R42, -OC(O)R41, -OCrC6- alkyl-C(O)OR41, -Od-Ce-alkyl-OR41, -S-d-C6-alkyl-C(O)OR41, -C2-C6-alkenyl- C(=O)OR41, -NR41-C(=0)-CrC6-alkyl-C(=O)OR41, -NR41-C(=O)-CrC6- alkenyl-C(=O)OR41 , -C(O)OR41, -C2-C6-alkenyl-C(=O)R41, =O, -NH-C(=0)-O-C Ce-alkyl, or -NH-C(=O)-C(=O)-O-d-Ce-alkyl,
•CrCe-alkyl, C2-C6-alkenyl or C2-C6-alkynyl, which may each optionally be substituted with one or more substituents selected from R43,
• aryl, aryloxy, aryloxycarbonyl, aroyl, arylsulfanyl, aryl-d-Cβ-alkoxy, aryl-d-Cβ-alkyl, aryl-C2-C6-alkenyl, aroyl-C2-C6-alkenyl, aryl-C2-C6-alkynyl, heteroaryl, heteroaryl-d- Ce-alkyl, heteroaryl-C2-C6-alkenyl or heteroaryl-C2-C6-alkynyl, wherein the cyclic moieties optionally may be substituted with one or more substituents selected from R44,
R41 and R42 are independently selected from hydrogen, -OH, d-Ce-alkyl, d-Ce-alkenyl, aryl- d-Cβ-alkyl or aryl, wherein the alkyl moieties may optionally be substituted with one or more substituents independently selected from R45, and the aryl moieties may optionally be substituted with one or more substituents independently selected from R46; R41 and R42 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
R43 is independently selected from halogen, -CN, -CF3, -OCF3, -OR41, and -NR41R42
R44 is independently selected from halogen, -C(O)OR41, -CH2C(O)OR41, -CH2OR41, -CN, -
CF3, -OCF3, -NO2, -OR41, -NR41R42 and d-Ce-alkyl,
R45 is independently selected from halogen, -CN, -CF3, -OCF3, -O-d-Cβ-alkyl, -C(O)-O-d-
Cβ-alkyl, -COOH and -NH2,
R46 is independently selected from halogen, -C(O)OCι-Cβ-alkyl, -COOH, -CN, -CF3, -OCF3, - NO2, -OH, -Od-Cβ-alkyl, -NH2, C(=O) or d-C6-alkyl,
Q is a valence bond, d-Cβ-alkylene, -CrC6-alkyl-O-, -d-Ce-alkyl-NH-, -NH-CrC6-alkyl, -NH-C(=O)-, -C(=O)-NH-, -O-d-Cβ-alkyl, -C(=O)-, or -d-C6-alkyl-C(=O)-N(R47)- wherein the alkyl moieties are optionally substituted with one or more substituents independently selected from R48, R47 and R48 are independently selected from hydrogen, d-Ce-alkyl, aryl optionally substituted with one or more R49,
R49 is independently selected from halogen and -COOH,
T is
• CrCe-alkylene, C2-C6-alkenylene , C2-C6-alkynylene, -CrC6-alkyloxy-carbonyl, wherein the alkylene, alkenylene and alkynylene moieties are optionally substituted with one or more substituents independently selected from R50,
• arylene, -aryloxy-, -aryloxy-carbonyl-, arylene-d-Cβ-alkylene, -aroyl-, arylene-d- C6-alkoxy-, arylene-C2-C6-alkenylene, arylene-C2-C6-alkynylene, heteroarylene, het- eroarylene-d-Cβ-alkylene, heteroarylene-C2-C6-alkenylene, heteroarylene-C2- Ce-alkynylene,
wherein any alkylene, alkenylene , alkynylene, arylene and heteroarylene moiety is optionally substituted with one or more substituents independently selected from R50,
R50 is CrCe-alkyl, CrC6-alkoxy, aryl, aryloxy, aryl-d-Cβ-alkoxy, -C(=O)-NH-CrC6-alkyl-aryl, heteroaryl, heteroaryl-d-Ce-alkoxy, -d-Cβ-alkyl-COOH, -O-d-Ce-alkyl-COOH, -S(O)2R51, -C2-C6-alkenyl-COOH, -OR51, -NO2, halogen, -COOH, -CF3, -CN, =O, -N(R5 R52), wherein the aryl or heteroaryl moieties are optionally substituted with one or more R53,
R51 and R52 are independently selected from hydrogen and d-Cβ-alkyl,
R53 is independently selected from C Cβ-alkyl, d-C6-alkoxy, -CrC6-alkyl-COOH, -C2- C6-alkenyl-COOH, -OR51, -NO2, halogen, -COOH, -CF3, -CN, or -N(R51R52),
or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base.
In another embodiment K is a valence bond, d-Ce-alkylene, -NH-C(=O)-U-, -d-Ce-alkyl-S-, -d-Ce-alkyl-O-, or -C(=O)-, wherein any d-C6-alkyl moiety is optionally substituted with R38. In another embodiment K is a valence bond, d-Ce-alkylene, -NH-C(=O)-U-, -d-Cβ-alkyl-S-, or -d-Ce-alkyl-O, wherein any d-Ce-alkyl moiety is optionally substituted with R38. In another embodiment K is a valence bond, Ci-C6-alkylene, or -NH-C(=O)-U, wherein any
CrCe-alkyl moiety is optionally substituted with R38.
In another embodiment K is a valence bond or d-Ce-alkylene, wherein any d-C6-alkyl moiety is optionally substituted with R38. In another embodiment K is a valence bond or -NH-C(=O)-U.
In another embodiment K is a valence bond.
In another embodiment U is a valence bond or -d-Cβ-alkyl-O-.
In another embodiment U is a valence bond
In another embodiment M is arylene or heteroarylene, wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from
R40.
In another embodiment M is ArG1 or Het1 , wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from R40.
In another embodiment M is ArG1 or Het2, wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from R40.
In another embodiment M is ArG1 or Het3, wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from R40.
In another embodiment M is phenylene optionally substituted with one or more substituents independently selected from R40. In another embodiment M is indolylene optionally substituted with one or more substituents independently selected from R40.
In another embodiment M is
Figure imgf000037_0001
In another embodiment M is carbazolylene optionally substituted with one or more substitu- ents independently selected from R40. In another embodiment M is
Figure imgf000037_0002
In another embodiment R40 is selected from • hydrogen, halogen, -CN, -CF3, -OCF3, -NO2, -OR41, -NR41R42, -SR41, -S(O)2R41, -NR41C(O)R42, -Od-Cβ-alkyl-C(O)NR41R42, -C2-C6-alkenyl-C(=O)OR41, -C(0)OR41, =O, -NH-C(=O)-O-CrC6-alkyl, or -NH-C(=O)-C(=O)-O-d-C6-alkyl,
d-Ce-alkyl or C2-C6- alkenyl which may each optionally be substituted with one or more substituents independently selected from R43,
• aryl, aryloxy, aryl-d-Cβ-alkoxy, aryl-d-Ce-alkyl, aryl-C2-C6-alkenyl, heteroaryl, het- eroaryl-d-C6-alkyl, or heteroaryl-C2-C6-alkenyl, wherein the cyclic moieties optionally may be substituted with one or more substituents selected from R44.
In another embodiment R40 is selected from
• hydrogen, halogen, -CN, -CF3, -OCF3, -NO2, -OR41, -NR41R42, -SR41, -S(O)2R41, -NR41C(O)R42, -OCrC6-alkyl-C(O)NR41R42, -C2-C6-alkenyl-C(=O)OR41, -C(O)OR41, =O, -NH-C(=O)-O-CrC6-alkyl, or -NH-C(=O)-C(=O)-O-CrC6-alkyl,
CrCe-alkyl or C2-Ce- alkenyl which may each optionally be substituted with one or more substituents independently selected from R43,
• ArG1 , ArG1-O-, ArG1-CrC6-alkoxy, ArG1-CrC6-alkyl, ArG1-C2-C6-alkenyl, Het3, Het3-CrC6-alkyl, or Het3-C2-Cβ-alkenyl, wherein the cyclic moieties optionally may be substituted with one or more substituents selected from R44. In another embodiment R40 is selected from
• hydrogen, halogen, -CF3, -NO2, -OR41, -NR41R42, -C(O)OR41, =O, or -NR41C(0)R42,
• CrCe-alkyl, •ArG1.
In another embodiment R40 is selected from
• Halogen, -NO2, -OR41, -NR41R42, -C(O)OR41, or -NR41C(O)R42,
• Methyl,
• Phenyl. In another embodiment R41 and R42 are independently selected from hydrogen, d-Ce-alkyl, or aryl, wherein the aryl moieties may optionally be substituted with halogen or -COOH. In another embodiment R41 and R42 are independently selected from hydrogen, methyl, ethyl, or phenyl, wherein the phenyl moieties may optionally be substituted with halogen or - COOH. In another embodiment Q is a valence bond, CrC6-alkylene, -d-Ce-alkyl-O-, -d- Ce-alkyl-NH-, -NH-d-C6-alkyl, -NH-C(=0)-, -C(=0)-NH-, -0-CrC6-alkyl, -C(=0)-, or -d- C6-alkyl-C(=0)-N(R47)- wherein the alkyl moieties are optionally substituted with one or more substituents independently selected from R48. In another embodiment Q is a valence bond, -CH2-, -CH2-CH2-, -CH2-0-, -CH2-CH2-0-, -CH2-NH-, -CH2-CH2-NH-, -NH-CH2-, -NH-CH2-CH2-, -NH-C(=0)-, -C(=0)-NH-, -0-CH2-, -O-CH2-CH2-, or -C(=O)-.
In another embodiment R47 and R48 are independently selected from hydrogen, methyl and phenyl. In another embodiment T is
• d-Cβ-alkylene optionally substituted with one or more substituents independently selected from R50,
• arylene, arylene-CrC6-alkylene, heteroarylene, wherein the alkylene, arylene and heteroarylene moieties are optionally substituted with one or more substituents inde- pendently selected from R50.
In another embodiment T is
• d-Cβ-alkylene optionally substituted with one or more substituents independently selected from R50,
• ArG1, ArGI-d-Ce-alkylene, Het3, wherein the alkyl, aryl and heteroaryl moieties are optionally substituted with one or more substituents independently selected from R50.
In another embodiment T is
• d-Cβ-alkylene, optionally substituted with one or more substituents independently selected from R50, »phenylene, phenylene-d-Cβ-alkylene, wherein the alkylene and phenylene moieties are optionally substituted with one or more substituents independently selected from R50.
In another embodiment R50 is d-Ce-alkyl, d-Ce-alkoxy, aryl, aryloxy, aryl-d-C6-alkoxy, -C(=O)-NH-d-C6-alkyl-aryl, heteroaryl, -CrC6-alkyl-COOH, -O-d-C6-alkyl-COOH, -S(0)2R51, -C2-Ce-alkenyl-COOH, -OR51, -N02, halogen, -COOH, -CF3, -CN, =0, -N(R51R52), wherein the aryl or heteroaryl moieties are optionally substituted with one or more R53. In another embodiment R50 is d-Cβ-alkyl, d-Ce-alkoxy, aryl, aryloxy, aryl-d-C6-alkoxy , -OR51, -N02, halogen, -COOH, -CF3, wherein any aryl moiety is optionally substituted with one or more R53. In another embodiment R50 is d-Ce-alkyl, aryloxy, aryl-d-C6-alkoxy , -OR51, halogen, -COOH, -CF3, wherein any aryl moiety is optionally substituted with one or more R53. In another embodiment R50 is d-Ce-alkyl, ArG1-O-, ArG1-d-C6-alkoxy , -OR51, halogen, -COOH, -CF3, wherein any aryl moiety is optionally substituted with one or more R53. In another embodiment R50 is phenyl, methyl or ethyl. In another embodiment R50 is methyl or ethyl. In another embodiment R51 is methyl. In another embodiment R53 is d-Ce-alkyl, d-C6-alkoxy, -OR51, halogen.or -CF3.
In another embodiment CGr is
Figure imgf000040_0001
wherein V is Ci-C6-alkylene, arylene, heteroarylene, arylene-d-e-alkylene or arylene-C2-6- alkenylene, wherein the alkylene or alkenylene is optionally substituted with one or more substituents independently selected from R54, and the arylene or heteroarylene is optionally substituted with one or more substituents independently selected from R55,
R54 is independently selected from halogen, -CN, -CF3, -OCF3, aryl, -COOH and -NH2, R55 is independently selected from
• hydrogen, halogen, -CN, -CH2CN, -CHF2, -CF3, -OCF3, -OCHF2, -OCH2CF3, -OCF2CHF2, -S(0)2CF3, -OS(0)2CF3, -SCF3, -N02, -OR56, -NR56R57, -SR56, -NR56S(0)2R57, -S(0)2NR56R57, -S(0)NR56R57, -S(0)R56, -S(0)2R56, -OS(0)2 R56, -C(0)NR56R57, -OC(0)NR56R57, -NR56C(0)R57, -CHzCCOJNR R57, -OCrC6- alkyl-C(0)NR56R57, -CH2OR56, -CH2OC(0)R56, -CH2NR56R57, -OC(0)R56, -OCι-C8- alkyl-C(0)OR56, -Od-Ce-alkyl-OR56, -Sd-C6-alkyl-C(0)OR56, -C2-C6-alkenyl- C(=0)OR56, -NR56-C(=0)-C1-Ce-alkyl-C(=0)OR56, -NR56-C(=0)-CrC6- alkenyl-C(=0)OR56 , -C(0)OR56, or -C2-C6-alkenyl-C(=0)R56,
• CrCe-alkyl, C2-C6-alkenyl or C2-C6-alkynyl,
which may optionally be substituted with one or more substituents selected from R58, • aryl, aryloxy, aryloxycarbonyl, aroyl, arylsulfanyl, aryl-d-Cβ-alkoxy, aryl-d-Cβ-alkyl, aryl-C2-C6-alkenyl, aroyl-C2-C6-alkenyl, aryl-C2-C6-alkynyl, heteroaryl, heteroaryl-d- Cβ-alkyl, heteroaryl-C2-C6-alkenyl or heteroaryl-C2-C6-alkynyl,
of which the cyclic moieties optionally may be substituted with one or more substituents selected from R59,
R56 and R57 are independently selected from hydrogen, OH, CF3, d-C12-alkyl, aryl-d-Ce- alkyl, -C(=0)-Ci-C6-alkyl or aryl, wherein the alkyl groups may optionally be substituted with one or more substituents independently selected from R60, and the aryl groups may optionally be substituted with one or more substituents independently selected from R61; R56 and R57 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
R58 is independently selected from halogen, -CN, -CF3, -OCF3, -OR56, and -NR56R57,
R59 is independently selected from halogen, -C(0)OR56, -CH2C(0)OR56, -CH2OR56, -CN, - CF3, -OCF3, -N02, -OR56, -NR56R57 and d-Cβ-alkyl,
R60 is independently selected from halogen, -CN, -CF3, -OCF3, -Od-C6-alkyl, -C(0)OCrC6- alkyl, -C(=0)-R62, -COOH and -NH2,
R61 is independently selected from halogen, -C(0)OCrC6-alkyl, -COOH, -CN, -CF3, -OCF3, - N02, -OH, -Od-Ce-alkyl, -NH2, C(=0) or d-Ce-alkyl,
R62 is d-Cβ-alkyl, aryl optionally substituted with one or more substituents independently selected from halogen, or heteroaryl optionally substituted with one or more d-C6-alkyl jnde- pendently, or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base.
In another embodiment V is arylene, heteroarylene, or arylene-d.Cβ-alkylene, wherein the alkylene is optionally substituted with one or more substituents independently selected R54, and the arylene or heteroarylene is optionally substituted with one or more substituents independently selected from R55.
In another embodiment V is arylene, Het1, or arylene-d-Cβ-alkylene, wherein the alkylene is optionally substituted with one or more substituents independently selected from R54, and the arylene or heteroarylene moiety is optionally substituted with one or more substituents independently selected from R55.
In another embodiment V is arylene, Het2, or arylene-d.C6-alkylene, wherein the alkylene is optionally substituted with one or more substituents independently selected from R54, and the arylene or heteroarylene moiety is optionally substituted with one or more substituents inde- pendently selected from R55.
In another embodiment V is arylene, Het3, or arylene-d.C6-alkylene, wherein the alkylene is optionally substituted with one or more substituents independently selected from R54, and the arylene or heteroarylene moiety is optionally substituted with one or more substituents independently selected from R55. In another embodiment V is arylene optionally substituted with one or more substituents independently selected from R55.
In another embodiment V is ArG1 optionally substituted with one or more substituents independently selected from R55. In another embodiment V is phenylene, naphthylene or anthranylene optionally substituted with one or more substituents independently selected from R55.
In another embodiment V is phenylene optionally substituted with one or more substituents independently selected from R55.
In another embodiment R55 is independently selected from
• halogen, d-Cβ-alkyl, -CN, -OCF3 ,-CF3, -N02, -OR56, -NR56R57, -NR56C(O)R57 -SR56, -Od-C8-alkyl-C(O)OR56, or -C(0)OR56,
• d-Ce-alkyl optionally substituted with one or more substituents independently selected from R58
• aryl, aryl-CrC6-alkyl, heteroaryl, or heteroaryl-d-Ce-alkyl of which the cyclic moieties optionally may be substituted with one or more substitu- ents independently selected from R59.
In another embodiment R55 is independently selected from
• halogen, d-Ce-alkyl, -CN, -OCF3 ,-CF3, -N02, -OR56, -NR56R57, -NR56C(O)R57 -SR56, -Od-C8-alkyl-C(0)OR56, or -C(0)OR56
• CrCe-alkyl optionally substituted with one or more substituents independently se- lected from R58 • ArG1, ArG1 -CrCe-alkyl, Het3, or Het3-d-C8-alkyl of which the cyclic moieties optionally may be substituted with one or more substituents independently selected from R59.
In another embodiment R55 is independently selected from halogen, -OR56, -NR56R57,
-C(0)OR56, -OC1-C8-alkyl-C(0)OR56, -NR56C(0)R57 or d-C6-alkyl.
In another embodiment R55 is independently selected from halogen, -OR56, -NR56R57,
-C(0)OR56,
Figure imgf000043_0001
-NR56C(0)R57, methyl or ethyl.
In another embodiment R56 and R57 are independently selected from hydrogen, CF3, d-C12-alkyl, or -C(=0)-C C6-alkyl; R56 and R57 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom.
In another embodiment R56 and R57 are independently selected from hydrogen or d-Ci2-alkyl, R56 and R57 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom. In another embodiment R56 and R57 are independently selected from hydrogen or methyl, ethyl, propyl butyl, R56 and R57 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom.
In another embodiment CGr is
Figure imgf000043_0002
wherein AA is d-Cβ-alkylene, arylene, heteroarylene, arylene-Ci-Cβ-alkylene or arylene-C2. Cβ-alkenylene, wherein the alkylene or alkenylene is optionally substituted with one or more substituents independently selected from R63, and the arylene or heteroarylene is optionally substituted with one or more substituents independently selected from R64,
R63 is independently selected from halogen, -CN, -CF3, -OCF3, aryl, -COOH and -NH2,
R64 is independently selected from
• hydrogen, halogen, -CN, -CH2CN, -CHF2, -CF3, -OCF3, -OCHF2, -OCH2CF3, -OCF2CHF2, -S(0)2CF3, -OS(0)2CF3, -SCF3, -N02, -OR65, -NR65R66, -SR65, -NR65S(0)2R66, -S(0)2NR65R66, -S(0)NR65R66, -S(0)R65, -S(0)2R65, -OS(0)2 R65, -C(0)NR65R66, -OC(0)NR65R66, -NR65C(0)R66, -CH2C(0)NR65R66, -OCrC6- alkyl-C(0)NR65R66, -CH2OR65, -CH2OC(0)R65, -CH2NR65R66, -OC(0)R65, -OCrC6- alkyl-C(0)OR65, -Od-Cβ-alkyl-OR65, -SCrC6-alkyl-C(0)OR65, -C2-C6-alkenyl- C(=0)OR65, -NR65-C(=0)-d-C6-alkyl-C(=0)OR65, -NR65-C(=0)-CrC6- alkenyl-C(=0)OR65 , -C(0)OR65, or -C2-C6-alkenyl-C(=0)R65,
• d-Cβ-alkyl, C2-C6-alkenyl or C2-C6-alkynyl, each of which may optionally be substituted with one or more substituents selected from R67,
• aryl, aryloxy, aryloxycarbonyl, aroyl, arylsulfanyl, aryl-CrC6-alkoxy, aryl-d-Ce-alkyl, aryl-C2-C6-alkenyl, aroyl-C2-C6-alkenyl, aryl-C2-C6-alkynyl, heteroaryl, heteroaryl-d- Cβ-alkyl, heteroaryl-C2-C6-alkenyl or heteroaryl-C2-C6-alkynyl,
of which the cyclic moieties optionally may be substituted with one or more substituents selected from R68,
R65 and R66 are independently selected from hydrogen, OH, CF3, d-C^-alkyl, aryl-Crdr alkyl, -C(=0)-R69, aryl or heteroaryl, wherein the alkyl groups may optionally be substituted with one or more substituents selected from R70, and the aryl and heteroaryl groups may optionally be substituted with one or more substituents independently selected from R71 ; R65 and R66 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
R67 is independently selected from halogen, -CN, -CF3, -OCF3, -OR65, and -NR65R66,
R68 is independently selected from halogen, -C(0)OR65, -CH2C(0)OR65, -CH2OR65, -CN, - CF3, -OCF3, -N02, -OR65, -NR65R66 and CrC6-alkyl,
R69 is independently selected from d-Ce-alkyl, aryl optionally substituted with one or more halogen, or heteroaryl optionally substituted with one or more d-Cβ-alkyl, R70 is independently selected from halogen, -CN, -CF3, -OCF3, -Od-Ce-alkyl, -C(0)OCrC6- alkyl, -COOH and -NH2,
R71 is independently selected from halogen, -C(0)Od-C6-alkyl, -COOH, -CN, -CF3, -OCF3, - N02, -OH, -OCrCβ-alkyl, -NH2, C(=0) or d-Cβ-alkyl,
or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base.
In another embodiment AA is arylene, heteroarylene or arylene-d-Ce-alkylene, wherein the alkylene is optionally substituted with one or more R63, and the arylene or heteroarylene is optionally substituted with one or more substituents independently selected from R64. In another embodiment AA is arylene or heteroarylene, wherein the arylene or heteroarylene is optionally substituted with one or more substituents independently selected from R64. In another embodiment AA is ArG1 or Het1 optionally substituted with one or more substitu- ents independently selected from R64.
In another embodiment AA is ArG1 or Het2 optionally substituted with one or more substituents independently selected from R64.
In another embodiment AA is ArG1 or Het3 optionally substituted with one or more substituents independently selected from R64. In another embodiment AA is phenylene, naphtylene, anthrylene, carbazolylene, thienylene, pyridylene, or benzodioxylene optionally substituted with one or more substituents independently selected from R64.
In another embodiment AA is phenylene or naphtylene optionally substituted with one or more substituents independently selected from R64. In another embodiment R64 is independently selected from hydrogen, halogen, -CF3, -OCF3, -OR65, -NR65R66, d-C6-alkyl, -OC(0)R65, -Od-C6-alkyl-C(0)OR65, aryl-C2-C6-alkenyl, aryloxy or aryl, wherein d-C6-alkyl is optionally substituted with one or more substituents independently selected from R67, and the cyclic moieties optionally are substituted with one or more substituents independently selected from R68. In another embodiment R64 is independently selected from halogen, -CF3, -OCF3, -OR65, -NR65R66, methyl, ethyl, propyl, -OC(O)R65, -OCH2-C(0)OR65, -OCH2-CH2-C(0)OR65, phenoxy optionally substituted with one or more substituents independently selected from R68. In another embodiment R65 and R66 are independently selected from hydrogen, CF3, d-C12-alkyl, aryl, or heteroaryl optionally substituted with one or more substituents independently selected from R71.
In another embodiment R85 and R66 are independently hydrogen, d-C12-alkyl, aryl, or het- eroaryl optionally substituted with one or more substituents independently selected from R71. In another embodiment R65 and R66 are independently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, ArG1 or Het1 optionally substituted with one or more substituents independently selected from R71. In another embodiment R65 and R66 are independently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, ArG1 or Het2 optionally substituted with one or more substituents independently selected from R71.
In another embodiment R65 and R66 are independently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, ArG1 or Het3 optionally substituted with one or more substituents independently selected from R71. In another embodiment R65 and R66 are independently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, phenyl, naphtyl, thiadiazolyl optionally substituted with one or more R71 independently; or isoxazoiyl optionally substituted with one or more substituents independently selected from R71. In another embodiment R71 is halogen or d-C6-alkyl. In another embodiment R71 is halogen or methyl.
In another embodiment Frg1 consists of 0 to 5 neutral amino acids independently selected from the group consisting of Gly, Ala, Thr, and Ser. In another embodiment Frg1 consists of 0 to 5 Gly. In another embodiment Frg1 consists of 0 Gly. In another embodiment Frg1 consists of 1 Gly. In another embodiment Frg1 consists of 2 Gly. In another embodiment Frg1 consists of 3 Gly. In another embodiment Frg 1 consists of 4 Gly. In another embodiment Frg1 consists of 5 Gly. In another embodiment GB is of the formula B1-B2-C(0)-, B1-B2-S02- or B1-B2-CH2-, wherein B1 and B2 are as defined in claim 1.
In another embodiment G8 is of the formula B1-B2-C(0)-, B1-B2-S02- or B1-B2-NH-, wherein B1 and B2 are as defined in claim 1. In another embodiment GB is of the formula B1-B2-C(0)-, B1-B2-CH2- or B1-B2-NH-, wherein B1 and B2 are as defined in claim 1. In another embodiment G8 is of the formula B1-B2-CH2-, B1-B2-Sθ2- or B1-B2-NH-, wherein B1 and B2 are as defined in claim 1.
In another embodiment G8 is of the formula B1-B2-C(0)- or B1-B2-S02-, wherein B1 and B2 are as defined in claim 1.
In another embodiment G8 is of the formula B1-B2-C(0)- or B1-B2-CH2-, wherein B1 and B2 are as defined in claim 1.
In another embodiment GB is of the formula B1-B2-C(0)- or B1-B2-NH-, wherein B1 and B2 are as defined in claim 1.
In another embodiment G8 is of the formula B1-B2-CH2- or B1-B2-S02- , wherein B1 and B2 are as defined in claim 1.
In another embodiment G8 is of the formula B1-B2-NH- or B1-B2-S02- , wherein B1 and B2 are as defined in claim 1.
In another embodiment G8 is of the formula B1-B2-CH2- or B -B2-NH- , wherein B1 and B2 are as defined in claim 1.
In another embodiment G8 is of the formula B1-B2-C(0)-.
In another embodiment G8 is of the formula B1-B2-CH2-.
In another embodiment G8 is of the formula B1-B2-S02-.
In another embodiment GB is of the formula B1-B -NH-.
In another embodiment B1 is a valence bond, -0-, or -S-. In another embodiment B1 is a valence bond, -0-, or -N(R6)-. In another embodiment B1 is a valence bond, -S-, or -N(R6)-. In another embodiment B1 is -O-, -S- or -N(R6)-. In another embodiment B1 is a valence bond or -O-. In another embodiment B1 is a valence bond or -S-. In another embodiment B1 is a valence bond or -N(R6)-. In another embodiment B1 is -O-or -S-. In another embodiment B1 is -0-or -N(R6)-. In another embodiment B1 is -S-or -N(R6)-. In another embodiment B1 is a valence bond. In another embodiment B1 is -0-. In another embodiment B1 is -S-. In another embodiment B1 is -N(R6)-. In another embodiment B2 is a valence bond, CrCι8-alkylene, C2-C18-alkenylene, C2-C18- alkynylene, arylene, heteroarylene, -CrCι8-alkyl-aryl-, -C(=0)-d-Ci8-alkyl-C(=0)-, -C(=0)- d-C18-alkyl-0-CrCi8-alkyl-C(=0)-, -C(=0)-CrC18-alkyl-S-CrC18-alkyl-C(=0)-, -C(=0)-d- C18-a!kyl-NR6-CrCi8-alkyl-C(=0)-; and the alkylene and arylene moieties are optionally substituted as defined in claim 1.
In another embodiment B2 is a valence bond, d-C18-alkylene, C2-C18-alkenylene, C2-d8- alkynylene, arylene, heteroarylene, -d-C18-alkyl-aryl-, -C(=0)-d-C18-alkyl-C(=0)-, -C(=O)- CrC18-alkyl-0-CrCi8-alkyl-C(=0)-, and the alkylene and arylene moieties are optionally substituted as defined in claim 1.
In another embodiment B2 is a valence bond, C C18-alkylene, C2-C18-alkenylene, C2-C18- alkynylene, arylene, heteroarylene, -CrC18-alkyl-aryl-, -C(=0)-CrC18-alkyl-C(=0)-, and the alkylene and arylene moieties are optionally substituted as defined in claim 1. In another embodiment Bz is a valence bond, d-C18-alkylene, arylene, heteroarylene, -C
C18-alkyl-aryl-, -C(=0)-d-Ci8-alkyl-C(=0)-, and the alkylene and arylene moieties are optionally substituted as defined in claim 1.
In another embodiment B2 is a valence bond, d-da-alkylene, arylene, heteroarylene, -d- C18-alkyl-aryl-, and the alkylene and arylene moieties are optionally substituted as defined in claim 1.
In another embodiment B2 is a valence bond, C C18-alkylene, arylene, -CrCι8-alkyl-aryl-, and the alkylene and arylene moieties are optionally substituted as defined in claim 1. In another embodiment B2 is a valence bond or -d-C18-alkylene, and the alkylene moieties are optionally substituted as defined in claim 1. In another embodiment Frg2 comprises 1 to 16 positively charged groups. In another embodiment Frg2 comprises 1 to 12 positively charged groups. In another embodiment Frg2 comprises 1 to 10 positively charged groups. In another embodiment Frg2 is a fragment containing basic amino acids independently selected from the group consisting of Lys and Arg and D-isomers of these. In another embodiment the basic amino acid is Arg. In another embodiment X is -OH or -NH2. In another embodiment X is -NH2.
In another embodiment the pharmaceutical preparation further comprises at least 3 phenolic molecules. In another embodiment the acid-stabilised insulin is selected from the group consisting of A21G
A21G, B28K, B29P A21G, B28D A21G, B28E A21G, B3K, B29E A21G, desB27
A21G, B9E
A21G, B9D
A21G, B10E In another embodiment zinc ions are present in an amount corresponding to 10 to 40 μg
Zn/100 U insulin
In another embodiment zinc ions are present in an amount corresponding to 10 to 26 μg
Zn/100 U insulin.
In another embodiment the ratio between insulin and the zinc-binding ligand of the invention is in the range from 99:1 to 1 :99.
In another embodiment the ratio between insulin and the zinc-binding ligand of the invention is in the range from 95:5 to 5:95
In another embodiment the ratio between between insulin and the zinc-binding ligand of the invention is in the range from 80:20 to 20:80 In another embodiment the ratio between between insulin and the zinc-binding ligand of the invention is in the range from 70:30 to 30:70
In another aspect the invention relates to a method of preparing a zinc-binding ligand of the invention comprising the steps of
• Identifying starter compounds that binds to the R-state His810-Zn2+ site • optionally attaching a fragment consisting of 0 to 5 neutral α- or β-amino acids
• attaching a fragment comprising 1 to 20 positively charged groups independently selected from amino or guanidino groups
In another aspect the invention relates to a method of prolonging the action of an acid- stabilised insulin preparation which comprises adding a zinc-binding ligand of the invention to the acid-stabilised insulin preparation.
In another aspect the invention relates to a method of treating type 1 or type 2 diabetes comprising administering to a patient in need thereof a theraputically effective amount of a pharmaceutical preparation comprising 1. Acid-stabilised insulin 2. Zinc ions
3. A zinc-binding ligand that binds to the R-state His8 0-Zn2+ site, where said ligand may be as described in the embodiments above.
In another aspect the invention provides an embodiment 1 , which is a pharmaceutical prepa- ration comprising 1. Acid-stabilised insulin
2. Zinc ions
3. A zinc-binding ligand of the following general formula (I)
CGr-Lnk-Frg1-Frg2-X (I)
wherein:
CGr is a chemical group which reversibly binds to a His810 Zn2+ site of an insulin hexamer;
Lnk is a linker selected from
• a valence bond
• a chemical group G8 of the formula -B1-B2-C(0)-, -B1-B2-S02-, -B1-B -CH2-, or -B1- B2-NH-; wherein B1 is a valence bond, -0-, -S-, or -NR68-, B2 is a valence bond, CrC18-alkylene, C2-Cι8-alkenylene, C2-C 8-alkynylene, arylene, heteroarylene, -d-C18-alkyl-aryl-, -C2-C18-alkenyl-aryl-, -C2-Cι8-alkynyl-aryl-, -C(=O)- d-C18-alkyl-C(=0)-, -C(=0)-d-C18-alkenyl-C(=0)-, -C(=0)-CrC18-alkyl-0-d-C18- alkyl-C(=0)-, -C(=0)- d-C18-alkyl-S-CrC18-alkyl-C(=0)-, -C(=0)-d-C18-alkyl-NR6-Cr C18-alkyl-C(=0)-, -C(=0)-aryl-C(=0)-, -C(=0)-heteroaryl-C(=0)-; wherein the alkylene, alkenylene, and alkynylene moieties are optionally substituted by -CN, -CF3, -OCF3, -OR68, or -NR68R78 and the arylene and heteroarylene moieties are optionally substituted by halogen, -C(0)OR68, -C(0)H, OCOR68, -S02, -CN, -CF3, -OCF3, -N02, -OR68, -NR6BR78, d-C18-alkyl, or d-C18-alkanoyl; R68and R78 are independently H, d-C4-alkyl;
Frg1 is a fragment consisting of 0 to 5 neutral α- or β-amino acids
Frg2 is a fragment comprising 1 to 20 positively charged groups independently selected from amino or guanidino groups; and
X is -OH, -NH2 or a diamino group,
or a salt thereof with a pharmaceutically acceptable acid or base, or any optical isomer or mixture of optical isomers, including a racemic mixture, or any tautomeric forms. Embodiment 2. A pharmaceutical preparation according to embodiment 1 wherein CGr is a chemical structure selected from the group consisting of carboxylates, dithiocarboxylates, . phenolates, thiophenolates, alkylthiolates, sulfonamides, imidazoles, triazoles, 4-cyano- 1 ,2,3-triazoles, benzimidazoles, benzotriazoles, purines, thiazolidinediones, tetrazoles, 5- mercaptotetrazoles, rhodanines, N-hydroxyazoles, hydantoines, thiohydantoines, barbiturates, naphthoic acids and salicylic acids.
Embodiment 3. A pharmaceutical preparation according to embodiment 2 wherein CGr is a chemical structure selected from the group consisting of benzotriazoles, 3-hydroxy 2- napthoic acids, salicylic acids, tetrazoles, thiazolidinediones, 5-mercaptotetrazoles, or 4- cyano-1 ,2,3-triazoles.
Embodiment 4. A pharmaceutical composition according to any one of the embodiments 1 to 3 wherein CGr is
Figure imgf000051_0001
wherein X is =0, =S or =NH Y is -S-, -O- or -NH-
R1, R1* and R4 are independently selected from hydrogen or d-Ce-alkyl, R2 and R2* are hydrogen or d-C6-alkyl or aryl, R1 and R2 may optionally be combined to form a double bond, R1* and R2* may optionally be combined to form a double bond,
R3, R3* and R5 are independently selected from hydrogen, halogen, aryl optionally substituted with one or more substituents independently selected from R16, d-C6-alkyl, or -C(0)NR11R12,
A, A1 and B are independently selected from d-Ce-alkyl, aryl, aryl-d-Ce-alkyl, -NR11-aryl, aryl-C2-C6-alkenyl or heteroaryl, wherein the alkyl or alkenyl is optionally substituted with one or more substituents independently selected from R6 and the aryl or heteroaryl is optionally substituted with up to four substituents R7, R8, R9, and R10, A and R3 may be connected through one or two valence bonds, B and R5 may be connected through one or two valence bonds,
R6 is independently selected from halogen, -CN, -CF3, -OCF3, aryl, -COOH and -NH2, R7, R8, R9 and R10 are independently selected from
• hydrogen, halogen, -CN, -CH2CN, -CHF2, -CF3, -OCF3, -OCHF2, -OCH2CF3, -OCF2CHF2, -S(0)2CF3, -OS(0)2CF3, -SCF3, -N02, -OR11, -NR11R12, -SR11, -NR11S(0)2R12, -S(0)2NR11R12, -S(0)NR 1R12, -S(0)R11, -S(0)2R11, -OS(0)2 R11,
-C(0)NR11R12, -OC(0)NR11R12, -NR11C(0)R12, -CH2C(0)NR 1R12,
-OCrC6-alkyl-C(0)NR11R12, -CH2OR11, -CH2OC(0)R11, -CH2NR11R12, -OC(O)R11, -Od-C15-alkyl-C(0)OR11, -Od-Cβ-alkyl-OR11, -Sd-C6-alkyl-C(0)OR11 -C2-Cβ-alkenyl-C(=0)OR11, -NR11-C(=0)-CrC6-alkyl-C(=0)OR11, -NR11-C(=O)-d-C6-alkenyl-C(=O)OR11 , -C(0)OR11, C(0)R11, or -C2-C6-alkenyl-
C(=0)R11, =0, or -C2-C6-alkenyl-C(=0)-NR11R12,
• CrCe-alkyl, C2-C6-alkenyl or C2-C6-alkynyl, each of which may optionally be substituted with one or more substituents independently selected from R13,
• aryl, aryloxy, aryloxycarbonyl, aroyl, arylsulfanyl, aryl-d-Cβ-alkoxy, aryl-d-Cβ-alkyl, aryl-C2-C6-alkenyl, aroyl-C2-C6-alkenyl, aryl-C2-Cβ-alkynyl, heteroaryl, heteroaryl-d- C6-alkyl, heteroaryl-C2-C6-alkenyl, heteroaryl-C2-Cβ-alkynyl, or C3-C6 cycloalkyl,
of which each cyclic moiety may optionally be substituted with one or more substituents independently selected from R14,
R11 and R12 are independently selected from hydrogen, OH, CrC20-alkyl, aryl-d-Ce-alkyl or aryl, wherein the alkyl groups may optionally be substituted with one or more substituents independently selected from R15, and the aryl groups may optionally be substituted one or more substituents independently selected from R16; R11 and R12 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
R13 is independently selected from halogen, -CN, -CF3, -OCF3, -OR11, -C(0)OR11 , -NR11R12, and -C(0)NR11R12,
R14 is independently selected from halogen, -C(0)OR11, -CH2C(0)OR11, -CH2OR11, -CN, - CF3, -OCF3, -N02, -OR11, -NR11R12, -NR11C(0)R11, -S(0)2R11, aryl and d-C6-alkyl, R15 is independently selected from halogen, -CN, -CF3, =0, -OCF3, -Od-Ce-alkyl, -C(0)Od- C6-alkyl, -COOH and -NH2,
R16 is independently selected from halogen, -C(0)Od-C6-alkyl, -COOH, -CN, -CF3, -OCF3, - N02, -OH, -Od-Cβ-alkyl, -NH2, C(=0) or CrC6-alkyl, or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base. Embodiment 5. A pharmaceutical composition according to embodiment 4 wherein X is =0 or =S.
Embodiment 6. A pharmaceutical composition according to embodiment 5 wherein X is =0. Embodiment 7. A pharmaceutical composition according to embodiment 5 wherein X is =S. Embodiment 8. A pharmaceutical composition according to any one of the embodiment s 4 to 7 wherein Y is -O- or -S-. Embodiment 9. A pharmaceutical composition according to embodiment 8 wherein Y is -O-. Embodiment 10. A pharmaceutical composition according to embodiment 8 wherein Y is -NH-.
Embodiment 11. A pharmaceutical composition according to embodiment 8 wherein Y is -S-. Embodiment 12. A pharmaceutical composition according to any one of the embodiment s 4 to 11 wherein A is aryl optionally substituted with up to four substituents, R7, R8, R9, and R10 which may be the same or different.
Embodiment 13. A pharmaceutical composition according to embodiment 12 wherein A is selected from ArG1 optionally substituted with up to four substituents, R7, R8, R9, and R10 which may be the same or different. Embodiment 14. A pharmaceutical composition according to embodiment 13 wherein A is phenyl or naphtyl optionally substituted with up to four substituents, R7, R8, R9, and R10 which may be the same or different. Embodiment 15. A pharmaceutical composition according to embodiment 14 wherein A is
Figure imgf000053_0001
16. A pharmaceutical composition according to embodiment 14 wherein A is phenyl. Embodiment 17. A pharmaceutical composition according to any one of the embodiment s 4 to 11 wherein A is heteroaryl optionally substituted with up to four substituents, R7, Ra, R9, and R10 which may be the same or different.
Embodiment 18. A pharmaceutical composition according to embodiment 17 wherein A is selected from Het1 optionally substituted with up to four substituents, R7, R8, R9, and R10 which may be the same or different.
Embodiment 19. A pharmaceutical composition according to embodiment 18 wherein A is selected from Het2 optionally substituted with up to four substituents, R7, R8, R9, and R10 which may be the same or different. Embodiment 20. A pharmaceutical composition according to embodiment 19 wherein A is selected from Het3 optionally substituted with up to four substituents, R7, R8, R9, and R10 which may be the same or different.
Embodiment 21. A pharmaceutical composition according to embodiment 20 wherein A is selected from the group consisting of indolyl, benzofuranyl, quinolyl, furyl, thienyl, or pyrrolyl, wherein each heteroaryl may optionally substituted with up to four substituents, R7, R8, R9, and R10 which may be the same or different.
Embodiment 22. A pharmaceutical composition according to embodiment 20 wherein A is benzofuranyl optionally substituted with up to four substituents R7, R8, R9, and R10 which may be the same or different. Embodiment 23. A pharmaceutical composition according to embodiment 22 wherein A is
Figure imgf000054_0001
24. A pharmaceutical composition according to embodiment 20 wherein A is carbazolyl optionally substituted with up to four substituents R7, R8, R9, and R10 which may be the same or different. Embodiment 25. A pharmaceutical composition according to embodiment 24 wherein A is
Figure imgf000054_0002
Embodiment 26. A pharmaceutical composition according to embodiment 20 wherein A is quinolyl optionally substituted with up to four substituents R7, R8, R9, and R10 which may be the same or different. Embodiment 27. A pharmaceutical composition according to embodiment 26 wherein A is
Figure imgf000055_0001
Embodiment 28. A pharmaceutical composition according to embodiment 20 wherein A is indolyl optionally substituted with up to four substituents R7, R8, R9, and R10 which may be the same or different. Embodiment 29. A pharmaceutical composition according to embodiment 28 wherein A is
Figure imgf000055_0002
Embodiment 30. A pharmaceutical composition according to any one of the embodiment s 4 to 29 wherein R1 is hydrogen.
Embodiment 31. A pharmaceutical composition according to any one of the embodiment s 4 to 30 wherein R2 is hydrogen. Embodiment 32. A pharmaceutical composition according to any one of the embodiment s 4 to 29 wherein R1 and R2 are combined to form a double bond.
Embodiment 33. A pharmaceutical composition according to any one of the embodiment s 4 to 32 wherein R3 is d-Ce-alkyl, halogen, or C(0)NR16R17.
Embodiment 34. A pharmaceutical composition according to embodiment 33 wherein R3 is d-Ce-alkyl or C(0)NR16R17.
Embodiment 35. A pharmaceutical composition according to embodiment 34 wherein R3 is methyl.
Embodiment 36. A pharmaceutical composition according to any one of the embodiment s 4 to 11 wherein B is phenyl optionally substituted with up to four substituents, R7, R8, R9, and R10 which may be the same or different.
Embodiment 37. A pharmaceutical composition according to any one of the embodiment s 4 to 11 or 36 wherein R4 is hydrogen. Embodiment 38. A pharmaceutical composition according to any one of the embodiment s 4 to 11 or 36 to 37 wherein R5 is hydrogen.
Embodiment 39. A pharmaceutical composition according to any one of the embodiment s 4 to 38 wherein R6 is aryl. Embodiment 40. A pharmaceutical composition according to embodiment 39 wherein R6 is phenyl.
Embodiment 41. A pharmaceutical composition according to any one of the embodiment s 4 to 40 wherein R7, R8, R9 and R10 are independently selected from
• hydrogen, halogen, -N02, -OR11, -NR11R12, -SR11, -NR11S(0)2R12, -S(0)2NR11R12,
-S(0)NR11R12, -S(0)R11, -S(0)2R11, -OS(0)2 R11, -NR 1C(0)R12, -CH2OR11, - CH2OC(0)R11, -CH2NR11R12, -OC(0)R11, -OCrCβ-alkyl-C(0)OR11, -Od-C6- alkyl-C(0)NR11R12, -Od-Cβ-alkyl-OR11, -Sd-C6-alkyl-C(0)OR11, -C2-C6-alkenyl- C(=0)OR11, -C(0)OR11, or -C2-Ce-alkenyl-C(=0)R11,
• CrCe-alkyl, C2-C6-alkenyl or C2-C6-alkynyl, which may each optionally be substituted with one or more substituents independently selected from R13
• aryl, aryloxy, aroyl, arylsulfanyl, aryl-d-Ce-alkoxy, aryl-d-Ce-alkyl, aryl-C2- Ce-alkenyl, aroyl-C2-C6-alkenyl, aryl-C2-C6-alkynyl, heteroaryl, heteroaryl-d-Ce-alkyl, wherein each of the cyclic moieties optionally may be substituted with one or more substituents independently selected from R14. Embodiment 42. A pharmaceutical composition according to embodiment 41 wherein R7, R8, R9 and R10 are independently selected from
• hydrogen, halogen, -N02, -OR11, -NR11R12, -SR11, -S(0)2R11, -OS(0)2 R11, - CH2OC(0)R11, -OC(0)R11, -OCrCβ-alkyl-C(0)OR11, -Od-C6-alkyl-OR11, -Sd-C6- alkyl-C(0)OR11, -C(0)OR11, or -C2-C6-alkenyl-C(=0)R11,
•CrCe-alkyl or d-Ce-alkenyl which may each optionally be substituted with one or more substituents independently selected from R13
• aryl, aryloxy, aroyl, aryl-d-Ce-alkoxy, aryl-d-Ce-alkyl, heteroaryl, of which each of the cyclic moieties optionally may be substituted with one or more substituents independently selected from R14. Embodiment 43. A pharmaceutical composition according to embodiment 42 wherein R?, R8, R9 and R10 are independently selected from
• hydrogen, halogen, -N02, -OR11, -NR11R12, -SR11, -S(0)2R11, -OS(0)2 R11, - CH2OC(0)R11, -OC(0)R11, -OCι-C6-alkyl-C(0)OR11, -Od-C6-alkyl-OR11, -SC Cβ- alkyl-C(0)OR11, -C(0)OR11, or -C2-C6-alkenyl-C(=0)R11,
βCrCe-alkyl or d-C6- which may each optionally be substituted with one or more substituents independently selected from R13
• aryl, aryloxy, aroyl, aryl-d-Ce-alkoxy, aryl-d-Ce-alkyl, heteroaryl,
of which each of the cyclic moieties optionally may be substituted with one or more substituents independently selected from R14. Embodiment 44. A pharmaceutical composition according to embodiment 43 wherein R7, R8, R9 and R10 are independently selected from
• hydrogen, halogen, -OR11, -Od-C6-alkyl-C(0)OR11, or -C(0)OR11,
• CrCe-alkyl which may each optionally be substituted with one or more substituents independently selected from R13
• aryl, aryloxy, aryl-d-Ce-alkoxy,
of which each of the cyclic moieties optionally may be substituted with one or more substituents independently selected from R14. Embodiment 45. A pharmaceutical composition according to embodiment 44 wherein R7, R8, R9 and R10 are independently selected from • hydrogen, halogen, -OR11, -Od-Ce-alkyl-C(0)OR11, or -C(0)OR11,
• d-Ce-alkyl which may each optionally be substituted with one or more substituents independently selected from R13
*ArG1 , ArGloxy, ArG1 -d-Ce-alkoxy, of which each of the cyclic moieties optionally may be substituted with one or more substituents independently selected from R14.
Embodiment 46. A pharmaceutical composition according to embodiment 45 wherein R7, R8, R9 and R10 are independently selected from
• hydrogen, halogen, -OR11, -OCrC6-alkyl-C(0)OR11, or -C(0)OR11,
• d-Ce-alkyl which may optionally be substituted with one or more substituents independently selected from R13
• phenyl, phenyloxy, phenyl-CrC6-alkoxy, wherein each of the cyclic moieties optionally may be substituted with one or more substituents independently selected from R14.
Embodiment 47. A pharmaceutical composition according to any one of the embodiment s 4 to 46 wherein R11 and R12 are independently selected from hydrogen, CrC20-alkyl, aryl or aryl-d-Ce-alkyl, wherein the alkyl groups may optionally be substituted with one or more substituents independently selected from R15, and the aryl groups may optionally be substituted one or more substituents independently selected from R16; R11 and R12 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitro- gen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds. Embodiment 48. A pharmaceutical composition according to embodiment 47 wherein R11 and R12 are independently selected from hydrogen, CrC20-alkyl, aryl or aryl-d-Ce-alkyl, wherein the alkyl groups may optionally be substituted with one or more substituents independently selected from R15, and the aryl groups may optionally be substituted one or more substituents independently selected from R16.
Embodiment 49. A pharmaceutical composition according to embodiment 48 wherein R11 and R12 are independently selected from phenyl or phenyl-d-Ce-alkyl. Embodiment 50. A pharmaceutical composition according to embodiment 48 wherein one or both of R11 and R12 are methyl.
Embodiment 51. A pharmaceutical composition according to any one of the embodiment s 4 to 50 wherein R13 is independently selected from halogen, CF3, OR11 or NR1 R12. Embodiment 52. A pharmaceutical composition according to embodiment 51 wherein R13 is independently selected from halogen or OR11. Embodiment 53. A pharmaceutical composition according to embodiment 52 wherein R13 is
OR11.
Embodiment 54. A pharmaceutical composition according to any one of the embodiment s 4 to 53 wherein R14 is independently selected from halogen, -C(0)OR11, -CN, -CF3, -OR11,
S(O)2R11, and d-C6-alkyl.
Embodiment 55. A pharmaceutical composition according to embodiment 54 wherein R14 is independently selected from halogen, -C(0)OR11, or -OR11.
Embodiment 56. A pharmaceutical composition according to any one of the embodiment s 4 to 55 wherein R15 is independently selected from halogen, -CN, -CF3, -C(0)Od-C6-alkyl,and
-COOH.
Embodiment 57. A pharmaceutical composition according to embodiment 56 wherein R15 is independently selected from halogen or -C(0)OCrC6-alkyl.
Embodiment 58. A pharmaceutical composition according to any one of the embodiment s 4 to 57 wherein R16 is independently selected from halogen, -C(0)OCrC6-alkyl, -COOH, -N02,
-Od-Ce-alkyl, -NH2, C(=0) or d-C6-alkyl.
Embodiment 59. A pharmaceutical composition according to embodiment 58 wherein R16 is independently selected from halogen, -C(0)OC Ce-alkyl, -COOH, -N02, or d-C6-alkyl.
Embodiment 60. A pharmaceutical composition according to any one of the embodiment s 1 to 3 wherein CGr is
Figure imgf000059_0001
wherein
R19 is hydrogen or d-C6-alkyl,
R20 is hydrogen or d-Ce-alkyl,
D, D1 and F are a valence bond, d-C6-alkylene or d-Cβ-alkenylene optionally substituted with one or more substituents independently selected from R72,
R is independently selected from hydroxy, d-C6-alkyl, or aryl, E is CrC6-alkyl, aryl or heteroaryl, wherein the aryl or heteroaryl is optionally substituted with up to three substituents R21, R22 and R23,
G and G1 are d-Cβ-alkyl, aryl or heteroaryl, wherein the aryl or heteroaryl is optionally sub- stituted with up to three substituents R24, R25 and R26,
R17, R18, R21, R22, R23, R24, R25 and R26 are independently selected from
• hydrogen, halogen, -CN, -CH2CN, -CHF2, -CF3, -OCF3, -OCHF2, -OCH2CF3, -OCF2CHF2, -S(0)2CF3, -SCF3, -N02, =0, -OR27, -NR27R28, -SR27, -NR27S(0)2R28,
-S(0)2NR27R28, -S(0)NR27R28, -S(0)R27, -S(0)2R27, -C(0)NR 7R28, -OC(0)NR27R28, -NR27C(0)R28, -NR27C(0)OR28, -CH2C(0)NR27R28, -OCH2C(0)NR27R28, -CH2OR27, -CH2NR27R28, -OC(0)R27, -OCrC6-alkyl-C(0)OR27, -Sd-Cβ-alkyl-C(0)OR27, -C2-C6- alkenyl-C(=0)OR27, -NR27-C(=0)-d-Ce-alkyl-C(=0)OR27, -NR27-C(=0)-C rdr alkenyl-C(=0)OR27, -C(=0)NR27-d-Cβ-alkyl-C(=0)OR27, -d-Ce-alkyl-C(=0)OR27,or
-C(0)OR27,
• d-Cβ-alkyl, C2-C6-alkenyl or C2-C6-alkynyl,
which may optionally be substituted with one or more substituents independently selected from R29,
• aryl, aryloxy, aryloxycarbonyl, aroyl, aryl-d-Cβ-alkoxy, aryl-d-Cβ-alkyl, aryl-C2- Cβ-alkenyl, aryl-C2-C6-alkynyl, heteroaryl, heteroaryl-d-Cβ-alkyl, heteroaryl-C2-C6- alkenyl or heteroaryl-C2-C6-alkynyl,
of which the cyclic moieties optionally may be substituted with one or more substituents selected from R30,
R27 and R28 are independently selected from hydrogen, d-Cβ-alkyl, aryl-d-Cβ-alkyl or aryl, or R27 and R28 when attached to the same nitrogen atom together with the said nitrogen atom may form a 3 to 8 membered heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds, R29 is independently selected from halogen, -CN, -CF3, -OCF3, -OR27, and -NR27R28,
R30 is independently selected from halogen, -C(0)OR27, -CN, -CF3, -OCF3, -N02, -OR27, -NR27R28 and d-Ce-alkyl, or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base.
Embodiment 61. A pharmaceutical composition according to embodiment 60 wherein D is a valence bond.
Embodiment 62. A pharmaceutical composition according to embodiment 60 wherein D is CrC6-alkylene optionally substituted with one or more hydroxy, d-Ce-alkyl, or aryl.
Embodiment 63. A pharmaceutical composition according to any one of the embodiment s 60 to 62 wherein E is aryl or heteroaryl, wherein the aryl or heteroaryl is optionally substituted with up to three substituents independently selected from R21, R22 and R23.
Embodiment 64. A pharmaceutical composition according to embodiment 63 wherein E is aryl optionally substituted with up to three substituents independently selected from R21, R22 and R23.
Embodiment 65. A pharmaceutical composition according to embodiment 64 wherein E is selected from ArG1 and optionally substituted with up to three substituents independently selected from R21, R22 and R23. Embodiment 66. A pharmaceutical composition according to embodiment 65 wherein E is phenyl optionally substituted with up to three substituents independently selected from R21,
R22 and R23.
Embodiment 67. A pharmaceutical composition according to embodiment 66 wherein CGr is
Figure imgf000061_0001
Embodiment 68. A pharmaceutical composition according to any one of the embodiment s 60 to 67 wherein R21, R22 and R23 are independently selected from
• hydrogen, halogen, -CHF2, -CF3, -OCF3, -OCHF2, -OCH2CF3, -OCF2CHF2, -SCF3, -
N02, -OR27, -NR27R28, -SR27, -C(0)NR27R28, -OC(0)NR27R28, -NR27C(0)R28, -NR27C(0)OR28, -CH2C(0)NR27R28, -OCH2C(0)NR27R28, -CH2OR27, -CH2NR27R28,
-OC(0)R27, -OCrC6-alkyl-C(0)OR27, -Sd-C6-alkyl-C(0)OR27, -C2-C6-alkenyl- C(=0)OR27, -NR27-C(=0)-CrC6-alkyl-C(=0)OR27, -NR27-C(=0)-CrC6- alkenyl-C(=0)OR27-, 'C(=0)NR27-d-C6-alkyl-C(=0)OR27, -d-C6-alkyl-C(=0)OR27, or -C(0)OR27,
•CrCe-alkyl, C2-C6-alkenyl or C2-C6-alkynyl,
which may optionally be substituted with one or more substituents independently selected from R29
*aryl, aryloxy, aryloxycarbonyl, aroyl, aryl-d-Ce-alkoxy, aryl-d-Ce-alkyl, aryl-C2-
Ce-alkenyl, aryl-C2-C6-alkynyl, heteroaryl, heteroaryl-CrC6-alkyl, heteroaryl-C2-C6- alkenyl or heteroaryl-C2-C6-alkynyl,
of which the cyclic moieties optionally may be substituted with one or more substitu- ents selected from R30.
Embodiment 69. A pharmaceutical composition according to embodiment 68 wherein R21, R22 and R23 are independently selected from
• hydrogen, halogen, -OCF3, -OR27, -NR27R28, -SR27, -NR27C(0)R28, -NR27C(0)OR28, -OC(0)R27, -Od-C6-alkyl-C(0)OR27, -Sd-C6-alkyl-C(0)OR27, -C2-C6-alkenyl-
C(=0)OR27, -C(=0)NR27-CrC6-alkyl-C(=0)OR27, -CrC6-alkyl-C(=0)OR27, or -C(0)OR27,
• d-C6-alkyl optionally substituted with one or more substituents independently se- lected from R29
• aryl, aryloxy, aroyl, aryl-d-Ce-alkoxy, aryl-d-Ce-alkyl, heteroaryl, heteroaryl-d-C6- alkyl,
of which the cyclic moieties optionally may be substituted with one or more substituents selected from R30. Embodiment 70. A pharmaceutical composition according to embodiment 69 wherein R21, R22 and R23 are independently selected from • hydrogen, halogen, -OCF3, -OR27, -NR27R28, -SR27, -NR27C(0)R28, -NR27C(0)OR28, -OC(0)R27, -OCrC6-alkyl-C(0)OR27, -SCrC6-alkyl-C(0)OR27, -C2-C6-alkenyl- C(=0)OR27, -C(=0)NR27-CrC6-alkyl-C(=0)OR27, -CrCβ-alkyl-C(=0)OR27, or -C(0)OR27,
• methyl, ethyl propyl optionally substituted with one or more substituents independently selected from R29
• aryl, aryloxy, aroyl, aryl-d-C6-alkoxy, aryl-d-Ce-alkyl, heteroaryl, heteroaryl-CrC6- alkyl of which the cyclic moieties optionally may be substituted with one or more substituents selected from R30.
Embodiment 71. A pharmaceutical composition according to embodiment 70 wherein R21,
R22 and R23 are independently selected from
• hydrogen, halogen, -OCF3, -OR27, -NR27R28, -SR27, -NR27C(0)R28, -NR27C(0)OR28, -OC(O)R27, -Od-C6-alkyl-C(0)OR27, -Sd-C6-alkyl-C(0)OR27, -C2-C6-alkenyl- C(=0)OR27, -C(=0)NR27-d-C6-alkyl-C(=0)OR27, -d-C6-alkyl-C(=0)OR27, or -C(0)OR27,
• methyl, ethyl propyl optionally substituted with one or more substituents independently selected from R29
• ArG1, ArG1-0-, ArG1-C(0)-, ArG1 -d-Ce-alkoxy, ArG1-d-C6-alkyl, Het3, Het3-d- C6-alkyl of which the cyclic moieties optionally may be substituted with one or more substituents selected from R30.
Embodiment 72. A pharmaceutical composition according to embodiment 71 wherein R21, R22 and R23 are independently selected from
• hydrogen, halogen, -OCF3, -OR27, -NR27R28, -SR27, -NR27C(0)R28, -NR 7C(0)0R28, -OC(0)R27, -Od-Ce-alkyl-C(0)OR27, -Sd-C6-alkyl-C(0)OR27, -C2-C6-alkenyl- C(=0)OR27, -C(=0)NR27-d-C6-alkyl-C(=0)OR27, -d-C6-alkyl-C(=0)OR27, or -C(0)OR27, • CrCe-alkyl optionally substituted with one or more substituents independently selected from R29
• phenyl, phenyloxy, phenyl-CrC6-alkoxy, phenyl-CrC6-alkyl, of which the cyclic moieties optionally may be substituted with one or more substituents selected from R30.
Embodiment 73. A pharmaceutical composition according to any one of the embodiment s 60 to 72 wherein R 9 is hydrogen or methyl.
Embodiment 74. A pharmaceutical composition according to embodiment 73 wherein R19 is hydrogen.
Embodiment 75. A pharmaceutical composition according to any one of the embodiment s 60 to 74 wherein R27 is Hydrogen, d-C6-alkyl or aryl.
Embodiment 76. A pharmaceutical composition according to embodiment 75 wherein R27 is hydrogen or d-C6-alkyl. Embodiment 77. A pharmaceutical composition according to any one of the embodiment s 60 to 76 wherein R28 is hydrogen or d-C6-alkyl.
Embodiment 78. A pharmaceutical composition according to embodiment 60 wherein F is a valence bond.
Embodiment 79. A pharmaceutical composition according to embodiment 60 wherein F is d- Cβ-alkylene optionally substituted with one or more hydroxy, CrC6-alkyl, or aryl.
Embodiment 80. A pharmaceutical composition according to any one of the embodiment s 60 or 78 to 79 wherein G is d-C6-alkyl or aryl, wherein the aryl is optionally substituted with up to three substituents R24, R25 and R26.
Embodiment 81. A pharmaceutical composition according to any one of the embodiment s 60 or 78 to 79 wherein G is d-Ce-alkyl or ArG1 , wherein the aryl is optionally substituted with up to three substituents R24, R25 and R26.
Embodiment 82. A pharmaceutical composition according to embodiment 80 wherein G is d-Ce-alkyl.
Embodiment 83. A pharmaceutical composition according to embodiment 82 wherein G is phenyl optionally substituted with up to three substituents R24, R25 and R26.
Embodiment 84. A pharmaceutical composition according to any one of the embodiment s 60 to 83 wherein R24, R25 and R26 are independently selected from
• hydrogen, halogen, -CHF2, -CF3, -OCF3, -OCHF2, -OCH2CF3, -OCF2CHF2, -SCF3, - N02, -OR27, -NR 7R28, -SR27, -C(0)NR27R28, -OC(0)NR27R28, -NR27C(0)R28, -NR27C(0)OR28, -CH2C(0)NR27R28, -OCH2C(0)NR27R28, -CH2OR27, -CH2NR27R28, -OC(0)R27, -Od-C6-alkyl-C(0)OR27, -Sd-C6-alkyl-C(0)OR27, -d-Ce-alkenyl- C(=0)OR27, -NR27-C(=0)-d-Ce-alkyl-C(=O)OR27, -NR27-C(=0)-C rdr alkenyl-C(=0)OR27-, -C(=0)NR27-d-C6-alkyl-C(=0)OR27, -CrC6-alkyl-C(=0)OR27, or -C(0)OR27,
• d-Ce-alkyl, C2-C6-alkenyl or C2-C6-alkynyl,
which may optionally be substituted with one or more substituents independently se- lected from R29
• aryl, aryloxy, aryloxycarbonyl, aroyl, aryl-CrC6-alkoxy, aryl-CrC6-alkyl, aryl-C2- Ce-alkenyl, aryl-C2-C6-alkynyl, heteroaryl, heteroaryl-d-Ce-alkyl, heteroaryl-C2-C6- alkenyl or heteroaryl-C2-C6-alkynyl,
of which the cyclic moieties optionally may be substituted with one or more substituents selected from R30.
Embodiment 85. A pharmaceutical composition according to embodiment 84 wherein R24,
R25 and R26 are independently selected from
• hydrogen, halogen, -OCF3, -OR27, -NR27R28, -SR27, -NR27C(0)R28, -NR27C(0)OR28, -OC(0)R27, -OCrCe-alkyl-C(0)OR27, -Sd-C6-alkyl-C(0)OR27, -C2-C6-alkenyl- C(=0)OR27, -C(=0)NR27-d-C6-alkyl-C(=0)OR27, -CrC6-alkyl-C(=0)OR27, or -C(0)OR27,
• d-Ce-alkyl, C2-C6-alkenyl or C2-C6-alkynyl,
which may optionally be substituted with one or more substituents independently selected from R29
• aryl, aryloxy, aryloxycarbonyl, aroyl, aryl-d-Cβ-alkoxy, aryl-Cι-C6-alkyl, aryl-C2- Ce-alkenyl, aryl-C2-C6-alkynyl, heteroaryl, heteroaryl-d-Ce-alkyl, heteroaryl-C2-C6- alkenyl or heteroaryl-C2-C6-alkynyl, of which the cyclic moieties optionally may be substituted with one or more substituents selected from R30.
Embodiment 86. A pharmaceutical composition according to embodiment 85 wherein R24,
R25 and R26 are independently selected from
• hydrogen, halogen, -OCF3, -OR27, -NR27R28, -SR27, -NR27C(0)R28, -NR27C(0)OR28, -OC(0)R27, -Od-Ce-alkyl-C(0)OR27, -Sd-C6-alkyl-C(0)OR27, -C2-C6-alkenyl- C(=0)OR27, -C(=0)NR27-CrC6-alkyl-C(=0)OR27, -d-Cβ-alkyl-C(=0)OR27, or -C(0)OR27,
• CrCe-alkyl optionally substituted with one or more substituents independently selected from R29
• aryl, aryloxy, aroyl, aryl-d-Ce-alkoxy, aryl-d-Ce-alkyl, heteroaryl, heteroaryl-CrC6- alkyl,
of which the cyclic moieties optionally may be substituted with one or more substituents selected from R30. Embodiment 87. A pharmaceutical composition according to embodiment 86 wherein R21, R22 and R23 are independently selected from
• hydrogen, halogen, -OCF3, -OR27, -NR27R28, -SR27, -NR27C(0)R28, -NR27C(0)OR28, -OC(0)R27, -OCrC6-alkyl-C(0)OR27, -SCrC6-alkyl-C(0)OR27, -C2-C6-alkenyl- C(=0)OR27, -C(=0)NR27-d-C6-alkyl-C(=0)OR27, -CrC6-alkyl-C(=0)OR27, or -C(0)OR27,
• methyl, ethyl propyl optionally substituted with one or more substituents independently selected from R29
»ArG1, ArG1-0-, ArGI-C(O)-, ArG1-d-C6-alkoxy, ArG1-d-C6-alkyl, Het3, Het3-d-
Ce-alkyl of which the cyclic moieties optionally may be substituted with one or more substituents selected from R30. Embodiment 88. A pharmaceutical composition according to embodiment 87 wherein R21, R22 and R23 are independently selected from • hydrogen, halogen, -OCF3, -OR27, -NR 7R28, -SR27, -NR27C(0)R28, -NR27C(0)OR28, -0C(0)R27, -OCrCe-alkyl-C(0)OR27, -Sd-C6-alkyl-C(0)OR27, -C2-C6-alkenyl- C(=0)OR27, -C(=0)NR27-d-C6-alkyl-C(=0)OR27, -CrC6-alkyl-C(=0)OR27, or -C(0)OR27,
• methyl, ethyl propyl optionally substituted with one or more substituents independently selected from R29
«ArG1, ArG1-0-, ArG1-C(0)-, ArG1 -d-Ce-alkoxy, ArG1 -d-Ce-alkyl, Het3, Het3-C
C6-alkyl of which the cyclic moieties optionally may be substituted with one or more substituents selected from R30. Embodiment 89. A pharmaceutical composition according to embodiment 88 wherein R21, R22 and R23 are independently selected from
• hydrogen, halogen, -OCF3, -OR27, -NR27R28, -SR27, -NR27C(0)R28, -NR27C(0)OR28, -OC(0)R27, -OCi-C6-alkyl-C(0)OR27, -Sd-C6-alkyl-C(0)OR27, -C2-C6-alkenyl- C(=0)OR27, -C(=0)NR27-Cι-Cβ-alkyl-C(=0)OR27, -d-C6-alkyl-C(=0)OR27, or -C(0)OR27,
• methyl, ethyl propyl optionally substituted with one or more substituents independently selected from R29
• ArG1, ArGI-O-, ArG1-CrC6-alkoxy, ArG1-d-C6-alkyl, of which the cyclic moieties optionally may be substituted with one or more substituents selected from R30.
Embodiment 90. A pharmaceutical composition according to any one of the embodiment s 60 or 78 to 89 wherein R20 is hydrogen or methyl.
Embodiment 91. A pharmaceutical composition according to embodiment 90 wherein R20 is hydrogen.
Embodiment 92. A pharmaceutical composition according to any one of the embodiment s 60 or 78 to 91 wherein R27 is hydrogen, d-d-alkyl or aryl.
Embodiment 93. A pharmaceutical composition according to embodiment 92 wherein R27 is hydrogen or d-Ce-alkyl or ArG1. Embodiment 94. A pharmaceutical composition according to embodiment 93 wherein R27 is hydrogen or d-C6-alkyl.
Embodiment 95. A pharmaceutical composition according to any one of the embodiment s 60 or 78 to 93 wherein R28 is hydrogen or d-C6-alkyl. Embodiment 96. A pharmaceutical composition according to embodiment 60 wherein Rl 7and R18 are independently selected from
• hydrogen, halogen, -CN, -CF3, -OCF3, -N02, -OR27, -NR27R28, -SR27, -S(O)R27, -S(0)2R27, -C(0)NR27R28, -CH2OR27, -OC(0)R27, -OCrC6-alkyl-C(0)OR27, -SCrC6- alkyl-C(0)OR27, or -C(0)OR27,
• CrCe-alkyl, C2-C6-alkenyl or C2-C6-alkynyl, optionally substituted with one or more substituents independently selected from R29
»aryl, aryloxy, aroyl, aryl-d-C6-alkoxy, aryl-CrC6-alkyl, heteroaryl, heteroaryl-d-Ce- alkyl,
of which the cyclic moieties optionally may be substituted with one or more substituents selected from R30. Embodiment 97. A pharmaceutical composition according to embodiment 96 wherein R17 and R18 are independently selected from
• hydrogen, halogen, -CN, -CF3, -N02, -OR27, -NR27R28, or -C(0)OR27,
"CrCe-alkyl optionally substituted with one or more substituents independently selected from R29
• aryl, aryloxy, aroyl, aryl-d-Ce-alkoxy, aryl-d-Ce-alkyl, heteroaryl, heteroaryl-d-Ce- alkyl,
of which the cyclic moieties optionally may be substituted with one or more substituents selected from R30. Embodiment 98. A pharmaceutical composition according to embodiment 97 wherein R17 and R18 are independently selected from • hydrogen, halogen, -CN, -CF3, -N02, -OR27, -NR27R28, or -C(0)OR27 • methyl, ethyl propyl optionally substituted with one or more substituents independently selected from R29
• aryl, aryloxy, aroyl, aryl-d-C6-alkoxy, aryl-d-Ce-alkyl, heteroaryl, heteroaryl-d-Ce- alkyl of which the cyclic moieties optionally may be substituted with one or more substituents selected from R30. Embodiment 99. A pharmaceutical composition according to embodiment 98 wherein R1 r and R18 are independently selected from
• hydrogen, halogen, -CN, -CF3, -N02, -OR27, -NR27R28, or -C(0)OR27 • methyl, ethyl propyl optionally substituted with one or more substituents independently selected from R29
• ArG1, ArG1-0-, ArG1-C(0)-, ArGI-d-Ce-alkoxy, ArG1-d-C6-alkyl, Het3, Het3-C Ce-alkyl of which the cyclic moieties optionally may be substituted with one or more substituents se- lected from R30.
Embodiment 100. A pharmaceutical composition according to embodiment 99 wherein R17 and R18 are independently selected from
• hydrogen, halogen, -CN, -CF3, -N02, -OR27, -NR27R28, or -C(0)OR27
• Crd-alkyl optionally substituted with one or more substituents independently se- lected from R29
• phenyl, phenyloxy, phenyl-CrC6-alkoxy, phenyl-d-Ce-alkyl, of which the cyclic moieties optionally may be substituted with one or more substituents selected from R30.
Embodiment 101. A pharmaceutical composition according to any one of the embodiment s 60 to 100 wherein R27 is hydrogen or d-Ce-alkyl.
Embodiment 102. A pharmaceutical composition according to embodiment 101 wherein R27 is hydrogen, methyl or ethyl.
Embodiment 103. A pharmaceutical composition according to any one of the embodiment s
60 to 102 wherein R28 is hydrogen or d-Ce-alkyl. Embodiment 104. A pharmaceutical composition according to embodiment 103 wherein R28 is hydrogen, methyl or ethyl.
Embodiment 105. A pharmaceutical composition according to any one of the embodiment s
60 to 104 wherein R72 is -OH or phenyl.
Embodiment 106. A pharmaceutical composition according to embodiment 60 wherein CGr is
Figure imgf000070_0001
Embodiment 107. A pharmaceutical composition according to any one of the embodiment s 1 to 3 wherein CGr is of the form H-l-J-
wherein H is
Figure imgf000070_0002
wherein the phenyl, naphthalene or benzocarbazole rings are optionally substituted with one or more substituents independently selected from R31
I is selected from
• a valence bond,
• -CH2N(R32)- or -S02N(R33)-,
Figure imgf000070_0003
is S(0)2 or CH2, Z2 is -NH-, -O-or -S-, and n is 1 or 2,
J is
• d-Cβ-alkyl, C2-C6-alkenyl or C2-C6-alkynyl, which may each optionally be substituted with one or more substituents selected from R34,
• Aryl, aryloxy, aryl-oxycarbonyl-, aroyl, aryl-d-Cβ-alkoxy-, aryl-d-Cβ-alkyl-, aryl-C2- Ce-alkenyl-, aryl-C2-C6-alkynyl-, heteroaryl, heteroaryl-d-Ce-alkyl-, heteroaryl-C2-C6- alkenyl- or heteroaryl-C2-C6-alkynyl-, wherein the cyclic moieties are optionally substituted with one or more substituents selected from R37,
• hydrogen,
R31 is independently selected from hydrogen, halogen, -CN, -CH2CN, -CHF2, -CF3, -OCF3
-OCHF2, -OCH2CF3, -OCF2CHF2, -S(0)2CF3, -SCF3, -N02, -OR3&, -C(0)R -.3~5 -NR J3J5SDR3J6, -SR J3Λ5, -NR35S(0)2R36, -S(0)2NR35R36, -S(0)NR35R36, -S(0)R35, -S(0)2R35, -C(0)NR35R36, -OC(0)NR35R36, -NR35C(0)R36, -CH2C(0)NR35R36, -OCH2C(0)NR35R36, -CH2OR35, -CH2NR35R36, -OCc.OJR35, -Od-C6-alkyl-C(0)OR35, -Sd-C6-alkyl-C(0)OR35 -C2-C6-alkenyl- C(=0)OR35, -NR35-C(=0)-Cι-Cβ-alkyl-C(=0)OR35, -NR35-C(=0)-d-C6-alkenyl-C(=0)OR35-, CrCe-alkyl, d-Cβ-alkanoyl or -C(0)OR35,
R32 and R33 are independently selected from hydrogen, d-C6-alkyl or d-Ce-alkanoyl,
R34 is independently selected from halogen, -CN, -CF3, -OCF3, -OR35, and -NR R36,
R35 and R36 are independently selected from hydrogen, d-Ce-alkyl, aryl-d-Ce-alkyl or aryl, or R35 and R36 when attached to the same nitrogen atom together with the said nitrogen atom may form a 3 to 8 membered heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
R37 is independently selected from halogen, -C(0)OR35, -C(0)H, -CN, -CF3, -OCF3, -NO2, - OR35, -NR35R36, Ci-C6-alkyl or C C6-alkanoyl,
or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base.
Embodiment 108. A pharmaceutical composition according to embodiment 107 wherein CGr is of the form H-l-J, wherein H is
Figure imgf000071_0001
wherein the phenyl, naphthalene or benzocarbazole rings are optionally substituted with one or more substituents independently selected from R31 ,
is selected from
• a valence bond,
• -CH2N(R32)- or -S02N(R33)-,
Figure imgf000072_0001
Z1 is S(0)2 or CH2, Z2 is N.-O-or -S-, and n is 1 or 2,
J is
• d-Cβ-alkyl, C d-alkenyl or C2-C6-alkynyl, which may each optionally be substituted with one or more substituents selected from R34,
• Aryl, aryloxy, aryl-oxycarbonyl-, aroyl, aryl-d-Ce-alkoxy-, aryl-d-Ce-alkyl-, aryl-C2- Ce-alkenyl-, aryl-C2-C6-alkynyl-, heteroaryl, heteroaryl-CrC6-alkyl-, heteroaryl-C2-C6- alkenyl- or heteroaryl-C2-C6-alkynyl-, wherein the cyclic moieties are optionally substituted with one or more substituents selected from R37, • hydrogen,
R31 is independently selected from hydrogen, halogen, -CN, -CH2CN, -CHF2, -CF3, -OCF3, -OCHF2, -OCH2CF3, -OCF2CHF2, -S(0)2CF3, -SCF3, -N02, -OR35, -C(0)R35, -NR35R3e, -SR35, -NR35S(0)2R36, -S(0)2NR35R36, -S(0)NR35R36, -S(0)R35, -S(0)2R35, -C(0)NR35R36, -OC(0)NR35R36, -NR35C(0)R36, -CH2C(0)NR35R36, -OCH2C(0)NR35R36, -CH2OR35, -CH2NR35R36, -OC(0)R35, -Od-C6-alkyl-C(0)OR35, -Sd-C6-alkyl-C(0)OR35 -C2-C6-alkenyl- C(=0)OR35, -NR35-C(=0)-d-Cβ-alkyl-C(=0)OR35, -NR35-C(=0)-d-C6-alkenyl-C(=0)OR35-, CrCe-alkyl, d-C6-alkanoyl or -C(0)OR35,
R32 and R33 are independently selected from hydrogen, CrCe-alkyl or d-Ce-alkanoyl,
R34 is independently selected from halogen, -CN, -CF3, -OCF3, -OR35, and -NR35R36,
R35 and R36 are independently selected from hydrogen, d-Ce-alkyl, aryl-d-C6-alkyl or aryl, or R35 and R36 when attached to the same nitrogen atom together with the said nitrogen atom may form a 3 to 8 membered heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
R37 is independently selected from halogen, -C(0)OR35, -C(0)H, -CN, -CF3, -OCF3, -N02, - OR35, -NR35R36, d-Ce-alkyl or C,-C6-alkanoyl, or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base, With the proviso that R31 and J cannot both be hydrogen.
Embodiment 109. A pharmaceutical composition according to any one of the embodiment s 107 or 108 wherein H is
Figure imgf000073_0001
Embodiment 110. A pharmaceutical composition according to embodiment 109 wherein H is
Figure imgf000073_0002
Embodiment 111. A pharmaceutical composition according to embodiment 109 wherein H is
Figure imgf000073_0003
Embodiment 112. A pharmaceutical composition according to any one of the embodiment s 107 to 111wherein I is a valence bond, -CH2N(R32)-, or -SO2N(R33)-. Embodiment 113. A pharmaceutical composition according to embodiment 112 wherein I is a valence bond. Embodiment 114. A pharmaceutical composition according to any one of the embodiment s 107 to 113 wherein J is
• hydrogen,
• d-Ce-alkyl, C2-C6-alkenyl or C2-C6-alkynyl, which may optionally be substituted with one or more substituents selected from halogen, -CN, -CF3, -OCF3, -OR35, and -NR35R36,
• aryl, or heteroaryl, wherein the cyclic moieties are optionally substituted with one or more substituents independently selected from R37.
Embodiment 115. A pharmaceutical composition according to embodiment 114 wherein J is
• hydrogen, • aryl or heteroaryl, wherein the cyclic moieties are optionally substituted with one or more substituents independently selected from R37.
Embodiment 116. A pharmaceutical composition according to embodiment 114 wherein J is
• hydrogen, *ArG1 or Het3, wherein the cyclic moieties are optionally substituted with one or more substituents independently selected from R37. Embodiment 117. A pharmaceutical composition according to embodiment 116 wherein J is
• hydrogen,
• phenyl or naphthyl optionally substituted with one or more substituents inde- pendently selected from R37.
Embodiment 118. A pharmaceutical composition according to embodiment 117 wherein J is hydrogen.
Embodiment 119. A pharmaceutical composition according to any one of the embodiment s
107 to 118 wherein R32 and R33 are independently selected from hydrogen or d-C6-alkyl. Embodiment 120. A pharmaceutical composition according to any one of the embodiment s
107 to 119 wherein R34 is hydrogen, halogen, -CN, -CF3, -OCF3, -SCF3, -N02, -OR35,
-C(O)R35, -NR35R36, -SR35, -C(O)NR35R36, -OC(0)NR35R36, -NR35C(0)R36, -OC(0)R35, -OC
C6-alkyl-C(0)OR35, -Sd-C6-alkyl-C(0)OR35 or -C(0)OR35.
Embodiment 121. A pharmaceutical composition according to embodiment 120 wherein R34 is hydrogen, halogen, -CF3, -N02, -OR35, -NR35R36, -SR35, -NR35C(0)R36, or -C(0)OR35.
Embodiment 122. A pharmaceutical composition according to embodiment 121 wherein R34 is hydrogen, halogen, -CF3, -N02, -OR35, -NR35R36, or -NR35C(0)R36.
Embodiment 123. A pharmaceutical composition according to embodiment 122 wherein R34 is hydrogen, halogen, or -OR35. Embodiment 124. A pharmaceutical composition according to any one of the embodiment s
107 to 123 wherein R35 and R36 are independently selected from hydrogen, d-Cβ-alkyl, or aryl.
Embodiment 125. A pharmaceutical composition according to embodiment 124 wherein R35 and R36 are independently selected from hydrogen or d-Ce-alkyl. Embodiment 126. A pharmaceutical composition according to any one of the embodiment s
107 to 125 wherein R37 is halogen, -C(0)OR35, -CN, -CF3, -OR35, -NR35R36, d-C6-alkyl or d-
C6-alkanoyl.
Embodiment 127. A pharmaceutical composition according to embodiment 126 wherein R37 is halogen, -C(0)OR35, -OR35, -NR35R36, d-C6-alkyl or d-C6-alkanoyl. Embodiment 128. A pharmaceutical composition according to embodiment 127 wherein R37 is halogen, -C(0)OR35 or -OR35.
Embodiment 129. A pharmaceutical composition according to any one of the embodiment s 1 to 3 wherein CGr is
Figure imgf000075_0001
wherein K is a valence bond, d-Ce-alkylene, -NH-C(=0)-U-, -CrC6-alkyl-S-, -d-Ce-alkyl-O-, -C(=0)-, or -C(=0)-NH-, wherein any d-C6-alkyt moiety is optionally substituted with R38,
U is a valence bond, d-Ce-alkenylene, -d-Ce-alkyl-O- or d-C6-alkylene wherein any d- Ce-alkyl moiety is optionally substituted with d-C6-alkyl,
R38 is CrCe-alkyl, aryl, wherein the alkyl or aryl moieties are optionally substituted with one or more substituents independently selected from R39,
R is independently selected from halogen, cyano, nitro, amino,
M is a valence bond, arylene or heteroarylene, wherein the aryl or heteroaryl moieties are optionally substituted with one or more substituents independently selected from R40,
340 is selected from
• hydrogen, halogen, -CN, -CH2CN, -CHF2, -CF3, -OCF3, -OCHF2, -OCH2CF3, -OCF2CHF2, -S(0)2CF3, -OS(0)2CF3l -SCF3, -N02, -OR41, -NR41R42, -SR41, -NR41S(0)2R42, -S(0)2NR41R42, -S(0)NR41R42, -S(0)R41, -S(0)2R41, -OS(0)2 R41, -C(0)NR41R42, -OC(0)NR41R42, -NR41C(0)R42, -CH2C(0)NR41R42, -Od-C6- alkyl-C(O)NR41R42, -CH2OR41, -CH2OC(0)R41, -CH2NR41R42, -OC(0)R41, -Od-C6- alkyl-C(0)OR41, -Od-C6-alkyl-OR41, -S-d-C6-alkyl-C(0)OR41, -C2-C6-alkenyl- C(=0)OR41, -NR4 -C(=0)-d-C6-alky)-C(=0)OR41, -NR41-C(=0)-d-C6- alkenyl-C(=0)OR41 , -C(0)OR41, -C2-C6-alkenyl-C(=0)R41, =0, -NH-C(=0)-0-d- Ce-alkyl, or -NH-C(=0)-C(=0)-0-d-C6-alkyl,
• d-Cβ-alkyl, C2-C6-alkenyl or C2-C6-alkynyl, which may each optionally be substituted with one or more substituents selected from R43, • aryl, aryloxy, aryloxycarbonyl, aroyl, arylsulfanyl, aryl-d-Ce-alkoxy, aryl-d-Ce-alkyl, aryl-C2-C6-alkenyl, aroyl-C2-Cβ-alkenyl, aryl-C2-Ce-alkynyl, heteroaryl, heteroaryl-d- Ce-alkyl, heteroaryl-d-Ce-alkenyl or heteroaryl-C2-C6-alkynyl, wherein the cyclic moieties optionally may be substituted with one or more substituents selected from
R44,
R41 and R42 are independently selected from hydrogen, -OH, d-Ce-alkyl, d-Ce-alkenyl, aryl- d-Ce-alkyl or aryl, wherein the alkyl moieties may optionally be substituted with one or more substituents independently selected from R45, and the aryl moieties may optionally be substituted with one or more substituents independently selected from R46; R41 and R42 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
R43 is independently selected from halogen, -CN, -CF3l -OCF3, -OR41, and -NR41R42
R44 is independently selected from halogen, -C(0)OR41, -CH2C(0)OR41, -CH2OR41, -CN, - CF3, -OCF3, -N02, -OR41, -NR41R42 and d-Ce-alkyl,
R45 is independently selected from halogen, -CN, -CF3, -OCF3, -0-d-C6-alkyl, -C(0)-0-C Ce-alkyl, -COOH and -NH2,
R46 is independently selected from halogen, -C(0)OCrC6-alkyl, -COOH, -CN, -CF3, -OCF3, - N02, -OH, -Od-Ce-alkyl, -NH2, C(=0) or d-C6-alkyl,
Q is a valence bond, d-Ce-alkylene, -d-Ce-alkyl-O-, -d-Ce-alkyl-NH-, -NH-Cι-Ce-alkyl, -NH-C(=0)-, -C(=0)-NH-, -0-d-Ce-alkyl, -C(=0)-, or -CrC6-alkyl-C(=0)-N(R47)- wherein the alkyl moieties are optionally substituted with one or more substituents independently selected from R48,
R47 and R48 are independently selected from hydrogen, d-Ce-alkyl, aryl optionally substituted with one or more R49,
R is independently selected from halogen and -COOH, T is
• hydrogen,
• CrCe-alkyl, C2-C6-alkenyl , C2-C6-alkynyl, d-Ce-alkyloxy-carbonyl, wherein the alkyl, alkenyl and alkynyl moieties are optionally substituted with one or more substituents independently selected from R50,
• aryl, aryloxy, aryloxy-carbonyl, aryl-CrC6-alkyl, aroyl, aryl-d-Ce-alkoxy, aryl-C2- C6-alkenyl, aryl-C2-C6-alkyny-, heteroaryl, heteroaryl-CrC6-alkyl, heteroaryl-C2- Ce-alkenyl, heteroaryl-C2-C6-alkynyl,
wherein any alkyl, alkenyl , alkynyl, aryl and heteroaryl moiety is optionally substituted with one or more substituents independently selected from R50,
R50 is CrCe-alkyl, d-Ce-alkoxy, aryl, aryloxy, aryl-d-Ce-alkoxy, -C(=0)-NH-CrC6-alkyl-aryl, -C(=O)-NR50A-C1-Ce-alkyl, -C(=0)-NH-(CH2CH20)rT,C1-C6-alkyl-COOH, heteroaryl, het- eroaryl-d-C6-alkoxy, -C C6-alkyl-COOH, -O-d-Ce-alkyl-COOH, -S(0)2R51,
-C2-C6-alkenyl-COOH, -OR51, -N02, halogen, -COOH, -CF3, -CN, =0, -N(R51R52), wherein m is 1 , 2, 3 or 4, and wherein the aryl or heteroaryl moieties are optionally substituted with one or more R53, and the alkyl moieties are optionally substituted with one or more R508. Embodiment R50A and R508 are independently selected from -C(0)OCrC6-alkyl, -COOH, -d- C6-alkyl-C(0)OCrC6-alkyl, -Ci-Ce-alkyl-COOH, or d-C6-alkyl, R51 and R52 are independently selected from hydrogen and d-Ce-alkyl, R53 is independently selected from CrC6-alkyl, d-Ce-alkoxy, -d-Ce-alkyl-COOH, -C2- Ce-alkenyl-COOH, -OR51, -N02, halogen, -COOH, -CF3, -CN, or -N(R51R52),
or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base.
Embodiment 130. A pharmaceutical composition according to embodiment 129 wherein K is a valence bond, d-C6-alkylene, -NH-C(=0)-U-, -d-C6-alkyl-S-, -d-Ce-alkyl-O-, or -C(=0)-, wherein any d-Ce-alkyl moiety is optionally substituted with R38.
Embodiment 131. A pharmaceutical composition according to embodiment 130 wherein K is a valence bond, d-Ce-alkylene, -NH-C(=0)-U-, -d-Ce-alkyl-S-, or -d-Ce-alkyl-O, wherein any d-d-alkyl moiety is optionally substituted with R38. Embodiment 132. A pharmaceutical composition according to embodiment 131 wherein K is a valence bond, d-C6-alkylene, or -NH-C(=0)-U, wherein any d-Ce-alkyl moiety is optionally substituted with R38.
Embodiment 133. A pharmaceutical composition according to embodiment 132 wherein K is a valence bond or C Cβ-alkylene, wherein any d-Ce-alkyl moiety is optionally substituted with R38.
Embodiment 134. A pharmaceutical composition according to embodiment 132 wherein K is a valence bond or -NH-C(=0)-U.
Embodiment 135. A pharmaceutical composition according to embodiment 133 wherein K is a valence bond.
Embodiment 136. A pharmaceutical composition according to any one of the embodiment s
129 to 135 wherein U is a valence bond or -CrC6-alkyl-0-.
Embodiment 137. A pharmaceutical composition according to embodiment 136 wherein U is a valence bond. Embodiment 138. A pharmaceutical composition according to any one of the embodiment s
129 to 137 wherein M is arylene or heteroarylene, wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from
R40.
Embodiment 139. A pharmaceutical composition according to embodiment 138 wherein M is ArG1 or Het1 , wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from R40.
Embodiment 140. A pharmaceutical composition according to embodiment 139 wherein M is
ArG1 or Het2, wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from R40. Embodiment 141. A pharmaceutical composition according to embodiment 140 wherein M is
ArG1 or Het3, wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from R40.
Embodiment 142. A pharmaceutical composition according to embodiment 141 wherein M is phenylene optionally substituted with one or more substituents independently selected from R40.
Embodiment 143. A pharmaceutical composition according to embodiment 141 wherein M is indolylene optionally substituted with one or more substituents independently selected from
R40.
Embodiment 144. A pharmaceutical composition according to embodiment 143 wherein M is
Figure imgf000079_0001
145. A pharmaceutical composition according to embodiment 141 wherein M is carbazolylene optionally substituted with one or more substituents independently selected from R40. Embodiment 146. A pharmaceutical composition according to embodiment 145 wherein is
Figure imgf000079_0002
Embodiment 147. A pharmaceutical composition according to any one of the embodiment s 129 to 146 wherein R40 is selected from
• hydrogen, halogen, -CN, -CF3, -OCF3, -N02, -OR41, -NR41R42, -SR41, -S(0)2R41, -NR41C(0)R42, -OCrC6-alkyl-C(0)NR4 R42, -C2-C6-alkenyl-C(=0)OR41, -C(0)OR41,
=O, -NH-C(=0)-0-d-C6-alkyl, or -NH-C(=0)-C(=0)-0-d-Ce-alkyl,
d-Cβ-alkyl or C2-C6- alkenyl which may each optionally be substituted with one or more substituents independently selected from R43,
• aryl, aryloxy, aryl-d-d-alkoxy, aryl-d-Ce-alkyl, aryl-C2-C6-alkenyl, heteroaryl, het- eroaryl-CrC6-alkyl, or heteroaryl-C2-C6-alkenyl, wherein the cyclic moieties optionally may be substituted with one or more substituents selected from R44.
Embodiment 148. A pharmaceutical composition according to embodiment 147 wherein R40 is selected from
• hydrogen, halogen, -CN, -CF3, -OCF3, -N02, -OR41, -NR41R42, -SR41, -S(0)2R41, -NR41C(0)R42, -OCrC6-alkyl-C(0)NR41R42, -C2-C6-alkenyl-C(=0)OR41 , -C(0)OR41, =O, -NH-C(=O)-0-d-Ce-alkyl, or -NH-C(=0)-C(=0)-0-CrCβ-alkyl,
CrCe-alkyl or C2-C6- alkenyl which may each optionally be substituted with one or more substituents independently selected from R43,
• ArG1 , ArGI-O-, ArG1-CrC6-alkoxy, ArG1-CrC6-alkyl, ArG1-C2-C6-alkenyl, Het3, Het3-CrC6-alkyl, or Het3-C2-C6-alkenyl, wherein the cyclic moieties optionally may be substituted with one or more substituents selected from R44. Embodiment 149. A pharmaceutical composition according to embodiment 148 wherein R40 is selected from
• hydrogen, halogen, -CF3, -N02, -OR41, -NR41R42, -C(0)OR41, =0, or -NR41C(0)R42,
• d-Ce-alkyl, .ArGI.
Embodiment 150. A pharmaceutical composition according to embodiment 149 wherein R40 is hydrogen.
Embodiment 151. A pharmaceutical composition according to embodiment 149 wherein R40 is selected from .halogen, -N02, -OR41, -NR41R42, -C(0)OR41, or -NR41C(0)R42,
• methyl,
• phenyl.
Embodiment 152. A pharmaceutical composition according to any one of the embodiment s 129 to 151 wherein R41 and R42 are independently selected from hydrogen, d-Ce-alkyl, or aryl, wherein the aryl moieties may optionally be substituted with halogen or -COOH.
Embodiment 153. A pharmaceutical composition according to embodiment 152 wherein R41 and R42 are independently selected from hydrogen, methyl, ethyl, or phenyl, wherein the phenyl moieties may optionally be substituted with halogen or -COOH. Embodiment 154. A pharmaceutical composition according to any one of the embodiment s 129 to 153 wherein Q is a valence bond, C C6-alkylene, -d-Ce-alkyl-O-, -C C6-alkyl-NH-, -NH-d-Ce-alkyl, -NH-C(=0)-, -C(=0)-NH-, -0-d-C6-alkyl, -C(=0)-, or -d- Ce-alkyl-C(=O)-N(R47)- wherein the alkyl moieties are optionally substituted with one or more substituents independently selected from R48. Embodiment 155. A pharmaceutical composition according to embodiment 154 wherein Q is a valence bond. -CH2-, -CH2-CH2-, -CH2-0-, -CH2-CH2-0-, -CH2-NH-, -CH2-CH2-NH-, -NH-CH2-, -NH-CH2-CH2-, -NH-C(=0)-, -C(=0)-NH-, -0-CH2-, -0-CH2-CH2-, or -C(=0)-. Embodiment 156. A pharmaceutical composition according to any one of the embodiment s 129 to 155 wherein R47 and R48 are independently selected from hydrogen, methyl and phenyl. Embodiment 157. A pharmaceutical composition according to any one of the embodiment s 129 to 156 wherein T is
• hydrogen,
• d-C6-alkyl optionally substituted with one or more substituents independently selected from R50, • aryl, aryl-d-Ce-alkyl, heteroaryl, wherein the alkyl, aryl and heteroaryl moieties are optionally substituted with one or more substituents independently selected from R50.
Embodiment 158. A pharmaceutical composition according to embodiment 157 wherein T is
• hydrogen, •CrCe-alkyl optionally substituted with one or more substituents independently selected from R50,
• ArG1 , ArG1 -d-Ce-alkyl, Het3, wherein the alkyl, aryl and heteroaryl moieties are optionally substituted with one or more substituents independently selected from R50.
Embodiment 159. A pharmaceutical composition according to embodiment 158 wherein T is •hydrogen,
• CrCe-alkyl, optionally substituted with one or more substituents independently selected from R50,
• phenyl, phenyl-CrC6-alkyl, wherein the alkyl and phenyl moieties are optionally substituted with one or more substituents independently selected from R50. Embodiment 160. A pharmaceutical composition according to embodiment 159 wherein T is phenyl substituted with R50.
Embodiment 161. A pharmaceutical composition according to any one of the embodiment s 129 to 160 wherein R50 is d-Ce-alkyl, d-Ce-alkoxy, aryl, aryloxy, aryl-d-Ce-alkoxy, -C(=O)-NH-CrC6-alkyl-aryl, -C(=O)-NR50A-CrCe-alkyl, -C(=0)-NH-(CH2CH20)mCrC6-alkyl- COOH, heteroaryl, -Crd-alkyl-COOH, -0-d-C6-alkyl-COOH, -S(0)2R51,
-C2-C6-alkenyl-COOH, -OR51, -N02, halogen, -COOH, -CF3, -CN, =0, -N(R51R52), wherein the aryl or heteroaryl moieties are optionally substituted with one or more R53. Embodiment 162. A pharmaceutical composition according to embodiment 161 wherein R50 is d-Ce-alkyl, d-C6-alkoxy, aryl, aryloxy, -C(=O)-NR50*-CrC6-alkyl, -C(=0)-NH-(CH2CH2θ)mCrC6-alkyl-COOH, aryl-d-C6-alkoxy , -OR51, -N02, halogen, -COOH, -CF3, wherein any aryl moiety is optionally substituted with one or more R53. Embodiment 163. A pharmaceutical composition according to embodiment 162 wherein R50 is d-Ce-alkyl, aryloxy, -C(=O)-NR50*-CrC6-alkyl, -C(=0)-NH-(CH2CH20)mCrC6-alkyl-COOH, aryl-d-Ce-alkoxy, -OR51, halogen, -COOH, -CF3, wherein any aryl moiety is optionally substi- tuted with one or more R53.
Embodiment 164. A pharmaceutical composition according to embodiment 163 wherein R50 is d-Ce-alkyl, ArG1-O-, -C(=O)-NR50*-d-C6-alkyl, -C(=0)-NH-(CH2CH20)md-C6-alkyl- COOH, ArG1 -d-Ce-alkoxy , -OR51, halogen, -COOH, -CF3, wherein any aryl moiety is optionally substituted with one or more R53. Embodiment 165. A pharmaceutical composition according to embodiment 164 wherein R50 is -C(=O)-NR50ACH2, -C(=0)-NH-(CH2CH20)2CH2l-COOH, or -C(=O)-NR50ACH2CH2.
Embodiment 166. A pharmaceutical composition according to embodiment 164 wherein R50 is phenyl, methyl or ethyl. Embodiment 167. A pharmaceutical composition according to embodiment 166 wherein R50 is methyl or ethyl.
Embodiment 168. A pharmaceutical composition according to any one of the embodiment s
129 to 167 wherein m is 1 or 2.
Embodiment 169. A pharmaceutical composition according to any one of the embodiment s 129 to 168 wherein R51 is methyl.
Embodiment 170. A pharmaceutical composition according to any one of the embodiment s
129 to 169 wherein R53 is d-Cβ-alkyl, d-C6-alkoxy, -OR51, halogen.or -CF3.
Embodiment 171. A pharmaceutical composition according to any one of the embodiment s
129 to 170 wherein R50* is -C(0)OCH3, -C(0)OCH2CH3 -COOH, -CH2C(O)OCH3, - CH2C(0)OCH2CH3, -CH2CH2C(0)OCH3, -CH2CH2C(0)OCH2CH3, -CH2COOH, methyl, or ethyl.
Embodiment 172. A pharmaceutical composition according to any one of the embodiment s
129 to 171 wherein R508 is -C(0)OCH3, -C(0)OCH2CH3 -COOH, -CH2C(O)OCH3, -
CH2C(0)OCH2CH3, -CH2CH2C(0)OCH3, -CH2CH2C(0)OCH2CH3, -CH2COOH, methyl, or ethyl.
Embodiment 173. A pharmaceutical composition according to any one of the embodiment s 1 to 3 wherein CGr is
Figure imgf000082_0001
wherein V is d-Ce-alkyl, aryl, heteroaryl, aryl-d-e-alkyl- or aryl-C2-e-alkenyl-, wherein the alkyl or alkenyl is optionally substituted with one or more substituents independently selected from R54, and the aryl or heteroaryl is optionally substituted with one or more substituents independently selected from R55,
R54 is independently selected from halogen, -CN, -CF3, -OCF3, aryl, -COOH and -NH2, R55 is independently selected from • hydrogen, halogen, -CN, -CH2CN, -CHF2, -CF3, -OCF3> -OCHF2, -OCH;>CF3, -OCF2CHF2, -S(0)2CF3, -OS(0)2CF3, -SCF3, -N02. -OR56, -NR56R57, -SR56, -NR56S(O)2R57, -StO^NR^R57, -S^NR∞R57, -S(0)R56, -S(0)2R56, -OS(0)2 R56, -C(0)NR56R57, -OC(0)NR56R57, -NR56C(0)R57, -CH2C(0)NR56R57, -Od-d- alkyl-C(0)NR56R57, -CH2OR56, -CH2OC(0)R56, -CHzNR∞R57, -OC(0)R56, -OCrd- alkyl-C(0)OR56, -OCrC6-alkyl-OR56, -Sd-C6-alkyl-C(0)OR56, -C2-C6-alkenyl- C(=0)OR56, -NR56-C(=0)-CrC6-alkyl-C(=0)OR56, -NR56-C(=0)-C1-C6- alkenyl-C(=0)OR56 , -Ct R56, or -C2-C6-alkenyl-C(=0)R56,
• d-Ce-alkyl, C2-C6-alkenyl or C2-C6-alkynyl,
which may optionally be substituted with one or more substituents selected from R58,
• aryl, aryloxy, aryloxycarbonyl, aroyl, arylsulfanyl, aryl-d-Cβ-alkoxy, aryl-d-Cβ-alkyl, aryl-C2-C6-alkenyl, aroyl-C2-Cβ-alkenyl, aryl-C2-C6-alkynyl, heteroaryl, heteroaryl-d- Ce-alkyl, heteroaryl-C2-C6-alkenyl or heteroaryl-C2-C6-alkynyl,
of which the cyclic moieties optionally may be substituted with one or more substituents selected from R59,
R56 and R57 are independently selected from hydrogen, OH, CF3, d-C12-alkyl, aryl-d-Ce- alkyl, -C(=0)-d-C6-alkyl or aryl, wherein the alkyl groups may optionally be substituted with one or more substituents independently selected from R60, and the aryl groups may optionally be substituted with one or more substituents independently selected from Rβ1; R56 and R57 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
R58 is independently selected from halogen, -CN, -CF3, -OCF3, -OR56, and -NR56R57,
R59 is independently selected from halogen, -C(0)OR56, -CH2C(0)OR56, -CH2OR56, -CN, - CF3, -OCF3, -N02, -OR56, -NR56R57 and d-Ce-alkyl, R60 is independently selected from halogen, -CN, -CF3, -OCF3, -Od-Cβ-alkyl, -C(0)OCrC6- alkyl, -C(=0)-R62, -COOH and -NH2,
R61 is independently selected from halogen, -C(0)OCrCe-alkyl, -COOH, -CN, -CF3, -OCF3, - N02, -OH, -Od-Ce-alkyl, -NH2, C(=0) or d-C6-alkyl,
R62 is CrCe-alkyl, aryl optionally substituted with one or more substituents independently selected from halogen, or heteroaryl optionally substituted with one or more d-C6-alkyl independently, or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base.
Embodiment 174. A pharmaceutical composition according to embodiment 173 wherein V is aryl, heteroaryl, or aryl-d-e-alkyl-, wherein the alkyl is optionally substituted with one or more substituents independently selected R54, and the aryl or heteroaryl is optionally substituted with one or more substituents independently selected from R55.
Embodiment 175. A pharmaceutical composition according to embodiment 174 wherein V is aryl, Het1 , or aryl-d-e-alkyl-, wherein the alkyl is optionally substituted with one or more substituents independently selected from R54, and the aryl or heteroaryl moiety is optionally substituted with one or more substituents independently selected from R55. Embodiment 176. A pharmaceutical composition according to embodiment 175 wherein V is aryl, Het2, or aryl-d-e-alkyl-, wherein the alkyl is optionally substituted with one or more substituents independently selected from R54, and the aryl or heteroaryl moiety is optionally substituted with one or more substituents independently selected from R55. Embodiment 177. A pharmaceutical composition according to embodiment 176 wherein V is aryl, Het3, or aryl-d-e-alkyl-, wherein the alkyl is optionally substituted with one or more substituents independently selected from R54, and the aryl or heteroaryl moiety is optionally substituted with one or more substituents independently selected from R55. Embodiment 178. A pharmaceutical composition according to embodiment 177 wherein V is aryl optionally substituted with one or more substituents independently selected from R55. Embodiment 179. A pharmaceutical composition according to embodiment 178 wherein V is ArG1 optionally substituted with one or more substituents independently selected from R55. Embodiment 180. A pharmaceutical composition according to embodiment 179 wherein V is phenyl, naphthyl or anthranyl optionally substituted with one or more substituents independently selected from R55. Embodiment 181. A pharmaceutical composition according to embodiment 180 wherein V is phenyl optionally substituted with one or more substituents independently selected from R55. Embodiment 182. A pharmaceutical composition according to any one of the embodiment s 173 to 181 wherein R55 is independently selected from •halogen, d-Ce-alkyl, -CN, -OCF3 ,-CF3, -N02, -OR56, -NR56R57, -NR56C(O)R57
-SR56, -Od-Cβ-alkyl-C OJOR56, or -dOJOR56,
• CrCe-alkyl optionally substituted with one or more substituents independently selected from R58
• aryl, aryl-d-C6-alkyl, heteroaryl, or heteroaryl-d-Ce-alkyl of which the cyclic moieties optionally may be substituted with one or more substituents independently selected from R59. Embodiment 183. A pharmaceutical composition according to embodiment 182 wherein R55 is independently selected from
• halogen, d-Ce-alkyl, -CN, -OCF3 ,-CF3, -N02, -OR56, -NR∞R57, -NR56C(O)R57 -SR56, -OC1-C8-alkyl-C(O)OR56, or -C(0)OR56
• d-d-alkyl optionally substituted with one or more substituents independently selected from R58
• ArG1, ArGI-d-Ce-alkyl, Het3, or Het3-C Ce-alkyl of which the cyclic moieties optionally may be substituted with one or more substitu- ents independently selected from R59.
Embodiment 184. A pharmaceutical composition according to embodiment 183 wherein R55 is independently selected from halogen, -OR56, -NR^R57, -C(0)OR56, -Od-C8- alkyl-C(0)OR56, -NR56C(0)R57 or d-C6-alkyl. Embodiment 185. A pharmaceutical composition according to embodiment 184 wherein R55 is independently selected from halogen, -OR56, -NR56R57, -C(0)OR56, -Od-C8- alkyl-C(0)OR56, -NR56C(0)R57, methyl or ethyl.
Embodiment 186. A pharmaceutical composition according to any one of the embodiment s 173 to 185 wherein R56 and R57 are independently selected from hydrogen, CF3, Cι-Cι2-alkyl, or -C(=0)-d-C6-alkyl; R56 and R57 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom.
Embodiment 187. A pharmaceutical composition according to embodiment 186 wherein R56 and R57 are independently selected from hydrogen or d-Cι2-alkyl, R56 and R57 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom. Embodiment 188. A pharmaceutical composition according to embodiment 187 wherein R56 and R57 are independently selected from hydrogen or methyl, ethyl, propyl butyl, R56 and R57 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom.
Embodiment 189. A pharmaceutical composition according to any one of the embodiment s 1 to 3 wherein CGr is
Figure imgf000086_0001
wherein AA is d-Ce-alkyl, aryl, heteroaryl, aryl-d-e-alkyl- or aryl-C2-6-alkenyl-, wherein the alkyl or alkenyl is optionally substituted with one or more substituents independently selected from R63, and the aryl or heteroaryl is optionally substituted with one or more substituents independently selected from R64,
R63 is independently selected from halogen, -CN, -CF3, -OCF3, aryl, -COOH and -NH2,
R64 is independently selected from
• hydrogen, halogen, -CN, -CH2CN, -CHF2, -CF3, -OCF3, -OCHF2, -OCH2CF3, -OCF2CHF2, -S(0)2CF3, -OS(0)2CF3, -SCF3, -N02, -OR65, -NR65R66, -SR65,
-NR65S(0)2R66, -S(0)2NR65R66, -S(0)NR65R66, -S(0)R65, -S(0)2R65, -OS(0)2 R65, -C(0)NR65R66, -OC(0)NR65R66, -NR65C(0)R66, -CH2C(0)NR65R66, -Od-C6- alkyl-C(0)NR65R66, -CH2OR65, -CH2OC(0)R65, -CH2NR65R66, -OC(0)R65, -Od-C6- alkyl-C(0)OR65, -Od-C6-alkyl-OR65, -SCrC6-alkyl-C(0)OR65, -C2-C6-alkenyl- C(=0)OR65, -NR65-C(=0)-d-C6-alkyl-C(=0)OR65, -NR65-C(=0)-d-Ce- alkenyl-C(=0)OR65 , -C(0)OR65, or -C2-Cβ-alkenyl-C(=0)R65,
• CrCe-alkyl, C2-C6-alkenyl or C2-C6-alkynyl, each of which may optionally be substituted with one or more substituents selected from R67, • aryl, aryloxy, aryloxycarbonyl, aroyl, arylsulfanyl, aryl-d-d-alkoxy, aryl-d-Ce-alkyl, aryl-C2-Ce-alkenyl, aroyl-C2-Cβ-alkenyl, aryl-C2-C6-alkynyl, heteroaryl, heteroaryl-d- Ce-alkyl, heteroaryl-C2-C6-alkenyl or heteroaryl-C2-C6-alkynyl,
of which the cyclic moieties optionally may be substituted with one or more substituents selected from R68,
R65 and R66 are independently selected from hydrogen, OH, CF3, C C12-alkyl, aryl-d-Ce- alkyl, -C(=0)-R69, aryl or heteroaryl, wherein the alkyl groups may optionally be substituted with one or more substituents selected from R70, and the aryl and heteroaryl groups may optionally be substituted with one or more substituents independently selected from R71 ; R65 and R66 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
R67 is independently selected from halogen, -CN, -CF3, -OCF3, -OR65, and -NR65R66,
R68 is independently selected from halogen, -C(0)OR65, -CH2C(0)OR65, -CH2OR65, -CN, - CF3, -OCF3, -N02, -OR65, -NR65R66 and d-Cβ-alkyl,
R69 is independently selected from d-Ce-alkyl, aryl optionally substituted with one or more halogen, or heteroaryl optionally substituted with one or more d-Ce-alkyl,
R70 is independently selected from halogen, -CN, -CF3, -OCF3, -Od-Ce-alkyl, -C(0)OCrC6- alkyl, -COOH and -NH2,
R71 is independently selected from halogen, -C(0)OCrC6-alkyl, -COOH, -CN, -CF3, -OCF3, - N02, -OH, -Od-Cβ-alkyl, -NH2, C(=0) or d-Ce-alkyl,
or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a, salt thereof with a pharmaceutically acceptable acid or base.
Embodiment 190. A pharmaceutical composition according to embodiment 189 wherein AA is aryl, heteroaryl or aryl-d-e-alkyl-, wherein the alkyl is optionally substituted with one or more R63, and the aryl or heteroaryl is optionally substituted with one or more substituents independently selected from R64.
Embodiment 191. A pharmaceutical composition according to embodiment 190 wherein AA is aryl or heteroaryl optionally substituted with one or more substituents independently se- lected from R64.
Embodiment 192. A pharmaceutical composition according to embodiment 191 wherein AA is ArG1 or Het1 optionally substituted with one or more substituents independently selected from R64. Embodiment 193. A pharmaceutical composition according to embodiment 192 wherein AA is ArG1 or Het2 optionally substituted with one or more substituents independently selected from R64.
Embodiment 194. A pharmaceutical composition according to embodiment 193 wherein AA is ArG1 or Het3 optionally substituted with one or more substituents independently selected from R64. Embodiment 195. A pharmaceutical composition according to embodiment 194 wherein AA is phenyl, naphtyl, anthryl, carbazolyl, thienyl, pyridyl, or benzodioxyl optionally substituted with one or more substituents independently selected from R64.
Embodiment 196. A pharmaceutical composition according to embodiment 195 wherein AA is phenyl or naphtyl optionally substituted with one or more substituents independently se- lected from R64.
Embodiment 197. A pharmaceutical composition according to any one of the embodiment s 189 to 196 wherein R64 is independently selected from hydrogen, halogen, -CF3, -OCF3, -OR65, -NR65R66, d-Ce-alkyl , -OC(0)R65, -OCrC6-alkyl-C(0)OR65, aryl-C2-C6-alkenyl, aryloxy or aryl, wherein d-d-alkyl is optionally substituted with one or more substituents inde- pendently selected from R67, and the cyclic moieties optionally are substituted with one or more substituents independently selected from R68.
Embodiment 198. A pharmaceutical composition according to embodiment 197 wherein R64 is independently selected from halogen, -CF3, -OCF3, -OR65, -NR65R66, methyl, ethyl, propyl, -OC(0)R65, -OCH2-C(0)OR65, -OCH2-CH2-C(0)OR65, phenoxy optionally substituted with one or more substituents independently selected from R68.
Embodiment 199. A pharmaceutical composition according to any one of the embodiment s 189 to 198 wherein R65 and R66 are independently selected from hydrogen, CF3, d-Cι2-alkyl, aryl, or heteroaryl optionally substituted with one or more substituents independently selected from R71. Embodiment 200. A pharmaceutical composition according to embodiment 199 wherein R65 and R66 are independently hydrogen, d-C12-alkyl, aryl, or heteroaryl optionally substituted with one or more substituents independently selected from R71.
Embodiment 201. A pharmaceutical composition according to embodiment 200 wherein R65 and R66 are independently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, ArG1 or Het1 optionally substituted with one or more substituents independently selected from R71.
Embodiment 202. A pharmaceutical composition according to embodiment 201 wherein R65 and R66 are independently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, ArG1 or Het2 optionally substituted with one or more substituents independently selected from R71. Embodiment 203. A pharmaceutical composition according to embodiment 202 wherein R65 and R66 are independently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, ArG1 or Het3 optionally substituted with one or more substituents independently selected from R71.
Embodiment 204. A pharmaceutical composition according to embodiment 203 wherein R65 and R66 are independently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, phenyl, naphtyl, thiadiazolyl optionally substituted with one or more R71 independently; or isoxazoiyl optionally substituted with one or more substituents independently selected from R7 .
Embodiment 205. A pharmaceutical composition according to any one of the embodiment s
189 to 204 wherein R71 is halogen or d-d-alkyl.
Embodiment 206. A pharmaceutical composition according to embodiment 205 wherein R71 is halogen or methyl.
Embodiment 207. A pharmaceutical preparation according to any one of the embodiment s 1 to 205 wherein Frg1 consists of 0 to 5 neutral amino acids independently selected from the group consisting of Gly, Ala, Thr, and Ser.
Embodiment 208. A pharmaceutical preparation according to embodiment 207 wherein Frg1 consists of 0 to 5 Gly.
Embodiment 209. A pharmaceutical preparation according to embodiment 208 wherein Frg1 consists of 0 Gly.
Embodiment 210. A pharmaceutical preparation according to embodiment 208 wherein Frg1 consists of 1 Gly. Embodiment 211. A pharmaceutical preparation according to embodiment 208 wherein Frg1 consists of 2 Gly.
Embodiment 212. A pharmaceutical preparation according to embodiment 208 wherein Frg1 consists of 3 Gly.
Embodiment 213. A pharmaceutical preparation according to embodiment 208 wherein Frg1 consists of 4 Gly. Embodiment 214. A pharmaceutical preparation according to embodiment 208 wherein Frg1 consists of 5 Gly.
Embodiment 215. A pharmaceutical preparation according to any one of the embodiment s 1 to 214 wherein GB is of the formula B1-B2-C(0)-, B -B2-S02- or B1-B2-CH2-, wherein B1 and B2 are as defined in embodiment 1.
Embodiment 216. A pharmaceutical preparation according to any one of the embodiment s 1 to 214 wherein GB is of the formula B1-B2-C(0)-, B1-B2-S02- or B1-B2-NH-, wherein B1 and B2 are as defined in embodiment 1.
Embodiment 217. A pharmaceutical preparation according to any one of the embodiment s 1 to 214 wherein GB is of the formula B -B -C(0)-, B1-B2-CH2- or B1-B2-NH-, wherein B1 and B2 are as defined in embodiment 1.
Embodiment 218. A pharmaceutical preparation according to any one of the embodiment s 1 to 214 wherein GB is of the formula B1-B2-CH2-, B1-B2-S02- or B1-B2-NH-, wherein B1 and B2 are as defined in embodiment 1. Embodiment 219. A pharmaceutical preparation according to any one of the embodiment s
215 or 216 wherein GB is of the formula B1-B2-C(0)- or B1-B2-S02-, wherein B1 and B2 are as defined in embodiment 1.
Embodiment 220. A pharmaceutical preparation according to any one of the embodiment s
215 or 217 wherein GB is of the formula B1-B2-C(0)- or B1-B2-CH2-, wherein B1 and B2 are as defined in embodiment 1.
Embodiment 221. A pharmaceutical preparation according to any one of the embodiment s
216 or 217 wherein GB is of the formula B1-B2-C(0)- or B1-B2-NH-, wherein B1 and B2 are as defined in embodiment 1.
Embodiment 222. A pharmaceutical preparation according to any one of the embodiment s 215 or 218 wherein G8 is of the formula B1-B -CH2- or B1-B2-S02- , wherein B1 and B2 are as defined in embodiment 1. Embodiment 223. A pharmaceutical preparation according to any one of the embodiment s
216 or 218 wherein GB is of the formula B1-B2-NH- or B1-B2-S02- , wherein B1 and B2 are as defined in embodiment 1. Embodiment 224. A pharmaceutical preparation according to any one of the embodiment s
217 or 218 wherein G8 is of the formula B1-B2-CH2- or B1-B2-NH- , wherein B1 and B2 are as defined in embodiment 1.
Embodiment 225. A pharmaceutical preparation according to any one of the embodiment s 219, 220, or 221 wherein GB is of the formula B1-B2-C(0)-. Embodiment 226. A pharmaceutical preparation according to any one of the embodiment s
220, 222 or 224 wherein GB is of the formula B1-B2-CH2-.
Embodiment 227. A pharmaceutical preparation according to any one of the embodiment s
220, 222 or 223 wherein G8 is of the formula B1-B2-S02-. Embodiment 228. A pharmaceutical preparation according to any one of the embodiment s
221, 223 or 224 wherein GB is of the formula B1-B2-NH-.
Embodiment 229. A pharmaceutical preparation according to any one of the embodiment s 1 to 228 wherein B1 is a valence bond, -0-, or -S-.
Embodiment 230. A pharmaceutical preparation according to any one of the embodiment s 1 to 228 wherein B1 is a valence bond, -0-, or -N(R68)-.
Embodiment 231. A pharmaceutical preparation according to any one of the embodiment s 1 to 228 wherein B1 is a valence bond, -S-, or -N(R68)-.
Embodiment 232. A pharmaceutical preparation according to any one of the embodiment s 1 to 228 wherein B1 is -0-, -S- or -N(R68)-. Embodiment 233. A pharmaceutical preparation according to any one of the embodiment s
229 or 230 wherein B1 is a valence bond or -0-.
Embodiment 234. A pharmaceutical preparation according to any one of the embodiment s
229 or 231 wherein B1 is a valence bond or -S-.
Embodiment 235. A pharmaceutical preparation according to any one of the embodiment s 230 or 231 wherein B1 is a valence bond or -N(R68)-.
Embodiment 236. A pharmaceutical preparation according to any one of the embodiment s
229 or 232 wherein B1 is -O-or -S-.
Embodiment 237. A pharmaceutical preparation according to any one of the embodiment s
230 or 232 wherein B1 is -O-or -N(R68)-. Embodiment 238. A pharmaceutical preparation according to any one of the embodiment s
231 or 232 wherein B1 is -S-or -N(R68)-.
Embodiment 239. A pharmaceutical preparation according to any one of the embodiment s
233, 234 or 235 wherein B1 is a valence bond.
Embodiment 240. A pharmaceutical preparation according to any one of the embodiment s 233, 236 or 237 wherein B1 is -0-.
Embodiment 241. A pharmaceutical preparation according to any one of the embodiment s
234, 236 or 238 wherein B1 is -S-.
Embodiment 242. A pharmaceutical preparation according to any one of the embodiment s
235, 237 or 238 wherein B1 is -N(R68)-. Embodiment 243. A pharmaceutical preparation according to any one of the embodiment s 1 to 242 wherein B2 is a valence bond, d-C18-alkylene, C2-C18-alkenylene, C2-C18-alkynylene, arylene, heteroarylene, -d-Ciβ-alkyl-aryl-, -C(=0)-CrC18-alkyl-C(=0)-, -C(=0)-d-C18-alkyl- 0-CrC18-alkyl-C(=0)-, -C(=0)-CrC18-alkyl-S-CrC18-alkyl-C(=0)-, -C(=0)-CrC18-alkyl-NR6- CrC18-alkyl-C(=0)-; and the alkylene and arylene moieties are optionally substituted as defined in embodiment 1.
Embodiment 244. A pharmaceutical preparation according to embodiment 243 wherein B2 is a valence bond, CrC18-alkylene, C2-C18-alkenylene, C2-C18-alkynylene, arylene, heteroarylene, -CrC18-alkyl-aryl-, -C(=0)-d-C18-alkyl-C(=0)-, -C(=0)-CrCi8-alkyl-0-d-C18-alkyl- C(=0)-, and the alkylene and arylene moieties are optionally substituted as defined in embodiment 1.
Embodiment 245. A pharmaceutical preparation according to embodiment 244 wherein B2 is a valence bond, d-Cι8-alkylene, C2-C18-alkenylene, C2-C18-alkynylene, arylene, heteroarylene, -CrC18-alkyl-aryl-, -C(=0)-CrC18-alkyl-C(=0)-, and the alkylene and arylene moieties are optionally substituted as defined in embodiment 1.
Embodiment 246. A pharmaceutical preparation according to embodiment 245 wherein B2 is a valence bond, d-C18-alkylene, arylene, heteroarylene, -d-C18-alkyl-aryl-, -C(=0)-d-C18- alkyl-C(=0)-, and the alkylene and arylene moieties are optionally substituted as defined in embodiment 1. Embodiment 247. A pharmaceutical preparation according to embodiment 246 wherein B2 is a valence bond, d-C18-alkylene, arylene, heteroarylene, -CrC18-alkyl-aryl-, and the alkylene and arylene moieties are optionally substituted as defined in embodiment 1. Embodiment 248. A pharmaceutical preparation according to embodiment 247 wherein B2 is a valence bond, C C18-alkylene, arylene, -d-C18-alkyl-aryl-, and the alkylene and arylene moieties are optionally substituted as defined in embodiment 1.
Embodiment 249. A pharmaceutical preparation according to embodiment 248 wherein B2 is a valence bond or -CrC18-alkylene, and the alkylene moieties are optionally substituted as defined in embodiment 1. Embodiment 250. A pharmaceutical preparation according to any one of the embodiment s 1 to 249 whereiin Frg2 comprises 1 to 16 positively charged groups.
Embodiment 251. A pharmaceutical preparation according to embodiment 250 wherein Frg2 comprises 1 to 12 positively charged groups.
Embodiment 252. A pharmaceutical preparation according to embodiment 251 wherein Frg2 comprises 1 to 10 positively charged groups. Embodiment 253. A pharmaceutical preparation according to any one of the embodiment s 1 to 249 wherein Frg2 comprises 10 to 20 positively charged groups. Embodiment 254. A pharmaceutical preparation according to embodiment 253 wherein Frg2 comprises 12 to 20 positively charged groups. Embodiment 255. A pharmaceutical preparation according to embodiment 254 wherein Frg2 comprises 16 to 20 positively charged groups.
Embodiment 256. A pharmaceutical preparation according to any one of the embodiment s 250 to 255 wherein the positively charged groups of Frg2 are basic amino acids independently selected from the group consisting of Lys and Arg and D-isomers of these. Embodiment 257. A pharmaceutical preparation according to embodiment 256 wherein the basic amino acids are all Arg.
Embodiment 258. A pharmaceutical preparation according to any one of the embodiment s 250 to 257, wherein Frg2 comprises one or more neutral amino acids independently selected from the group consisting of Gly, Ala, Thr, and Ser. Embodiment 259. A pharmaceutical preparation according to embodiment 258, wherein Frg2 comprises one or more Gly.
Embodiment 260. A pharmaceutical preparation according to any one of the embodiment s 1 to 259 wherein X is -OH or -NH2. Embodiment 261. A pharmaceutical preparation according to embodiment 260 wherein X is - NH2.
Embodiment 262. A pharmaceutical preparation according to any one of the embodiment s 1 to 261 which further comprises at least 3 phenolic molecules per putative insulin hexamer. Embodiment 263. A pharmaceutical preparation according to any one of the embodiment s 1 to 262 wherein the acid-stabilised insulin is an analogue of human insulin wherein A21 is Ala, Gin, Glu, Gly, His, He, Leu, Met, Phe, Ser, Thr, Trp, Tyr, Val, and hSer.
Embodiment 264. A pharmaceutical preparation according to embodiment 263 wherein the acid-stabilised insulin is an analogue of human insulin wherein A21 is Ala, Gly, He, Leu, Phe, Ser, Thr, Val, and hSer. Embodiment 265. A pharmaceutical preparation according to embodiment 264 wherein the acid-stabilised insulin is an analogue of human insulin wherein A21 is Ala or Gly.
Embodiment 266. A pharmaceutical preparation according to embodiment 265 wherein the acid-stabilised insulin is an analogue of human insulin wherein A21 is Gly. Embodiment 267. A pharmaceutical preparation according to any one of the embodiment s 263 to 266 wherein the acid-stabilised insulin is an analogue of human insulin further modi- fied by exchange or deletion of one or more amino acid residues according to the following: B3 is selected from Thr, Ser, Lys or Ala
Alδ is GIn
B28 is Lys, Asp or Glu
B29 is Pro or Glu B9 is Glu or Asp
BIO is Glu
B25 is deleted
B30 is deleted.
Embodiment 268. A pharmaceutical preparation according to embodiment 267 wherein the acid-stabilised insulin is an analogue of human insulin further modified by exchange of B28 to Lys or Asp.
Embodiment 269. A pharmaceutical preparation according to embodiment 268 wherein the acid-stabilised insulin is an analogue of human insulin further modified by exchange of B28 to Asp. Embodiment 270. A pharmaceutical preparation according to embodiment 268 wherein the acid-stabilised insulin is an analogue of human insulin further modified by exchange of B28 to Lys.
Embodiment 271. A pharmaceutical preparation according any one of the embodiment s 263 to 270 wherein the acid-stabilised insulin is an analogue of human insulin further modified by exchange of B29 to Pro.
Embodiment 272. A pharmaceutical preparation according any one of the embodiment s 263 to 271 wherein the acid-stabilised insulin is an analogue of human insulin further modified by exchange of B3 to Lys or Ala.
Embodiment 273. A pharmaceutical preparation according any one of the embodiment s 263 to 272 wherein the acid-stabilised insulin is an analogue of human insulin further modified by exchange of A18 to Gin.
Embodiment 274. A pharmaceutical preparation according any one of the embodiment s 263 to 273 wherein the acid-stabilised insulin is an analogue of human insulin further modified by deletion of B25. Embodiment 275. A pharmaceutical preparation according any one of the embodiment s 263 to 274 wherein the acid-stabilised insulin is an analogue of human insulin further modified by deletion of B30.
Embodiment 276. A pharmaceutical preparation according to embodiment 263 wherein the acid-stabilised insulin is selected from the group A21G A21G, B28K, B29P
A21G, B28D
A21G, B28E
A21G, B3K, B29E A21G, desB27
A21G, B9E
A21G, B9D
A21G, B10E
A21G, desB25 A21G, desB30
A21G, B28K, B29P, desB30
A21G, B28D, desB30
A21G, B28E, desB30
A21G, B3K, B29E, desB30 A21G, desB27, desB30
A21G, B9E, desB30
A21G, B9D, desB30
A21G, B10E, desB30
A21G, desB25, desB30. Embodiment 277. A pharmaceutical preparation according to any one of the embodiment s 1 to 276 wherein zinc ions are present in an amount corresponding to 10 to 40 μg Zn/100 U insulin.
Embodiment 278. A pharmaceutical preparation according to embodiment 277 wherein zinc ions are present in an amount corresponding to 10 to 26 μg Zn/100 U insulin. Embodiment 279. A pharmaceutical preparation according to any one of the embodiment s 1 to 278 wherein the ratio between insulin and the zinc-binding ligand according to any one of the embodiment s 1 to 261 is in the range from 99:1 to 1.99.
Embodiment 280. A pharmaceutical preparation according to embodiment 279 wherein the ratio between insulin and the zinc-binding ligand according to any one of the embodiment s 1 to 261 is in the range from 95:5 to 5:95.
Embodiment 281. A pharmaceutical preparation according to embodiment 280 wherein the ratio between between insulin and the zinc-binding ligand according to any one of the embodiment s 1 to 261 is in the range from 80:20 to 20:80. Embodiment 282. A pharmaceutical preparation according to embodiment 281 wherein the ratio between between insulin and the zinc-binding ligand according to any one of the embodiment s 1 to 261 is in the range from 70:30 to 30:70.
Embodiment 283. A pharmaceutical preparation according to any one of the embodiment s 1 to 282 wherein the concentration of insulin is 60 to 3000 nmol/ml.
Embodiment 284. A pharmaceutical preparation according to embodiment 283 wherein the concentration of insulin is 240 to 1200 nmol/ml.
Embodiment 285. A pharmaceutical preparation according to embodiment 284 wherein the concentration of insulin is about 600 nmol/ml. Embodiment 286. A method of preparing a zinc-binding ligand according to embodiment 1 comprising the steps of
• Identifying starter compounds that binds to the R-state HisB10-Zn2+ site
• optionally attaching a fragment consisting of 0 to 5 neutral α- or β-amino acids
• attaching a fragment comprising 1 to 20 positively charged groups independently se- lected from amino or guanidino groups.
Embodiment 287. Method of prolonging the action of an acid-stabilised insulin preparation which comprises adding a zinc-binding ligand according to any of embodiment s 1 to 261 to the acid-stabilised insulin preparation.
Embodiment 288. A method of treating type 1 or type 2 diabetes comprising administering to a patient in need thereof a theraputically effective amount of a pharmaceutical preparation according to any one of the embodiment s 1 to 282.
Embodiment 289. Use of a preparation according to any one of the embodiment s 1 to 282 for the preparation of a medicament for treatment of type 1 or type 2 diabetes.
PHARMACEUTICAL PREPARATIONS
The present invention also relates to a pharmaceutical preparation for the treatment of diabetes in a patient in need of such a treatment comprising an R-state hexamer of insulin according to the invention together with a pharmaceutically acceptable carrier. In one embodiment of the invention the insulin preparation comprises 60 to 3000 nmol/ml of insulin.
In another embodiment of the invention the insulin preparation comprises 240 to 1200 nmol/ml of insulin. In another embodiment of the invention the insulin preparation comprises about 600 nmol/ml of insulin.
Zinc ions may be present in an amount corresponding to 10 to 40 μg Zn/100 U insulin, more preferably 10 to 26 μg Zn/100 U insulin. Insulin formulations of the invention are usually administered from multi-dose containers where a preservative effect is desired. Since phenolic preservatives also stabilize the R-state hexamer the formulations may contain up to 50 mM of phenolic molecules. The phenolic molecules in the insulin formulation may be selected from the group consisting of phenol, m- cresol, chloro-cresol, thymol, 7-hydroxyindole or any mixture thereof. In one embodiment of the invention 0.5 to 4.0 mg/ml of phenolic compound may be employed.
In another embodiment of the invention 0.6 to 4.0 mg/ml of m-cresol may be employed.
In another embodiment of the invention 0.5 to 4.0 mg/ml of phenol may be employed.
In another embodiment of the invention 1.4 to 4.0 mg/ml of phenol may be employed.
In another embodiment of the invention 0.5 to 4.0 mg/ml of a mixture of m-cresol or phenol may be employed.
In another embodiment of the invention 1.4 to 4.0 mg/ml of a mixture of m-cresol or phenol may be employed.
The pharmaceutical preparation may further comprises a buffer substance, such as a TRIS, phosphate, glycine or glycylglycine (or another zwitterionic substance) buffer, an isotonicity agent, such as NaCl, glycerol, mannitol and/or lactose. Chloride would be used at moderate concentrations (e.g. up to 50 mM) to avoid competition with the zinc-site ligands of the present invention.
The action of insulin may further be slowed down in vivo by the addition of physiologically acceptable agents that increase the viscosity of the pharmaceutical preparation. Thus, the pharmaceutical preparation according to the invention may furthermore comprise an agent which increases the viscosity, such as polyethylene glycol, polypropylene glycol, copolymers thereof, dextrans and/or polylactides.
In a particular embodiment the insulin preparation of the invention comprises between 0.001
% by weight and 1 % by weight of a non-ionic surfactant, for example tween 20 or Polox 188. A nonionic detergent can be added to stabilise insulin against fibrillation during storage and handling.
The insulin preparation of the present invention may have a pH value in the range of 2.5 to
4.5, more preferably pH 3 to 4. In one embodiment the preparations of the invention are used in connection with insulin pumps. The insulin pumps may be prefilled and disposable, or the insulin preparations may be supplied from a reservoir which is removable. Insulin pumps may be skin-mounted or carried, and the path of the insulin preparation from the storage compartment of the pump to the patient may be more or less tortuous. Non-limiting examples of insulin pumps are disclosed in US 5,957,895, US 5,858,001 , US 4,468,221, US 4,468,221 , US 5,957,895, US 5,858,001 , US 6,074,369, US 5,858,001 , US 5,527,288, and US 6,074,369.
In another embodiment the preparations of the invention are used in connection with pen-like injection devices, which may be prefilled and disposable, or the insulin preparations may be supplied from a reservoir which is removable. Non-limiting examples of pen-like injection devices are FlexPen®, InnoLet®, InDuo™, Innovo®.
In a further embodiment preparations of the invention are used in connection with devices for pulmonary administration of aqueous insulin preparations, a non-limiting example of which is the AerX® device.
COMBINATION TREATMENT
The invention furthermore relates to treatment of a patient in which the pharmaceutical preparation of the invention, i.e. comprising zinc ions, acid-stabilised insulin analogue and a ligand for the R-state His810 Zn2+ site, is combined with another form of treatment.
In one aspect of the invention, treatment of a patient with the pharmaceutical preparation of the invention is combined with diet and/or exercise. In another aspect of the invention the pharmaceutical preparation of the invention is administered in combination with one or more further active substances in any suitable ratios. Such further active substances may e.g. be selected from antiobesity agents, antidiabetics, antihypertensive agents, agents for the treatment of complications resulting from or associated with diabetes and agents for the treatment of complications and disorders resulting from or associated with obesity.
Thus, in a further aspect of the invention the pharmaceutical preparation of the invention may be administered in combination with one or more antiobesity agents or appetite regulating agents. Such agents may be selected from the group consisting of CART (cocaine amphetamine regulated transcript) agonists, NPY (neuropeptide Y) antagonists, MC4 (melano- cortin 4) agonists, MC3 (melanocortin 3) agonists, orexin antagonists, TNF (tumor necrosis factor) agonists, CRF (corticotropin releasing factor) agonists, CRF BP (corticotropin releas- ing factor binding protein) antagonists, urocortin agonists, β3 adrenergic agonists such as CL-316243, AJ-9677, GW-0604, LY362884, LY377267 or AZ-40140, MSH (melanocyte- stimulating hormone) agonists, MCH (melanocyte-concentrating hormone) antagonists, CCK (cholecystokinin) agonists, serotonin re-uptake inhibitors such as fluoxetine, seroxat or cita- lopram, serotonin and noradrenaline re-uptake inhibitors, mixed serotonin and noradrenergic compounds, 5HT (serotonin) agonists, bombesin agonists, galanin antagonists, growth hormone, growth factors such as prolactin or placental lactogen, growth hormone releasing compounds, TRH (thyreotropin releasing hormone) agonists, UCP 2 or 3 (uncoupling protein 2 or 3) modulators, leptin agonists, DA agonists (bromocriptin, doprexin), lipase/amylase inhibitors, PPAR (peroxisome proliferator-activated receptor) modulators, RXR (retinoid X re- ceptor) modulators, TR β agonists, AGRP (Agouti related protein) inhibitors, H3 histamine antagonists, opioid antagonists (such as naltrexone), exendin-4, GLP-1 and ciliary neurotro- phic factor.
In one embodiment of the invention the antiobesity agent is leptin.
In another embodiment the antiobesity agent is dexamphetamine or amphetamine. In another embodiment the antiobesity agent is fenfluramine or dexfenfluramine.
In still another embodiment the antiobesity agent is sibutramine.
In a further embodiment the antiobesity agent is orlistat.
In another embodiment the antiobesity agent is mazindol or phentermine.
In still another embodiment the antiobesity agent is phendimetrazine, diethylpropion, fluoxetine, bupropion, topiramate or ecopipam.
The orally active hypoglycemic agents comprise imidazolines, sulphonylureas, biguanides, meglitinides, oxadiazolidinediones, thiazolidinediones, insulin sensitizers, insulin secretagogues such as glimepride, α-glucosidase inhibitors, agents acting on the ATP- dependent potassium channel of the β-cells eg potassium channel openers such as those disclosed in WO 97/26265, WO 99/03861 and WO 00/37474 (Novo Nordisk A/S) which are incorporated herein by reference, or mitiglinide, or a potassium channel blocker, such as BTS-67582, nateglinide, glucagon antagonists such as those disclosed in WO 99/01423 and WO 00/39088 (Novo Nordisk A/S and Agouron Pharmaceuticals, Inc.), which are incorpo- rated herein by reference, GLP-1 agonists such as those disclosed in WO 00/42026 (Novo Nordisk A/S and Agouron Pharmaceuticals, Inc.), which are incorporated herein by reference, DPP-IV (dipeptidyl peptidase-IV) inhibitors, PTPase (protein tyrosine phosphatase) inhibitors, inhibitors of hepatic enzymes involved in stimulation of gluconeogenesis and/or glycogenolysis, glucose uptake modulators, GSK-3 (glycogen synthase kinase-3) inhibitors, compounds modifying the lipid metabolism such as antilipidemic agents, compounds lowering food intake, PPAR (peroxisome proliferator-activated receptor) and RXR (retinoid X receptor) agonists, such as ALRT-268, LG-1268 or LG-1069.
In a further embodiment of the invention the pharmaceutical preparation of the invention is administered in combination with a sulphonylurea e.g. tolbutamide, chlorpropa- mide, tolazamide, glibenclamide, glipizide, glimepiride, glicazide or glyburide.
In another embodiment of the invention the pharmaceutical preparation of the invention is administered in combination with a biguanide, e.g. metformin.
In yet another embodiment of the invention the pharmaceutical preparation of the invention is administered in combination with a meglitinide eg repaglinide or nateglinide. In still another embodiment of the invention the pharmaceutical preparation of the invention is administered in combination with a thiazolidinedione insulin sensitizer, e.g. trogli- tazone, ciglitazone, pioglitazone, rosiglitazone, isaglitazone, darglitazone, englitazone, CS- 011/CI-1037 or T 174 or the compounds disclosed in WO 97/41097, WO 97/41119, WO 97/41120, WO 00/41121 and WO 98/45292 (Dr. Reddy's Research Foundation), which are incorporated herein by reference.
In still another embodiment of the invention the pharmaceutical preparation of the invention may be administered in combination with an insulin sensitizer, e.g. such as GI 262570, YM-440, MCC-555, JTT-501, AR-H039242, KRP-297, GW-409544, CRE-16336, AR-H049020, LY510929, MBX-102, CLX-0940, GW-501516 or the compounds disclosed in WO 99/19313, WO 00/50414, WO 00/63191 , WO 00/63192, WO 00/63193 (Dr. Reddy's Research Foundation) and WO 00/23425, WO 00/23415, WO 00/23451, WO 00/23445, WO 00/23417, WO 00/23416, WO 00/63153, WO 00/63196, WO 00/63209, WO 00/63190 and WO 00/63189 (Novo Nordisk A S), which are incorporated herein by reference.
In a further embodiment of the invention the pharmaceutical preparation of the in- vention is administered in combination with an α-glucosidase inhibitor, e.g. voglibose, emigli- tate, miglitol or acarbose.
In another embodiment of the invention the pharmaceutical preparation of the invention is administered in combination with an agent acting on the ATP-dependent potassium channel of the β-cells, e.g. tolbutamide, glibenclamide, glipizide, glicazide, BTS-67582 or re- paglinide. In yet another embodiment of the invention the pharmaceutical preparation of the invention may be administered in combination with nateglinide.
In still another embodiment of the invention the pharmaceutical preparation of the invention is administered in combination with an antilipidemic agent, e.g. cholestyramine, colestipol, clofibrate, gemfibrozil, lovastatin, pravastatin, simvastatin, probucol or dextrothyroxine.
In another aspect of the invention, the pharmaceutical preparation of the invention is administered in combination with more than one of the above-mentioned compounds, e.g. in combination with metformin and a sulphonylurea such as glyburide; a sulphonylurea and acarbose; nateglinide and metformin; acarbose and metformin; a sulphonylurea, metformin and troglitazone; metformin and a sulphonylurea; etc.
Furthermore, the pharmaceutical preparation of the invention may be administered in combination with one or more antihypertensive agents. Examples of antihypertensive agents are β-blockers such as alprenolol, atenolol, timolol, pindolol, propranolol and metoprolol, ACE (angiotensin converting enzyme) inhibitors such as benazepril, captopril, enalapril, fosinopril, lisinopril, quinapril and ramipril, calcium channel blockers such as nifedipine, felodipine, nicardipine, isradipine, nimodipine, diltiazem and verapamil, and α- blockers such as doxazosin, urapidil, prazosin and terazosin. The pharmaceutical preparation of the invention may also be combined with NEP inhibitors such as candoxatril. Further reference can be made to Remington: The Science and Practice of Pharmacy, 19th Edition, Gennaro, Ed., Mack Publishing Co., Easton, PA, 1995.
It should be understood that any suitable combination of the compounds according to the invention with diet and/or exercise, one or more of the above-mentioned compounds and optionally one or more other active substances are considered to be within the scope of the present invention.
EXAMPLES
The following examples and general procedures refer to intermediate compounds and final products identified in the specification and in the synthesis schemes. The preparation of the compounds of the present invention is described in detail using the following examples, but the chemical reactions described are disclosed in terms of their general applicability to the preparation of compounds of the invention. Occasionally, the reaction may not be applicable as described to each compound included within the disclosed scope of the invention. The compounds for which this occurs will be readily recognised by those skilled in the art. In these cases the reactions can be successfully performed by conventional modifications known to those skilled in the art, that is, by appropriate protection of interfering groups, by changing to other conventional reagents, or by routine modification of reaction conditions. Alternatively, other reactions disclosed herein or otherwise conventional will be applicable to the preparation of the corresponding compounds of the invention. In all preparative methods, all starting materials are known or may easily be prepared from known starting materials. All temperatures are set forth in degrees Celsius and unless otherwise indicated, all parts and percentages are by weight when referring to yields and all parts are by volume when refer- ring to solvents and eluents.
HPLC-MS (Method A)
The following instrumentation was used:
• Hewlett Packard series 1100 G1312A Bin Pump • Hewlett Packard series 1100 Column compartment
• Hewlett Packard series 1100 G13 15A DAD diode array detector
• Hewlett Packard series 1100 MSD
The instrument was controlled by HP Chemstation software. The HPLC pump was connected to two eluent reservoirs containing:
A: 0.01% TFA in water
B: 0.01 % TFA in acetonitrile
The analysis was performed at 40 °C by injecting an appropriate volume of the sample (preferably 1 μL) onto the column, which was eluted with a gradient of acetonitrile.
The HPLC conditions, detector settings and mass spectrometer settings used are given in the following table.
Figure imgf000103_0001
HPLC-MS (Method B)
The following instrumentation was used: Sciex AP1 100 Single quadropole mass spectrometer Perkin Elmer Series 200 Quard pump Perkin Elmer Series 200 autosampler Applied Biosystems 785A UV detector
Sedex 55 evaporative light scattering detector
A Valco column switch with a Valco actuator controlled by timed events from the pump.
The Sciex Sample control software running on a Macintosh PowerPC 7200 computer was used for the instrument control and data acquisition.
The HPLC pump was connected to four eluent reservoirs containing:
Acetonitrile
Water
0.5% TFA in water
0.02 M ammonium acetate
The requirements for samples are that they contain approximately 500 μg/mL of the com- pound to be analysed in an acceptable solvent such as methanol, ethanol, acetonitrile, THF, water and mixtures thereof. (High concentrations of strongly eluting solvents will interfere with the chromatography at low acetonitrile concentrations.)
The analysis was performed at room temperature by injecting 20 μL of the sample solution on the column, which was eluted with a gradient of acetonitrile in either 0.05% TFA or 0.002 M ammonium acetate. Depending on the analysis method varying elution conditions were used.
The eluate from the column was passed through a flow splitting T-connector, which passed approximately 20 μL/min through approx. 1 m. 75 μ fused silica capillary to the API interface of API 100 spectrometer.
The remaining 1.48 mL/min was passed through the UV detector and to the ELS detector.
During the LC-analysis the detection data were acquired concurrently from the mass spectrometer, the UV detector and the ELS detector.
The LC conditions, detector settings and mass spectrometer settings used for the different methods are given in the following table.
Figure imgf000104_0001
HPLC-MS (Method C) The following instrumentation is used:
• Hewlett Packard series 1100 G1312A Bin Pump
• Hewlett Packard series 1100 Column compartment
• Hewlett Packard series 1100 G1315A DAD diode array detector
• Hewlett Packard series 1100 MSD
• Sedere 75 Evaporative Light Scattering detector The instrument is controlled by HP Chemstation software.
The HPLC pump is connected to two eluent reservoirs containing:
0.01% TFA in water
B 0.01% TFA in acetonitrile The analysis is performed at 40 °C by injecting an appropriate volume of the sample (preferably 1 μl) onto the column which is eluted with a gradient of acetonitrile. The HPLC conditions, detector settings and mass spectrometer settings used are given in the following table.
Figure imgf000105_0001
After the DAD the flow is divided yielding approximately 1 ml/min to the ELS and 0.5 ml/min to the MS.
HPLC-MS (Method D) The following instrumentation was used:
Sciex API 150 Single Quadropole mass spectrometer
Hewlett Packard Series 1100 G1312A Bin pump
Gilson 215 micro injector
Hewlett Packard Series 1100 G1315A DAD diode array detector Sedex 55 evaporative light scattering detector
A Valco column switch with a Valco actuator controlled by timed events from the pump.
The Sciex Sample control software running on a Macintosh Power G3 computer was used for the instrument control and data acquisition.
The HPLC pump was connected to two eluent reservoirs containing:
A: Acetonitrile containing 0.05% TFA
B: Water containing 0.05% TFA
The requirements for the samples are that they contain approximately 500 μg/ml of the compound to be analysed in an acceptable solvent such as methanol, ethanol, acetonitrile, THF, water and mixtures thereof. (High concentrations of strongly eluting solvents will interfere with the chromatography at low acetonitrile concentrations.) The analysis was performed at room temperature by injecting 20 μl of the sample solution on the column, which was eluted with a gradient of acetonitrile in 0.05% TFA The eluate from the column was passed through a flow splitting T-connector, which passed approximately 20 μl/min through approx. 1 m 75 μ fused silica capillary to the API interface of API 150 spectrometer.
The remaining 1.48 ml/min was passed through the UV detector and to the ELS detector. During the LC-analysis the detection data were acquired concurrently from the mass spectrometer, the UV detector and the ELS detector.
The LC conditions, detector settings and mass spectrometer settings used for the different methods are given in the following table.
Figure imgf000106_0001
EXAMPLES
0102-0000-0273Example 1 HBOL IH-Benzotriazole
Figure imgf000107_0001
0102-0000-0274Example 2 HBOL 5,6-Dimethyl-1 H-benzotriazole
Figure imgf000107_0002
0102-0000-0275Example 3 HBOL 1 rV-Benzotriazole-5-carboxylic acid
Figure imgf000107_0003
0102-0000-0280Example 4 HBOL 4-Nitro-1 H-benzotriazole
Figure imgf000107_0004
0102-0000-0284Example 5 HBOL 5-Amino-1 --benzotriazole
Figure imgf000107_0005
0102-0000-0287Example 6 HBOL 5-Chloro-1 H-benzotriazole
Figure imgf000108_0001
0102-0000-0286Example 7 HBOL 5-Nitro-1 H-benzotriazole
Figure imgf000108_0002
0102-0000-3015Example 8 PEM
4-[(1 H-Benzotriazole-5-carbonyl)amino]benzoic acid
Figure imgf000108_0003
4-[(1H-Benzotriazole-5-carbonyl)amino]benzoic acid methyl ester (5.2 g, 17.6 mmol) was dissolved in THF (60 mL) and methanol (10 mL) was added followed by 1 N sodium hydroxide (35 mL). The mixture was stirred at room temperature for 16 hours and then 1 N hydrochloric acid (45 mL) was added. The mixture was added water (200 mL) and extracted with ethyl acetate (2 x 500 mL). The combined organic phases were evaporated in vacuo to afford 0.44 g of 4-[(1H-benzotriazole-5-carbonyl)amino]benzoic acid. By filtration of the aqueous phase a further crop of 4-[(1H-benzotriazole-5-carbonyl)amino]benzoic acid was isolated (0.52 g).
1H-NMR (DMSO-dβ): δ 7.97 (4H, s), 8.03 (2H, m), 8.66 (1 H, bs), 10.7 (1 H, s), 12.6 (1 H, bs); HPLC-MS (Method A): m/z: 283 (M+1); Rt = 1.85 min. General procedure (A) for preparation of compounds of general formula :
Figure imgf000109_0001
wherein D, E and R19 are as defined above, and E is optionally substituted with up to three substituents R21, R22 and R23 independently as defined above.
The carboxylic acid of 1 H-benzotriazole-5-carboxylic acid is activated, ie the OH functionality is converted into a leaving group L (selected from eg fluorine, chlorine, bromine, iodine, 1- imidazolyl, 1 ,2,4-triazolyl, 1-benzotriazolyloxy, 1-(4-aza benzotriazolyl)oxy, pentafluoro- phenoxy, N-succinyloxy 3,4-dihydro-4-oxo-3-(1 ,2,3-benzotriazinyl)oxy, benzotriazole 5-COO, or any other leaving group known to act as a leaving group in acylation reactions. The activated benzotriazole-5-carboxylic acid is then reacted with R2-(CH2)n-B' in the presence of a base. The base can be either absent (i.e. R2-(CH2)n-B' acts as a base) or triethylamine, N- ethyl-N,N.-diisopropylamine, N-methylmorpholine, 2,6-lutidine, 2,2,6,6-tetramethylpiperidine, potassium carbonate, sodium carbonate, caesium carbonate or any other base known to be useful in acylation reactions. The reaction is performed in a solvent solvent such as THF, di- oxane, toluene, dichloromethane, DMF, NMP or a mixture of two or more of these. The reaction is performed between 0 °C and 80 °C, preferably between 20 °C and 40 °C. When the acylation is complete, the product is isolated by extraction, filtration, chromatography or other methods known to those skilled in the art.
The general procedure (A) is further illustrated in the following example: 0102-0000-1020Example 9 (General Procedure (A))PEM 1H-Benzotriazole-5-carboxylic acid phenylamide
Figure imgf000109_0002
Benzotriazole-5-carboxylic acid (856 mg), HOAt (715 mg) and EDAC (1.00 g) were dissolved in DMF (17.5 mL) and the mixture was stirred at room temperature 1 hour. A 0.5 mL aliqot of this mixture was added to aniline (13.7 //L, 0.15 mmol) and the resulting mixture was vigorously shaken at room temperature for 16 hours. 1N hydrochloric acid (2 mL) and ethyl acetate (1 mL) were added and the mixture was vigorously shaken at room temperature for 2 hours. The organic phase was isolated and concentrated in vacuo to afford the title compound.
HPLC-MS (Method B): m/z: 239 (M+1); Rt = 3.93 min.
The compounds in the following examples were similarly made. Optionally, the compounds may be isolated by filtration or by chromatography.
0102-0000-1019Example 10 (General Procedure (A))PEM 1 H-Benzotriazole-5-carboxylic acid (4-methoxyphenyl)amide
Figure imgf000110_0001
HPLC-MS (Method A): m/z: 269 (M+1) & 291 (M+23); Rt = 2.41 min HPLC-MS (Method B): m/z: 239 (M+1); Rt = 3.93 min.
0102-0000-1021 Example 11 (General Procedure (A))PEM
{4-[(1H-Benzotriazole-5-carbonyl)amino]phenyl}carbamic acid tert-butyl ester
Figure imgf000110_0002
HPLC-MS (Method B): m/z: 354 (M+1 ); Rt = 4.58 min.
0102-0000-1022Example 12 (General Procedure (A))PEM
1H-Benzotriazole-5-carboxylic acid (4-acetylaminophenyl)amide
Figure imgf000110_0003
HPLC-MS (Method B): m/z: 296 (M+1 ); Rt = 3.32 min.
0102-0000-1023Example 13 (General Procedure (A))PEM 1 H-Benzotriazole-5-carboxylic acid (3-fluorophenyl)amide
Figure imgf000111_0001
HPLC-MS (Method B): m/z: 257 (M+1); Rt = 4.33 min.
0102-0000-1024Example 14 (General Procedure (A))PEM lH-Benzotriazole-5-carboxylic acid (2-chlorophenyl)amide
Figure imgf000111_0002
HPLC-MS (Method B): m/z: 273 (M+1); Rt = 4.18 min.
0102-0000-1025Example 15 (General Procedure (A))PEM 4-[(1H-Benzotriazole-5-carbonyl)amino]benzoic acid methyl ester
Figure imgf000111_0003
HPLC-MS (Method A):m/z: 297 (M+1); Rt : 2,60 min. HPLC-MS (Method B): m/z: 297 (M+1); Rt = 4.30 min.
0102-0000-1026Example 16 (General Procedure (A))PEM 1H-Benzotriazole-5-carboxylic acid (4-butylphenyl)amide
Figure imgf000111_0004
HPLC-MS (Method B): m/z: 295 (M+1); Rt = 5.80 min.
0102-0000-1027Example 17 (General Procedure (A))PEM 1 H-Benzotriazole-5-carboxylic acid (1 -phenylethyl)amide
O CH3
HPLC-MS (Method B): m/z: 267 (M+1); Rt = 4.08 min. 0102-0000-1028Example 18 (General Procedure (A))PEM 1 H-Benzotriazole-5-carboxylic acid benzylamide
Figure imgf000112_0001
HPLC-MS (Method B): m/z: 253 (M+1); Rt = 3.88 min.
0102-0000-1029Example 19 (General Procedure (A))PEM 1 H-Benzotriazole-5-carboxylic acid 4-chlorobenzylamide
Figure imgf000112_0002
HPLC-MS (Method B): m/z: 287 (M+1); Rt = 4.40 min.
0102-0000-1030Example 20 (General Procedure (A))PEM 1 H-Benzotriazole-5-carboxylic acid 2-chlorobenzylamide
Figure imgf000112_0003
HPLC-MS (Method B): m/z: 287 (M+1); Rt = 4.25 min.
0102-0000-1031 Example 21 (General Procedure (A))PEM 1 H-Benzotriazole-5-carboxylic acid 4-methoxybenzylamide
Figure imgf000112_0004
HPLC-MS (Method B): m/z: 283 (M+1 ); Rt = 3.93 min.
0102-0000-1032Example 22 (General Procedure (A))PEM 1 H-Benzotriazole-5-carboxylic acid 3-methoxybenzylamide
Figure imgf000112_0005
HPLC-MS (Method B): m/z: 283 (M+1 ); Rt = 3.97 min. 0102-0000-1033Example 23 (General Procedure (A))PEM 1H-Benzotriazole-5-carboxylic acid (1 ,2-diphenylethyl)amide
Figure imgf000113_0001
HPLC-MS (Method B): m/z: 343 (M+1); Rt = 5.05 min.
0102-0000-1034Example 24 (General Procedure (A))PEM 1H-Benzotriazole-5-carboxylic acid 3-bromobenzylamide
Figure imgf000113_0002
HPLC-MS (Method B): m/z: 331 (M+1); Rt = 4.45 min.
0102-0000-1035Example 25 (General Procedure (A))PEM 4-{[(1 H-Benzotriazole-5-carbonyl)amino]methyl}benzoic acid
Figure imgf000113_0003
HPLC-MS (Method B): m/z: 297 (M+1); Rt = 3.35 min.
0102-0000-1036Example 26 (General Procedure (A))PEM 1H-Benzotriazole-5-carboxylic acid phenethylamide
Figure imgf000113_0004
HPLC-MS (Method B): m/z: 267 (M+1); Rt = 4.08 min.
0102-0000-1037Example 27 (General Procedure (A))PEM 1H-Benzotriazole-5-carboxylic acid [2-(4-chlorophenyl)ethyl]amide
Figure imgf000113_0005
HPLC-MS (Method B): m/z: 301 (M+1); Rt = 4.50 min. 0102-0000-1038Example 28 (General Procedure (A))PEM
1 H-Benzotriazole-5-carboxylic acid [2-(4-methoxyphenyl)ethyl]amide
Figure imgf000114_0001
HPLC-MS (Method B): m/z: 297 (M+1); Rt = 4.15 min.
0102-0000-1039Example 29 (General Procedure (A))PEM
1 H-Benzotriazole-5-carboxylic acid [2-(3-methoxyphenyl)ethyl]amide
Figure imgf000114_0002
HPLC-MS (Method B): m/z: 297 (M+1 ); Rt = 4.13 min.
0102-0000-1040Example 30 (General Procedure (A))PEM
1 H-Benzotriazole-5-carboxylic acid [2-(3-chlorophenyl)ethyl]amide
Figure imgf000114_0003
HPLC-MS (Method B): m/z: 301 (M+1); Rt = 4.55 min.
0102-0000-1041 Example 31 (General Procedure (A))PEM 1 H-Benzotriazole-5-carboxylic acid (2,2-diphenylethyl)amide
Figure imgf000114_0004
HPLC-MS (Method B): m/z: 343 (M+1 ); Rt = 5.00 min.
0102-0000-1042Example 32 (General Procedure (A))PEM
1 H-Benzotriazole-5-carboxylic acid (3,4-dichlorophenyl)methylamide
Figure imgf000114_0005
HPLC-MS (Method B): m/z: 321 (M+1); Rt = 4.67 min.
0102-0000-1043Example 33 (General Procedure (A))PEM 1H-Benzotriazole-5-carboxylic acid methylphenylamide
Figure imgf000115_0001
HPLC-MS (Method B): m/z: 253 (M+1); Rt = 3.82 min.
0102-0000-1044Example 34 (General Procedure (A))PEM 1H-Benzotriazole-5-carboxylic acid benzylmethylamide
Figure imgf000115_0002
HPLC-MS (Method B): m/z: 267 (M+1); Rt = 4.05 min.
0102-0000-1045Example 35 (General Procedure (A))PEM
1 H-Benzotriazole-5-carboxylic acid [2-(3-chloro-4-methoxyphenyl)ethyl]methyl-amide
Figure imgf000115_0003
HPLC-MS (Method B): m/z: 345 (M+1 ); Rt = 4.37 min.
0102-0000-1046Example 36 (General Procedure (A))PEM 1H-Benzotriazole-5-carboxylic acid methylphenethylamide
Figure imgf000115_0004
HPLC-MS (Method B): m/z: 281 (M+1); Rt = 4.15 min.
0102-0000-1047Example 37 (General Procedure (A))PEM
1 H-Benzotriazole-5-carboxylic acid [2-(3,4-dimethoxyphenyl)ethyl]methylamide
Figure imgf000116_0001
HPLC-MS (Method B): m/z: 341 (M+1); Rt = 3.78 min;
0102-0000-1048Example 38 (General Procedure (A))PEM 1 H-Benzotriazole-5-carboxylic acid (2-hydroxy-2-phenylethyl)methylamide
Figure imgf000116_0002
HPLC-MS (Method B): m/z: 297 (M+1); Rt = 3.48 min.
Example 39 (General procedure (A)) 1 H-Benzotriazole-5-carboxylic acid (3-bromophenyl)amide
Figure imgf000116_0003
HPLC-MS (Method A): m/z: 317 (M+1); Rt = 3.19 min.
Example 40 (General procedure (A)) 1 H-Benzotriazole-5-carboxylic acid (4-bromophenyl)amide
Figure imgf000116_0004
HPLC-MS (Method A): m/z: 317 (M+1 ); Rt = 3.18 min.
Example 41 (General procedure (A)) {4-[(1 H-Benzotriazole-5-carbonyl)amino]benzoylamino}acetic acid
Figure imgf000116_0005
HPLC-MS (Method A): m/z: 340 (M+1); Rt = 1.71 min. Example 42 (General procedure (A))
{4-[(1 H-Benzotriazole-5-carbonyl)amino]phenyl}acetic acid
Figure imgf000117_0001
HPLC-MS (Method A): m/z: 297 (M+1 ); Rt = 2.02 min.
Example 43 (General procedure (A)) 3-{4-[(1 H-Benzotriazole-5-carbonyl)amino]phenyl}acrylic acid
Figure imgf000117_0002
HPLC-MS (Method A): m/z: 309 (M+1); Rt = 3.19 min.
Example 44 (General procedure (A))
{3-[(1 H-Benzotriazole-5-carbonyl)amino]phenyl}acetic acid
Figure imgf000117_0003
HPLC-MS (Method A): m/z: 297 (M+1 ); Rt = 2.10 min.
Example 45 (General procedure (A))
2-{4-[(1 H-Benzotriazole-5-carbonyl)amino]phenoxy}-2-methylpropionic acid
O
VN TJO&Ϊ 4:)H .N^^ ^N^^π3° ^
H HPLC-MS (Method A): m/z: 341 (M+1 ); Rt = 2.42 min.
Example 46 (General procedure (A)) 3-{4-[(1 H-Benzotriazole-5-carbonyl)amino]benzoylamino}propionic acid
Figure imgf000118_0001
HPLC-MS (Method A): m/z: 354 (M+1 ); Rt = 1.78 min.
Example 47 (General procedure (A)) 3-{4-[(1 H-Benzotriazole-5-carbonyl)amino]phenyl}propionic acid
Figure imgf000118_0002
HPLC-MS (Method A): m/z: 311 (M+1 ); Rt = 2.20 min.
Example 48 (General procedure (A)) 1H-Benzotriazole-5-carboxylic acid (4-benzyloxyphenyl)amide
Figure imgf000118_0003
HPLC-MS (Method A): m/z: 345 (M+1); Rt = 3.60 min.
Example 49 (General procedure (A)) 1 H-Benzotriazole-5-carboxylic acid (3-chloro-4-methoxyphenyl)amide
Figure imgf000118_0004
HPLC-MS (Method A): m/z: 303 (M+1); Rt = 2.88 min.
Example 50 (General procedure (A)) 1 H-Benzotriazole-5-carboxylic acid (4-phenoxyphenyl)amide
Figure imgf000119_0001
HPLC-MS (Method A): m/z: 331 (M+1); Rt = 3.62 min.
Example 51 (General procedure (A)) 1 H-Benzotriazole-5-carboxylic acid (4-butoxyphenyl)amide
Figure imgf000119_0002
HPLC-MS (Method A): m/z: 311 (M+1 ); Rt = 3.59 min.
Example 52 (General procedure (A)) 1 H-Benzotriazole-5-carboxylic acid (3-bromo-4-trifluoromethoxyphenyl)amide
Figure imgf000119_0003
HPLC-MS (Method A): m/z: 402 (M+1 ); Rt = 3.93 min.
Example 53 (General procedure (A)) 1 H-Benzotriazole-5-carboxylic acid (3,5-dichloro-4-hydroxyphenyl)amide
Figure imgf000119_0004
HPLC-MS (Method A): m/z: 323 (M+1); Rt = 2.57 min.
Example 54 (General procedure (A)) 4-{[(1 H-Benzotriazole-5-carbonyl)amino]methyl}benzoic acid
Figure imgf000120_0001
HPLC-MS (Method A): m/z: 297 (M+1); Rt = 1.86 min.
Example 55 (General procedure (A)) {4-[(1H-Benzotriazole-5-carbonyl)amino]phenylsulfanyl}acetic acid
Figure imgf000120_0002
HPLC-MS (Method A): m/z: 329 (M+1 ); Rt = 2.34 min.
Example 56 N-(1 H-Benzotriazol-5-yl)acetamide
Figure imgf000120_0003
HPLC-MS (Method A): m/z: 177 (M+1); Rt = 0.84 min.
Example 57 (General Procedure (A)) 1 H-Benzotriazole-5-carboxylic acid 4-nitrobenzylamide
Figure imgf000120_0004
The following compound is prepared according to general procedure (N) as described below: Example 58 (General procedure (N)) 1H-Benzotriazole-5-carboxylic acid 4-chlorobenzylamide
Figure imgf000120_0005
HPLC-MS (Method B): m/z: 287 (M+1 ); Rt = 4.40 min. Example 59 2-[(1 H-Benzotriazol-5-ylimino)methyl]-4,6-dichlorophenol
Figure imgf000121_0001
Example 60 Diethyl 2-[(1H-benzotriazol-6-ylamino)methylidene]malonate
Figure imgf000121_0002
Example 61 N1-(1H-Benzotriazol-5-yl)-3-chlorobenzamide
Figure imgf000121_0003
Example 62 N1-(1H-Benzotriazol-5-yl)-3,4,5-trimethoxybenzamide
Figure imgf000121_0004
Example 63 N2-(1 H-Benzotriazol-5-yl)-3-chlorobenzo[b]thiophene-2-carboxamide
Figure imgf000122_0001
Example 64 6-Bromo-1 H-benzotriazole
Figure imgf000122_0002
Example 65 2-[(1H-Benzotriazol-5-ylimino)methyl]-4-bromophenol
Figure imgf000122_0003
General procedure (B) for preparation of compounds of general formula l2:
Figure imgf000122_0004
wherein X, Y, A and R3 are as defined above and A is optionally substituted with up to four substituents R7, R8, R9, and R10 as defined above.
The chemistry is well known (eg Lohray et al., J. Med. Chem., 1999, 42, 2569-81) and is generally performed by reacting a carbonyl compound (aldehyde or ketone) with the heterocyclic ring (eg thiazolidine-2,4-dione (X = O; Y = S), rhodanine (X = Y = S) and hydantoin (X = O; Y = NH) in the presence of a base, such as sodium acetate, potassium acetate, ammonium acetate, piperidinium benzoate or an amine (eg piperidine, triethylamine and the like) in a solvent (eg acetic acid, ethanol, methanol, DMSO, DMF, NMP, toluene, benzene) or in a mixture of two or more of these solvents. The reaction is performed at room temperature or at elevated temperature, most often at or near the boiling point of the mixture. Optionally, azeotropic removal of the formed water can be done.
This general procedure (B) is further illustrated in the following example: Example 66 (General procedure (B)) 5-(3-Phenoxybenzylidene)thiazolidine-2,4-dione
Figure imgf000123_0001
A solution of thiazolidine-2,4-dione (90%, 78 mg, 0.6 mmol) and ammonium acetate (92 mg, 1.2 mmol) in acetic acid (1 mL) was added to 3-phenoxybenzaldehyde (52 μl, 0.6 mmol) and the resulting mixture was shaken at 115 °C for 16 hours. After cooling, the mixture was con- centrated in vacuo to afford the title compound.
HPLC-MS (Method A): m/z: 298 (M+1); Rt = 4.54 min.
The compounds in the following examples were similarly prepared. Optionally, the com- pounds can be further purified by filtration and washing with water, ethanol and / or heptane instead of concentration in vacuo. Also optionally the compounds can be purified by washing with ethanol, water and/or heptane, or by chromatography, such as preparative HPLC. Example 67 (General procedure (B)) 5-(4-Dimethylaminobenzylidene)thiazolidine-2,4-dione
Figure imgf000123_0002
HPLC-MS (Method C): m/z: 249 (M+1); Rt = 4.90 min
Example 68 (General procedure (B)) 5-Naphthalen-1-ylmethylenethiazolidine-2,4-dione
Figure imgf000124_0001
HPLC-MS (Method A): m/z: 256 (M+1); Rt = 4,16 min.
Example 69 (General procedure (B)) 5-Benzylidene-thiazolidine-2,4-dione
Figure imgf000124_0002
HPLC-MS (Method A): m/z: 206 (M+1 ); Rt = 4,87 min.
Example 70 (General procedure (B)) 5-(4-Diethylaminobenzylidene)thiazolidine-2,4-dione
Figure imgf000124_0003
HPLC-MS (Method A): m/z: 277 (M+1); Rt = 4.73 min.
Example 71 (General procedure (B)) 5-(4-Methoxy-benzylidene)-thiazolidine-2,4-dione
Figure imgf000124_0004
HPLC-MS (Method A): m/z: 263 (M+1); Rt = 4,90 min.
Example 72 (General procedure (B)) 5-(4-Chloro-benzylidene)-thiazolidine-2,4-dione
Figure imgf000125_0001
HPLC-MS (Method A): m/z: 240 (M+1); Rt = 5,53 min.
Example 73 (General procedure (B)) 5-(4-Nitro-benzylidene)-thiazolidine-2,4-dione
Figure imgf000125_0002
HPLC-MS (Method A): m/z: 251 (M+1); Rt = 4,87 min.
Example 74 (General procedure (B)) 5-(4-Hydroxy-3-methoxy-benzylidene)-thiazolidine-2,4-dione
Figure imgf000125_0003
HPLC-MS (Method A): m/z: 252 (M+1); Rt = 4,07 min.
Example 75 (General procedure (B)) 5-(4-Methylsulfanylbenzylidene)thiazolidine-2,4-dione
Figure imgf000125_0004
HPLC-MS (Method A): m/z: 252 (M+1); Rt = 5,43 min.
Example 76 (General procedure (B)) 5-(2-Pentyloxybenzylidene)thiazolidine-2,4-dione
Figure imgf000126_0001
HPLC-MS (Method C): m/z: 292 (M+1); Rt = 4.75 min.
1H NMR (DMSO-de): δ = 0.90 (3H, t), 1.39 (4H, m), 1.77 (2H, p), 4.08 (2H, t), 7.08 (1 H, t),
7.14 (1 H, d), 7.43 (2H, m), 8.03 (1 H, s), 12.6 (1 H, bs).
Example 77 (General procedure (B)) 5-(3-Fluoro-4-methoxybenzylidene)thiazolidine-2,4-dione
Figure imgf000126_0002
HPLC-MS (Method A): m/z: 354 (M+1); Rt = 4,97 min.
Example 78 (General procedure (B)) 5-(4-tert-Butylbenzylidene)thiazolidine-2,4-dione
Figure imgf000126_0003
HPLC-MS (Method A): m/z: 262 (M+1); Rt = 6,70 min.
Example 79 (General procedure (B)) Λ/-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenyl]acetamide
Figure imgf000126_0004
HPLC-MS (Method A): m/z: 263 (M+1); Rt = 3,90 min. Example 80 (General procedure (B)) 5-Biphenyl-4-ylmethylene-thiazolidine-2,4-dione
Figure imgf000127_0001
HPLC-MS (Method A): m/z: 282 (M+1); Rt = 4,52 min.
Example 81 (General procedure (B)) 5-(4-Phenoxy-benzylidene)-thiazolidine-2,4-dione
Figure imgf000127_0002
HPLC-MS (Method A): m/z: 298 (M+1); Rt = 6,50 min.
Example 82 (General procedure (B)) 5-(3-Benzyloxybenzylidene)thiazolidine-2,4-dione
Figure imgf000127_0003
HPLC-MS (Method A): m/z: 312 (M+1); Rt = 6,37 min.
Example 83 (General procedure (B)) 5-(3-p-Tolyloxybenzylidene)thiazolidine-2,4-dione
Figure imgf000127_0004
HPLC-MS (Method A): m/z: 312 (M+1); Rt = 6,87 min.
Example 84 (General procedure (B)) 5-Naphthalen-2-ylmethylene-thiazolidine-2,4-dione
Figure imgf000127_0005
HPLC-MS (Method A): m/z: 256 (M+1); Rt - 4.15 min.
Example 85 (General procedure (B)) 5-Benzo[1,3]dioxol-5-ylmethylenethiazolidine-2,4-dione
Figure imgf000128_0001
HPLC-MS (Method A): m/z: 250 (M+1), Rt = 3.18 min.
Example 86 (General procedure (B)) 5-(4-Chlorobenzylidene)-2-thioxothiazolidin-4-one
Figure imgf000128_0002
HPLC-MS (Method A): m/z: 256 (M+1); Rt = 4,51 min.
Example 87 (General procedure (B)) 5-(4-Dimethylaminobenzylidene)-2-thioxothiazolidin-4-one
Figure imgf000128_0003
HPLC-MS (Method A): m/z: 265 (M+1); Rt = 5,66 min.
Example 88 (General procedure (B)) 5-(4-Nitrobenzylidene)-2-thioxothiazolidin-4-one
Figure imgf000128_0004
HPLC-MS (Method A): m/z: 267 (M+1); Rt = 3,94 min. Example 89 (General procedure (B))
5-(4-Methylsulfanylbenzylidene)-2-thioxothiazolidin-4-one
Figure imgf000129_0001
HPLC-MS (Method A): m/z: 268 (M+1 ); Rt = 6,39 min.
Example 90 (General procedure (B)) 5-(3-Fluoro-4-methoxybenzylidene)-2-thioxothiazolidin-4-one
Figure imgf000129_0002
HPLC-MS (Method A): m/z: 270 (M+1); Rt = 5,52 min.
Example 91 (General procedure (B)) 5-Naphthalen-2-ylmethylene-2-thioxothiazolidin-4-one
Figure imgf000129_0003
HPLC-MS (Method A): m/z: 272 (M+1); Rt = 6,75 min.
Example 92 (General procedure (B)) 5-(4-Diethylaminobenzylidene)-2-thioxothiazolidin-4-one
Figure imgf000129_0004
HPLC-MS (Method A): m/z: 293 (M+1); Rt = 5,99 min.
Example 93 (General procedure (B)) 5-Biphenyl-4-ylmethylene-2-thioxothiazolidin-4-one
Figure imgf000130_0001
HPLC-MS (Method A): m/z: 298 (M+1); Rt = 7,03 min.
Example 94 (General procedure (B)) 5-(3-Phenoxybenzylidene)-2-thioxothiazolidin-4-one
Figure imgf000130_0002
HPLC-MS (Method A): m/z: 314 (M+1); Rt = 6,89 min.
Example 95 (General procedure (B)) 5-(3-Benzyloxybenzylidene)-2-thioxothiazolidin-4-one
Figure imgf000130_0003
HPLC-MS (Method A): m/z: 328 (M+1); Rt = 6,95 min.
Example 96 (General procedure (B)) 5-(4-Benzyloxybenzylidene)-2-thioxothiazolidin-4-one
Figure imgf000130_0004
HPLC-MS (Method A): m/z: 328 (M+1 ); RT = 6,89 min.
Example 97 (General procedure (B)) 5-Naphthalen-1 -ylmethylene-2-thioxothiazolidin-4-one
Figure imgf000130_0005
HPLC-MS (Method A): m/z: 272 (M+1); Rt = 6,43 min. Example 98 (General procedure (B)) 5-(3-Methoxybenzyl)thiazolidine-2,4-dione
Figure imgf000131_0001
HPLC-MS (Method A): m/z: 236 (M+1 ); Rt = 3,05 min.
Example 99 (General procedure (D)) 4-[2-Chloro-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]butyric acid ethyl ester
Figure imgf000131_0002
HPLC-MS (Method A): m/z: 392 (M+23), Rt = 4.32 min.
Example 100 (General procedure (D)) 4-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)-phenoxy]-butyric acid
Figure imgf000131_0003
HPLC-MS (Method A): m/z: 410 (M+23); Rt = 3,35 min.
Example 101 (General procedure (B)) 5-(3-Bromobenzylidene)thiazolidine-2,4-dione
Figure imgf000131_0004
HPLC-MS (Method A): m/z: 285 (M+1 ); Rt = 4.01 min.
Example 102 (General procedure (B)) 5-(4-Bromobenzylidene)thiazolidine-2,4-dione
Figure imgf000132_0001
HPLC-MS (Method A): m/z: 285 (M+1); Rt = 4.05 min.
Example 103 (General procedure (B)) 5-(3-Chlorobenzylidene)thiazolidine-2,4-dione
Figure imgf000132_0002
HPLC-MS (Method A): m/z: 240 (M+1); Rt = 3.91 min.
Example 104 (General procedure (B)) 5-Thiophen-2-ylmethylenethiazolidine-2,4-dione
Figure imgf000132_0003
HPLC-MS (Method A): m/z: 212 (M+1); Rt = 3.09 min.
Example 105 (General procedure (B)) 5-(4-Bromothiophen-2-ylmethylene)thiazolidine-2,4-dione
Figure imgf000132_0004
HPLC-MS (Method A): m/z: 291 (M+1); Rt = 3.85 min.
Example 106 (General procedure (B)) 5-(3,5-Dichlorobenzylidene)thiazolidine-2,4-dione
Figure imgf000133_0001
HPLC-MS (Method A): m/z: 274 (M+1 ); Rt = 4.52 min.
Example 107 (General procedure (B))
5-(1 -Methyl-1 H-indol-3-ylmethylene)thiazolidine-2,4-dione
Figure imgf000133_0002
HPLC-MS (Method A): m/z: 259 (M+1); Rt = 3.55 min.
Example 108 (General procedure (B)) 5-(1 H-lndol-3-ylmethylene)thiazolidine-2,4-dione
Figure imgf000133_0003
HPLC-MS (Method A): m/z: 245 (M+1); Rt = 2.73 min.
Example 109 (General procedure (B)) 5-Fluoren-9-ylidenethiazolidine-2,4-dione
Figure imgf000133_0004
HPLC-MS (Method A): m/z: 280 (M+1); Rt = 4.34 min. Example 110 (General procedure (B)) 5-(1-Phenylethylidene)thiazolidine-2,4-dione
Figure imgf000134_0001
HPLC-MS (Method A): m/z: 220 (M+1); Rt = 3,38 min.
Example 111 (General procedure (B)) 5-[1-(4-Methoxyphenyl)-ethylidene]-thiazolidine-2,4-dione
Figure imgf000134_0002
HPLC-MS (Method A): m/z: 250 (M+1); Rt = 3.55 min.
Example 112 (General procedure (B)) 5-(1-Naphthalen-2-yl-ethylidene)-thiazolidine-2,4-dione
Figure imgf000134_0003
HPLC-MS (Method A): m/z: 270 (M+1); Rt = 4,30 min.
Example 113 (General procedure (B)) 5-[1-(4-Bromophenyl)-ethylidene]-thiazolidine-2,4-dione
Figure imgf000134_0004
HPLC-MS (Method A): m/z: 300 (M+1); Rt = 4,18 min. Example 114 (General procedure (B)) 5-(2,2-Diphenylethylidene)-thiazolidine-2,4-dione
Figure imgf000135_0001
HPLC-MS (Method A): m/z: 296 (M+1); Rt = 4,49 min.
Example 115 (General procedure (B)) 5-[1-(3-Methoxyphenyl)-ethylidene]-thiazolidine-2,4-dione
Figure imgf000135_0002
HPLC-MS (Method A): m/z: 250 (M+1); Rt = 3,60 min.
Example 116 (General procedure (B)) 5-[1-(6-Methoxynaphthalen-2-yl)-ethylidene]-thiazolidine-2,4-dione
Figure imgf000135_0003
HPLC-MS (Method A): m/z: 300 (M+1); Rt = 4,26 min.
Example 117 (General procedure (B))
5-[1-(4-Phenoxyphenyl)-ethylidene]-thiazolidine-2,4-dione
Figure imgf000135_0004
HPLC-MS (Method A): m/z: 312 (M+1); Rt = 4,68 min.
Example 118 (General procedure (B)) 5-[1-(3-Fluoro-4-methoxyphenyl)ethylidene]thiazolidine-2,4-dione
Figure imgf000136_0001
HPLC-MS (Method A): m/z: 268 (M+1); Rt = 3,58 min.
Example 119 (General procedure (B)) 5-[1-(3-Bromophenyl)-ethylidene]-thiazolidine-2,4-dione
Figure imgf000136_0002
HPLC-MS (Method A): m/z: 300 (M+1); Rt = 4,13 min.
Example 120 (General procedure (B)) 5-Anthracen-9-ylmethylenethiazolidine-2,4-dione
Figure imgf000136_0003
HPLC-MS (Method A): m/z: 306 (M+1); Rt = 4,64 min.
Example 121 (General procedure (B)) 5-(2-Methoxynaphthalen-1-ylmethylene)-thiazolidine-2,4-dione
Figure imgf000136_0004
HPLC-MS (Method A): m/z: 286 (M+1); Rt = 4,02 min.
Example 122 (General procedure (B)) 5-(4-Methoxynaphthalen-1-ylmethylene)-thiazolidine-2,4-dione
Figure imgf000137_0001
HPLC-MS (Method A): m/z: 286 (M+1 ); Rt = 4,31 min.
Example 123 (General procedure (B)) 5-(4-Dimethylaminonaphthalen-1-ylmethylene)-thiazolidine-2,4-dione
Figure imgf000137_0002
HPLC-MS (Method A): m/z: 299 (M+1); Rt = 4,22 min.
Example 124 (General procedure (B)) 5-(4-Methylnaphthalen-1-ylmethylene)-thiazolidine-2,4-dione
Figure imgf000137_0003
HPLC-MS (Method A): m/z: 270 (M+1 ); Rt = 4,47 min.
Example 125 (General procedure (B)) 5-Pyridin-2-ylmethylene-thiazolidine-2,4-dione
Figure imgf000137_0004
Example 126 5-Pyridin-2-ylmethyl-thiazolidine-2,4-dione
Figure imgf000138_0001
5-Pyridin-2-ylmethylene-thiazolidine-2,4-dione (5 g) in tetrahydrofuran (300 ml) was added 10% Pd/C (1 g) and the mixture was hydrogenated at ambient pressure for 16 hours. More 10% Pd/C (5 g) was added and the mixture was hydrogenated at 50 psi for 16 hours. After filtration and evaporation in vacuo, the residue was purified by column chromatography eluting with a mixture of ethyl acetate and heptane (1:1). This afforded the title compound (0.8 g, 16%) as a solid.
TLC: Rf = 0.30 (Si02; EtOAc: heptane 1 :1)
Example 127 (General procedure (B)) 5-(1H-lmidazol-4-ylmethylene)-thiazolidine-2,4-dione
Figure imgf000138_0002
Example 128 (General procedure (B))
5-(4-Benzyloxy-benzylidene)-thiazolidine-2,4-dione
Figure imgf000138_0003
HPLC-MS (Method A): m/z: 6,43 min ; 99 % (2A)
Example 129 (General procedure (B))
5-[4-(4-Fluorobenzyloxy)benzylidene]-2-thioxothiazolidin-4-one
Figure imgf000138_0004
Example 130 (General procedure (B)) 5-(4-Butoxybenzylidene)-2-thioxothiazolidin-4-one
Figure imgf000139_0001
Example 131 (General procedure (B)) 5-(3-Methoxybenzylidene)thiazolidine-2,4-dione
Figure imgf000139_0002
HPLC-MS (Method A): m/z: 236 (M+1 ); Rt = 4,97 min
Example 132 (General procedure (B)) 5-(3-Methoxybenzylidene)imidazolidine-2,4-dione
Figure imgf000139_0003
HPLC-MS (Method A): m/z: 219 (M+1); Rt = 2.43 min.
Example 133 (General procedure (B)) 5-(4-Methoxybenzylidene)imidazolidine-2,4-dione
Figure imgf000139_0004
HPLC-MS (Method A): m/z: 219 (M+1); Rt = 2.38 min. Example 134 (General procedure (B)) 5-(2,3-Dichlorobenzylidene)thiazolidine-2,4-dione
Figure imgf000140_0001
Example 135 (General procedure (B)) 5-Benzofuran-7-ylmethylenethiazolidine-2,4-dione
Figure imgf000140_0002
HPLC-MS (Method C): m/z: 247 (M+1); Rt = 4,57 min.
Example 136 (General procedure (B))
5-Benzo[1,3]dioxol-4-ylmethylenethiazolidine-2,4-dione
Figure imgf000140_0003
HPLC-MS (Method C): m/z: 250 (M+1); Rt = 4,00 min.
Example 137 (General procedure (B))
5-(4-Methoxy-2,3-dimethylbenzylidene)thiazolidine-2,4-dione
Figure imgf000140_0004
HPLC-MS (Method C): m/z: 264 (M+1); Rt = 5,05 min.
Example 138 (General procedure (B))
5-(2-Benzyloxy-3-methoxybenzylidene)thiazolidine-2,4-dione
Figure imgf000141_0001
HPLC-MS (Method C): m/z: 342 (M+1); Rt = 5,14 min.
Example 139 (General procedure (B)) 5-(2-Hydroxybenzylidene)thiazolidine-2,4-dione
Figure imgf000141_0002
HPLC-MS (Method C): m/z: 222 (M+1); Rt = 3,67 min.
Example 140 (General procedure (B)) 5-(2,4-Dichlorobenzylidene)thiazolidine-2,4-dione
Figure imgf000141_0003
Η-NMR (DMSO-de): 7.60 (2H, "s"), 7.78 (1H, s), 7.82 (1H, s).
Example 141 (General procedure (B)) 5-(2-Chlorobenzylidene)thiazolidine-2,4-dione
HN 3 δ ci
1H-NMR (DMSO-de): 7.40 (1H, t), 7.46 (1H, t), 7.57 (1H, d), 7.62 (1H, d), 7.74 (1H, s).
Example 142 (General procedure (B)) 5-(2-Bromobenzylidene)thiazolidine-2,4-dione
Figure imgf000141_0004
Η-NMR (DMSO-de): 7.33 (1H, t), 7.52 (1H, t), 7.60 (1H, d), 7.71 (1H, s), 7.77 (1H, d). Example 143 (General procedure (B)) 5-(2,4-Dimethoxybenzylidene)thiazolidine-2,4-dione
Figure imgf000142_0001
HPLC-MS (Method C): m/z: 266 (M+1 ) Rt = 4,40 min.
Example 144 (General procedure (B)) 5-(2-Methoxybenzylidene)thiazolidine-2,4-dione
Figure imgf000142_0002
HPLC-MS (Method C): m/z: 236 (M+1); Rt = 4,17 min.
Example 145 (General procedure (B)) 5-(2,6-Difluorobenzylidene)thiazolidine-2,4-dione
Figure imgf000142_0003
HPLC-MS (Method C): m/z: 242 (M+1); Rt = 4,30 min.
Example 146 (General procedure (B)) 5-(2,4-Dimethylbenzylidene)thiazolidine-2,4-dione O^ ^CH3
HN Jϋ O CH3 HPLC-MS (Method C): m/z: 234 (M+1 ); Rt = 5,00 min.
Example 147 (General procedure (B)) 5-(2,4,6-Trimethoxybenzylidene)thiazolidine-2,4-dione
Figure imgf000143_0001
HPLC-MS (Method C): m/z: 296 (M+1); Rt = 4,27 min.
Example 148 (General procedure (B)) 5-(4-Hydroxy-2-methoxybenzylidene)thiazolidine-2,4-dione
Figure imgf000143_0002
HPLC-MS (Method C): m/z: 252 (M+1); Rt = 3,64 min.
Example 149 (General procedure (B)) 5-(4-Hydroxynaphthalen-1 -ylmethylene)thiazolidine-2,4-dione
Figure imgf000143_0003
Η-NMR (DMSO-dβ): δ = 7.04 (1H, d), 7.57 (2H, m), 7.67 (1H, t), 8.11 (1H, d), 8.25 (1H, d), 8.39 (1 H, s) 11.1 (1H, s), 12.5 (1H, bs). HPLC-MS (Method C): m/z: 272 (M+1); Rt = 3.44 min.
Example 150 (General procedure (B)) 5-(2-Trifluoromethoxybenzylidene)thiazolidine-2,4-dione
Figure imgf000143_0004
HPLC-MS (Method C): m/z: 290 (M+1); Rt = 4,94 min.
Example 151 (General procedure (B)) 5-Biphenyl-2-ylmethylenethiazolidine-2,4-dione
Figure imgf000144_0001
HPLC-MS (Method C): m/z: 282 (M+1); Rt = 5,17 min.
Example 152 (General procedure (B)) 5-(2-Benzyloxybenzylidene)thiazolidine-2,4-dione
Figure imgf000144_0002
HPLC-MS (Method C): m/z: 312 (M+1); Rt = 5,40 min.
Example 153 (General procedure (B)) 5-Adamantan-2-ylidenethiazolidine-2,4-dione
Figure imgf000144_0003
HPLC-MS (Method A): m/z: 250 (M+1); Rt = 4,30 min.
Example 154 (General Procedure (B)) 5-[3-(4-Nitrophenyl)allylidene]thiazolidine-2,4-dione
Figure imgf000144_0004
HPLC-MS (Method C): m/z: 277 (M+1); Rt = 3.63 min.
Example 155 (General Procedure (B)) 5-[3-(2-Methoxyphenyl)allylidene]thiazolidine-2,4-dione
Figure imgf000145_0001
HPLC-MS (Method C): m/z: 262 (M+1); Rt = 3.81 min.
Example 156 (General Procedure (B)) 5-[3-(4-Methoxyphenyl)allylidene]thiazolidine-2,4-dione
Figure imgf000145_0002
HPLC-MS (Method C): m/z: 262 (M+1); Rt = 3.67 min.
Example 157 (General procedure (B)) 5-(4-Hydroxybenzylidene)thiazolidine-2,4-dione
Figure imgf000145_0003
Example 158 (General procedure (B)) 5-(4-Dimethylaminobenzylidene)pyrimidine-2,4,6-trione
Figure imgf000145_0004
HPLC-MS (Method C): m/z = 260 (M+1 ) Rt = 2,16 min.
Example 159 (General procedure (B)) 5-(9-Ethyl-9H-carbazol-2-ylmethylene)-pyrimidine-2,4,6-trione
Figure imgf000146_0001
HPLC-MS (Method C): m/z = 334 (M+1); Rt = 3,55 min.
Example 160 (General procedure (B)) 5-(4-Hexyloxynaphthalen-1 -ylmethylene)thiazolidine-2,4-dione
Figure imgf000146_0002
HPLC-MS (Method C): m/z = 356 (M+1); Rt = 5.75 min.
Example 161 (General procedure (B)) 5-(4-Decyloxynaphthalen-1-ylmethylene)thiazolidine-2,4-dione
Figure imgf000146_0003
HPLC-MS (Method C): m/z = 412 (M+1); Rt = 6.44 min.
Example 162 (General procedure (B)) 5-[4-(2-Aminoethoxy)-naphthalen-1 -ylmethylene]-thiazolidine-2,4-dione
Figure imgf000147_0001
HPLC-MS (Method C): m/z = 315 (M+1 ); Rt = 3,24 min.
Example 163 (General procedure (B)) 5-(2,4-Dimethyl-9H-carbazol-3-ylmethylene)-pyrimidine-2,4,6-trione
Figure imgf000147_0002
HPLC-MS (Method C): m/z = 334 (M+1 ); Rt = 3,14 min.
Example 164 (General procedure (B)) 4-(4-Hydroxy-3-methoxybenzylidine)hydantoin
Figure imgf000147_0003
Example 165 (General procedure (B)) 5-Benzylidenehydantoin
Figure imgf000147_0004
General procedure (C) for preparation of compounds of general formula l2:
Figure imgf000148_0001
l2 wherein X, Y, A, and R3 are as defined above and A is optionally substituted with up to four substituents R7, R8, R9, and R10 as defined above.
This general procedure (C) is quite similar to general procedure (B) and is further illustrated in the following example:
Example 166 (General procedure (C)) 5-(3,4-Dibromobenzylidene)thiazolidine-2,4-dione
Figure imgf000148_0002
A mixture of thiazolidine-2,4-dione (90%, 65 mg, 0.5 mmol), 3,4-dibromobenzaldehyde (132 mg, 0.5 mmol), and piperidine (247 μL, 2.5 mmol) was shaken in acetic acid (2 mL) at 11 0 °C for 16 hours. After cooling, the mixture was concentrated to dryness in vacuo . The resulting crude product was shaken with water, centrifuged, and the supernatant was discarded. Subsequently the residue was shaken with ethanol, centrifuged, the supernatant was discarded and the residue was further evaporated to dryness to afford the title compound.
Η NMR (Acetone-dβ): δH 7.99 (d,1H), 7.90 (d,1 H), 7.70 (s,1 H), 7.54 (d,1H); HPLC-MS (Method A): m/z: 364 (M+1 ); Rt = 4.31 min.
The compounds in the following examples were similarly prepared. Optionally, the compounds can be further purified by filtration and washing with water instead of concentration in vacuo. Also optionally the compounds can be purified by washing with ethanol, water and/or heptane, or by preparative HPLC.
Example 167 (General procedure (C)) 5-(4-Hydroxy-3-iodo-5-methoxybenzylidene)thiazolidine-2,4-dione
Figure imgf000149_0001
Mp = 256 °C; Η NMR (DMSO-d6) δ = 12.5 (s,broad,1 H), 10.5 (s,broad,1 H), 7.69 (s,1H), 7.51 (d,1 H), 7.19 (d,1H)3.88 (s,3H), 13C NMR (DMSO-dβ) δc = 168.0, 167.7 , 149.0, 147.4, 133.0, 131.2, 126.7, 121.2, 113.5, 85.5, 56.5; HPLC-MS (Method A): m/z: 378 (M+1); Rt = 3.21 min.
Example 168 (General procedure (C)) 5-(4-Hydroxy-2,6-dimethylbenzylidene)thiazolidine-2,4-dione
Figure imgf000149_0002
HPLC-MS (Method C): m/z: 250 (M+1 ); Rt.= 2.45 min.
Example 169 (General procedure (C)) 4-[5-Bromo-6-(2,4-dioxothiazolidin-5-ylidenemethyl)-naphthalen-2-yloxymethyl]-benzoic acid
Figure imgf000149_0003
HPLC-MS (Method C): m/z: 506 (M+23); Rt.= 4.27 min.
Example 170 (General procedure (C)) 5-(4-Bromo-2,6-dichlorobenzylidene)thiazolidine-2,4-dione
Figure imgf000149_0004
HPLC-MS (Method C): m/z: 354 (M+1); Rt.= 4.36 min. Example 171 (General procedure (C)) 5-(6-Hydroxy-2-naphthylmethylene) thiazolidine-2,4-dione
Figure imgf000150_0001
Mp 310-314 °C, Η NMR (DMSO-d6): δH = 12.5 (s,broad,1 H), 8.06(d,1 H), 7.90- 7.78(m,2H),7.86 (s,1 H), 7.58 (dd,1 H),7.20 7.12 (m,2H). 13C NMR (DMSO-d6): δc = 166.2, 165.8 , 155.4, 133.3, 130.1 , 129.1, 128.6, 125.4, 125.3, 125.1 , 124.3, 120.0, 117.8, 106.8; HPLC-MS (Method A): m/z: 272 (M+1); Rt = 3.12 min.
Preparation of the starting material, 6-hydroxy-2-naphtalenecarbaldehyde:
6-Cyano-2-naphthalenecarbaldehyde (1.0 g, 5.9 mmol) was dissolved in dry hexane (15 L) under nitrogen. The solution was cooled to -60 °C and a solution of diisobutyl aluminium hydride (DIBAH) (15 mL, 1 M in hexane) was added dropwise. After the addition, the solution was left at room temperature overnight. Saturated ammonium chloride solution (20 mL) was added and the mixture was stirred at room temperature for 20 min, subsequently aqueous H2S04 (10% solution, 15 mL) was added followed by water until all salt was dissolved. The resulting solution was extracted with ethyl acetate (3x), the combined organic phases were dried with MgS04, evaporated to dryness to afford 0.89 g of 6-hydroxy-2- naphtalenecarbaldehyde.
Mp.: 153.5-156.5 °c; HPLC-MS (Method A): m/z: 173 (M+1); Rt = 2.67 min; 1 H NMR (DMSO- de): δH = 10.32(s,1 H), 8.95 (d,1 H), 10.02 (s,1 H), 8.42 (s,broad,1 H), 8.01 (d,1 H), 7.82-7.78 (m,2H), 7.23-7.18 (m,2H).
Alternative preparation of 6-hydroxy-2-naphtalenecarbaldehyde:
To a stirred cooled mixture of 6-bromo-2-hydroxynaphthalene (25.3 g, 0.113 mol) in THF (600 mL) at -78 °C was added n-BuLi (2.5 M, 100 mL, 0.250 mol) dropwise. The mixture turned yellow and the temperature rose to -64 °C. After ca 5 min a suspension appeared!. After addition, the mixture was maintained at -78 °C. After 20 minutes, a solution of DMF (28.9 mL, 0.373 mol) in THF (100 mL) was added over 20 minutes. After addition, the mixture was allowed to warm slowly to room temperature. After 1 hour, the mixture was poured in ice/water (200 mL). To the mixture citric acid was added to a pH of 5. The mixture was. stirred for 0.5 hour. Ethyl acetate (200 mL) was added and the organic layer was separated and washed with brine (100 mL), dried over Na2S04and concentrated. To the residue w^s added heptane with 20% ethyl acetate (ca 50 mL) and the mixture was stirred for 1 hour. The mixture was filtered and the solid was washed with ethyl acetate and dried in vacuo to afford 16 g of the title compound.
Example 172 (General procedure (C)) 5-(3-lodo-4-methoxybenzylidene)thiazolidiene-2,4-dione
Figure imgf000151_0001
Η NMR (DMSO-de): δH 12.55 (s,broad,1H), 8.02 (d,1H), 7.72 (s,1H), 7.61 (d,1H)7.18(d,1 H), 3.88 (s,3H); 13C NMR (DMSO-de): δc 168.1, 167.7 , 159.8, 141.5, 132.0, 130.8, 128.0, 122.1 , 112.5, 87.5, 57.3. HPLC-MS (Method A): m/z: 362 (M+1); Rt = 4.08 min.
Preparation of the starting material, 3-iodo-4-methoxybenzaldehyde:
4-Methoxybenzaldehyde (0.5 g, 3.67 mmol) and silver trifluoroacetate (0.92 g, 4.19 mmol) were mixed in dichloromethane (25 mL). Iodine (1.19 g, 4.7 mmol) was added in small portions and the mixture was stirred overnight at room temperature under nitrogen. The mixture was subsequently filtered and the residue washed with DCM. The combined filtrates were treated with an acqueous sodium thiosulfate solution (1 M) until the colour disappeared. Subsequent extraction with dichloromethane (3 x 20 mL) followed by drying with MgS04 and evaporation in vacuo afforded 0.94 g of 3-iodo-4-methoxybenzaldehyde.
Mp 104-107 °C; HPLC-MS (Method A): m/z:263 (M+1); Rt = 3.56 min.;Η NMR (CDCI3): δH = 8.80 (s,1H), 8.31 (d,1 H), 7.85 (dd,1 H) 6.92 (d,1 H), 3.99 (s, 3H).
Example 173 (General procedure (C)) 5-(1-Bromonaphthalen-2-ylmethylene)thiazolidine-2,4-dione
Figure imgf000151_0002
HPLC-MS (Method A): m/z: =336 (M+1); Rt = 4.46 min. Example 174 (General procedure (C))
1-[5-(2,4-Dioxothiazolidin-5-ylidenemethyl)thiazol-2-yl]piperidine-4-carboxylic acid ethyl ester
Figure imgf000152_0001
Η NMR (DMSO-de): δH = 7.88 (s,1 H), 7.78 (s,1 H), 4.10 (q,2H), 4.0-3.8 (m,2H), 3.40-3.18 (m,2H), 2.75-2.60 (m, 1 H), 2.04-1.88 (m,2H), 1.73-1.49 (m,2H) , 1.08 (t,3H); HPLC-MS (Method A): m/z: 368 (M+1 ); Rt = 3.41 min.
Example 175 (General procedure (C))
5-(2-Phenyl-[1 ,2,3]triazol-4-ylmethylene) thiazolidine-2,4-dione
Figure imgf000152_0002
Η NMR (DMSO-de): δH = 12.6 (s,broad,1H), 8.46 (s,1H), 8.08 (dd,2H), 7.82 (s,1 H), 7.70-7.45 (m, 3H). HPLC-MS (Method A): m/z: 273 (M+1); Rt = 3.76 min.
Example 176 (General procedure (C)) 5-(Quinolin-4-ylmethylene)thiazolidine-2,4-dione
Figure imgf000152_0003
HPLC-MS (Method A): m/z: 257 (M+1 ); Rt = 2.40 min.
Example 177 (General procedure (C)) 5-(6-Methylpyridin-2-ylmethylene)thiazolidine-2,4-dione
Figure imgf000153_0001
1H NMR (DMSO-dβ): δH = 12.35 (s,broad,1H), 7.82 (t,1H), 7.78 (s,1H), 7.65 (d,1H), 7.18 (d,1H), 2.52 (s,3 H); HPLC-MS (Method A): m/z: 221 (M+1); Rt = 3.03 min.
Example 178 (General procedure (C))
5-(2,4-dioxothiazolidin-5-ylidenemethyl)-furan-2-ylmethylacetate
Figure imgf000153_0002
Η NMR (DMSO-de): δH = 12.46 (s,broad,1H), 7.58 (s,1H), 7.05 (d,1H), 6.74 (s,1 H), 5.13 (s,2H), 2.10 (s,3H). HPLC-MS (Method A): m/z: 208 (M-CH3COO); Rt = 2.67 min.
Example 179 (General procedure (C)) 5-(2,4-Dioxothiazolidin-5-ylidenemethyl)furan-2-sulfonic acid
Figure imgf000153_0003
HPLC-MS (Method A): m/z:276 (M+1); Rt = 0.98 min.
Example 180 (General procedure (C)) 5-(5-Benzyloxy-1H-pyrrolo[2,3-c]pyridin-3-ylmethylene)-thiazolidine-2,4-dione
Figure imgf000153_0004
HPLC-MS (Method A): m/z: 352 (M+1); Rt = 3.01 min.
Example 181 (General procedure (C)) 5-(Quinolin-2-ylmethylene)thiazolidine-2,4-dione
Figure imgf000154_0001
HPLC-MS (Method A): m/z: 257 (M+1); Rt = 3.40 min.
Example 182 (General procedure (C)) 5-(2,4-Dioxothiazolidin-5-ylidenemethyl)thiophene-2-carboxylic acid
Figure imgf000154_0002
HPLC-MS (Method A): m/z: 256 (M+1); Rt = 1.96 min.
Example 183 (General procedure (C)) 5-(2-Phenyl-1 H-imidazol-4-ylmethylene)thiazolidine-2,4-dione
Figure imgf000154_0003
HPLC-MS (Method A): m/z: 272 (M+1); Rt = 2.89 min.
Example 184 (General procedure (C)) 5-(4-lmidazol-1-yl-benzylidene)thiazolidine-2,4-dione
Figure imgf000154_0004
HPLC-MS (Method A): m/z: 272 (M+1); Rt = 1.38 min.
Example 185 (General procedure (C)) 5-(9-Ethyl-9H-carbazol-3-ylmethylene)thiazolidine-2,4-dione
Figure imgf000154_0005
HPLC-MS (Method A): m/z: 323 (M+1); Rt = 4.52 min.
Example 186 (General procedure (C))
5-(1,4-Dimethyl-9H-carbazol-3-ylmethylene)thiazolidine-2,4-dione
Figure imgf000155_0001
HPLC-MS (Method A): m/z: 323 (M+1); Rt = 4.35 min.
Example 187 (General procedure (C)) 5-(2-Methyl-1H-indol-3-ylmethylene)thiazolidine-2,4-dione
Figure imgf000155_0002
HPLC-MS (Method A): m/z: 259 (M+1); Rt = 3.24 min.
Example 188 (General procedure (C)) 5-(2-Ethylindol-3-ylmethylene)thiazolidine-2,4-dione
Figure imgf000155_0003
2-Methylindole (1.0 g, 7.6mmol) dissolved in diethyl ether (100 mL) under nitrogen was treated with n-Butyl lithium (2 M in pentane, 22.8 mmol) and potassium tert-butoxide (15.2 mmol) with stirring at RT for 30 min. The temperature was lowered to -70 C and methyl lo- dide (15.2 mmol) was added and the resulting mixture was stirred at -70 for 2 h. Then 5 drops of water was added and the mixture allowed to warm up to RT. Subsequently, the mixture was poured into water (300 mL), pH was adjusted to 6 by means of 1 hydrochloric acid and the mixture was extracted with diethyl ether. The organic phase was dried with Na2S04 and evaporated to dryness. The residue was purified by column chromatography on silica gel using heptane/ether( 4/1) as eluent. This afforded 720 mg (69 %) of 2-ethylindole. 1H NMR (DMSO-de ): δ = 10.85 (1 H,s); 7.39 (1 H,d); 7.25 (1 H,d); 6.98(1 H,t); 6.90(1H,t); 6.10 (1H,s); 2.71 (2H,q); 1.28 (3H,t).
2-Ethylindole (0.5 g, 3.4mmol) dissolved in DMF (2 mL) was added to a cold (0 °C) premixed (30 minutes) mixture of DMF (1.15 mL) and phosphorous oxychloride (0.64 g, 4.16 mmol). After addition of 2-ethylindole, the mixture was heated to 40 °C for 1 h, water (5 mL) was added and the pH adjusted to 5 by means of 1 N sodium hydroxide.The mixture was subsequently extracted with diethyl ether, the organic phase isolated, dried with MgS04 and evaporated to dryness affording 2-ethylindole-3-carbaldehyde (300 mg ).
HPLC-MS (Method C): m/z: 174 (M+1); Rt. =2.47 min.
2-Ethylindole-3-carbaldehyde (170 mg) was treated with thiazolidine-2,4-dione using the general procedure (C) to afford the title compound (50 mg).
HPLC-MS (Method C):m/z: 273 (M+1); Rt.= 3.26 min.
Example 189 (General procedure (C))
5-[2-(4-Bromophenylsulfanyl)-1-methyl-1H-indol-3-ylmethylene]thiazolidine-2,4-dione
Figure imgf000156_0001
HPLC-MS (Method A): m/z: 447 (M+1); Rt = 5.25 min.
Example 190 (General procedure (C)) 5-[2-(2,4-Dichlorobenzyloxy)-naphthalen-1-ylmethylene]thiazolidine-2,4-dione
Figure imgf000157_0001
HPLC-MS (Method A): (anyone 1) m/z: 430 (M+1); Rt = 5.47 min.
Example 191 (General procedure (C)) 5-{4-[3-(4-Bromophenyl)-3-oxopropenyl]-benzylidene}thiazolidine-2,4-dione
Figure imgf000157_0002
HPLC-MS (Method A): m/z: 416 (M+1); Rt = 5.02 min.
Example 192 (General procedure (C)) 5-(4-Pyridin-2-ylbenzylidene)thiazolidine-2,4-dione
Figure imgf000157_0003
HPLC-MS (Method A): m/z: 283 (M+1 ), Rt = 2.97 min.
Example 193 (General procedure (C)) 5-(3,4-Bisbenzyloxybenzylidene)thiazolidine-2,4-dione
Figure imgf000157_0004
HPLC-MS (Method A): m/z: 418 (M+1); Rt = 5.13 min.
Example 194 (General procedure (C)) 5-[4-(4-Nitrobenzyloxy)-benzylidene]thiazolidine-2,4-dione
Figure imgf000158_0001
HPLC-MS (Method A): m/z: 357 (M+1); Rt = 4.45 min.
Example 195 (General procedure (C)) 5-(2-Phenyl-1 H-indol-3-ylmethylene)thiazolidine-2,4-dione
Figure imgf000158_0002
HPLC-MS (Method A): m/z: 321 (M+1); Rt = 3.93 min.
Example 196 (General procedure (C)) 5-(5-Benzyloxy-1 H-indol-3-ylmethylene)thiazolidine-2,4-dione
Figure imgf000158_0003
HPLC-MS (Method A): m/z: 351 (M+1); Rt = 4.18 min.
Example 197 (General procedure (C)) 5-(4-Hydroxybenzylidene)thiazolidine-2,4-dione
Figure imgf000158_0004
HPLC-MS (Method A): m/z: 222 (M+1); Rt = 2.42 min. Example 198 (General procedure (C)) 5-(1 -Methyl-1 H-indol-2-ylmethylene)thiazolidine-2,4-dione
Figure imgf000159_0001
1H NMR (DMSO-de): cJH = 12.60 (s,broad,1H), 7.85 (s,1H), 7.68 (dd,1H), 7.55 (dd,1H), 7.38 (dt, 1H), 7.11 (dt,1H) 6.84 (s,1H), 3.88 (s,3H); HPLC-MS (Method A): m/z: 259 (M+1); Rt = 4.00 min.
Example 199 (General procedure (C)) 5-(5-Nitro-1H-indol-3-ylmethylene)thiazolidine-2,4-dione
Figure imgf000159_0002
.Mp 330-333 °C, 1H NMR (DMSO-d6): δH = 12.62 (s,broad,1H), 8.95 (d,1H), 8.20 (s,1H), 8.12 (dd,1H), 7.98 (s,broad,1H), 7.68 (d,1H); HPLC-MS (Method A): m/z: 290 (M+1); Rt = 3.18 min.
Example 200 (General procedure (C)) 5-(6-Methoxynaphthalen-2-ylmethylene)thiazolidine-2,4-dione
Figure imgf000159_0003
HPLC-MS (Method A): m/z: 286 (M+1); Rt = 4.27 min.
Example 201 (General procedure (C)) 5-(3-Bromo-4-methoxybenzylidene)thiazolidine-2,4-dione
Figure imgf000159_0004
HPLC-MS (Method A): m/z: 314 (M+1), Rt = 3.96 min.
Example 202 (General procedure (C)) 3-{(2-Cyanoethyl)-[4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenyl]amino}propionitrile
Figure imgf000160_0001
HPLC-MS (Method A): m/z: 327 (M+1); Rt = 2.90 min.
Example 203 (General procedure (C)) 3-(2,4-Dioxothiazolidin-5-ylidenemethyl)indole-6-carboxylic acid methyl ester
Figure imgf000160_0002
HPLC-MS (Method A): m/z: 303 (M+1); Rt = 3.22-3-90 min.
Example 204 3-(2,4-Dioxothiazolidin-5-ylidenemethyl)indole-6-carboxylic acid pentyl ester.
Figure imgf000160_0003
3-(2,4-Dioxothiazolidin-5-ylidenemethyl)indole-6-carboxylic acid methyl ester (example 203, 59 mg; 0.195mmol) was stirred in pentanol (20 mL) at 145 °C for 16 hours. The mixture was evaporated to dryness affording the title compound (69 mg).
HPLC-MS (Method C): m/z: 359 (M+1); Rt.= 4.25 min.
Example 205 (General procedure (C)) 3-(2,4-Dioxothiazolidin-5-ylidenemethyl)indole-7-carboxylic acid
Figure imgf000160_0004
HPLC-MS (Method A): m/z: 289 (M+1); Rt = 2.67 min.
Example 206 (General procedure (C)) 5-(1-Benzylindol-3-ylmethylene)thiazolidine-2,4-dione
Figure imgf000161_0001
HPLC-MS (Method A): m/z: 335 (M+1); Rt = 4.55 min.
Example 207 (General procedure (C)) 5-(1-Benzenesulfonylindol-3-ylmethylene)thiazolidine-2,4-dione
Figure imgf000161_0002
HPLC-MS (Method A): m/z: = 385 (M+1); Rt = 4.59 min.
Example 208 (General procedure (C)) 5-(4-[1 ,2,3]Thiadiazol-4-ylbenzylidene)thiazolidine-2,4-dione
Figure imgf000161_0003
HPLC-MS (Method A): m/z: 290 (M+1); Rt = 3.45 min.
Example 209 (General procedure (C)) 5-[4-(4-Nitrobenzyloxy)-benzylidene]thiazolidine-2,4-dione
Figure imgf000161_0004
HPLC-MS (Method A): m/z: 357 (M+1); Rt = 4.42 min. Example 210 (General procedure (C))
3-(2,4-Dioxothiazolidin-5-ylidenemethyl)indole-1 -carboxylic acid ethyl ester
Figure imgf000162_0001
HPLC-MS (Method A): m/z: 317 (M+1); Rt = 4.35 min.
Example 211 (General procedure (C)) 5-[2-(4-Pentylbenzoyl)-benzofuran-5-ylmethylene]thiazolidine-2,4-dione
Figure imgf000162_0002
HPLC-MS (Method A): m/z: 420 (M+1); Rt = 5.92 min.
Example 212 (General procedure (C)) 5-[1-(2-Fluorobenzyl)-4-nitroindol-3-ylmethylene]thiazolidine-2,4-dione
Figure imgf000162_0003
HPLC-MS (Method A): (Anyone 1) m/z: 398 (M+1 ); Rt = 4.42 min.
Example 213 (General procedure (C)) 5-(4-Benzyloxyindol-3-ylmethylene)thiazolidine-2,4-dione
Figure imgf000162_0004
HPLC-MS (Method A): m/z: 351 (M+1); Rt = 3.95 min. Example 214 (General procedure (C)) 5-(4-lsobutylbenzylidene)-thiazolidine-2,4-dione
Figure imgf000163_0001
HPLC-MS (Method A): m/z: 262 (M+1); Rt = 4.97 min.
Example 215 (General procedure (C))
Trifluoromethanesulfonic acid 4-(2,4-dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yl ester
Figure imgf000163_0002
HPLC-MS (Method A): m/z: 404 (M+1 ); Rt = 4.96 min.
Preparation of starting material:
4-Hydroxy-1-naphthaldehyde (10 g, 58 mmol) was dissolved in pyridin (50 ml) and the mixture was cooled to 0-5 °C. With stirring, trifluoromethanesulfonic acid anhydride (11.7 ml, 70 mmol) was added drop-wise. After addition was complete, the mixture was allowed to warm up to room temperature, and diethyl ether (200 ml) was added. The mixture was washed with water (2 x 250 ml), hydrochloric acid (3N, 200 ml), and saturated aqueous sodium chloride (100 ml). After drying (MgS04), filtration and concentration in vacuo, the residue was purified by column chromatography on silica gel eluting with a mixture of ethyl acetate and heptane (1 :4). This afforded 8.35 g (47%) trifluoromethanesulfonic acid 4-formylnaphthalen-1-yl ester, mp 44-46.6 °C.
Example 216 (General procedure (C)) 5-(4-Nitroindol-3-ylmethylene)-thiazolidine-2,4-dione
Figure imgf000163_0003
HPLC-MS (Method A): m/z: 290 (M+1); Rt = 3.14 min. Example 217 (General procedure (C)) 5-(3,5-Dibromo-4-hydroxy-benzylidene)thiazolidine-2,4-dione
Figure imgf000164_0001
1H NMR (DMSO-de): <5H = 12.65 (broad,1H), 10.85 (broad,1H), 7.78 (s,2H), 7.70 (s,1H); HPLC-MS (Method A): m/z: 380 (M+1); Rt = 3.56 min.
Example 218 (General procedure (C))
Figure imgf000164_0002
HPLC-MS (Method A): m/z: 385 (M+1); Rt = 5.08 min.
General procedure for preparation of starting materials for examples 218 - 221 : lndole-3-carbaldehyde (3.8 g, 26 mmol) was stirred with potassium hydroxide (1.7 g) in acetone (200 mL) at RT until a solution was obtained indicating full conversion to the indole potassium salt. Subsequently the solution was evaporated to dryness in vacuo. The residue was dissolved in acetone to give a solution containing 2.6 mmol/20 mL.
20 mL portions of this solution were mixed with equimolar amounts of arylmethylbromides in acetone (10 mL). The mixtures were stirred at RT for 4 days and subsequently evaporated to dryness and checked by HPLC-MS. The crude products, 1-benzylated indole-3- carbaldehydes, were used for the reaction with thiazolidine-2,4-dione using the general procedure C.
Example 219 (General procedure (C)) 4-[3-(2,4-Dioxothiazolidin-5-ylidenemethyl)indol-1-ylmethyl]benzoic acid methyl ester
Figure imgf000165_0001
HPLC-MS (Method A): m/z: 393 (M+1); Rt = 4.60 min.
Example 220 (General procedure (C))
5-[1-(9,10-Dioxo-9,10-dihydroanthracen-2-ylmethyl)-1H-indol-3-ylmethylene]thiazolidine-2,4- dione
Figure imgf000165_0002
HPLC-MS (Method A): m/z: 465 (M+1); Rt = 5.02 min.
Example 221 (General procedure (C)) 4'-[3-(2,4-Dioxothiazolidin-5-ylidenemethyl)indol-1-ylmethyl]biphenyl-2-carbonitrile
Figure imgf000165_0003
HPLC-MS (Method A): m/z: 458 (M+23); Rt = 4.81 min.
Example 222 (General procedure (C)) 3-[3-(2,4-Dioxothiazolidin-5-ylidenemethyl)-2-methylindol-1-ylmethyl]benzonitrile.
Figure imgf000165_0004
2-Methylindole-3-carbaldehyde (200 mg, 1.26 mmol) was added to a slurry of 3- bromomethylbenzenecarbonitrile (1.26 mmol) followed by sodium hydride, 60%, (1.26 mmol) in DMF (2 mL). The mixture was shaken for 16 hours, evaporated to dryness and washed with water and ethanol. The residue was treated with thiazolidine-2,4-dione following the general procedure C to afford the title compound (100 mg).
HPLC-MS (Method C): m/z: 374 (M+1 ); Rt. = 3.95 min.
Example 223 (General procedure (C)) 5-(1-Benzyl-2-methylindol-3-ylmethylene)thiazolidine-2,4-dione.
Figure imgf000166_0001
This compound was prepared in analogy with the compound described in example 222 from benzyl bromide and 2-methylindole-3-carbaldehyde, followed by reaction with thiazolidine- 2,4-dione resulting in 50 mg of the title compound.
HPLC-MS (Method C): m/z: 349 (M+1); Rt. = 4.19 min.
Example 224
4-[3-(2,4-Dioxothiazolidin-5-ylidenemethyl)-2-methylindol-1-ylmethyl]benzoic acid methyl ester
Figure imgf000166_0002
This compound was prepared in analogy with the compound described in example 222 from 4-(bromomethyl)benzoic acid methyl ester and 2-methylindole-3-carbaldehyde, followed by reaction with thiazolidine-2,4-dione.
HPLC-MS (Method C): m/z: 407 (M+1); Rt.= 4.19 min. Example 225 (General procedure (C))
5-(2-Chloro-1 -methyl-1 H-indol-3-ylmethylene)thiazolidine-2,4-dione
Figure imgf000167_0001
HPLC-MS (Method A): m/z: 293 (M+1); Rt = 4.10 min.
Example 226 (General procedure (C)) 5-(4-Hydroxy-3,5-diiodo-benzylidene)-thiazolidine-2,4-dione
Figure imgf000167_0002
HPLC-MS (Method A): m/z: 474 (M+1); Rt = 6.61 min.
Example 227 (General procedure (C)) 5-(4-Hydroxy-3-iodobenzylidene)thiazolidine-2,4-dione
Figure imgf000167_0003
HPLC-MS (Method C): m/z: 348 (M+1); Rt. = 3.13 min
1H-NMR: (DMSO-dβ ): 11.5 (1 H.broad); 7.95(1 H,d); 7.65(1 H,s); 7.45 (1 H.dd); 7.01 (1 H.dd);
3.4 (IH.broad).
Example 228 (General procedure (C))
5-(2,3,6-Trichlorobenzylidene)thiazolidine-2,4-dione
Figure imgf000168_0001
H PLC-MS (Method C): m/z: 309 (M+1); Rt.= 4.07 min
Example 229 (General procedure (C)) 5-(2,6-Dichlorobenzylidene)thiazolidine-2,4-dione
Figure imgf000168_0002
Mp. 152-154°C.
HPLC-MS (Method C): m/z: 274 (M+1), Rt.= 3.70 min
1 H-NMR: (DMSO-dβ): 12.8 (1H, broad); 7.72 (1H,s); 7.60 (2H,d); 7.50 (1H,t).
Example 230 (General procedure (C))
5-[1-(2,6-Dichloro-4-trifluoromethylphenyl)-2,5-dimethyl-1H-pyrrol-3-ylmethylene]thiazolidine-
2,4-dione
Figure imgf000168_0003
HPLC-MS (Method C): m/z: 436 (M+1); Rt. 4.81 min
Example 231 (General procedure (C))
5-[1-(3,5-Dichlorophenyl)-5-(4-methanesulfonylphenyl)-2-methyl-1H-pyrrol-3-ylmethylene]- thiazolidine-2,4-dione
Figure imgf000169_0001
HPLC-MS (Method C): m/z: 508 (M+1); Rt. = 4.31 min
Example 232 (General procedure (C))
5-[1-(2,5-Dimethoxyphenyl)-5-(4-methanesulfonylphenyl)-2-methyl-1H-pyrrol-3-ylmethylene]- thiazolidine-2,4-dione
Figure imgf000169_0002
HPLC-MS (Method C): m/z: 499 (M+1); Rt. = 3.70 min
Example 233 (General procedure (C))
4-[3-(2,4-Dioxothiazolidin-5-ylidenemethyl)-2,5-dimethylpyrrol-1-yl]benzoic acid
Figure imgf000169_0003
HPLC-MS (Method C): m/z:342 (M+1 ); Rt .= 3.19 min
Example 234 (General procedure (C))
5-(4-Hydroxy-2,6-dimethoxybenzylidene)thiazolidine-2,4-dione
Figure imgf000169_0004
HPLC-MS (Method C): m/z:282( M+1); Rt.= 2.56, mp=331-333 °C Example 235 (General procedure (C)) 5-(2,6-Dimethylbenzylidene)thiazolidine-2,4-dione
Figure imgf000170_0001
M.p: 104-105 °C
HPLC-MS (Method C): m/z: 234 (M+1); Rt.= 3.58 min,
Example 236 (General procedure (C)) 5-(2,6-Dimethoxybenzylidene)thiazolidine-2,4-dione
Figure imgf000170_0002
Mp: 241-242 °C
HPLC-MS (Method C): m/z: 266 (M+1); Rt.= 3.25 min;
Example 237 (General procedure (C)) 5-[4-(2-Fluoro-6-nitrobenzyloxy)-2,6-dimethoxybenzylidene]thiazolidine-2,4-dione
Figure imgf000170_0003
Mp: 255-256 °C
HPLC-MS (Method C): m/z: 435 (M+1 ), Rt 4.13 min,
Example 238 (General procedure (C)) 5-Benzofuran-2-ylmethylenethiazolidine-2,4-dione
Figure imgf000170_0004
HPLC-MS (Method C): m/z:246 (M+1 ); Rt.= 3.65 min, mp = 265-266 °C Example 239 (General procedure (C)) 5-[3-(4-Dimethylaminophenyl)allylidene]thiazolidine-2,4-dione
Figure imgf000171_0001
HPLC-MS (Method C): m/z:276(M+1 ); Rt.= 3.63, mp = 259-263 °C
1H-NMR: (DMSO-dβ ) δ= 12.3 (IH.broad); 7.46 (2H,d); 7.39 (1 H,d); 7.11 (1 H,d); 6.69 (2H,d);
6.59 (1H, dd); 2.98 (3H,s).
Example 240 (General procedure (C)) 5-(2-Methyl-3-phenylallylidene)thiazolidine-2,4-dione
Figure imgf000171_0002
Mp: 203-210 °C
HPLC-MS (Method C): m/z: 246 (M+1); Rt = 3.79 min.
Example 241 (General procedure (C)) 5-(2-Chloro-3-phenylallylidene)thiazolidine-2,4-dione
Figure imgf000171_0003
Mp: 251-254 °C
HPLC-MS (Method C): m/z: 266 (M+1 ; Rt = 3.90 min
Example 242 (General procedure (C)) 5-(2-Oxo-1 ,2-dihydroquinolin-3-ylmethylene)thiazolidine-2,4-dione
Mp: 338-347 °C HPLC-MS (Method C): m/z: 273 (M+1 ); Rt. = 2.59 min.
Example 243 (General procedure (C)) 5-(2,4,6-Tribromo-3-hydroxybenzylidene)thiazolidine-2,4-dione.
Figure imgf000172_0001
HPLC-MS (Method C): m/z: 459 (M+1);Rt .= 3.65 min.
Example 244 (General procedure (C)) 5-(5-Bromo-2-methylindol-3-ylmethylene)thiazolidine-2,4-dione.
Figure imgf000172_0002
HPLC-MS (Method C): m/z: 339 (M+1); Rt = 3.37min.
Example 245 (General procedure (C)) 5-(7-Bromo-2-methylindol-3-ylmethylene)thiazolidine-2,4-dione.
Figure imgf000172_0003
HPLC-MS (Method C): m/z: 319 (M+1); Rt = 3.48min.
Example 246 (General procedure (C)) 5-(6-Bromoindol-3-ylmethylene)thiazolidine-2,4-dione.
Figure imgf000172_0004
HPLC-MS (Method C): m/z: 325 (M+1); Rt = 3.54 min.
Example 247 (General procedure (C)) 5-(8-Methyl-2-oxo-1,2-dihydroquinolin-3-ylmethylene)thiazolidine-2,4-dione.
Figure imgf000173_0001
HPLC-MS (Method C): m/z: 287 (M+1); Rt = 2.86 min.
Example 248 (General procedure (C)) 5-(6-Methoxy-2-oxo-1 ,2-dihydroquinolin-3-ylmethylene)thiazolidine-2,4-dione.
Figure imgf000173_0002
HPLC-MS (Method C): m/z: 303 (M+1); Rt = 2.65 min.
Example 249 (General procedure (C)) 5-Quinolin-3-ylmethylenethiazolidine-2,4-dione.
Figure imgf000173_0003
HPLC-MS (Method C): m/z: 257 (M+1); Rt = 2.77 min.
Example 250 (General procedure (C))
5-(8-Hydroxyquinolin-2-ylmethylene)thiazolidine-2,4-dione.
Figure imgf000173_0004
HPLC-MS (Method C): m/z: 273 (M+1); Rt = 3.44 min.
Example 251 (General procedure (C))
5-Quinolin-8-ylmethylenethiazolidine-2,4-dione.
Figure imgf000173_0005
HPLC-MS (Method C): m/z: 257 (M+1); Rt = 3.15 min. Example 252 (General procedure (C)) 5-(1-Bromo-6-methoxynaphthalen-2-ylmethylene)thiazolidine-2,4-dione.
Figure imgf000174_0001
HPLC-MS (Method C): m/z: 366 (M+1 ); Rt = 4.44 min.
Example 253 (General procedure (C)) 5-(6-Methyl-2-oxo-1,2-dihydroquinolin-3-ylmethylene)thiazolidine-2,4-dione.
Figure imgf000174_0002
HPLC-MS (Method C): m/z: 287 (M+1 ); Rt. = 2.89 min.
Example 254 (General procedure (D)) 5-(2,6-Dichloro-4-dibenzylaminobenzylidene)thiazolidine-2,4-dione.
Figure imgf000174_0003
HPLC-MS (Method C): m/z: 469 (M+1 ); Rt = 5.35 min.
Example 255 (General Procedure (C))
7-(2,4-Dioxothiazolidin-5-ylidenemethyl)-4-methoxybenzofuran-2-carboxylic acid
Figure imgf000174_0004
HPLC-MS (Method C): m/z: 320 (M+1 ); Rt = 2.71 min.
Preparation of the intermediate, 7-formyl-4-methoxybenzofuran-2-carboxylic acid: A mixture of 2-hydroxy-6-methoxybenzaldehyde (6.4 g, 42 mmol), ethyl bromoacetate (14.2 mL, 128 mmol) and potassium carbonate (26 g, 185 mmol) was heated to 130 °C. After 3 h the mixture was cooled to room temperature and acetone (100 mL) was added, the mixture was subsequently filtered and concentrated in vacuo. The residue was purified by column chromatography on silica gel eluting with a mixture of ethyl acetate and heptane (1 :4). This afforded 7.5 g (55%) of ethyl 4-methoxybenzofuran-2-carboxylate.
A solution of ethyl 4-methoxybenzofuran-2-carboxylate (6.9 g, 31.3 mmol) in dichloromethane (70 ml) was cooled to 0 °C and a solution of titanium tetrachloride (13.08 g, 69 mmol) was added drop wise. After 10 minutes dichloromethoxymethane (3.958 g, 34 mmol) was added over 10 minutes. After addition, the mixture was warmed to room temperature for 18 hours and the mixture poured into hydrochloric acid (2N, 100 mL). The mixture was stirred for 0.5 hour and then extracted with a mixture of ethyl acetate and toluene (1:1). The organic phase was dried over Na2S04 and concentrated in vacuo. The residue was purified by col- umn chromatography on silica gel eluting with a mixture of ethyl acetate and heptane (1:4). This afforded 5.8 g (80%) of ethyl 7-formyl-4-methoxybenzofuran-2-carboxylate.
7-formyl-4-methoxybenzofuran-2-carboxylate (5.0 g, 21.5 mmol) and sodium carbonate (43 mmol) in water (100 mL) was refluxed until a clear solution appeared (about 0.5 hour). The solution was filtered and acidified to pH =1 with hydrochloric acid (2 N), the resulting product was filtered off and washed with ethyl acetate and ethanol and dried to afford 3.5 g (74%) of 7-formyl-4-methoxybenzofuran-2-carboxylic acid as a solid.
1H NMR (DMSO-d6): δ = 10.20 (s, 1 H) ; 8.07 (d, 1H) ; 7.70 (s, 1 H) ; 7.17 (d, 1 H) ; 4.08 (s, 3H).
Example 256 (General Procedure (C)) 5-(4-Methoxybenzofuran-7-ylmethylene)thiazolidine-2,4-dione
Figure imgf000175_0001
HPLC-MS (Method C): m/z: 267 (M+1 ); Rt = 3.30 min. Preparation of the intermediate, 4-methoxybenzofuran-7-carbaldehyde:
A mixture of 7-formyl-4-methoxybenzofuran-2-carboxylic acid (3.0 g, 13.6 mmol) and Cu (0.6 g, 9.44 mmol) in quinoline (6 mL) was refluxed. After 0.5 h the mixture was cooled to room temperature and water (100 mL) and hydrochloric acid (10 N, 20 mL) were added. The mixture was extracted with a mixture of ethyl acetate and toluene (1 :1), filtered through celite and the organic layer separated and washed with a sodium carbonate solution, dried over Na2S04 and concentrated in vacuo to afford 1.5 g crude product. Column chromatography SiO2, EtOAc/heptanes=1/4 gave 1.1 g (46%) of 4-methoxybenzofuran-7-carbaldehyde as a solid.
1H NMR (CDCI3): δ: 10.30 (s,1H) ; 7.85 (d,1H) ; 7.75 (d,1H) ; 6.98 (d,1H) ; 6.87 (d,1H) ; 4.10 (s,3H). HPLC-MS (Method C) :m/z: 177 (M+1); Rt. = 7.65 min.
Example 257 (General Procedure (C))
5-(4-Hydroxybenzofuran-7-ylmethylene)thiazolidine-2,4-dione
Figure imgf000176_0001
HPLC-MS (Method C): m/z: = 262 (M+1 ); Rt 2.45 min.
Preparation of the intermediate, 4-hydroxybenzofuran-7-carbaldehyde
A mixture of 4-methoxybenzofuran-7-carbaldehyde (1.6 g, 9.1 mmol) and pyridine hydrochloride (4.8 g, 41.7mmol) in quinoline (8 mL) was refluxed. After 8 h the mixture was cooled to room temperature and poured into water (100 mL) and hydrochloric acid (2 N) was added to pH = 2. The mixture was extracted with a mixture of ethyl acetate and toluene (1:1), washed with a sodium carbonate solution, dried with Na2SO4 and concentrated in vacuo to afford 0.8 g crude product. This was purified by column chromatography on silica gel, eluting with a mixture of ethyl acetate and heptane (1:3). This afforded 250 mg of 4-hydroxybenzofuran-7- carbaldehyde as a solid. 1H NMR (DMSO-d6): δ = 11.35 (s, broad,1H) ; 10.15 (s, 1H) ; 8.05 (d, 1H) ; 7.75 (d, 1H) ; 7.10 (d, 1H); 6.83 (d, 1H). HPLC-MS (Method C): m/z: 163 (M+1); Rt. = 6.36 min.
Example 258 (General Procedure (C)) 5-(5-Bromo-2,3-dihydrobenzofuran-7-ylmethylene)thiazolidine-2,4-dione
Figure imgf000177_0001
HPLC-MS (Method C): m/z: 328 (M+1); Rt = 3.66 min.
Preparation of the intermediate, 5-bromo-2,3-dihydrobenzofuran-7-carbaldehyde: To a cooled (15 °C) stirred mixture dihydrobenzofuran (50.9 g, 0.424 mol) in acetic acid (500 mL), a solution of bromine (65.5 mL, 1.27 mol) in acetic acid (200 mL) was added drop wise over 1 hour. After stirring for 18 hours, a mixture of Na2S2O5 (150 g) in water (250 mL) was added carefully, and the mixture was concentrated in vacuo. Water (200 mL) was added and the mixture was extracted with ethyl acetate containing 10% heptane, dried over Na2SO4 and concentrated in vacuo to give crude 5,7-dibromo-2,3-dihydrobenzofuran which was used as such for the following reaction steps. To a cooled solution (-78 °C) of crude 5,7-dibromo-2,3- dihydrobenzofuran (50.7 g, 0.182 mol) in THF (375 mL) a solution of n-Buϋ (2.5 M, 80 mL, 0.200 mol) in hexane was added. After addition, the mixture was stirred for 20 min. DMF (16 mL) was then added drop wise at -78 °C. After addition, the mixture was stirred at room tem- perature for 3 h and then the mixture was poured into a mixture of ice water, (500 mL) and hydrochloric acid (10 N, 40 mL) and extracted with toluene, dried over Na2SO4 and concentrated in vacuo. Column chromatography on silica gel eluting with a mixture of ethyl acetate and heptane (1:4) afforede 23 g of 5-bromo-2,3-dihydrobenzofuran-7-carbaldehyde as a solid.
1H NMR (CDCI3): <J:10.18 (s,1H) ; 7.75 (d,1H) ;7.55 (d,1H) ; 4.80 (t,2H) ; 3.28 (t,2H).
Example 259 (General Procedure (C)) 5-(4-Cyclohexylbenzylidene)thiazolidine-2,4-dione
Figure imgf000178_0001
HPLC-MS (Method C): m/z: 288 (M+1); Rt = 5.03 min.
Preparation of the intermediate, 4-cyclohexylbenzaldehyde:
This compound was synthesized according to a modified literature procedure (J. Org. Chem., 37, No.24, (1972), 3972-3973).
Cyclohexylbenzene (112.5 g, 0.702 mol) and hexamethylenetetramine (99.3 g, 0.708 mol) were mixed in TFA (375 mL). The mixture was stirred under nitrogen at 90 °C for 3 days. Af- ter cooling to room temperature the red-brown mixture was poured into ice-water (3600 ml) and stirred for 1 hour. The solution was neutralized with Na2CO3 (2 M solution in water) and extracted with dichloromethane (2.5 L). The organic phase was dried (Na2SO4) and the solvent was removed in vacuo. The remaining red-brown oil was purified by fractional distillation to afford the title compound (51 g, 39%). 1H NMR (CDCI3): (59.96 (s, 1 H), 7.80 (d, 2H), 7.35 (d, 2H), 2.58 (m, 1H), 1.94-1.70 (m, 5 H), 1.51-1.17 (m, 5H)
Other ligands of the invention include
3',5,-Dichloro-4'-(2,4-dioxothiazolidin-5-ylidenemethyl)biphenyl-4-carboxylic acid:
Figure imgf000178_0002
Example 260 (General procedure (C)) 5-(1-Bromo-6-hydroxynaphthalen-2-ylmethylene)-thiazolidine-2,4-dione
Figure imgf000179_0001
HPLC-MS (Method C): m/z = 350 (M+1); Rt. = 3.45 min.
Example 261 (General procedure (C))
5-[4-(2-Bromoethoxy)-naphthalen-1-ylmethylene]-thiazolidine-2,4-dione
Figure imgf000179_0002
HPLC-MS (Method C): m/z = 380 (M+1); Rt = 3.52 min.
Example 262 (General procedure (C)) 5-(2-Methyl-5-nitro-1H-indol-3-ylmethylene)-thiazolidine-2,4-dione
Figure imgf000179_0003
HPLC-MS (Method C): m/z = 304 (M+1); Rt = 2.95 min.
Example 263 (General procedure (C)) 5-(4-Naphthalen-2-yl-thiazol-2-ylmethylene)-thiazolidine-2,4-dione
Figure imgf000180_0001
HPLC-MS (Method C): m/z = 339 (M+1 ); Rt .= 4.498 min.
Example 264 (General procedure (C)) 5-[4-(4-Methoxy-naphthalen-1-yl)-thiazol-2-ylmethylene]-thiazolidine-2,4-dione
Figure imgf000180_0002
HPLC-MS (Method C): m/z = 369 (M+1); Rt.= 4.456 min.
Example 265 (General procedure (C)) 5-(2-Pyridin-4-yl-1 H-indol-3-ylmethylene)-thiazolidine-2,4-dione
Figure imgf000180_0003
HPLC-MS (Method C): m/z = 322 (M+1); Rt. = 2.307 min. Example 266 (General procedure (C)) 5-[5-(4-Chlorophenyl)-1H-pyrazol-4-ylmethylene]-thiazolidine-2,4-dione
Figure imgf000181_0001
HPLC-MS (Method C): m/z = 306 (M+1); Rt.= 3.60 min.
Example 267 (General procedure (C)) 5-[5-(2,5-Dimethylphenyl)-1 H-pyrazol-4-ylmethylene]-thiazolidine-2,4-dione
Figure imgf000181_0002
HPLC-MS (Method C): m/z = 300 (M+1); Rt. = 3.063 min.
Example 268 (General procedure (C)) 5-(2-Phenyl-benzo[d]imidazo[2,1-b]thiazol-3-ylmethylene)-thiazolidine-2,4-dione
Figure imgf000181_0003
HPLC-MS (Method C): m/z = 378 (M+1 ); Rt = 3.90 min. Example 269 (General procedure (C)) N-{4-[2-(2,4-Dioxothiazolidin-5-ylidenemethyl)-phenoxy]-phenyl}-acetamide
Figure imgf000182_0001
HPLC-MS (Method C): m/z = 355 (M+1); Rt 3.33 min.
Example 270 (General procedure (C))
5-(2-Phenyl-imidazo[1,2-a]pyridin-3-ylmethylene)-thiazolidine-2,4-dione
Figure imgf000182_0002
HPLC-MS (Method C): m/z = 322 (M+1); Rt. = 2.78 min.
Example 271 (General procedure (C)) 5-(2-Naphthalen-2-yl-imidazo[1,2-a]pyridin-3-ylmethylene)-thiazolidine-2,4-dione
Figure imgf000182_0003
HPLC-MS (Method C): m/z = 372 (M+1); Rt. = 2.78 min.
Example 272 (General procedure (C)) 5-[6-Bromo-2-(3-methoxyphenyl)-imidazo[1,2-a]pyridin-3-ylmethylene]-thiazolidine-2,4-dione
Figure imgf000183_0001
HPLC-MS (Method C): m/z = 431 (M+1); Rt.= 3.30 min.
Example 273 (General procedure (C))
5-(1 ,2,3,4-Tetrahydrophenanthren-9-ylmethylene)thiazolidine-2,4-dione
Figure imgf000183_0002
HPLC-MS (Method C): m/z = 310 (M+1); Rt.= 4.97 min.
Example 274 (General procedure (C))
5-(3,5,5,8,8-Pentamethyl-5,6,7,8-tetrahydro-naphthalen-2-ylmethylene)thiazolidine-2,4-dione
Figure imgf000183_0003
HPLC-MS (Method C): m/z = 330 (M+1); Rt.= 5.33 min. Example 275 (General procedure (C)) 5-[6-(2,4-Dichloro-phenyl)-imidazo[2,1-b]thiazol-5-ylmethylene]-thiazolidine-2,4-dione
Figure imgf000184_0001
HPLC-MS (Method C): m/z = 396 (M+1); Rt. = 3.82 min.
Example 276 (General procedure (C)) 5-(5-Bromobenzofuran-7-ylmethylene)-thiazolidine-2,4-dione
Figure imgf000184_0002
HPLC-MS (Method C): m/z = 324 (M+1); Rt. = 3.82 min.
Example 277 (General procedure (C)) 4-[3-(2,4-Dioxothiazolidin-5-ylidenemethyl)-1,4-dimethylcarbazol-9-ylmethyl]-benzoic acid
Figure imgf000184_0003
HPLC-MS (Method C): m/z = 457 (M+1); Rt = 4,23 min. Preparation of intermediary aldehyde:
1 ,4 Dimethylcarbazol-3-carbaldehyde (0.68 g, 3.08 mmol) was dissolved in dry DMF (15 mL), NaH (diethyl ether washed) (0.162 g, 6.7 mol) was slowly added under nitrogen and the mixture was stirred for 1 hour at room temperature. 4-Bromomethylbenzoic acid (0.73 g, 3.4 mmol) was slowly added and the resulting slurry was heated to 40 °C for 16 hours. Water (5 mL) and hydrochloric acid (6N, 3 mL) were added. After stirring for 20 min at room temperature, the precipitate was filtered off and washed twice with acetone to afford after drying 0.38 g (34%) of 4-(3-formyl-1 ,4-dimethylcarbazol-9-ylmethyl)benzoic acid.
HPLC-MS (Method C) : m/z = 358 (M+1 ), RT. = 4.15 min.
Example 278 (General procedure (C)) 4-[7-(2,4-Dioxothiazolidin-5-ylidenemethyl)-benzofuran-5-yl]-benzoic acid
Figure imgf000185_0001
Starting aldehyde commercially available (Syncom BV, NL) HPLC-MS (Method C): m/z = 366 (M+1); Rt. = 3.37 min.
Example 279 (General procedure (C))
4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)-2-nitrophenoxy]-benzoic acid methyl ester
Figure imgf000185_0002
HPLC-MS (Method C): m/z = 401 (M+1 ); Rt. = 4.08 min. Example 280 (General procedure (C))
3',5'-Dichloro-4'-(2,4-dioxothiazolidin-5-ylidenemethyl)-biphenyl-4-carboxylic acid
Figure imgf000186_0001
Starting aldehyde commercially available (Syncom BV, NL)
HPLC-MS (Method C): m/z = 394 (M+1); Rt. = 3.71 min.
Example 281 (General procedure (C))
Figure imgf000186_0002
HPLC-MS (Method C): m/z = 232( M+1); Rt .= 3.6 min.
Example 282 5-(2-Methyl-1H-indol-3-ylmethyl)-thiazolidine-2,4-dione
Figure imgf000186_0003
5-(2-Methyl-1H-indol-3-ylmethylene)thiazolidine-2,4-dione (prepared as described in example 187, 1.5 g, 5.8 mmol) was dissolved in pyridine (20 mL) and THF (50 mL), LiBH4 (2 M in THF, 23.2 mmol) was slowly added with a syringe under cooling on ice. The mixture was heated to 85 °C for 2 days. After cooling, the mixture was acidified with concentrated hydro- chloric acid to pH 1. The aquous layer was extracted 3 times with ethyl acetate, dried with MgSO treated with activated carbon, filtered and the resulting filtrate was evaporated in vacuo to give 1.3 g (88%) of the title compound.
HPLC-MS (Method C): m/z = 261 (M+1 ); Rt. = 3.00 min.
Example 283 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1-yloxy]butyric acid
Figure imgf000187_0001
4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxyjbutyric acid (4.98 g, 13.9 mmol, prepared as described in example 469) was dissolved in dry THF (50 mL) and added dry pyridine (50 mL) and, in portions, lithium borohydride (2.0 M, in THF, 14 mL). The resulting slurry was refluxed under nitrogen for 16 hours, added (after cooling) more lithium borohydride (2.0 M, in THF, 7 mL). The resulting mixture was refluxed under nitrogen for 16 hours. The mixture was cooled and added more lithium borohydride (2.0 M, in THF, 5 mL). The resulting mixture was refluxed under nitrogen for 16 hours. After cooling to 5 °C, the mixture was added water (300 mL) and hydrochloric acid (150 mL). The solid was isolated by filtration, washed with water (3 x 500 mL) and dried. Recrystallization from acetonitrile (500 mL) afforded2.5 g of the title compound.
1H-NMR (DMSO-de. selected peaks): δ = 3.42 (1 H, dd), 3.90 (1 H, dd), 4.16 (2H, "t"), 4.95 (1 H, dd), 6.92 (1H, d), 7.31 (1 H, d), 7.54 (1 H, t), 7.62 (1H, t), 8.02 (1 H, d), 8.23 (1H, d), 12.1 (1 H, bs), 12.2 (1 H, bs). HPLC-MS (Method C): m/z = 382 (M+23); Rt = 3,23 min.
Example 284 5-Naphthalen-1-ylmethylthiazolidine-2,4-dione
Figure imgf000188_0001
5-Naphthalen-1-ylmethylenethiazolidine-2,4-dione (1.08 g, 4.2 mmol, prepared as described in example 68) was dissolved in dry THF (15 mL) and added dry pyridine (15 mL) and, in portions, lithium borohydride (2.0 M, in THF, 4.6 mL). The resulting mixture was refluxed under nitrogen for 16 hours. After cooling to 5 °C, the mixture was added water (100 mL), and, in portions, concentrated hydrochloric acid (40 mL). More water (100 mL) was added, and the mixture was extracted with ethyl acetate (200 mL). The organic phase was washed with water (3 x 100 mL), dried and concentrated in vacuo. The residue was dissolved in ethyl acetate (50 mL) added activated carbon, filtered and concentrated in vacuo and dried to afford 0.82 g (75%) of the title compound.
1H-NMR (DMSO-de): δ = 3.54 (1H, dd), 3.98 (1 H, dd), 5.00 (1H, dd), 7.4-7.6 (4H, m), 7.87 (1 H, d), 7.96 (1 H, d), 8.11 (1 H, d), 12.2 (1 H, bs). HPLC-MS (Method C): m/z = 258 (M+1 ); Rt = 3,638 min.
The following preferred compounds of the invention may be prepared according to procedures similar to those described in the three examples above:
Figure imgf000188_0002
Figure imgf000189_0001
Figure imgf000190_0001
Figure imgf000191_0001
Figure imgf000192_0001
Figure imgf000193_0001
Figure imgf000194_0001
Figure imgf000195_0001
Figure imgf000196_0001
Figure imgf000197_0001
Figure imgf000198_0001
Figure imgf000199_0001
Figure imgf000200_0001
The following compounds are commercially available and may be prepared using general procedures (B) and / or (C). Example 380 5-(5-Bromo-1H-indol-3-ylmethylene)thiazolidine-2,4-dione
Figure imgf000200_0002
Example 381
5-Pyridin-4-ylmethylenethiazolidine-2,4-dione O !V~S r^ my o
Example 382 5-(3-Bromo-4-methoxybenzylidene)thiazolidine-2,4-dione
Figure imgf000201_0001
Example 383 5-(3-Nitrobenzylidene)thiazolidine-2,4-dione
Figure imgf000201_0002
Example 384 5-Cyclohexylidene-1,3-thiazolidine-2,4-dione
Figure imgf000201_0003
Example 385 5-(3,4-Dihydroxybenzylidene)thiazolidine-2,4-dione
Figure imgf000201_0004
Example 386
5-(3-Ethoxy-4-hydroxybenzylidene)thiazolidine-2,4-dione
Figure imgf000202_0001
Example 387 5-(4-Hydroxy-3-methoxy-5-nitrobenzylidene)thiazolidine-2,4-dione
Figure imgf000202_0002
Example 388
5-(3-Ethoxy-4-hydroxybenzylidene)thiazolidine-2,4-dione
Figure imgf000202_0003
Example 389 5-(4-Hydroxy-3,5-dimethoxybenzylidene)thiazolidine-2,4-dione
Figure imgf000202_0004
Example 390
5-(3-Bromo-5-ethoxy-4-hydroxybenzylidene)thiazolidine-2,4-dione
Figure imgf000202_0005
Example 391 5-(3-Ethoxy-4-hydroxy-5-nitrobenzylidene)thiazolidine-2,4-dione
Figure imgf000203_0001
Example 392
Figure imgf000203_0002
Example 393
Figure imgf000203_0003
Example 394
Figure imgf000203_0004
Example 395
Figure imgf000203_0005
Example 396
Figure imgf000204_0001
Example 397
Figure imgf000204_0002
Example 398
Figure imgf000204_0003
Example 399
Figure imgf000204_0004
Example 400
Figure imgf000204_0005
Example 401
Figure imgf000205_0001
Example 402
Figure imgf000205_0002
Example 403
Figure imgf000205_0003
Example 404
Figure imgf000205_0004
Example 405 5-(3-Hydroxy-5-methyl-phenylamino)-thiazolidine-2,4-dione
Figure imgf000206_0001
Example 406
Figure imgf000206_0002
Example 407
Figure imgf000206_0003
Example 408
Example 409
Figure imgf000207_0001
Example 410
Figure imgf000207_0002
Example 411
Figure imgf000207_0003
Example 412
Figure imgf000207_0004
Example 413
Figure imgf000208_0001
Example 414
Figure imgf000208_0002
Example 415
Figure imgf000208_0003
Example 416
Figure imgf000208_0004
Example 417
Figure imgf000209_0001
Example 418
Figure imgf000209_0002
Example 419
Figure imgf000209_0003
Example 420
Figure imgf000209_0004
Example 421
Figure imgf000209_0005
Example 422
Figure imgf000210_0001
Example 423
Figure imgf000210_0002
Example 424
Figure imgf000210_0003
Example 425
Figure imgf000210_0004
Example 426
Figure imgf000210_0005
Example 427
Figure imgf000211_0001
Example 428
Figure imgf000211_0002
Example 430
Figure imgf000211_0003
Example 431 5-(4-Diethylamino-2-methoxy-benzylidene)-imidazolidine-2,4-dione
Figure imgf000212_0001
Example 432
Figure imgf000212_0002
Example 433
Figure imgf000212_0003
Example 434
Figure imgf000212_0004
Example 435
Figure imgf000213_0001
Example 436
Figure imgf000213_0002
Example 437
Figure imgf000213_0003
Example 438
Figure imgf000213_0004
Example 439
Figure imgf000214_0001
Example 440
Figure imgf000214_0002
Example 441
Figure imgf000214_0003
Example 442
Figure imgf000214_0004
Example 443
Figure imgf000215_0001
Example 444
Figure imgf000215_0002
Example 445
Figure imgf000215_0003
Example 446
Figure imgf000215_0004
Example 447
Figure imgf000216_0001
Figure imgf000216_0002
Example 449
Figure imgf000216_0003
Figure imgf000216_0004
Example 451
Figure imgf000217_0001
Example 452
Figure imgf000217_0002
Example 453
Figure imgf000217_0003
Example 454 5-(4-Diethylamino-benzylidene)-2-imino-thiazolidin-4-one
Figure imgf000217_0004
Figure imgf000217_0005
Example 456
Figure imgf000218_0001
Figure imgf000218_0002
Example 458
Figure imgf000218_0003
Example 459
Figure imgf000218_0004
General procedure (D) for preparation of compounds of general formula l3:
Figure imgf000219_0001
Step 3
Figure imgf000219_0002
wherein X, Y, and R3 are as defined above, n is 1 or 3-20,
E is arylene or heterarylene (including up to four optional substituents, R13, R14, R15, and R15A as defined above),
R' is a standard carboxylic acid protecting group, such as Ci-Cβ-alkyI or benzyl and Lea is a leaving group, such as chloro, bromo, iodo, methanesulfonyloxy, toluenesulfonyloxy or the like.
Step 1 is an alkylation of a phenol moiety. The reaction is preformed by reacting R10-C(=O)- E-OH with an ω-bromo-alkane-carboxylic acid ester (or a synthetic equivalent) in the presence of a base such as sodium or potassium carbonate, sodium or potassium hydroxide, sodium hydride, sodium or potassium alkoxide in a solvent, such as DMF, NMP, DMSO, acetone, acetonitrile, ethyl acetate or isopropyl acetate. The reaction is performed at 20 - 160 °C, usually at room temperature, but when the phenol moiety has one or more substituents heating to 50 °C or more can be beneficial, especially when the substituents are in the ortho position relatively to the phenol. This will readily be recognised by those skilled in the art.
Step 2 is a hydrolysis of the product from step 1. Step 3 is similar to general procedure (B) and (C).
This general procedure (D) is further illustrated in the following examples:
Example 460 (General procedure (D)) 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenoxy]butyric acid
Figure imgf000220_0001
Step l : A mixture of 4-hydroxybenzaldehyde (9.21 g, 75 mmol), potassium carbonate (56 g, 410 mmol) and 4-bromobutyric acid ethyl ester (12.9 mL, 90 mmol) in Λ/,Λ/-dimethylformamide (250 mL) was stirred vigorously for 16 hours at room temperature. The mixture was filtered and concentrated in vacuo to afford 19.6 g (100%) of 4-(4-formylphenoxy)butyric acid ethyl ester as an oil. 1H-NMR (DMSO-d6): δ 1.21 (3H, t), 2.05 (2H, p), 2.49 (2H, t), 4.12 (4H, m), 7.13 (2H, d), 7.87 (2H, d), 9.90 (1H, s). HPLC-MS (Method A): m/z = 237 (M+1); Rt = 3.46 min.
Step 2:
4-(4-Formylphenoxy)butyric acid ethyl ester (19.6 g, 75 mmol) was dissolved in methanol
(250 mL) and 1N sodium hydroxide (100 mL) was added and the resulting mixture was stirred at room temperature for 16 hours. The organic solvent was evaporated in vacuo (40 °C, 120 mBar) and the residue was acidified with 1 N hydrochloric acid (110 mL). The mixture was filtered and washed with water and dried in vacuo to afford 14.3 g (91%) 4-(4- formylphenoxy)butyric acid as a solid. 1H-NMR (DMSO-d6): δ 1.99 (2H, p), 2.42 (2H, t), 4.13 (2H, t), 7.14 (2H, d), 7.88 (2H, d), 9.90 (1 H, s), 12.2 (1 H, bs). HPLC-MS (Method A): m/z = 209 (M+1); R, = 2.19 min.
Step 3:
Thiazolidine-2,4-dione (3.55 g, 27.6 mmol), 4-(4-formylphenoxy)butyric acid (5.74 g, 27.6 mmol), anhydrous sodium acetate (11.3 g, 138 mmol) and acetic acid (100 mL) was refluxed for 16 h. After cooling, the mixture was filtered and washed with acetic acid and water. Drying in vacuo afforded 2.74 g (32%) of 4-[4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]butyric acid as a solid. 1H-NMR (DMSO-d6): 1.97 (2H, p), 2.40 (2H. t), 4.07 (2H, t), 7.08 (2H, d), 7.56 (2H, d), 7.77 (1 H, s), 12.2 (1 H, bs), 12.5 (1 H, bs); HPLC-MS (Method A): m/z: 308 (M+1); Rt = 2.89 min.
Example 461 (General procedure (D)) [3-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenoxy]acetic acid
Figure imgf000221_0001
Step 3:
Thiazolidine-2,4-dione (3.9 g, 33 mmol), 3-formylphenoxyacetic acid (6.0 g, 33 mmol), anhydrous sodium acetate (13.6 g, 165 mmol) and acetic acid (100 mL) was refluxed for 16 h. Af- ter cooling, the mixture was filtered and washed with acetic acid and water. Drying in vacuo afforded 5.13 g (56%) of [3-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]acetic acid as a solid.
Η-NMR (DMSO-d6): δ 4.69 (2H, s), 6.95 (1 H, dd), 7.09 (1 H, t), 7.15 (1H, d), 7.39 (1 H, t),7.53 (1H, s); HPLC-MS (Method A): m/z = 280 (M+1) (poor ionisation); Rt = 2.49 min.
The compounds in the following examples were similarly prepared.
Example 462 (General procedure (D)) 3-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenyl]acrylic acid
Figure imgf000221_0002
1H-NMR (DMSO-d6): 6.63 (1 H, d), 7.59-7.64 (3H, m), 7.77 (1 H, s), 7.83 (2H, m).
Example 463 (General procedure (D)) [4-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenoxy]acetic acid
Figure imgf000222_0001
Triethylamine salt: 1H-NMR (DMSO-d6): δ 4.27 (2H, s), 6.90 (2H, d), 7.26 (1H, s), 7.40 (2H, d).
Example 464 (General procedure (D)) 4-(2,4-Dioxothiazolidin-5-ylidenemethyl)benzoic acid
Figure imgf000222_0002
Example 465 (General procedure (D))
3-(2,4-Dioxothiazolidin-5-ylidenemethyl)benzoic acid
Figure imgf000222_0003
1H-NMR (DMSO-d6): δ 7.57 (1H, s), 7.60 (1H, t), 7.79 (1H, dt), 7.92 (1H, dt), 8.14 (1H, t).
Example 466 (General procedure (D))
4-[2-Chloro-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]butyric acid
Figure imgf000222_0004
1H-NMR (DMSO-d6): δ 2.00 (2H, p), 2.45 (2H, t), 4.17 (2H, t), 7.31 (1H, d), 7.54 (1H, dd), 7.69 (1H, d), 7.74 (1H, s), 12.2 (1H, bs), 12.6 (1H, bs). HPLC-MS (Method A): m/z: 364 (M+23); Rt = 3.19 min.
Example 467 (General procedure (D)) 4-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]butyric acid
Figure imgf000223_0001
1H-NMR (DMSO-de): δ 1.99 (2H, p), 2.46 (2H, t), 4.17 (2H, t), 7.28 (1 H, d), 7.57 (1H, dd), 7.25 (1 H, s), 7.85 (1H, d), 12.2 (1 H, bs), 12.6 (1H, bs). HPLC-MS (Method A): m/z: 410 (M+23); Rt = 3.35 min.
Example 468 (General procedure (D))
4-[2-Bromo-4-(4-oxo-2-thioxothiazolidin-5-ylidenemethyl)phenoxy]butyric acid
Figure imgf000223_0002
1H-NMR (DMSO-d6): δ 1.99 (2H, p), 2.45 (2H, t), 4.18 (2H, t), 7.28 (1H, d), 7.55 (1H, dd), 7.60 (1 H, s), 7.86 (1H, d), 12.2 (1 H, bs), 13.8 (1H, bs). HPLC-MS (Method A): m/z: 424 (M+23); Rt = 3.84 min. HPLC-MS (Method A): m/z: 424 (M+23); Rt = 3,84 min
Example 469 (General procedure (D)) 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxyjbutyric acid
Figure imgf000223_0003
1H-NMR (DMSO-d6): δ 2.12 (2H, p), 2.5 (below DMSO), 4.28 (2H, t), 7.12 (1 H, d), 7.6-7.7 (3H, m), 8.12 (1H, d), 8.31 (1H, d), 8.39 (1H, s), 12.2 (1H, bs), 12.6 (1H, bs). HPLC-MS (Method A): m/z: 380 (M+23); Rt = 3.76 min.
Example 470 (General procedure (D)) 5-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]pentanoic acid
Figure imgf000224_0001
HPLC-MS (Method A): m/z: 394 (M+23); Rt = 3.62 min.
Η-NMR (DMSO-d6): δ 1.78 (2H, m), 1.90 (2H, m), 2.38 (2H, t), 4.27 (2H, t), 7.16 (1 H, d),
7.6-7.75 (3H, m), 8.13 (1H, d), 8.28 (1H, d), 8.39 (1H, s), 12.1 (1H, bs), 12.6 (1H, bs).
Example 471 5-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]pentanoic acid.
Figure imgf000224_0002
5-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)-naphthalen-1-yloxy]pentanoic acid (example 470, 185 mg, 0.5 mmol) was treated with an equimolar amount of bromine in acetic acid (10 mL). Stirring at RT for 14 days followed by evaporation to dryness afforded a mixture of the bro- minated compound and unchanged starting material. Purification by preparative HPLC on a C18 column using acetonitrile and water as eluent afforded 8 mg of the title compound.
HPLC-MS (Method C): m/z: 473 (M+23), Rt. = 3.77 min
Example 472 4-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]butyric acid.
Figure imgf000224_0003
Starting with 4-[4-(2,4-dioxothiazolidin-5-ylidenemethyl)-naphthalen-1-yloxy]-butyric acid (example 469, 0.5 mmol) using the same method as in example 471 afforded 66 mg of the title compound.
HPLC-MS (Method C): m/z: 459 (M+23) ; Rt. = 3.59 min. Example 473 (General procedure (D))
[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]acetic acid
Figure imgf000225_0001
1H-NMR (DMSO-d6): δ 4.90 (2H, s), 7.12 (1H, d), 7.52 (1H, dd), 7.65 (1H, s) 7.84 (1H, d).HPLC-MS (Method A): m/z: not observed; Rt = 2.89 min.
Example 474 (General procedure (D)) 4-[3-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenoxy]butyric acid
Figure imgf000225_0002
1H-NMR (DMSO-d6): δ 1.98 (2H, p), 2.42 (2H, t), 4.04 (2H, t), 7.05 (1H, dd), 7.15 (2H, m), 7.45 (1 H, t), 7.77 (1 H, s), 12.1 (1H, bs), 12.6 (1H, bs). HPLC-MS (Method A): m/z: 330 (M+23); Rt = 3.05 min.
Example 475 (General procedure (D))
[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)-3-methoxyphenoxy]acetic acid
Figure imgf000225_0003
HPLC-MS (Method B): m/z: 310 (M+1); Rt = 3,43 min.
Example 476 (General procedure (D)) [4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]acetic acid
Figure imgf000226_0001
HPLC-MS (Method A): m/z: 330 (M+1); Rt = 3.25 min.
Example 477 (General procedure (D)) 8-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalene-1 -carboxylic acid
Figure imgf000226_0002
HPLC-MS (Method A): m/z: 299 (M+1 ); Rt = 2,49 min.
Example 478 (General procedure (D)) [3-(2,4-Dioxothiazolidin-5-ylidenemethyl)indol-1 -yljacetic acid
Figure imgf000226_0003
HPLC-MS (Method A): m/z: 303 (M+1); Rt = 2.90 min.
Preparation of starting material: 3-Formylindol (10 g, 69 mmol) was dissolved in Λ/,Λ/-dimethylformamide (100 mL) and under an atmosphere of nitrogenand with external cooling, keeping the temperature below 15 °C, sodium hydride (60% in mineral oil, 3.0 g, 76 mmol) was added in portions. Then a solution of ethyl bromoacetate (8.4 mL, 76 mmol) in Λ/,Λ/-dimethylformamide (15 mL) was added dropwise over 30 minutes and the resulting mixture was stirred at room temperature for 16 hours. The mixture was concentrated in vacuo and the residue was partitioned between water (300 mL) and ethyl acetate (2 x 150 mL). The combined organic extracts were washed with a saturated aqueous solution of ammonium chloride (100 mL), dried (MgS0 ) and con- centrated in vacuo to afford 15.9 g (quant.) of (3-formylindol-1-yl)acetic acid ethyl ester as an oil.
1H-NMR (CDCI3): δH = 1.30 (3H, t), 4.23 (2H, q), 4.90 (2H, s), 7.3 (3H, m), 7.77 (1H, s), 8.32 (1 H, d), 10.0 (1 H, s).
(3-Formylindol-1-yl)acetic acid ethyl ester (15.9 g 69 mmol) was dissolved in 1 ,4-dioxane (100 mL) and 1N sodium hydroxide (10 mL) was added and the resulting mixture was stirred at room temperature for 4 days. Water (500 mL) was added and the mixture was washed with diethyl ether (150 mL). The aqueous phase was acidified with 5N hydrochloric acid and extracted with ethyl acetate (250 + 150 mL). The combined organic extracts were dried (MgS04) and concentrated in vacuo to afford 10.3 g (73%) of (3-formylindol-1-yl)acetic acid as a solid.
1H-NMR (DMSO-dβ): δH = 5.20 (2H, s), 7.3 (2H, m), 7.55 (1 H, d), 8.12 (1H, d), 8.30 (1 H, s), 9.95 (1 H, s), 13.3 (1H, bs).
Example 479 (General procedure (D))
3-[3-(2,4-Dioxothiazolidin-5-ylidenemethyl)indol-1-yl]propionic acid
Figure imgf000227_0001
HPLC-MS (Method A): m/z: 317 (M+1); Rt = 3.08 min.
Preparation of starting material:
A mixture of 3-formylindol (10 g, 69 mmol), ethyl 3-bromopropionate (10.5 mL, 83 mmol) and potassium carbonate (28.5 g, 207 mmol) and acetonitrile (100 mL) was stirred vigorously at refux temperature for 2 days. After cooling, the mixture was filtered and the filtrate was concentrated in vacuo to afford 17.5 g (quant.) of 3-(3-formylindol-1-yl)propionic acid ethyl ester as a solid. 1H-NMR (DMSO-dβ): δH = 1.10 (3H, t), 2.94 (2H, t), 4.02 (2H, q), 4.55 (2H, t), 7.3 (2H, m), 7.67 (1 H, d), 8.12 (1 H, d), 8.30 (1 H, s), 9.90 (1H, s).
3-(3-Formylindol-1-yl)propionic acid ethyl ester (17.5 g 69 mmol) was hydrolysed as de- scribed above to afford 12.5 g (83%) of 3-(3-formylindol-1-yl)propionic acid as a solid.
H-NMR (DMSO-dβ): δH = 2.87 (2H, t), 4.50 (2H, t), 7.3 (2H, m), 7.68 (1H, d), 8.12 (1 H, d), 8.31 (1H, s), 9.95 (1H, s), 12.5 (1H, bs).
Example 480 (General procedure (D))
{5-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)benzylidene]-4-oxo-2-thioxothiazolidin-3-yl}acetic acid
Figure imgf000228_0001
HPLC-MS (Method A): m/z: 429 (M+23); Rt = 3.89 min.
Example 481 (General procedure (D))
6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxyoctanoic acid
Figure imgf000228_0002
HPLC-MS (Method C): m/z: 436 (M+23); Rt.= 4.36 min
The intermediate aldehyde for this compound was prepared by a slightly modified procedure: 6-Hydroxynaphthalene-2-carbaldehyde (1.0 g, 5.8 mmol) was dissolved in DMF (10 mL) and sodium hydride 60% (278 mg) was added and the mixture stirred at RT for 15 min. 8- Bromooctanoic acid (0.37 g, 1.7 mmol) was converted to the sodium salt by addition of sodium hydride 60% and added to an aliquot (2.5 mL) of the above naphtholate solution and the resulting mixture was stirred at RT for 16 hours. Aqueous acetic acid (10 %) was added and the mixture was extracted 3 times with diethyl ether. The combined organic phases were dried with MgSO4 and evaporated to dryness affording 300 mg of 8-(6-formylnaphthalen-2- yloxy)octanoic acid.
HPLC-MS (Method C): m/z 315 (M+1); Rt. = 4.24 min.
Example 482 (General procedure (D)) 12-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]dodecanoic acid.
Figure imgf000229_0001
HPLC-MS (Method C): m/z: 492 (M+23); Rt .= 5.3 min.
The intermediate aldehyde was prepared similarly as described in example 481.
Example 483 (General procedure (D))
11 -[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]undecanoic acid.
Figure imgf000229_0002
HPLC-MS (Method C): m/z:478 (M+23); Rt.= 5.17 min.
The intermediate aldehyde was prepared similarly as described in example 481.
Example 484 (General procedure (D)) 15-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]pentadecanoic acid.
Figure imgf000229_0003
HPLC-MS (Method C): m/z: 534 (M+23); Rt.= 6.07 min.
The intermediate aldehyde was prepared similarly as described in example 481.
Example 485 (General procedure (D)) 6-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]hexanoic acid.
Figure imgf000230_0001
HPLC-MS (Method C): m/z: 408 (M+23); Rt.= 3.71 min.
Example 486 (General procedure (D)) 4-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]butyric acid.
Figure imgf000230_0002
HPLC-MS (Method C): m/z: 380 (M+23); Rt.= 3.23 min.
Example 487 (General procedure (D)) 6-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]hexanoic acid ethyl ester.
Figure imgf000230_0003
HPLC-MS (Method C): m/z: 436 (M+23); Rt .= 4.64 min.
Example 488 (General procedure (D)) 4-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]butyric acid ethyl ester.
Figure imgf000230_0004
HPLC-MS (Method C): m/z: 408 (M+23); Rt .= 4.28 min.
Example 489 (General procedure (D)) 2-{5-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]pentyl}malonic acid
Figure imgf000231_0001
HPLC-MS (Method C): m/z = 444 (M+1); Rt = 3,84 min.
Example 490 (General procedure (D)
2-{5-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]pentyl}malonic acid diethyl ester
Figure imgf000231_0002
HPLC-MS (Method C): m/z = 500 (M+1); Rt = 5.18 min.
Example 491 (General procedure (D))
4-[4-(2,4,6-Trioxotetrahydropyrimidin-5-ylidenemethyl)naphthalen-1 -yloxyjbutyric acid
Figure imgf000231_0003
HPLC-MS (Method C): m/z = 369 (M+1); Rt = 2,68 min.
Example 492
N-(3-Aminopropyl)-4-[4-(2,4-dioxothiazolidin-5-ylidenemethyl)-naphthalen-1-yloxy]- butyramide
Figure imgf000232_0001
To a mixture of 4-[4-(2,4-dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxyjbutyric acid (example 469, 5.9 g, 16.5 mmol) and 1-hydroxybenzotriazole (3.35 g, 24.8 mmol) in DMF (60 mL) was added 1-ethyl-3-(3'-dimethylaminopropyl)carbodiimide hydrochloride (4.75 g, 24.8 mmol) and the resulting mixture was stirred at room temperature for 2 hours. Λ/-(3-amino- propylcarbamic acid tert-butyl ester (3.45 g, 19.8 mmol) was added and the resulting mixture was stirred at room temperature for 16 hours. The mixture was concentrated in vacuo and ethyl acetate and dichloromethane were added to the residue. The mixture was filtered, washed with water and dried in vacuo to afford 4.98 g (59%) of (3-{4-[4-(2,4-dioxothiazolidin- 5-ylidenemethyl)naphthalen-1-yloxy]butyrylamino}propyl)carbamic acid tert-butyl ester.
HPLC-MS (Method C): m/z: 515 (M+1); Rt = 3.79 min.
(3-{4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]butyrylamino}- propyl)carbamic acid tert-butyl ester (4.9 g, 9.5 mmol) was added dichloromethane (50 mL) and trifluoroacetic acid (50 mL) and the resulting mixture was stirred at room temperature for 45 minutes. The mixture was concentrated in vacuo and co-evaporated with toluene. To the residue was added ethyl acetate (100 mL) and the mixture was filtered and dried in vacuo to afford the title compound as the trifluoroacetic acid salt.
HPLC-MS (Method C): m/z: 414 (M+1 ); Rt = 2,27 min.
Compounds of the invention includes:
Example 493
Figure imgf000233_0001
Example 494
Figure imgf000233_0002
Example 495
Figure imgf000233_0003
Example 496
Figure imgf000233_0004
Example 497
Figure imgf000233_0005
Example 498
Figure imgf000233_0006
Example 499
Figure imgf000234_0001
Example 500
Figure imgf000234_0002
Example 501
Figure imgf000234_0003
Example 502
Figure imgf000234_0004
Example 503
Figure imgf000234_0005
Example 504 (Prepared analogously to General Procedure (D)) 2-{5-[4-(2,4-Thiazolidindion-5-ylidenemethyl)naphthalen-1-yloxy]pentyl}malonic acid
Figure imgf000235_0001
A solution of 4-hydroxy-1-naphtaldehyde (1.0 g, 5.81 mmol), 2-(5-bromopentyl)malonic acid diethyl ester (2.07 g, 6.68 mmol) and potassium carbonate (4.01 g, 29 mmol) in DMF (50 mL) was stirred at 100° C for 3 hours. The mixture was cooled and the salt was filtered off. The solvent was then removed under reduced pressure to afford 2.9 g of crude 2-[5-(4- formylnaphtalen-1-yloxy)pentyl]malonic acid diethyl ester which was used for the next reaction without further purification.
HPLC-MS (Method C): m/z: 401 (M+1); Rt = 5.16 min. 1 H-NMR (DMSO-d6): δ = 1.18 (t, 6 H), 1 ,39 (m, 2 H), 1.55 (m, 2 H), 1.87 (m, 4 H), 3.48 (t, 1 H), 4.13 (m, 4 H), 4.27 (t, 2 H), 7.17 (d, 1 H), 7.64(t, 1 H), 7.75 (t, 1 H), 8.13 (d, 1 H), 8.29 (d, 1 H), 9.24 (d, 1 H), 10.19 (s, 1 H).
1.4 g (3.5 mmol) of crude 2-[5-(4-formylnaphtalen-1-yloxy)pentyl]malonic acid diethyl ester was treated with aqueous sodium hydroxide (1N, 8.75 mL, 8.75 mmol) and methanol (50 mL). The solution was stirred at 70° C for 5 hours and the mixture was concentrated under reduced pressure. Hydrochloric acid (6 N) was added until pH <2. The resulting slurry was stirred untill it solidified. The crystals were filtered off, washed with water and then dried in vacuo to afford 1.1 g (92%) of 2-[5-(4-formylnaphtalen-1-yloxy)pentyl]malonic acid. The product was used in the next step without further purification.
HPLC-MS (Method C): m/z: 345 (M+1); Rt = 3.52 min. 1H-NMR(DMSO-d6): δ = 1 ,40 (m, 2 H), 1.55 (m, 2 H), 1.80 (m, 2 H), 1.90 (m, 2 H), 3.24 (t, 1 H), 4.29 (t, 2 H), 7.19 (d, 1 H), 7.64(t, 1 H), 7.75 (t, 1 H), 8.14 (d, 1 H), 8.30 (d, 1 H), 9.23 (d, 1 H), 10.18 (s, 1 H), 12.69 (s, 2 H).
To a solution of 2-[5-(4-formylnaphtalen-1-yloxy) pentyljmalonic acid (0.36 g, 1.05 mmol) in acetic acid (10 mL) was added 2,4-thiazolidindione (0.16 g,1.36 mmol) and piperidine (0.52 mL, 5.25 mmol). The solution was heated to 105 °C for 24 hours. After cooling to room temperature, the solvents were removed in vacuo. Water was added to the residue. The precipi- tate was filtered off and washed with water. Recrystalisation from acetonitrile afforded 200 mg (43%) of the title compound as a solid.
HPLC-MS (Method C): m/z: 422 (M-CO2+Na); Rt = 4.08 min. 1H-NMR(DMSO-d6): δ = 1,41 (m, 2 H), 1.55 (m, 4 H), 1.88 (m, 2 H), 2.23 (t, 1 H), 4.24 (t, 2 H), 7.61-7.74 (m, 3 H), 8.12 (d, 1 H), 8.28 (d, 1 H), 8.38 (s, 1 H), 12.00 (s, 1 H), 12.59 (s, 2 H).
The following compounds are commercially available and may be prepared according to general procedure (D): Example 505
Figure imgf000236_0001
Example 506
Figure imgf000236_0002
Example 507
Figure imgf000236_0003
Example 508
Figure imgf000237_0001
Example 509
Figure imgf000237_0002
Example 510
Figure imgf000237_0003
Example 511
Figure imgf000237_0004
The following salicylic acid derivatives do all bind to the His B10 Zn2+ site of the insulin hexamer: Example 512
Salicylic acid
Figure imgf000238_0001
Example 513
Thiosalicylic acid (or: 2-Mercaptobenzoic acid)
Figure imgf000238_0002
Example 514 2-Hydroxy-5-nitrobenzoic acid
Figure imgf000238_0003
Example 515 3-Nitrosalicyclic acid
Figure imgf000238_0004
Example 516
5,5'-Methylenedisalicylic acid
Figure imgf000238_0005
Example 517
2-Amino-5-trifluoromethylbenzoesyre
Figure imgf000239_0001
Example 518
2-Amino-4-chlorobenzoic acid
Figure imgf000239_0002
Example 519
2-Amino-5-methoxybenzoesyre
Figure imgf000239_0003
Example 520
Figure imgf000239_0004
Example 521
Figure imgf000239_0005
Example 522
Figure imgf000240_0001
Example 523
Figure imgf000240_0002
Example 524
Figure imgf000240_0003
Example 525
Figure imgf000240_0004
Example 526 5-lodosalicylic acid
Figure imgf000240_0005
Example 527 5-Chlorosalicylic acid
Figure imgf000241_0001
Example 528
1-Hydroxy-2-naphthoic acid
Figure imgf000241_0002
Example 529
3,5-Dihydroxy-2-naphthoic acid
Figure imgf000241_0003
Example 530
3-Hydroxy-2-naphthoic acid
Figure imgf000241_0004
Example 531
3,7-Dihydroxy-2-naphthoic acid
Figure imgf000241_0005
Example 532 2-Hydroxybenzo[a]carbazole-3-carboxylic acid
Figure imgf000242_0001
Example 533
7-Bromo-3-hydroxy-2-naphthoic acid
Figure imgf000242_0002
This compound was prepared according to Murphy et al., J. Med. Chem. 1990, 33, 171-8. HPLC-MS (Method A): m/z: 267 (M+1); Rt: = 3.78 min.
Example 534
1 ,6-Dibromo-2-hydroxynaphthalene-3-carboxylic acid
Figure imgf000242_0003
This compound was prepared according to Murphy et al., J. Med. Chem. 1990, 33, 171-8. HPLC-MS (Method A): m/z: 346 (M+1); Rt: = 4,19 min.
Example 535
7-Formyl-3-hydroxynaphthalene-2-carboxylic Acid
Figure imgf000242_0004
A solution of 7-bromo-3-hydroxynaphthalene-2-carboxylic acid (15.0 g, 56.2 mmol) (example 533) in tetrahydrofuran (100 mL) was added to a solution of lithium hydride (893 mg, 112 mmol) in tetrahydrofuran (350 mL). After 30 minutes stirring at room temperature, the resulting solution was heated to 50 °C for 2 minutes and then allowed to cool to ambient temperature over a period of 30 minutes. The mixture was cooled to -78 °C, and butyllithium (1.6 M in hexanes, 53 mL, 85 mmol) was added over a period of 15 minutes. Λ/,Λ/-Dimethylformamide (8.7 mL, 8.2 g, 112 mmol) was added after 90 minutes additional stirring. The cooling was discontinued, and the reaction mixture was stirred at room temperature for 17 hours before it was poured into 1 N hydrochloric acid (aq.) (750 mL). The organic solvents were evaporated in vacuo, and the resulting precipitate was filtered off and rinsed with water (3 x 100 mL) to yield the crude product (16.2 g). Purification on silica gel (dichloromethane / methanol / acetic acid = 90:9:1) furnished the title compound as a solid.
1H-NMR (DMSO-de): 11.95 (1H, bs), 10.02 (1H, s), 8.61 (1H, s), 8.54 (1H, s), 7.80 (2H, bs), 7.24 (1H, s); HPLC-MS (Method (A)): m/z: 217 (M+1); Rt = 2.49 min.
Example 536
3-Hydroxy-7-methoxy-2-naphthoic acid
Figure imgf000243_0001
Example 537
4-Amino-2-hydroxybenzoic acid
Figure imgf000243_0002
Example 538
5-Acetylamino-2 -hydroxybenzoic acid
Figure imgf000243_0003
Example 539 2-Hydroxy-5-methoxybenzoic acid
Figure imgf000244_0001
The following compounds were prepared as described below: Example 540
4-Bromo-3-hydroxynaphthalene-2-carboxylic acid
Figure imgf000244_0002
3-Hydroxynaphthalene-2-carboxylic acid (3.0 g, 15.9 mmol) was suspended in acetic acid (40 mL) and with vigorous stirring a solution of bromine (817 μL, 15.9 mmol) in acetic acid (10 mL) was added drop wise during 30 minutes. The suspension was stirred at room temperature for 1 hour, filtered and washed with water. Drying in vacuo afforded 3.74 g (88%) of 4-bromo-3-hydroxynaphthalene-2-carboxylic acid as a solid.
1H-NMR (DMSO-d6): δ 7.49 (1 H, t), 7.75 (1H, t), 8.07 (2H, "t"), 8.64 (1H, s). The substitution pattern was confirmed by a COSY experiment, showing connectivities between the 3 (4 hydrogen) "triplets". HPLC-MS (Method A): m/z: 267 (M+1); Rt = 3.73 min.
Example 541
3-Hydroxy-4-iodonaphthalene-2-carboxylic acid
Figure imgf000244_0003
3-Hydroxynaphthalene-2-carboxylic acid (0.5 g, 2.7 mmol) was suspended in acetic acid (5 mL) and with stirring iodine monochloride (135 μL, 2.7 mml) was added. The suspension was stirred at room temperature for 1 hour, filtered and washed with water. Drying afforded 0.72 g (85%) of 4-iodo-3-hydroxynaphthalene-2-carboxylic acid as a solid. 1H-NMR (DMSO-d6): δ 7.47 (1H, t), 7.73 (1H, t), 7.98 (1H, d), 8.05 (1H, d), 8.66 (1H, s). HPLC-MS (Method A): m/z: 315 (M+1); Rt = 3.94 min. Example 542
2-Hydroxy-5-[(4-methoxyphenylamino)methyl]benzoic acid
Figure imgf000245_0001
p-Anisidine (1.3 g, 10.6 mmol) was dissolved in methanol (20 mL) and 5-formylsalicylic acid (1.75 g, 10.6 mmol)was added and the resulting mixture was stirred at room temperature for 16 hours. The solid formed was isolated by filtration, re-dissolved in N-methyl pyrrolidone (20 mL) and methanol (2 mL). To the mixture was added sodium cyanoborohydride (1.2 g) and the mixture was heated to 70 °C for 3 hours. To the cooled mixture was added ethyl acetate (100 mL) and the mixture was extracted with water (100 mL) and saturated aqueous ammonium chloride (100 mL). The combined aqueous phases were concentrated in vacuo and a 2 g aliquot was purified by SepPac chromatography eluting with mixtures of aetonitrile and water containing 0.1% trifluoroacetic acid to afford the title compound.
HPLC-MS (Method A): m/z: 274 (M+1); Rt = 1.77 min.
1H-NMR (methanol-d4): δ 3.82 (3H, s), 4.45 (2H, s), 6.96 (1 H, d), 7.03 (2H, d), 7.23 (2H, d),
7.45 (1H, dd), 7.92 (1 H, d).
Example 543
2-Hydroxy-5-(4-methoxyphenylsulfamoyl)benzoic acid H.
Figure imgf000245_0002
A solution of 5-chlrosulfonylsalicylic acid (0.96 g, 4.1 mmol) in dichloromethane (20 mL) and triethylamine (1.69 mL, 12.2 mmol) was added p-anisidine (0.49 g, 4.1 mmol) and the resulting mixture was stirred at room temperature for 16 hours. The mixture was added dichloromethane (50 mL) and was washed with water (2 x 100 mL). Drying (MgS04) of the organic phase and concentration in vacuo afforded 0.57 g crude product. Purification by column chromatography on silica gel eluting first with ethyl acetate:heptane (1 :1) then with methanol afforded 0.1 g of the title compound.
HPLC-MS (Method A): m z: 346 (M+23); Rt = 2.89 min.
1H-NMR (DMSO-de): δ 3.67 (3H, s), 6.62 (1H, d), 6.77 (2H, d), 6.96 (2H, d), 7.40 (1 H, dd),
8.05 (1H, d), 9.6 (1 H, bs).
General procedure (E) for preparation of compounds of general formula l4:
Figure imgf000246_0001
wherein Lea is a leaving group such as Cl, Br, I or OS02CF3, R is hydrogen or CrC6-alkyl, optionally the two R-groups may together form a 5-8 membered ring, a cyclic boronic acid ester, and J is as defined above.
An analogous chemical transformation has previously been described in the literature (Bumagin et al., Tetrahedron, 1997, 53, 14437-14450). The reaction is generally known as the Suzuki coupling reaction and is generally performed by reacting an aryl halide or triflate with an arylboronic acid or a heteroarylboronic acid in the presence of a palladium catalyst and a base such as sodium acetate, sodium carbonate or sodium hydroxide. The solvent can be water, acetone, DMF, NMP, HMPA, methanol, ethanol toluene or a mixture of two or more of these solvents. The reaction is performed at room temperature or at elevated temperature.
The general procedure (E) is further illustrated in the following example: Example 544 (General Procedure (E))
7-(4-Acetylphenyl)-3-hydroxynaphthalene-2-carboxylic Acid
Figure imgf000246_0002
To 7-bromo-3-hydroxynaphthalene-2-carboxylic acid (100 mg, 0.37 mmol) (example 533) was added a solution of 4-acetylphenylboronic acid (92 mg, 0.56 mmol) in acetone (2.2 mL) followed by a solution of sodium carbonate (198 mg, 1.87 mmol) in water (3.3 mL). A suspension of palladium(ll) acetate (4 mg, 0.02 mmol) in acetone (0.5 mL) was filtered and added to the above solution. The mixture was purged with N2 and stirred vigorously for 24 hours at room temperature. The reaction mixture was poured into 1 N hydrochloric acid (aq.) (60 mL) and the precipitate was filtered off and rinsed with water (3 x 40 mL). The crude product was dissolved in acetone (25 mL) and dried with magnesium sulfate (1 h). Filtration followed by concentration furnished the title compound as a solid (92 mg). 1H-NMR (DMSO-d6): 12.60 (1 H, bs), 8.64 (1H, s), 8.42 (1 H, s), 8.08 (2H, d), 7.97 (2H, d), 7.92 (2H, m), 7.33 (1 H, s), 2.63 (3H, s); HPLC-MS (Method (A): m/z: 307 (M+1); Rt = 3.84 min.
The compounds in the following examples were prepared in a similar fashion. Optionally, the compounds can be further purified by recrystallization from e.g. ethanol or by chromatography.
Example 545 (General Procedure (E)) 3-Hydroxy-7-(3-methoxyphenyl)naphthalene-2-carboxylic acid
Figure imgf000247_0001
HPLC-MS (Method (A)): m/z: 295 (M+1); Rt = 4.60 min.
Example 546 (General Procedure (E)) 3-Hydroxy-7-phenylnaphthalene-2-carboxylic acid
Figure imgf000247_0002
HPLC-MS (Method (A)): m/z: 265 (M+1); Rt = 4.6 min.
Example 547 (General Procedure (E)) 3-Hydroxy-7-p-tolylnaphthalene-2-carboxylic acid
Figure imgf000248_0001
HPLC-MS (Method (A)): m/z: 279 (M+1); Rt = 4.95 min.
Example 548 (General Procedure (E)) 7-(4-Formylphenyl)-3-hydroxynaphthalene-2-carboxylic acid
Figure imgf000248_0002
HPLC-MS (Method (A)): m/z: 293 (M+1); Rt = 4.4 min.
Example 549 (General Procedure (E)) 6-Hydroxy-[1 ,2]binaphthalenyl-7-carboxylic acid
Figure imgf000248_0003
HPLC-MS (Method (A)): m/z: 315 (M+1); Rt = 5.17 min.
Example 550 (General Procedure (E)) 7-(4-Carboxy-phenyl)-3-hydroxynaphthalene-2-carboxylic acid
Figure imgf000248_0004
HPLC-MS (Method (A)): m/z: 309 (M+1); Rt = 3.60 min.
Example 551 (General Procedure (E)) 7-Benzofuran-2-yl-3-hydroxynaphthalene-2-carboxylic acid
Figure imgf000249_0001
HPLC-MS (Method (A)): m/z: 305 (M+1); Rt = 4.97 min.
Example 552 (General Procedure (E)) 3-Hydroxy-7-(4-methoxyphenyl)-naphthalene-2-carboxylic acid
Figure imgf000249_0002
HPLC-MS (Method (A)): m/z: 295 (M+1); Rt = 4.68 min.
Example 553 (General Procedure (E)) 7-(3-Ethoxyphenyl)-3-hydroxynaphthalene-2-carboxylic acid
Figure imgf000249_0003
HPLC-MS (Method (A)): m/z: 309 (M+1); Rt = 4.89 min.
Example 554 (General Procedure (E))
7-Benzo[1 ,3]dioxol-5-yl-3-hydroxynaphthalene-2-carboxylic acid
Figure imgf000249_0004
HPLC-MS (Method (A)): m/z: 309 (M+1); Rt = 5.61 min.
Example 555 (General Procedure (E)) 7-Biphenyl-3-yl-3-hydroxynaphthalene-2-carboxylic acid
Figure imgf000249_0005
HPLC-MS (Method (A)): m/z: 341 (M+1); Rt = 5.45 min.
General procedure (F) for preparation of compounds of general formula l5:
Figure imgf000250_0001
wherein R is hydrogen or d-Cβ-alkyl and T is as defined above
This general procedure (F) is further illustrated in the following example:
Example 556 (General procedure (F))
3-Hydroxy-7-[(4-(2-propyl)phenylamino)methyl]naphthalene-2-carboxylic Acid
Figure imgf000250_0002
7-Formyl-3-hydroxynaphthalene-2-carboxylic acid (40 mg, 0.19 mmol) (example 535) was suspended in methanol (300 μL). Acetic acid (16 μL, 17 mg, 0.28 mmol) and 4-(2- propyl)aniline (40 μL, 40 mg, 0.30 mmol) were added consecutively, and the resulting mixture was stirred vigorously at room temperature for 2 hours. Sodium cyanoborohydride (1.0 M in tetrahydrofuran, 300 μL, 0.3 mmol) was added, and the stirring was continued for another 17 hours. The reaction mixture was poured into 6 N hydrochloric acid (aq.) (6 mL), and the precipitate was filtered off and rinsed with water (3 x 2 mL) to yield the title compound (40 mg) as its hydrochloride salt. No further purification was necessary.
1H-NMR (DMSO-d6): £ 10.95 (1H, bs), 8.45 (1 H, s), 7.96 (1H, s), 7.78 (1H, d), 7.62 (1H, d), 7.32 (1 H, s), 7.13 (2H, bd), 6.98 (2H, bd), 4.48 (2H, s), 2.79 (1H, sept), 1.14 (6H, d); HPLC- MS (Method (A)): m/z: 336 (M+1 ); Rt = 3.92 min. The compounds in the following examples were made using this general procedure (F).
Example 557 (General procedure (F))
7-{[(4-Bromophenyl)amino]methyl}-3-hydroxynaphthalene-2-carboxylic Acid
Figure imgf000251_0001
HPLC-MS (Method C): m/z: 372 (M+1); Rt = 4.31 min.
Example 558 (General procedure (F)) 7-{[(3,5-Dichlorophenyl)amino]methyl}-3-hydroxynaphthalene-2-carboxylic Acid
Figure imgf000251_0002
HPLC-MS (Method C): m/z: 362 (M+1); Rt = 4.75 min.
Example 559 (General procedure (F)) 7-{[(Benzothiazol-6-yl)amino]methyl}-3-hydroxynaphthalene-2-carboxylic Acid
Figure imgf000251_0003
HPLC-MS (Method C): m/z: 351 (M+1); Rt = 3.43 min.
Example 560 (General procedure (F)) 3-Hydroxy-7-{[(quinolin-6-yl)amino]methyl}naphthalene-2-carboxylicAcid
Figure imgf000251_0004
HPLC-MS (Method C): m/z: 345 (M+1); Rt = 2.26 min.
Example 561 (General procedure (F)) 3-Hydroxy-7-{[(4-methoxyphenyl)amino]methyl}naphthalene-2-carboxylic Acid
Figure imgf000252_0001
HPLC-MS (Method C): m/z: 324 (M+1); Rt = 2.57min.
Example 562 (General procedure (F))
7-{[(2,3-Dihydrobenzofuran-5-ylmethyl)amino]methyl}-3-hydroxynaphthalene-2-carboxylic Acid
Figure imgf000252_0002
HPLC-MS (Method C): m/z: 350 (M+1); Rt = 2.22 min.
Example 563 (General procedure (F))
7-{[(4-Chlorobenzyl)amino]methyl}-3-hydroxynaphthalene-2-carboxylic Acid
Figure imgf000252_0003
HPLC-MS (Method C): m/z: 342 (M+1); Rt = 2.45 min.
Example 564 (General procedure (F))
3-Hydroxy-7-{[(naphthalen-1-ylmethyl)amino]methyl}naphthalene-2-carboxylic Acid
Figure imgf000252_0004
HPLC-MS (Method C): m/z: 357 (M+1); Rt = 2.63 min.
Example 565 (General procedure (F))
7-{[(Biphenyl-2-ylmethyl)amino]methyl}-3-hydroxynaphthalene-2-carboxylic Acid
Figure imgf000252_0005
HPLC-MS (Method C): m/z: 384 (M+1); Rt = 2.90 min.
Example 566 (General procedure (F))
3-Hydroxy-7-{[(4-phenoxybenzyl)amino]methyl}naphthalene-2-carboxylic Acid
Figure imgf000253_0001
HPLC-MS (Method C): m/z: 400 (M+1 ); Rt = 3.15 min.
Example 567 (General procedure (F))
3-Hydroxy-7-{[(4-methoxybenzyl)amino]methyl}naphthalene-2-carboxylic Acid
Figure imgf000253_0002
HPLC-MS (Method C): m/z: 338 (M+1); Rt = 2.32 min.
General procedure (G) for preparation of compounds of general formula lβ:
Figure imgf000253_0003
wherein J is as defined above and the moiety (Ci-C6-alkanoyl)20 is an anhydride.
The general procedure (G) is illustrated by the following example: Example 568 (General procedure (G))
Λ/-Acetyl-3-hydroxy-7-[(4-(2-propyl)phenylamino)methyl]naphthalene-2-carboxylic Acid
Figure imgf000253_0004
3-Hydroxy-7-[(4-(2-propyl)phenylamino)methyl]naphthalene-2-carboxylic acid (25 mg, 0.07 mmol) (example 556) was suspended in tetrahydrofuran (200 μL). A solution of sodium hydrogencarbonate (23 mg, 0.27 mmol) in water (200 μL) was added followed by acetic anhydride (14 μL, 15 mg, 0.15 mmol). The reaction mixture was stirred vigorously for 65 hours at room temperature before 6 N hydrochloric acid (4 mL) was added. The precipitate was filtered off and rinsed with water (3 x 1 mL) to yield the title compound (21 mg). No further purification was necessary.
1H-NMR (DMSO-de): £10.96 (1 H, bs), 8.48 (1H, s), 7.73 (1 H, s), 7.72 (1 H, d), 7.41 (1 H, dd), 7.28 (1H, s), 7.23 (2H, d), 7.18 (2H, d), 4.96 (2H, s), 2.85 (1 H, sept), 1.86 (3H, s), 1.15 (6H, d); HPLC-MS (Method (A)): m/z: 378 (M+1); Rt = 3.90 min.
The compounds in the following examples were prepared in a similar fashion.
Example 569 (General procedure (G))
Λ/-Acetyl-7-{[(4-bromophenyl)amino]methyl}-3-hydroxynaphthalene-2-carboxylic Acid
Figure imgf000254_0001
HPLC-MS (Method C): m/z: 414 (M+1); Rt = 3.76 min.
Example 570 (General procedure (G))
Λ/-Acetyl-7-{[(2,3-dihydrobenzofuran-5-ylmethyl)amino]methyl}-3-hydroxynaphthalene-2- carboxylic Acid
Figure imgf000254_0002
HPLC-MS (Method C): m/z: 392 (M+1); Rt = 3.26 min.
Example 571 (General procedure (G))
Λ/-Acetyl-7-{[(4-chlorobenzyl)amino]methyl}-3-hydroxynaphthalene-2-carboxylic Acid
Figure imgf000254_0003
HPLC-MS (Method C): m/z: 384 (M+1); Rt = 3.67 min.
Compounds of the invention may also include tetrazoles: Example 572 5-(3-(Naphthalen-2-yloxymethyl)-phenyl)-1 H-tetrazole
Figure imgf000255_0001
To a mixture of 2-naphthol (10 g, 0.07 mol) and potassium carbonate (10 g, 0.073 mol) in acetone (150 mL), alpha-bromo-m-tolunitril (13.6 g, 0.07 mol) was added in portions. The reaction mixture was stirred at reflux temperature for 2.5 hours. The cooled reaction mixture was filtered and evaporated in vacuo affording an oily residue (19 g) which was dissolved in diethyl ether (150 mL) and stirred with a mixture of active carbon and MgS04 for 16 hours.
The mixture was filtered and evaporated in vacuo affording crude 18.0 g (100 %) of 3-
(naphthalen-2-yloxymethyl)-benzonitrile as a solid.
12 g of the above benzonitrile was recrystallised from ethanol (150 mL) affording 8.3 g (69 %) of 3-(naphthalen-2-yloxymethyl)-benzonitrile as a solid.
M.p. 60 - 61 °C. Calculated for C18H13NO: C, 83.37 %; H, 5.05 %; N, 5.40 %; Found C, 83.51 %; H, 5.03 %; N, 5.38 %.
To a mixture of sodium azide (1.46 g, 22.5 mmol) and ammonium chloride (1.28 g, 24.0 mmol) in dry dimethylformamide (20 mL) under an atmosphere of nitrogen, 3-(naphthalen-2- yloxymethyl)-benzonitrile (3.9 g, 15 mmol) was added and the reaction mixture was stirred at 125 °C for 4 hours. The cooled reaction mixture was poured on to ice water (300 mL) and acidified to pH = 1 with 1 N hydrochloric acid. The precipitate was filtered off and washed with water, dried at 100 °C for 4 hours affording 4.2 g (93 %) of the title compound.
M.p. 200 - 202 °C. Calculated for C18H14N40:
C, 71.51 %; H, 4.67 %; N, 18.54 %; Found C, 72.11 %; H, 4.65 %; N, 17.43 %. 1H NMR (400 MHz, DMSO-d6) δH 5.36 (s, 2H), 7.29 (dd, 1H), 7.36 (dt, 1H), 7.47 (m, 2H), 7.66 (t, 1 H), 7.74 (d, 1 H), 7.84 (m, 3H), 8.02 (d, 1 H), 8.22 (s, 1 H).
Example 573
N-(3-(Tetrazol-5-yl)phenyl)-2-naphtoic acid amide
Figure imgf000256_0001
2-Naphtoic acid (10 g, 58 mmol) was dissolved in dichloromethane (100 mL) and N,N- dimethylformamide (0.2 mL) was added followed by thionyl chloride (5.1 ml, 70 mmol). The mixture was heated at reflux temperature for 2 hours. After cooling to room temperature, the mixture was added dropwise to a mixture of 3-aminobenzonitril (6.90 g, 58 mmol) and triethyl amine (10 mL) in dichloromethane (75 mL). The resulting mixture was stirred at room temperature for 30 minutes. Water (50 mL) was added and the volatiles was exaporated in. vacuo. The resulting mixture was filtered and the filter cake was washed with water followed by heptane (2 x 25 mL). Drying in vacuo at 50 °C for 16 hours afforded 15.0 g (95 %) of N-(3- cyanophenyl)-2-naphtoic acid amide.
M.p. 138-140 °C
The above naphthoic acid amide (10 g, 37 mmol) was dissolved in N,N-dimethylformamide (200 mL) and sodium azide (2.63 g, 40 mmol) and ammonium chloride (2.16 g, 40 mmol) were added and the mixture heated at 125 °C for 6 hours. Sodium azide (1.2 g) and ammonium chloride (0.98 g) were added and the mixture heated at 125 °C for 16 hours. After cool- ing, the mixture was poured into water (1.5 I) and stirred at room temperature for 30 minutes. The solid formed was filtered off, washed with water and dried jn vacuo at 50 °C for 3 days affording 9.69 g (84 %) of the title compound as a solid which could be further purified by treatment with ethanol at reflux temperature.
1H NMR (200 MHz, DMSO-d6): δH 7.58-7.70 (m, 3H), 7.77 (d, 1 H), 8.04-8.13 (m, 5H), 8.65 (d, 1 H), 10.7 (s, 1 H). Calculated for C18H13N50, 0.75 H20:
C, 65.74 %; H, 4.44 %; N, 21.30 %. Found:
C, 65.58 %; H, 4.50 %; N, 21.05 %.
Example 574 5-[3-(Biphenyl-4-yloxymethyl)phenyl]-1 H-tetrazole
Figure imgf000257_0001
To a solution of 4-phenylphenol (10.0 g, 59 mmol) in dry N,N-dimethyl-formamide (45 mL) kept under an atmosphere of nitrogen, sodium hydride (2.82 g, 71 mmol, 60 % dispersion in oil) was added in portions and the reaction mixture was stirred until gas evolution ceased. A solution of m-cyanobenzyl bromide (13 g, 65 mmol) in dry N,N-dimethylformamide (45 mL) was added dropwise and the reaction mixture was stirred at room temperature for 18 hours.
The reaction mixture was poured on to ice water (150 mL). The precipitate was filtered of and washed with 50 % ethanol (3 x 50 mL), ethanol (2 x 50 mL), diethyl ether (80 mL), and dried in vacuo at
50 °C for 18 hours affording crude 17.39 g of 3-(biphenyl-4-yloxymethyl)-benzonitrile as a solid.
1H NMR (200 MHz, CDCI3) δH 5.14 (s, 2H), 7.05 (m, 2H), 7.30 - 7.78 (m, 11H).
To a mixture of sodium azide (2.96 g, 45.6 mmol) and ammonium chloride (2.44 g, 45.6 mmol) in dry N,N-dimethylformamide (100 mL) under an atmosphere of nitrogen, 3-(biphenyl- 4-yloxymethyl)-benzonitrile (10.0 g, 35.0 mmol) was added and the reaction mixture was stirred at 125 °C for 18 hours. The cooled reaction mixture was poured on to a mixture of 1 N hydrochloric acid (60 mL) and ice water (500 mL). The precipitate was filtered off and washed with water (3 x 100 mL), 50 % ethanol (3 x 100 mL), ethanol (50 mL), diethyl ether (50 mL), ethanol (80 mL), and dried in vacuo at 50 °C for 18 hours affording 8.02 g (70 %) of the tjtje compound.
1H NMR (200 MHz, DMSO-d6) δH 5.31 (s, 2H), 7.19 (m, 2H), 7.34 (m, 1 H), 7.47 (m, 2H), 7.69 (m, 6H), 8.05 (dt, 1 H), 8.24 (s, 1H). Example 575 5-(3-Phenoxymethyl)-phenyl)-tetrazole
Figure imgf000258_0001
3-Bromomethylbenzonitrile (5.00 g, 25.5 mmol) was dissolved in N,N-dimethylformamide (50 mL), phenol (2.40 g, 25.5 mmol) and potassium carbonate (10.6 g, 77 mmol) were added. The mixture was stirred at room temperature for 16 hours. The mixture was poured into water (400 mL) and extracted with ethyl acetate (2 x 200 mL). The combined organic extracts were washed with water (2 x 100 mL), dried (MgS0 ) and evaporated in vacuo to afford 5.19 g (97 %) 3-(phenoxymethyl)benzonitrile as an oil.
TLC: Rf = 0.38 (Ethyl acetate/heptane = 1 :4)
The above benzonitrile (5.19 g, 24.8 mmol) was dissolved in N,N-dimethylformamide (100 mL) and sodium azide (1.93 g, 30 mmol) and ammonium chloride (1.59 g, 30 mmol) were added and the mixture was heated at 140 °C for 16 hours. After cooling, the mixture was poured into water (800 mL). The aqeous mixture was washed with ethyl acetate (200 mL).
The pH of the aqueous phase was adjusted to 1 with 5 N hydrochloric acid and stirred at room temperature for 30 minutes. Filtration, washing with water and drying in vacuo at 50 °C afforded 2.06 g (33 %) of the title compound as a solid.
1H NMR (200 MHz, CDCI3 + DMSO-d6) δH 5.05 (s, 2H), 6.88 (m, 3H), 7.21 (m, 2H), 7.51 (m,
2H), 7.96 (dt, 1H), 8.14 (s, 1H).
Example 576 5-[3-(Biphenyl-4-ylmethoxy)phenyl]-1 H-tetrazole
Figure imgf000258_0002
To a solution of 3-cyanophenol (5.0 g, 40.72 mmol) in dry N,N-dimethylformamide (100 mL) kept under an atmosphere of nitrogen, sodium hydride (2 g, 48.86 mmol, 60 % dispersion in oil) was added in portions and the reaction mixture was stirred until gas evolution ceased, p- Phenylbenzyl chloride (9.26 g, 44.79 mmol) and potassium iodide (0.2 g, 1.21 mmol) were added and the reaction mixture was stirred at room temperature for 60 hours. The reaction mixture was poured on to a mixture of saturated sodium carbonate (100 mL) and ice water (300 mL). The precipitate was filtered of and washed with water (3 x 100 mL), n-hexane (2 x 80 mL) and dried in vacuo at 50 °C for 18 hours affording 11.34 g (98 %) of 3-(biphenyl-4- ylmethoxy)-benzonitrile as a solid.
To a mixture of sodium azide (2.37 g, 36.45 mmol) and ammonium chloride (1.95 g, 36.45 mmol) in dry N,N-dimethylformamide (100 mL) under an atmosphere of nitrogen, 3-(biphenyl- 4-ylmethoxy)-benzonitrile (8.0 g, 28.04 mmol) was added and the reaction mixture was stirred at
125 °C for 18 hours. To the cooled reaction mixture water (100 mL) was added and the reaction mixture stirred for 0.75 hour. The precipitate was filtered off and washed with water, 96 % ethanol (2 x 50 mL), and dried in vacuo at 50°C for 18 hours affording 5.13 g (56 %) of the title compound.
1H NMR (200 MHz, DMSO-d6) δH 5.29 (s, 2H), 7.31 (dd, 1H), 7.37 - 7.77 (m, 12H).
Example 577 5-[4-(Biphenyl-4-ylmethoxy)-3-methoxyphenyl]-1 H-tetrazol
Figure imgf000259_0001
This compound was made similarly as described in example 576.
Figure imgf000259_0002
Example 579 5-(2-Naphtylmethyl)-1 H-tetrazole
Figure imgf000260_0001
This compound was prepared similarly as described in example 572, step 2.
Example 580 5-(1 -Naphtylmethyl)-1 H-tetrazole
Figure imgf000260_0002
This compound was prepared similarly as described in example 572, step 2.
Example 581 5-[4-(Biphenyl-4-yloxymethyl)phenyl]-1 H-tetrazole
N-ι
OO KB
A solution of alpha-bromo-p-tolunitrile (5.00 g, 25.5 mmol), 4-phenylphenol (4.56 g, 26.8 mmol), and potassium carbonate (10.6 g, 76.5 mmol) in N,N-dimethylformamide (75 mL) was stirred vigorously for 16 hours at room temperature. Water (75 mL) was added and the mix- ture was stirred at room temperature for 1 hour. The precipitate was filtered off and washed with thoroughly with water. Drying in vacuo over night at 50 °C afforded 7.09 g (97 %) of 4- (biphenyl-4-yloxymethyl)benzonitrile as a solid.
The above benzonitrile (3.00 g, 10.5 mmol) was dissolved in N,N-dimethylformamide (50 mL), and sodium azide (1.03 g, 15.8 mmol) and ammonium chloride (0.84 g, 15.8 mmol) were added and the mixture was stirred 16 hours at 125 °C. The mixture was cooled to room temperature and water (50 mL) was added. The suspension was stirred overnight, filtered, washed with water and dried in vacuo at 50 °C for 3 days to give crude 3.07 g (89 %) of the title compound. From the mother liquor crystals were colected and washed with water, dried by suction to give 0.18 g
(5 %) of the title compound as a solid. 1H NMR (200 MHz, DMSO-d6): δH 5.21 (s, 2H), 7.12 (d, 2H), 7.30 (t, 1 H), 7.42 (t, 2H), 7.56- 7.63 (m, 6H), 8.03 (d, 2H).
Calculated for C20H16N4O, 2H20: C, 65.92 %; H, 5.53 %; N, 15.37 %. Found: C, 65.65 %; H, 5.01 %; N, 14.92 %.
Example 582
Figure imgf000261_0001
This compound was prepared similarly as described in example 576.
Example 583
Figure imgf000261_0002
Example 584
Figure imgf000261_0003
Example 585
Figure imgf000262_0001
Example 586 5-(3-(Biphenyl-4-yloxymethyl)-benzyl)-1 H-tetrazole
Figure imgf000262_0002
Example 587
5-(1-Naphthyl)-1 H-tetrazole
Figure imgf000262_0003
This compound was prepared similarly as described in example 572, step 2.
Example 588 5-[3-Methoxy-4-(4-methylsulfonylbenzyloxy)phenyl]-1 H-tetrazole
Figure imgf000262_0004
This compound was made similarly as described in example 576.
Example 589 5-(2-Naphthyl)-1 H-tetrazole
Figure imgf000263_0001
This compound was prepared similarly as described in example 572, step 2.
Example 590 2-Amino-N-(1 H-tetrazol-5-yl)-benzamide
Figure imgf000263_0002
Example 591 5-(4-Hydroxy-3-methoxyphenyl)-1 H-tetrazole
Figure imgf000263_0003
This compound was prepared similarly as described in example 572, step 2.
Example 592
4-(2H-Tetrazol-5-ylmethoxy)benzoic acid
Figure imgf000263_0004
To a mixture of methyl 4-hydroxybenzoate (30.0 g, 0.20 mol), sodium iodide (30.0 g, 0.20 mol) and potassium carbonate (27.6 g, 0.20 mol) in acetone (2000 mL) was added chloroacetonitrile (14.9 g , 0.20 mol). The mixture was stirred at RT for 3 days. Water was added and the mixture was acidified with 1 N hydrochloric acid and the mixture was extracted with diethyl ether. The combined organic layers were dried over Na2SO4 and concentrated in vacuo. The residue was dissolved in acetone and chloroacetonitrile (6.04 g,0.08 mol), so- dium iodide (12.0 g, 0.08 mol) and potassium carbonate (11.1 g, 0.08 mol) were added and the mixture was stirred for 16 hours at RT and at 60 °C. More chloroacetonitrile was added until the conversion was 97%. Water was added and the mixture was acidified with 1 N hydrochloric acid and the mixture was extracted with diethyl ether. The combined organic lay- ers were dried over Na2S04 and concentrated in vacuo to afford methyl 4- cyanomethyloxybenzoate in quantitative yield. This compound was used without further purification in the following step.
A mixture of methyl 4-cyanomethyloxybenzoate (53.5 g,0.20 mol), sodium azide (16.9 g, 0.26 mol) and ammonium chloride (13.9 g, 0.26 mol) in DMF 1000 (mL) was refluxed overnight under N2. After cooling, the mixture was concentrated in vacuo. The residue was suspended in cold water and extracted with ethyl acetate. The combined organic phases were washed with brine, dried over Na2S04 and concentrated in vacuo, to afford methyl 4-(2H-tetrazol-5- ylmethoxy)benzoate. This compound was used as such in the following step.
Methyl 4-(2H-Tetrazol-5-ylmethoxy)-benzoate was refluxed in 3N sodium hydroxide. The reaction was followed by TLC (DCM:MeOH = 9:1). The reaction mixture was cooled, acidified and the product filtered off. The impure product was washed with DCM, dissolved in MeOH, filtered and purified by column chromatography on silica gel (DCM: MeOH = 9:1).The result- ing product was recrystallised from DCM:MeOH=95:5. This was repeated until the product was pure. This afforded 13.82 g (30 %) of the title compound.
1H-NMR (DMSO-dβ): 4.70 (2H, s), 7.48 (2H, d), 7.73 (2H, d), 13 (1 H, bs).
Example 593
4-(2H-Tetrazol-5-ylmethylsulfanyl)benzoic acid
Figure imgf000264_0001
To a solution of sodium hydroxide (10.4 g, 0.26 mol) in degassed water (600 mL) was added 4-mercaptobenzoic acid (20.0 g, 0.13 mol). This solution was stirred for 30 minutes. To a solution of potassium carbonate (9.0 g, 65 mmol) in degassed water (400 mL) was added chloroacetonitrile (9.8 g, (0.13 mol) portion-wise. These two solutions were mixed and stirred for 48 hours at RT under N2. The mixture was filtered and washed with heptane. The aque- ous phase was acidified with 3N hydrochloric acid and the product was filtered off, washed with water and dried, affording 4-cyanomethylsulfanylbenzoic acid (27.2 g, 88%). This compound was used without further purification in the following step.
A mixture of 4-cyanomethylsulfanylbenzoic acid (27.2 g, 0.14 mol), sodium azide (11.8 g, 0,18 mol) and ammonium chloride (9.7 g, 0.18 mol) in DMF (1000 mL) was refluxed overnight under N2. The mixture was concentrated in vacuo. The residue was suspended in cold water and extracted with diethyl ether. The combined organic phases were washed with brine, dried over Na2S04 and concentrated in vacuo. Water was added and the precipitate was filtered off. The aqueous layer was concentrated in vacuo, water was added and the precipitate filtered off. The combined impure products were purified by column chromatography using DCM:MeOH = 9:1 as eluent, affording the title compound (5.2 g, 16%).
1H-NMR (DMSO-d6): 5.58 (2H, s), 7.15 (2H, d), 7.93 (2H, d), 12.7 (1 H, bs).
Example 594 3-(2H-Tetrazol-5-yl)-9H-carbazole
Figure imgf000265_0001
3-Bromo-9H-carbazole was prepared as described by Smith et al. in Tetrahedron 1992, 48, 7479-7488.
A solution of 3-bromo-9H-carbazole (23.08 g, 0.094 mol) and cuprous cyanide (9.33 g, 0.103 mol) in Λ/-methyl-pyrrolidone (300 ml) was heated at 200 °C for 5 h. The cooled reaction mixture was poured on to water (600 ml) and the precipitate was filtered off and washed with ethyl acetate (3 x 50 ml). The filtrate was extracted with ethyl acetate (3 x 250 ml) and the combined ethyl acetate extracts were washed with water (150 ml), brine (150 ml), dried
(MgS04) and concentrated in vacuo. The residue was crystallised from heptanes and recrystallised from acetonitrile (70 ml) affording 7.16 g (40 %) of 3-cyano-9H-carbazole as a solid. M.p. 180 - 181 °C.
3-Cyano-9H-carbazole (5.77 g, 30 mmol) was dissolved in A/,Λ/-dimethylformamide (150 ml), and sodium azide (9.85 g, 152 mmol), ammonium chloride (8.04 g, 150 mmol) and lithium chloride (1.93 g, 46 mmol) were added and the mixture was stirred for 20 h at 125 °C. To the reaction mixture was added an additional portion of sodium azide (9.85 g, 152 mmol) and ammonium chloride (8.04 g, 150 mmol) and the reaction mixture was stirred for an additional 24 h at 125 °C. The cooled reaction mixture was poured on to water (500 ml). The suspen- sion was stirred for 0.5 h, and the precipitate was filtered off and washed with water (3 x 200 ml) and dried in vacuo at 50 °C. The dried crude product was suspended in diethyl ether (500 ml) and stirred for 2 h, filtered off and washed with diethyl ether (2 x 200 ml) and dried in vacuo at 50 °C affording 5.79 g (82 %) of the title compound as a solid.
1H-NMR (DMSO-d6): £ 11.78 (1 H, bs), 8.93 (1H, d), 8.23 (1 H, d), 8.14 (1 H, dd), 7.72 (1H, d), 7.60 (1 H, d), 7.49 (1 H, t), 7.28 (1H, t); HPLC-MS (Method C): m/z: 236 (M+1); Rt = 2.77 min.
The following commercially available tetrazoles do all bind to the His B10 Zn2+ site of the insulin hexamer:
Example 595 5-(3-Tolyl)-1 H-tetrazole
Figure imgf000266_0001
Example 596
5-(2-Bromophenyl)tetrazole
Figure imgf000266_0002
Example 597 5-(4-Ethoxalylamino-3-nitrophenyl)tetrazole
Figure imgf000267_0001
Example 598
Figure imgf000267_0002
Example 599
Figure imgf000267_0003
Example 600
Figure imgf000267_0004
Example 601
Figure imgf000268_0001
Example 602
Tetrazole
Figure imgf000268_0002
Example 603 5-Methyltetrazole
Figure imgf000268_0003
Example 604 5-Benzyl-2H-tetrazole
Figure imgf000268_0004
Example 605
4-(2H-Tetrazol-5-yl)benzoic acid
Figure imgf000268_0005
Example 606 5-Phenyl-2H-tetrazole
Figure imgf000268_0006
Example 607
5-(4-Chlorophenylsulfanylmethyl)-2H-tetrazole
Figure imgf000269_0001
Example 608 5-(3-Benzyloxyphenyl)-2H-tetrazole
Figure imgf000269_0002
Example 609
2-Phenyl-6-(1H-tetrazol-5-yl)-chromen-4-one
Figure imgf000269_0003
Example 610
Figure imgf000269_0004
Example 611
Figure imgf000270_0001
Example 612
Figure imgf000270_0002
Example 613
Figure imgf000270_0003
Example 614
Figure imgf000270_0004
Example 615
5-(4-Bromo-phenyl)-1 H-tetrazole
Figure imgf000271_0001
Example 616
Figure imgf000271_0002
Example 617
Figure imgf000271_0003
Example 618
Figure imgf000271_0004
Example 619
Figure imgf000271_0005
Example 620
Figure imgf000272_0001
Example 621
Figure imgf000272_0002
Example 622
Figure imgf000272_0003
Example 623
Figure imgf000272_0004
Example 624
Figure imgf000273_0001
Example 625
Figure imgf000273_0002
Example 626
Figure imgf000273_0003
Example 627
Figure imgf000273_0004
Example 628
Figure imgf000273_0005
Example 629
Figure imgf000274_0001
Example 630
Figure imgf000274_0002
Example 631
Figure imgf000274_0003
Example 632
Figure imgf000274_0004
Example 633
Figure imgf000275_0001
Example 634
Figure imgf000275_0002
Example 635
Figure imgf000275_0003
Example 636
Figure imgf000275_0004
Example 637
Figure imgf000276_0001
Example 638
Figure imgf000276_0002
Example 639
Figure imgf000276_0003
Example 640
Figure imgf000276_0004
Example 641
Figure imgf000277_0001
Example 642
Figure imgf000277_0002
Example 643
Figure imgf000277_0003
Example 644
Figure imgf000277_0004
Example 645
Figure imgf000278_0001
Example 646 5-(2,6-Dichlorobenzyl)-2H-tetrazole
Figure imgf000278_0002
General procedure (H) for preparation of compounds of general formula l7:
NaCBH,
Figure imgf000278_0003
wherein K, M, and T are as defined above.
The reaction is generally known as a reductive alkylation reaction and is generally performed by stirring an aldehyde with an amine at low pH (by addition of an acid, such as acetic acid or formic acid) in a solvent such as THF, DMF, NMP, methanol, ethanol, DMSO, dichloromethane, 1 ,2-dichloroethane, trimethyl orthoformate, triethyl orthoformate, or a mixture of two or more of these. As reducing agent sodium cyano borohydride or sodium triacetoxy borohydride may be used. The reaction is performed between 20°C and 120°C, preferably at room temperature.
When the reductive alkylation is complete, the product is isolated by extraction, filtration, chromatography or other methods known to those skilled in the art.
The general procedure (H) is further illustrated in the following example 647: Example 647 (General procedure (H)) Biphenyl-4-ylmethyl-[3-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000279_0001
A solution of 5-(3-aminophenyl)-2H-tetrazole (example 874, 48 mg, 0.3 mmol) in DMF (250 μL) was mixed with a solution of 4-biphenylylcarbaldehyde (54 mg, 0.3 mmol) in DMF (250 μL) and acetic acid glacial (250 μL) was added to the mixture followed by a solution of sodium cyano borohydride (15 mg, 0.24 mmol) in methanol (250 μL). The resulting mixture was shaken at room temperature for 2 hours. Water (2 mL) was added to the mixture and the resulting mixture was shaken at room temperature for 16 hours. The mixture was centrifugated (6000 rpm, 10 minutes) and the supernatant was removed by a pipette. The residue was washed with water (3 mL), centrifugated (6000 rpm, 10 minutes) and the supernatant was removed by a pipette. The residue was dried in vacuo at 40 °C for 16 hours to afford the title compound as a solid.
HPLC-MS (Method C): m/z: 328 (M+1 ), 350 (M+23); Rt = 4.09 min.
Example 648 (General procedure (H)) Benzyl-[3-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000279_0002
HPLC-MS (Method D): m/z: 252 (M+1 ); Rt = 3,74 min.
Example 649 (General procedure (H)) (4-Methoxybenzyl)-[3-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000279_0003
HPLC-MS (Method D): m/z: 282,2 (M+1 ); Rt = 3,57min. Example 650 (General procedure (H)) 4-{[3-(2H-Tetrazol-5-yl)phenylamino]methyl}phenol
Figure imgf000280_0001
HPLC-MS (Method D): m/z: 268,4 (M+1); Rt = 2,64 min.
Example 651 (General procedure (H)) (4-Nitrobenzyl)-[3-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000280_0002
HPLC-MS (Method D): m/z: 297,4 (M+1 ); Rt = 3,94 min.
Example 652 (General procedure (H)) (4-Chlorobenzyl)-[3-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000280_0003
HPLC-MS (Method D): m/z: 287,2 (M+1); Rt = 4,30 min.
Example 653 (General procedure (H)) (2-Chlorobenzyl)-[3-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000280_0004
HPLC-MS (Method D): m/z: 286 (M+1 ); Rt = 4,40 min.
Example 654 (General procedure (H)) (4-Bromobenzyl)-[3-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000281_0001
HPLC-MS (Method D): m/z:332 (M+1); Rt = 4,50 min.
Example 655 (General procedure (H)) (3-Benzyloxybenzyl)-[3-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000281_0002
HPLC-MS (Method D): m/z: 358 (M+1); Rt = 4,94 min.
Example 656 (General procedure (H)) Naphthalen-1 -ylmethyl-[3-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000281_0003
HPLC-MS (Method D): m/z: 302 (M+1); Rt = 4,70 min.
Example 657 (General procedure (H)) Naphthalen-2-ylmethyl-[3-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000281_0004
HPLC-MS (Method D): m/z: 302 (M+1); Rt = 4,60 min.
Example 658 (General procedure (H)) 4-{[3-(2H-Tetrazol-5-yl)phenylamino]methyl}benzoic acid
Figure imgf000281_0005
HPLC-MS (Method D): m/z: 296 (M+1); Rt = 3,24 min. Example 659 (General procedure (H)) [3-(2H-Tetrazol-5-yl)-phenyl]-[3-(3-trifluoromethyl-phenoxy)benzyl]amine
Figure imgf000282_0001
HPLC-MS (Method D): m/z: 412 (M+1 ); Rt = 5,54 min.
Example 660 (General procedure (H)) (3-Phenoxybenzyl)-[3-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000282_0002
HPLC-MS (Method D): m/z: 344 (M+1); Rt = 5,04 min.
Example 661 (General procedure (H)) (4-Phenoxy-benzyl)-[3-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000282_0003
HPLC-MS (Method D): m/z: 344 (M+1 ); Rt = 5,00 min.
Example 662 (General procedure (H)) (4-{[3-(2H-Tetrazol-5-yl)phenylamino]methyl}phenoxy)acetic acid
Figure imgf000282_0004
HPLC-MS (Method D): m/z: 326 (M+1); Rt = 3,10 min.
Example 663 (General procedure (H)) (4-Benzyloxybenzyl)-[3-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000283_0001
HPLC-MS (Method D): m/z: 358 (M+1); Rt = 4,97 min.
Example 664 (General procedure (H)) 3-(4-{[3-(2H-Tetrazol-5-yl)phenylamino]methyl}phenyl)acrylic acid
Figure imgf000283_0002
HPLC-MS (Method D): m/z: 322 (M+1); Rt = 3,60 min.
Example 665 (General procedure (H)) Dimethyl-(4-{[3-(2H-tetrazol-5-yl)phenylamino]methyl}naphthalen-1 -yl)amine
Figure imgf000283_0003
HPLC-MS (Method D): m/z: 345 (M+1); Rt = 3,07 min.
Example 666 (General procedure (H)) (4'-Methoxybiphenyl-4-ylmethyl)-[3-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000283_0004
HPLC-MS (Method D): m/z: 358 (M+1); Rt = 4,97 min.
Example 667 (General procedure (H)) (2'-Chlorobiphenyl-4-ylmethyl)-[3-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000284_0001
HPLC-MS (Method D): m/z: 362 (M+1); Rt = 5,27 min.
Example 668 (General procedure (H)) Benzyl-[4-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000284_0002
For preparation of starting material, see example 875. HPLC-MS (Method D): m/z: 252 (M+1); Rt = 3,97 min.
Example 669 (General procedure (H))
(4-Methoxybenzyl)-[4-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000284_0003
HPLC-MS (Method D): m/z: 282 (M+1); Rt = 3,94 min.
Example 670 (General procedure (H))
4-{[4-(2H-Tetrazol-5-yl)phenylamino]methyl}phenol
Figure imgf000284_0004
HPLC-MS (Method D): m/z: 268 (M+1); Rt = 3,14 min.
Example 671 (General procedure (H))
(4-Nitrobenzyl)-[4-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000285_0001
HPLC-MS (Method D): m/z: (M+1); Rt = 3,94 min.
Example 672 (General procedure (H)) (4-Chlorobenzyl)-[4-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000285_0002
HPLC-MS (Method D): m/z: (M+1); Rt = 4,47 min.
Example 673 (General procedure (H)) (2-Chlorobenzyl)-[4-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000285_0003
HPLC-MS (Method D): m/z: 286 (M+1); Rt = 4,37 min.
Example 674 (General procedure (H)) (4-Bromobenzyl)-[4-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000285_0004
HPLC-MS (Method D): m/z: 331 (M+1); Rt = 4,57 min.
Example 675 (General procedure (H)) (3-Benzyloxybenzyl)-[4-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000285_0005
HPLC-MS (Method D): m/z: 358 (M+1); Rt = 5,07min.
Example 676 (General procedure (H)) Naphthalen-1-ylmethyl-[4-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000286_0001
HPLC-MS (Method D): m/z: 302 (M+1); Rt = 4,70 min.
Example 677 (General procedure (H)) Naphthalen-2-ylmethyl-[4-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000286_0002
HPLC-MS (Method D): m/z: 302 (M+1); Rt = 4,70 min.
Example 678 (General procedure (H)) Bipheny!-4-ylmethyl-[4-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000286_0003
HPLC-MS (Method D): m/z: 328 (M+1); Rt = 5,07 min.
Example 679 (General procedure (H)) 4-{[4-(2H-Tetrazol-5-yl)phenylamino]methyl}benzoic acid
Figure imgf000286_0004
HPLC-MS (Method D): m/z: 296 (M+1); Rt = 3,34 min. Example 680 (General procedure (H)) [4-(2H-Tetrazol-5-yl)phenyl]-[3-(3-trifluoromethylphenoxy)benzyl]amine
Figure imgf000287_0001
HPLC-MS (Method D): m/z: 412 (M+1); Rt = 5,54 min.
Example 681 (General procedure (H)) (3-Phenoxybenzyl)-[4-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000287_0002
HPLC-MS (Method D): m/z: 344 (M+1); Rt = 5,07 min.
Example 682 (General procedure (H)) (4-Phenoxybenzyl)-[4-(2H-tetrazol-5-yl)-phenyl]-amine
Figure imgf000287_0003
HPLC-MS (Method D): m/z: 344 (M+1); Rt = 5,03 min.
Example 683 (General procedure (H)) 3-{[4-(2H-Tetrazol-5-yl)phenylamino]methyl}benzoic acid
Figure imgf000287_0004
HPLC-MS (Method D): m/z: 286 (M+1); Rt = = 3,47 min
Example 684 (General procedure (H)) (4-{[4-(2H-Tetrazol-5-yl)phenylamino]methyl}phenoxy)acetic acid
Figure imgf000288_0001
HPLC-MS (Method D): m/z: 326 (M+1); Rt = 3,40 min.
Example 685 (General procedure (H)) (4-Benzyloxybenzyl)-[4-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000288_0002
HPLC-MS (Method D): m/z: 358 (M+1); Rt = 5,14 min.
Example 686 (General procedure (H)) 3-(4-{[4-(2H-Tetrazol-5-yl)phenylamino]methyl}phenyl)acrylic acid
Figure imgf000288_0003
HPLC-MS (Method D): m/z: 322 (M+1); Rt = 3,66 min.
Example 687 (General procedure (H)) Dimethyl-(4-{[4-(2H-tetrazol-5-yl)phenylamino]methyl}naphthalen-1 -yl)amine
Figure imgf000288_0004
HPLC-MS (Method D): m/z: 345 (M+1); Rt = 3,10 min.
Example 688 (General procedure (H)) (4'-Methoxybiphenyl-4-ylmethyl)-[4-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000288_0005
HPLC-MS (Method D): m/z: 358 (M+1); Rt = 5,04 min. Example 689 (General procedure (H)) (2'-Chlorobiphenyl-4-ylmethyl)-[4-(2H-tetrazol-5-yl)-phenyl]-amine
Figure imgf000289_0001
HPLC-MS (Method D): m/z: 362 (M+1); Rt = 5,30 min.
General procedure (I) for preparation of compounds of general formula l8:
HOAt
Figure imgf000289_0002
wherein K, M and T are as defined above.
This procedure is very similar to general procedure (A), the only difference being the carboxylic acid is containing a tetrazole moiety. When the acylation is complete, the product is isolated by extraction, filtration, chromatography or other methods known to those skilled in the art.
The general procedure (I) is further illustrated in the following example 690:
Example 690 (General procedure (I)) 4-[4-(2H-Tetrazol-5-yl)benzoylamino]benzoic acid
Figure imgf000289_0003
To a solution of 4-(2H-tetrazol-5-yl)benzoic acid (example 605, 4 mmol) and HOAt (4.2 mmol) in DMF (6 mL) was added 1-ethyl-3-(3'-dimethylaminopropyl)carbodiimide hydrochloride (4.2 mmol) and the resulting mixture was stirred at room temperature for 1 hour. An alquot of this HOAt-ester solution (0.45 mL) was mixed with 0.25 mL of a solution of 4- aminobenzoic acid (1.2 mmol in 1 mL DMF). (Anilines as hydrochlorides can also be utilised, a slight excess of triethylamine was added to the hydrochloride suspension in DMF prior to mixing with the HOAt-ester.) The resulting mixture was shaken for 3 days at room temperature. 1 N hydrochloric acid (2 mL) was added and the mixture was shaken for 16 hours at room temperature. The solid was isolated by centrifugation (alternatively by filtration or ex- traction) and was washed with water (3 mL). Drying in vacuo at 40 °C for 2 days afforded the title compound.
HPLC-MS (Method D): m/z: 310 (M+1); Rt = 2.83 min.
Example 691 (General procedure (I))
3-[4-(2H-Tetrazol-5-yl)benzoylamino]benzoic acid
Figure imgf000290_0001
HPLC-MS (Method D): m/z: 310 (M+1); Rt = 2.89 min.
Example 692 (General procedure (I))
3-{4-[4-(2H-Tetrazol-5-yl)benzoylamino]phenyl}acrylic acid
Figure imgf000290_0002
HPLC-MS (Method D): m/z: 336 (M+1); Rt = 3.10 min.
Example 693 (General procedure (I))
3-{4-[4-(2H-Tetrazol-5-yl)benzoylamino]phenyl}propionic acid
Figure imgf000290_0003
HPLC-MS (Method D): m/z: 338 (M+1); Rt = 2.97 min.
Example 694 (General procedure (I))
3-Methoxy-4-[4-(2H-tetrazol-5-yl)benzoylamino]benzoic acid
Figure imgf000291_0001
HPLC-MS (Method D): m/z: 340 (M+1); Rt = 3.03 min.
Example 695 (General procedure (I)) rv-(4-Benzyloxyphenyl)-4-(2H-tetrazol-5-yl)benzamide
Figure imgf000291_0002
HPLC-MS (Method D): m/z: 372 (M+1); Rt = 4.47 min.
Example 696 (General procedure (I)) Λ/-(4-Phenoxyphenyl)-4-(2H-tetrazol-5-yl)benzamide
Figure imgf000291_0003
HPLC-MS (Method D): m/z: 358 (M+1); Rt = 4.50 min.
Example 697 (General procedure (I)) Λ-(9H-Fluoren-2-yl)-4-(2H-tetrazol-5-yl)benzamide
Figure imgf000291_0004
HPLC-MS (Method D): m/z: 354 (M+1); Rt = 4.60 min.
Example 698 (General procedure (I)) Λ/-(9-Ethyl-9H-carbazol-2-yl)-4-(2H-tetrazol-5-yl)benzamide
Figure imgf000292_0001
HPLC-MS (Method D): m/z: 383 (M+1); Rt = 4.60 min.
Example 699 (General procedure (I)) Λ/-Phenyl-4-(2H-tetrazol-5-yl)benzamide
Figure imgf000292_0002
HPLC-MS (Method D): m/z: 266 (M+1); Rt = 3.23 min.
Example 700 (General procedure (I)) 4-[4-(2H-Tetrazol-5-ylmethoxy)benzoylamino]benzoic acid
Figure imgf000292_0003
The starting material was prepared as described in example 592. HPLC-MS (Method D): m/z: 340 (M+1); Rt = 2.83 min.
Example 701 (General procedure (I))
3-[4-(2H-Tetrazol-5-ylmethoxy)benzoylamino]benzoic acid
Figure imgf000292_0004
HPLC-MS (Method D): m/z: 340 (M+1); Rt = 2.90 min.
Example 702 (General procedure (I))
3-{4-[4-(2H-Tetrazol-5-ylmethoxy)benzoylamino]phenyl}acrylic acid
Figure imgf000293_0001
HPLC-MS (Method D): m/z: 366 (M+1); Rt= 3.07 min.
Example 703 (General procedure (I)) 3-{4-[4-(2H-Tetrazol-5-ylmethoxy)benzoylamino]phenyl}propionic acid
Figure imgf000293_0002
HPLC-MS (Method D): m/z: 368 (M+1); Rt = 2.97 min.
Example 704 (General procedure (I)) 3-Methoxy-4-[4-(2H-tetrazol-5-ylmethoxy)benzoylamino]benzoic acid
Figure imgf000293_0003
HPLC-MS (Method D): m/z: 370 (M+1); Rt = 3.07 min.
Example 705 (General procedure (I)) Λ/-(4-Benzyloxyphenyl)-4-(2H-tetrazol-5-ylmethoxy)benzamide
Figure imgf000293_0004
HPLC-MS (Method D): m/z: 402 (M+1); Rt = 4.43 min.
Example 706 (General procedure (I)) Λ/-(4-Phenoxyphenyl)-4-(2H-tetrazol-5-ylmethoxy)benzamide
Figure imgf000293_0005
HPLC-MS (Method D): m/z: 388 (M+1); Rt = 4.50 min.
Example 707 (General procedure (I)) Λ/-(9H-Fluoren-2-yl)-4-(2H-tetrazol-5-ylmethoxy)benzamide
Figure imgf000294_0001
HPLC-MS (Method D): m/z: 384 (M+1); Rt = 4.57 min.
Example 708 (General procedure (I)) Λ/-(9-Ethyl-9H-carbazol-2-yl)-4-(2H-tetrazol-5-ylmethoxy)benzamide
Figure imgf000294_0002
HPLC-MS (Method D): m/z: 413 (M+1); Rt = 4.57 min.
Example 709 (General procedure (I)) Λ/-Phenyl-4-(2H-tetrazol-5-ylmethoxy)benzamide
Figure imgf000294_0003
HPLC-MS (Method D): m/z: 296 (M+1); Rt = 3.23 min.
Example 710 (General procedure (I))
4-[4-(2H-Tetrazol-5-ylmethylsulfanyl)benzoylamino]benzoic acid
Figure imgf000294_0004
The starting material was prepared as described in example 593. HPLC-MS (Method D): m/z: 356 (M+1); Rt = 2.93 min. Example 711 (General procedure (I))
3-[4-(2H-Tetrazol-5-ylmethylsulfanyl)benzoylamino]benzoic acid
Figure imgf000295_0001
HPLC-MS (Method D): m/z: 356 (M+1); Rt = 3.00 min.
Example 712 (General procedure (I))
3-{4-[4-(2H-Tetrazol-5-ylmethylsulfanyl)benzoylamino]phenyl}acrylic acid
Figure imgf000295_0002
HPLC-MS (Method D): m/z: 382 (M+1); Rt = 3.26 min.
Example 713 (General procedure (I)) 3-{4-[4-(2H-Tetrazol-5-ylmethylsulfanyl)benzoylamino]phenyl}propionic acid
Figure imgf000295_0003
HPLC-MS (Method D): m/z: 384 (M+1); Rt = 3.10 min.
Example 714 (General procedure (I)) 3-Methoxy-4-[4-(2H-tetrazol-5-ylmethylsulfanyl)benzoylamino]benzoic acid
Figure imgf000295_0004
HPLC-MS (Method D): m/z: 386 (M+1); Rt = 3.20 min.
Example 715 (General procedure (I)) Λ/-(4-Benzyloxyphenyl)-4-(2H-tetrazol-5-ylmethylsulfanyl)benzamide
Figure imgf000296_0001
HPLC-MS (Method D): m/z: 418 (M+1); Rt = 4.57 min.
Example 716 (General procedure (I)) Λ/-(4-Phenoxyphenyl)-4-(2H-tetrazol-5-ylmethylsulfanyl)benzamide
Figure imgf000296_0002
HPLC-MS (Method D): m/z: 404 (M+1); Rt = 4.60 min.
Example 717 (General procedure (I)) Λ-(9H-Fluoren-2-yl)-4-(2H-tetrazol-5-ylmethylsulfanyl)benzamide
Figure imgf000296_0003
HPLC-MS (Method D): m/z: 400 (M+1); Rt = 4.67 min.
Example 718 (General procedure (I)) Λ/-(9-Ethyl-9H-carbazol-2-yl)-4-(2H-tetrazol-5-ylmethylsulfanyl)benzamide
Figure imgf000296_0004
HPLC-MS (Method D): m/z: 429 (M+1); Rt = 4.67 min.
Example 719 (General procedure (I)) Λ/-Phenyl-4-(2H-tetrazol-5-ylmethylsulfanyl)benzamide
Figure imgf000296_0005
HPLC-MS (Method D): m/z: 312 (M+1); Rt = 3.40 min.
General procedure (J) for solution phase preparation of amides of general formula lg:
Figure imgf000297_0001
wherein T is as defined above.
This general procedure (J) is further illustrated in the following example.
Example 720 (General procedure (J)). 9-(3-Chlorobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000297_0002
3-(2H-Tetrazol-5-yl)-9H-carbazole (example 594, 17 g, 72.26 mmol) was dissolved in N,N- dimethylformamide (150 mL). Triphenylmethyl chloride (21.153 g, 75.88 mmol) and triethylamine (20.14 mL, 14.62 g, 144.50 mmol) were added consecutively. The reaction mixture was stirred for 18 hours at room temperature, poured into water (1.5 L) and stirred for an additional 1 hour. The crude product was filtered off and dissolved in dichloromethane (500 mL). The organic phase was washed with water (2 x 250 mL) and dried with magnesium sulfate (1 h). Filtration followed by concentration yielded a solid which was triturated in heptanes (200 mL). Filtration furnished 3-[2-(triphenylmethyl)-2H-tetrazol-5-yl]-9H-carbazole (31.5 g) which was used without further purification.
1H-NMR (CDCI3): £8.87 (1H, d), 8.28 (1H, bs), 8.22 (1H, dd), 8.13 (1H, d), 7.49 (1H, d), 7.47- 7.19 (18H, m); HPLC-MS (Method C): m/z: 243 (triphenylmethyl); Rt = 5.72 min. 3-[2-(Triphenylmethyl)-2H-tetrazol-5-yl]-9H-carbazole (200 mg, 0.42 mmol) was dissolved in methyl sulfoxide (1.5 mL). Sodium hydride (34 mg, 60 %, 0.85 mmol) was added, and the resulting suspension was stirred for 30 min at room temperature. 3-Chlorobenzyl chloride (85 μL, 108 mg, 0.67 mmol) was added, and the stirring was continued at 40 °C for 18 hours. The reaction mixture was cooled to ambient temperature and poured into 0.1 N hydrochloric acid (aq.) (15 mL). The precipitated solid was filtered off and washed with water (3 x 10 rnL) to furnish 9-(3-chlorobenzyl)-3-[2-(triphenylmethyl)-2H-tetrazol-5-yl]-9H-carbazole, which was dissolved in a mixture of tetrahydrofuran and 6 N hydrochloric acid (aq.) (9:1 ) (10 mL) and stirred at room temperature for 18 hours. The reaction mixture was poured into water (1O0 mL). The solid was filtered off and rinsed with water (3 x 10 mL) and dichloromethane (3 x 10 mL) to yield the title compound (127 mg). No further purification was necessary. 1H-NMR (DMSO-d6): £8.89 (1H, d), 8.29 (1 H, d), 8.12 (1H, dd), 7.90 (1H, d), 7.72 (1 H, d ), 7.53 (1H, t), 7.36-7.27 (4H, m), 7.08 (1 H, bt), 5.78 (2H, s); HPLC-MS (Method B): m/z: 360 (M+1); Rt = 5.07 min.
The compounds in the following examples were prepared in a similar fashion. Optionally, the compounds can be further purified by recrystallization from e.g. aqueous sodium hydroxide (1 N) or by chromatography.
Example 721 (General Procedure (J)).
9-(4-Chlorobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000298_0001
HPLC-MS (Method C): m/z: 360 (M+1); Rt = 4.31 min.
Example 722 (General Procedure (J)).
9-(4-Methylbenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000298_0002
HPLC-MS (Method C): m/z: 340 (M+1); Rt = 4.26 min.
Example 723 (General Procedure (J)). 3-(2H-Tetrazol-5-yl)-9-(4-trifluoromethylbenzyl)-9H-carbazole
Figure imgf000299_0001
HPLC-MS (Method C): m/z: 394 (M+1 ); Rt = 4.40 min.
Example 724 (General Procedure (J)). 9-(4-Benzyloxybenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000299_0002
HPLC-MS (Method C): m/z: 432 (M+1); Rt = 4.70 min.
Example 725 (General Procedure (J)). 9-(3-Methylbenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000299_0003
,
HPLC-MS (Method C): m/z: 340 (M+1 ); Rt = 4.25 min.
Example 726 (General Procedure (J)). 9-Benzyl-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000300_0001
1H-NMR (DMSO-de): £8.91 (1H, dd), 8.30 (1H, d), 8.13 (1 H, dd), 7.90 (1H, d), 7.73 (1H, d), 7.53 (1 H, t), 7.36-7.20 (6H, m), 5.77 (2H, s).
Example 727 (General Procedure (J)). 9-(4-Phenylbenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000300_0002
1H-NMR (DMSO-d6): £8.94 (1 H, s), 8.33 (1 H, d), 8.17 (1 H, dd), 7.95 (1 H, d), 7.77 (1H, d), 7.61-7.27 (11H, m), 5.82 (2H, s).
Example 728 (General Procedure (J)). 9-(3-Methoxybenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000300_0003
HPLC-MS (Method C): m/z: 356 (M+1); Rt = 3.99 min.
Example 729 (General Procedure (J)). 9-(Naphthalen-2-ylmethyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000301_0001
HPLC-MS (Method C): m/z: 376 (M+1); Rt = 4.48 min.
Example 730 (General Procedure (J)). 9-(3-Bromobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000301_0002
HPLC-MS (Method C): m/z: 404 (M+1); Rt = 4.33 min.
Example 731 (General Procedure (J)). 9-(Biphenyl-2-ylmethyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000301_0003
HPLC-MS (Method C): m/z: 402 (M+1); Rt = 4.80 min.
Example 732 (General Procedure (J)). 3-(2H-Tetrazol-5-yl)-9-[4-(1 ,2,3-thiadiazol-4-yl)benzyl]-9H-carbazole
Figure imgf000302_0001
Example 733 (General Procedure (J)).
9-(2'-Cyanobiphenyl-4-ylmethyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000302_0002
1H-NMR (DMSO-de): £8.91 (1H, d), 8.31 (1H, d), 8.13 (1H, dd), 7.95 (1 H, d), 7.92 (1H, d), 7.78 (1H, d), 7.75 (1H, dt), 7.60-7.47 (5H, m), 7.38-7.28 (3H, m), 5.86 (2H, s); HPLC-MS (Method C): m/z: 427 (M+1 ); Rt = 4.38 min.
Example 734 (General Procedure (J)).
9-(4-lodobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000302_0003
HPLC-MS (Method C): m/z: 452 (M+1); Rt = 4.37 min.
Example 735 (General Procedure (J)).
9-(3,5-Bis(trifluoromethyl)benzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000303_0001
HPLC-MS (Method C): m/z: 462 (M+1); Rt = 4.70 min.
Example 736 (General Procedure (J)). 9-(4-Bromobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000303_0002
1H-NMR (DMSO-d6): £8.89 (1H, d), 8.29 (1 H, d), 8.11 (1H, dd), 7.88 (1 H, d), 7.70 (1H, d), 7.52 (1 H, t), 7.49 (2H, d), 7.31 (1 H, t), 7.14 (2H, d), 5.74 (2H, s); HPLC-MS (Method C): m/z: 404 (M+1); Rt = 4.40 min.
Example 737 (General Procedure (J)). 9-(Anthracen-9-ylmethyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000303_0003
HPLC-MS (Method C): m/z: 426 (M+1); Rt = 4.78 min.
Example 738 (General Procedure (J)). 9-(4-Carboxybenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000304_0001
3.6 fold excess sodium hydride was used.
1H-NMR (DMSO-de): £ 12.89 (1H, bs), 8.89 (1H, d), 8.30 (1H, d), 8.10 (1H, dd), 7.87 (1H, d), 7.86 (2H, d), 7.68 (1H, d), 7.51 (1H, t), 7.32 (1 H, t), 7.27 (2H, d), 5.84 (2H, s); HPLC-MS (Method C): m/z: 370 (M+1 ); Rt = 3.37 min.
Alternative mode of preparation of 9-(4-Carboxybenzyl)-3-(2H-tetrazol-5-yl)-9H- carbazole:
Carbazole (52.26 g, 0.30 mol) was dissolved in dichloromethane (3 L) and silicagel (60 mesh, 600 g) was added to the mixture and the mixture was cooled to 10 °C. A mixture of Λ/-bromosuccinimide (NBS, 55 g, 0.30 mol) in dichloromethane (400 mL) was added at 10 °C. After addition, the mixture was allowed to reach room temperature. After standing for 42 hours, the mixture was filtered, and the solid was washed with dichloromethane (4 x 200 mL), the combined filtrates were washed with water (300 mL) and dried over Na2S04 . Evaporation in vacuo to dryness afforded 77 g of crude product. Recrystallization from 2-propanol (800 mL) afforded 71 % 3-bromocarbazole.
To a stirred solution of 3-bromocarbazole (63 g, 0.256 mol) in Λ/-methylpyrrolidone
(900 mL) was added cuprous cyanide (CuCN, 25.22 g, 0.28 mol) and the mixture was heated to 190 °C. After 9 hours of heating, the mixture was cooled to room temperature. The mixture was concentrated by bulb-to-bulb distillation (100 °C, 0.1 mm Hg). The residue was treated with NH4OH (25%, 300 mL) and subsequently extracted with ethyl acetate (10%) in toluene. The organic layer was dried over Na2S04 and concentrated by bulb-to-bulb distillation (100 °C, 0.1 mm Hg) to give 34 g (70%) of 3-cyanocarbazole.
Sodium hydride 55-60% in mineral oil (3.7 g, 0.093 mol) was added in portions to a stirred, cooled (5 °C) mixture of 3-cyanocarbazole (17.5 g, 0.091 mol) in N,N- dimethylformamide (200 mL). After 0.5 hours, a solution of methyl 4-bromomethylbenzoate (22.9 g, 100 mmol) in Λ/,Λ/-dimethylformamide (80 mL) was added dropwise to the cooled mixture. The mixture was subsequently slowly warmed to room temperature and stirred overnight. The mixture was poured into ice water and extracted with dichloromethane (2 x 200 mL), the organic layer was washed several times with water, dried over Na2S04 and concentrated in vacuo. A mixture of ethyl acetate and heptane (1/1 , 50 mL) was added to the concentrate and the solid was product filtered off. Yield 24 g (78%) of 4-(3-cyanocarbazol-9- ylmethyl)benzoic acid methyl ester.
Sodium azide (7.8 g, 0.12 mol) and ammonium chloride (6.42 g , 0.12 mol) were added to a stirred mixture of 4-(3-cyanocarbazol-9-ylmethyl)benzoic acid methyl ester (24.8 g, 0.073 mol) in Λ/,Λ/-dimethylformamide (130 mL) and the mixture was heated to 110 °C. After 48 hours, the mixture was cooled to room temperature and poured into water (500 mL) and cooled to 5 °C. Hydrochloric acid (10 N) was then added to pH = 2. After stirring for 1 hour at 5 °C the precipitate was filtered off and washed with water. The solid obtained was air dried. Yield 27.9 g of 4-[3-(1H-tetrazol-5-yl)carbazol-9-ylmethyl]benzoic acid methyl ester.
31.1 g of 4-[3-(1H-tetrazol-5-yl)carbazol-9-ylmethyl]benzoic acid methyl ester was added to a solution of sodium hydroxide (8.76 g, 0.219 mol) in water (150 mL) and the mixture was heated to 80 °C, after 0.5 h activated carbon (0.5 g) was added and the mixture was filtered through celite. The filtrate was treated with hydrochloric acid (10 N) to pH = 1 and the formed precipitate was filtered off and air dried. This procedure was repeated as the first treatment did not give complete hydrolysis of the ester. Finally the product was dissolved in 2-propanol, the filtered the mother liquor was concentrated to approximately 100 mL and the product was isolated by filtration to afford 19 g of the title compound. After evaporation of the mother liquor to dryness and re-treatment with 2-propanol further 8 g of product was isolated resulting in a yield of 90 %.
Example 739 (General Procedure (J)). 9-(2-Chlorobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000305_0001
HPLC-MS (Method B): m/z: 360 (M+1 ); Rt = 5.30 min. Example 740 (General Procedure (J)). 9-(4-Fluorobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000306_0001
1H-NMR (DMSO-de): £8.88 (1 H, d), 8.28 (1 H, d), 8.10 (1 H, dd), 7.89 (1 H, d), 7.72 (1H, d), 7.52 (1H, t), 7.31 (1 H, t), 7.31-7.08 (4H, m), 5.74 (2H, s); HPLC-MS (Method C): m/z: 344 (M+1); Rt = 4.10 min.
Example 741 (General Procedure (J)). 9-(3-Fluorobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000306_0002
1H-NMR (DMSO-de): £8.89 (1 H, d), 8.29 (1 H, d), 8.12 (1 H, dd), 7.90 (1 H, d), 7.72 (1 H, d), 7.53 (1 H, t), 7.37-7.27 (2H, m), 7.12-7.02 (2H, m), 6.97 (1 H, d), 5.78 (2H, s); HPLC-MS (Method C): m/z: 344 (M+1); Rt = 4.10 min.
Example 742 (General Procedure (J)). 9-(2-lodobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000306_0003
HPLC-MS (Method C): m/z: 452 (M+1); Rt = 4.58 min.
Example 743 (General Procedure (J)). 9-(3-Carboxybenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000307_0001
3.6 fold excess sodium hydride was used.
1H-NMR (DMSO-d6): £ 12.97 (1 H, bs), 8.90 (1 H, bs), 8.30 (1 H, d), 8.12 (1H, bd), 7.89 (1H, d), 7.82 (1 H, m), 7.77 (1 H, bs), 7.71 (1 H, d), 7.53 (1 H, t), 7.46-7.41 (2H, m), 7.32 (1 H, t), 5.84 (2H, s); HPLC-MS (Method C): m/z: 370 (M+1); Rt = 3.35 min.
Example 744 (General Procedure (J)). 9-[4-(2-Propyl)benzyl]-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000307_0002
1H-NMR (DMSO-de): £8.87 (1H, d), 8.27 (1H, d), 8.10 (1 H, dd), 7.87 (1H, d), 7.71 (1 H, d), 7.51 (1H, t), 7.31 (1H, t), 7.15 (2H, d), 7.12 (2H, d), 5.69 (2H, s), 2.80 (1 H, sept), 1.12 (6H, d); HPLC-MS (Method C): m/z: 368 (M+1); Rt = 4.73 min.
Example 745 (General Procedure (J)).
9-(3,5-Dimethoxybenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000307_0003
HPLC-MS (Method C): m/z: 386 (M+1); Rt = 4.03 min. Example 746 (General Procedure (J)). 3-(2H-Tetrazol-5-yl)-9-(2,4,5-trifluorobenzyl)-9H-carbazole
Figure imgf000308_0001
HPLC-MS (Method B): m/z: 380 (M+1); Rt = 5.00 min.
Example 747 (General Procedure (J)). Λ/-Methyl-Λ/-phenyl-2-[3-(2H-tetrazol-5-yl)carbazol-9-yl]acetamide
Figure imgf000308_0002
HPLC-MS (Method B): m/z: 383 (M+1); Rt = 4.30 min.
Example 748 (General Procedure (J)). 9-(4-Methoxybenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000308_0003
1H-NMR (DMSO-de): £8.86 (1 H, d), 8.26 (1H, d), 8.10 (1 H, dd), 7.90 (1 H, d), 7.73 (1 H, d), 7.51 (1H, t), 7.30 (1 H, t), 7.18 (2H, d), 6.84 (2H, d), 5.66 (2H, s), 3.67 (3H, s); HPLC-MS (Method B): m/z: 356 (M+1); Rt = 4.73 min.
Example 749 (General Procedure (J)). 9-(2-Methoxybenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000309_0001
1H-NMR (DMSO-de): £8.87 (1 H, d), 8.27 (1 H, d), 8.09 (1 H, dd), 7.77 (1H, d), 7.60 (1 H, d), 7.49 (1 H, t), 7.29 (1 H, t), 7.23 (1H, bt), 7.07 (1H, bd), 6.74 (1 H, bt), 6.61 (1H, bd), 5.65 (2H, s), 3.88 (3H, s); HPLC-MS (Method B): m/z: 356 (M+1); Rt = 4.97 min.
Example 750 (General Procedure (J)). 9-(4-Cyanobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000309_0002
HPLC-MS (Method C): m/z: 351 (M+1 ); Rt = 3.74 min.
Example 751 (General Procedure (J)). 9-(3-Cyanobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000309_0003
HPLC-MS (Method C): m/z: 351 (M+1 ); Rt = 3.73 min.
Example 752 (General Procedure (J)). 9-(5-Chloro-2-methoxybenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000310_0001
1H-NMR (DMSO-de): £8.87 (1H, d), 8.35 (1H, d), 8.10 (1H, dd), 7.73 (1H, d), 7.59 (1H, d), 7.49 (1H, t), 7.29 (1H, t), 7.27 (1H, dd), 7.11 (1H, d), 6.51 (1H, d), 5.63 (2H, s), 3.88 (3H, s); HPLC-MS (Method C): m/z: 390 (M+1); Rt = 4.37 min.
Example 753 (General Procedure (J)). Λ/-Phenyl-2-[3-(2H-tetrazol-5-yl)carbazol-9-yl]acetamide
Figure imgf000310_0002
1H-NMR (DMSO-de): £10.54 (1H, s), 8.87 (1H, bs), 8.27 (1H, d), 8.12 (1H, bd), 7.83 (1H, d), 7.66 (1H, d), 7.61 (2H, d), 7.53 (1H,t), 7.32 (1H, t), 7.32 (2H, t), 7.07 (1H, t), 5.36 (2H, s); HPLC-MS (Method C): m/z: 369 (M+1); Rt = 3.44 min.
Example 754 (General Procedure (J)). Λ/-Butyl-2-[3-(2H-tetrazol-5-yl)carbazol-9-yl]acetamide
Figure imgf000310_0003
1H-NMR (DMSO-de): £8.85 (1H, d), 8.31 (1H, t), 8.25 (1H, d), 8.10 (1H, dd), 7.75 (1H, d), 7.58 (1H, d), 7.52 (1H, t), 7.30 (1H, t), 5.09 (2H, s), 3.11 (2H, q), 1.42 (2H, quint), 1.30 (2H, sext), 0.87 (3H, t); HPLC-MS (Method C): m/z: 349 (M+1); Rt = 3.20 min. Example 755 (General Procedure (J)). 9-(2,4-Dichlorobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000311_0001
1H-NMR (DMSO-d6): £8.92 (1 H, d), 8.32 (1 H, d), 8.09 (1 H, dd), 7.76 (1H, d), 7.74 (1 H, d), 7.58 (1 H, d), 7.51 (1 H, t), 7.33 (1 H, t), 7.23 (1 H, dd), 6.42 (1 H, d), 5.80 (2H, s); HPLC-MS (Method B): m/z: 394 (M+1); Rt = 5.87 min.
Example 756 (General Procedure (J)). 9-(2-Methylbenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000311_0002
1H-NMR (DMSO-de): £8.92 (1H, d), 8.32 (1H, d), 8.08 (1H, dd), 7.72 (1H, d), 7.55 (1H, d), 7.48 (1 H, t), 7.32 (1 H, t), 7.26 (1 H, d), 7.12 (1 H, t), 6.92 (1 H, t), 6.17 (1H, d), 5.73 (2H, s), 2.46 (3H, s); HPLC-MS (Method B): m/z: 340 (M+1); Rt = 5.30 min.
Example 757 (General Procedure (J)).
9-(3-Nitrobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000311_0003
HPLC-MS (Method C): m/z: 371 (M+1); Rt = 3.78 min.
Example 758 (General Procedure (J)).
9-(3,4-Dichlorobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000312_0001
HPLC-MS (Method B): m/z: 394 (M+1); Rt = 5.62 min.
Example 759 (General Procedure (J)). 9-(2,4-Difluorobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000312_0002
1H-NMR (DMSO-de): £8.89 (1H, d), 8.29 (1 H, d), 8.11 (1 H, dd), 7.88 (1H, d), 7.69 (1 H, d), 7.52 (1 H, t), 7.36-7.24 (2H, m), 7.06-6.91 (2H, m), 5.78 (2H, s); HPLC-MS (Method B): m/z: 362 (M+1 ); Rt = 5.17 min.
Example 760 (General Procedure (J)). 9-(3,5-Difluorobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000312_0003
1H-NMR (DMSO-de): £8.90 (1H, bs), 8.31 (1H, d), 8.13 (1H, bd), 7.90 (1H, d), 7.73 (1H, d), 7.54 (1 H, t), 7.34 (1H, t), 7.14 (1 H, t), 6.87 (2H, bd), 5.80 (2H, s); HPLC-MS (Method B): m/z: 362 (M+1); Rt = 5.17 min.
Example 761 (General Procedure (J)). 9-(3,4-Difluorobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000313_0001
1H-NMR (DMSO-de): £8.89 (1 H, bs), 8.29 (1 H, d), 8.12 (1H, bd), 7.92 (1H, d), 7.74 (1H, d), 7.54 (1 H, t), 7.42-7.25 (3H, m), 6.97 (1 H, bm), 5.75 (2H, s); HPLC-MS (Method B): m/z: 362 (M+1); Rt = 5.17 min.
Example 762 (General Procedure (J)). 9-(3-lodobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000313_0002
HPLC-MS (Method B): m/z: 452 (M+1 ); Rt = 5.50 min.
Example 763 (General Procedure (J)). 3-(2H-Tetrazol-5-yl)-9-[3-(trifluoromethyl)benzyl]-9H-carbazole
Figure imgf000313_0003
1H-NMR (DMSO-d6): £8.89 (1 H, d), 8.30 (1 H, d), 8.11 (1H, dd), 7.90 (1 H, d), 7.72 (1H, d), 7.67 (1H, bs), 7.62 (1 H, bd), 7.53 (1 H, t), 7.50 (1H, bt), 7.33 (1 H, bd), 7.32 (1 H, t), 5.87 (2H, s); HPLC-MS (Method B): m/z: 394 (M+1 ); Rt = 5.40 min.
Example 764 (General Procedure (J)). Λ/-(4-Carboxyphenyl)-2-[3-(2H-tetrazol-5-yl)carbazol-9-yl]acetamide
Figure imgf000314_0001
3.6 fold excess sodium hydride was used.
HPLC-MS (Method B): m/z: 413 (M+1 ); Rt = 3.92 min.
Example 765 (General Procedure (J)). Λ/-(2-Propyl)-2-[3-(2H-tetrazol-5-yl)carbazol-9-yl]acetamide
Figure imgf000314_0002
HPLC-MS (Method B): m/z: 335 (M+1 ); Rt = 3.70 min.
Example 766 (General Procedure (J)). Λ/-Benzyl-Λ/-phenyl-2-[3-(2H-tetrazol-5-yl)carbazol-9-yl]acetamide
Figure imgf000314_0003
HPLC-MS (Method B): m/z: 459 (M+1 ); Rt = 5.37 min.
Example 767 (General Procedure (J)). Λ/-[4-(2-Methyl-2-propyl)phenyl]-2-[3-(2H-tetrazol-5-yl)carbazol-9-yl]acetamide
Figure imgf000315_0001
HPLC-MS (Method B): m/z: 425 (M+1); Rt = 5.35 min.
Example 768 (General Procedure (J)). Λ/-Phenethyl-2-[3-(2H-tetrazol-5-yl)carbazol-9-yl]acetamide
Figure imgf000315_0002
HPLC-MS (Method C): m/z: 397 (M+1); Rt = 3.43 min.
Example 769 (General Procedure (J)). 3-(2H-Tetrazol-5-yl)-9-[2-(trifluoromethyl)benzyl]-9H-carbazole
Figure imgf000315_0003
HPLC-MS (Method C): m/z: 394 (M+1); Rt = 4.44 min.
Example 770 (General Procedure (J)). 9-[2-Fluoro-6-(trifluoromethyl)benzyl]-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000316_0001
HPLC-MS (Method C): m/z: 412 (M+1); Rt = 4.21 min.
Example 771 (General Procedure (J)). 9-[2,4-Bis(trifluoromethyl)benzyl)]-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000316_0002
HPLC-MS (Method C): m/z: 462 (M+1 ); Rt = 4.82 min.
Example 772 (General Procedure (J)). 3-(2H-Tetrazol-5-yl)-9-(2,4,6-trimethylbenzyl)-9H-carbazole
Figure imgf000316_0003
HPLC-MS (Method C): m/z: 368 (M+1); Rt = 4.59 min.
Example 773 (General Procedure (J)). 9-(2,3,5,6-Tetramethylbenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000316_0004
HPLC-MS (Method C): m/z: 382 (M+1); Rt = 4.47 min.
Example 774 (General Procedure (J)). 9-[(Naphthalen-1-yl)methyl]-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000317_0001
HPLC-MS (Method C): m/z: 376 (M+1); Rt = 4.43 min.
Example 775 (General Procedure (J)). 9-[Bis(4-fluorophenyl)methyl]-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000317_0002
HPLC-MS (Method C): m/z: 438 (M+1); Rt = 4.60 min.
Example 776 (General Procedure (J)). 9-(2-Bromobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000317_0003
HPLC-MS (Method C): m/z: 404 (M+1 ); Rt = 4.50 min.
Example 777 (General Procedure (J)). 9-(2-Fluorobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000318_0001
HPLC-MS (Method C): m/z: 344 (M+1 ); Rt = 4.09 min.
Example 778 (General Procedure (J)). 9-(4-Carboxy-2-methylbenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000318_0002
In this preparation, a 3.6-fold excess of sodium hydride was used. HPLC-MS (Method C): m/z: 384 (M+1 ); Rt = 3.56 min.
Example 779 (General Procedure (J)). 9-(2-Phenylethyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
HPLC-MS (Method C): m/z: 340 (M+1); Rt = 4.08 min.
Example 780 (General Procedure (J)).
9-[2-Fluoro-5-(trifluoromethyl)benzyl]-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000319_0001
HPLC-MS (Method C): m/z: 412 (M+1); Rt = 4.34 min.
Example 781 (General Procedure (J)). 9-(4-Carboxy-2-fluorobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000319_0002
3-Fluoro-4-methylbenzoic acid (3.0 g, 19.5 mmol) and benzoyl peroxide (0.18 g, 0.74 mmol) were suspended in benzene. The mixture was purged with N2 and heated to reflux. N- Bromosuccinimide (3.47 g, 19.5 mmol) was added portionwise, and reflux was maintained for 18 hours. The reaction mixture was concentrated, and the residue was washed with water (20 mL) at 70 °C for 1 hour. The crude product was isolated by filtration and washed with additional water (2 x 10 mL). The dry product was recrystallized from heptanes. Filtration furnished 4-bromomethyl-3-fluorobenzoic acid (1.92 g) which was used in the following step according to General Procedure (J). In this preparation, a 3.6-fold excess of sodium hydride was used. HPLC-MS (Method C): m/z: 388 (M+1); Rt = 3.49 min.
Example 782 (General Procedure (J)). 5-{4-[[(3-(2H-Tetrazol-5-yl)carbazol-9-yl)methyl]naphthalen-1-yl]oxy}pentanoic Acid
Figure imgf000319_0003
5-[(4-Formylnaphthalen-1-yl)oxy]pentanoic acid intermediate obtained in example 470(3.0 g, 11.0 mmol) was dissolved in a mixture of methanol and tetrahydrofuran (9:1) (100 mL), and sodium borohydride (1.67 g, 44.1 mmol) was added portionwise at ambient temperature. After 30 minutes, the reaction mixture was concentrated to 50 mL and added to hydrochloric acid (0.1 N, 500 mL). Additional hydrochloric acid (1 N, 40 mL) was added, and 5-[(4- hydroxymethyl-naphthalen-1-yl)oxy]pentanoic acid (2.90 g) was collected by filtration. To the crude product was added concentrated hydrochloric acid (100 mL), and the suspension was stirred vigorously for 48 hours at room temperature. The crude product was filtered off and washed with water, until the pH was essentially neutral. The material was washed with heptanes to furnish 5-[(4-chloromethylnaphthalen-1-yl)oxy]pentanoic acid (3.0 g) which was used in the following step according to General Procedure (J).
In this preparation, a 3.6-fold excess of sodium hydride was used. HPLC-MS (Method C): m/z: 492 (M+1); Rt = 4.27 min.
Example 783 (General procedure (J)) 9-(2,3-Difluorobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole fit
Figure imgf000320_0001
HPLC-MS (Method C): m/z= 362 (M+1); Rt = 4.13 min.
Example 784 (General procedure (J)) 9-(2,5-Difluorobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000320_0002
HPLC-MS (Method C): m/z = 362 (M+1); Rt = 4.08 min. Example 785 (General procedure (J)) 9-Pentafluorophenylmethyl-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000321_0001
HPLC-MS (Method C): m/z = 416 (M+1); Rt = 4.32 min.
Example 786 (General procedure (J)) 9-(2,6-Difluorobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000321_0002
HPLC-MS (Method C): m/z = 362 (M+1); Rt = 3.77 min.
Further compounds of the invention that may be prepared according to general procedure (J), and includes:
Figure imgf000322_0001
The following compounds of the invention may be prepared eg. from 9-(4-bromobenzyl)-3- (2H-tetrazol-5-yl)-9H-carbazole (example 736) or from 9-(3-bromobenzyl)-3-(2H-tetrazol-5- yl)-9H-carbazole (example 730) and aryl boronic acids via the Suzuki coupling reaction eg as described in Littke, Dai & Fu J. Am. Chem. Soc, 2000, 122, 4020-8 (or references cited therein), or using the methodology described in general procedure (E), optionally changing the palladium catalyst to bis(tri-.ert-butylphosphine)palladium (0). Example 800 Example 801 Example 802
Figure imgf000323_0001
Example 803 Example 804 Example 805
Figure imgf000323_0002
General procedure (K) for preparation of compounds of general formula l10:
Figure imgf000323_0003
wherein T is as defined above.
The general procedure (K) is further illustrated by the following example: Example 806 (General procedure (K)). 1 -Benzyl-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000323_0004
5-Cyanoindole (1.0 g, 7.0 mmol) was dissolved in Λ/,Λ/-dimethylformamide (14 mL) and cooled in an ice-water bath. Sodium hydride (0.31 g, 60 %, 7.8 mmol) was added, and the resulting suspension was stirred for 30 min. Benzyl chloride (0.85 mL, 0.94 g, 7.4 mmol) was added, and the cooling was discontinued. The stirring was continued for 65 hours at room temperature. Water (150 mL) was added, and the mixture was extracted with ethyl acetate (3 x 25 mL). The combined organic phases were washed with brine (30 mL) and dried with sodium sulfate (1 hour). Filtration and concentration yielded the crude material. Purification by flash chromatography on silica gel eluting with ethyl acetate/heptanes = 1 :3 afforded 1.60 g 1 -benzyl-1 H-indole-5-carbonitrile.
HPLC-MS (Method C): m/z: 233 (M+1); Rt = 4.17 min.
1 -Benzyl-1 H-indole-5-carbonitrile was transformed into 1 -benzyl-5-(2H-tetrazol-5-yl)-1 H- indole by the method described in general procedure (J) and in example 594. Purification was done by flash chromatography on silica gel eluting with dichloromethane/methanol = 9:1.
HPLC-MS (Method C): m/z: 276 (M+1 ); Rt = 3.35 min.
The compounds in the following examples were prepared by the same procedure.
Example 807 (General procedure (K)).
1 -(4-Bromobenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000324_0001
HPLC-MS (Method C): m/z: 354 (M+1); Rt = 3.80 min.
Example 808 (General procedure (K)).
1 -(4-Phenylbenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000324_0002
1H-NMR (200 MHz, DMSO-d6): δ = 5.52 (2H, s), 6.70 (1 H, d), 7.3-7.45 (6H, m), 7.6 (4H, m), 7.7-7.8 (2H, m), 7.85(1 H, dd), 8.35 (1 H, d). Calculated for C22H17N5, H20: 73.32% C; 5.03% H; 19.43% N. Found: 73.81% C; 4.90% H; 19.31% N.
Example 809 4'-[5-(2H-Tetrazol-5-yl)indol-1-ylmethyl]biphenyl-4-carboxylic acid
Figure imgf000325_0001
5-(2H-Tetrazol-5-yl)-1 H-indole (Syncom BV, Groningen, NL) (1.66g, 8.9 mmol) was treated with trityl chloride (2.5 g, 8.9 mmol) and triethyl amine (2.5 mL, 17.9 mmol) in DMF(25 mL) by stirring at RT overnight. The resulting mixture was treated with water. The gel was isolated, dissolved in methanol, treated with activated carbon; filtered and evaporated to dryness in vacuo. This afforded 3.6 g (94%) of crude 5-(2-trityl-2H-tetrazol-5-yl)-1 H-indole.
HPLC-MS (Method C): m/z = 450 (M+23); Rt. = 5.32 min.
4-MethyIphenylbenzoic acid (5 g, 23.5 mmol) was mixed with CCI4 (100 mL) and under an atmosphere of nitrogen, the slurry was added Λ/-Bromosuccinimide (4.19 g, 23.55 mmol) and dibenzoyl peroxide (0.228 g, 0.94 mmol). The mixture was subsequently heated to reflux for 0.5 hour. After cooling, DCM and water (each 30 mL) were added. The resulting precipitate was isolated, washed with water and a small amount of methanol. The solid was dried in vacuo to afford 5.27 g (77%) of 4'-bromomethylbiphenyl-4-carboxylic acid.
HPLC-MS (Method C): m/z = 291 (M+1 ); Rt. = 3.96 min.
5-(2-Trityl-2H-tetrazol-5-yl)-1 H-indole (3.6 g, 8.4 mmol) was dissolved in DMF (100 mL). Under nitrogen, NaH (60 % suspension in mineral oil, 34 mmol) was added slowly. 4'- Bromomethylbiphenyl-4-carboxylic acid (2.7 g, 9.2 mmol) was added over 5 minutes and the resulting slurry was heated at 40 °C for 16 hours. The mixture was poured into water (100mL) and the precipitate was isolated by filtration and treated with THF/6N HCI (9/1) (70 mL) at room temperature for 16 hours. The mixture was subsequently evaporated to dryness in vacuo, the residue was treated with water and the solid was isolated by filtration and washed thoroughly 3 times with DCM. The solid was dissolved in hot THF (400 mL) treated with activated carbon and filtered. The filtrate was evaporated in vacuo to dryness. This afforded 1.6 g (50%) of the title compound.
HPLC-MS (Method C): m/z = 396 (M+1); Rt. = 3.51 min.
Example 810 (General procedure (K)). 5-(2H-Tetrazol-5-yl)-1 H-indole
Figure imgf000326_0001
5-(2H-Tetrazol-5-yl)-1 H-indole was prepared from 5-cyanoindole according to the method described in example 594.
HPLC-MS (Method C): m/z: 186 (M+1); Rt = 1.68 min.
Example 811 (General procedure (K)). 1 -Benzyl-4-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000326_0002
1 -Benzyl-1 H-indole-4-carbonitrile was prepared from 4-cyanoindole according to the method described in example 806.
HPLC-MS (Method C): m/z: 233 (M+1); Rt = 4.24 min.
1-Benzyl-4-(2H-tetrazol-5-yl)-1 H-indole was prepared from 1 -benzyl-1 H-indole-4-carbonitrile according to the method described in example 594. HPLC-MS (Method C): m/z: 276 (M+1 ); Rt = 3.44 min.
General procedure (L) for preparation of compounds of general formula In:
Pol-CI/ DIPEA i: LiHMDS / DMSO
DMF / DCM ii: X-CH7-T-H; X = Cl, Br
Figure imgf000327_0001
Figure imgf000327_0002
Figure imgf000327_0003
wherein T is as defined above and
Pol- is a polystyrene resin loaded with a 2-chlorotrityl linker, graphically shown below:
Figure imgf000327_0004
This general procedure (L) is further illustrated by the following example: Example 812 (General procedure (L)). 5-(2H-Tetrazol-5-yl)-1 -[3-(trifluoromethyl)benzyl]-1 H-indole
Figure imgf000327_0005
2-Chlorotritylchloride resin (100 mg, 0.114 mmol active chloride) was swelled in dichloromethane (2 mL) for 30 min. The solvent was drained, and a solution of 5-(2H-tetrazol-5-yl)- 1 H-indole (example 810) (63 mg, 0.34 mmol) in a mixture of Λ/,Λ/-dimethylformamide, dichloromethane and Λ/,Λ/-di(2-propyl)ethylamine (DIPEA) (5:5:2) (1.1 mL) was added. The reaction mixture was shaken at room temperature for 20 hours. The solvent was removed by filtration, and the resin was washed consecutively with Λ/,Λ/-dimethylformamide (2 x 4 mL), dichloromethane (6 x 4 mL) and methyl sulfoxide (2 x 4 mL). Methyl sulfoxide (1 mL) was added, followed by the addition of a solution of lithium bis(trimethylsilyl)amide in tetrahydrofuran (1.0 M, 0.57 mL, 0.57 mmol). The mixture was shaken for 30 min at room temperature, before 3-(trifluoromethyl)benzyl bromide (273 mg, 1.14 mmol) was added as a solution in methyl sulfoxide (0.2 mL). The reaction mixture was shaken for 20 hours at room tempera- ture. The drained resin was washed consecutively with methyl sulfoxide (2 x 4 mL), dichloromethane (2 x 4 mL), methanol (2 x 4 mL), dichloromethane (2 x 4 mL) and tetrahydrofuran (4 mL). The resin was treated with a solution of hydrogen chloride in tetrahydrofuran, ethyl ether and ethanol = 8:1:1 (0.1 M, 3 mL) for 6 hours at room temperature. The resin was drained and the filtrate was concentrated in vacuo. The crude product was re-suspended in dichloromethane (1.5 mL) and concentrated three times to afford the title compound (35 mg). No further purification was necessary.
HPLC-MS (Method B): m/z: 344 (M+1 ); Rt = 4.35 min.
1H-NMR (DMSO-de): £8.29 (1H, s), 7.80 (1H, dd), 7.72 (2H, m), 7.64 (2H, bs), 7.56 (1H, t), 7.48 (1 H, d), 6.70 (1 H, d), 5.62 (2H, s).
The compounds in the following examples were prepared in a similar fashion. Optionally, the compounds can be further purified by recrystallization or by chromatography.
Example 813 (General procedure (L)).
1 -(4-Chlorobenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000328_0001
HPLC-MS (Method B): m/z: 310 (M+1 ); Rt = 4.11 min.
Example 814 (General procedure (L)).
1 -(2-Chlorobenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000329_0001
HPLC-MS (Method B): m/z: 310 (M+1); Rt = 4.05 min.
Example 815 (General procedure (L)).
1 -(4-Methoxybenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000329_0002
HPLC-MS (Method B): m/z: 306 (M+1); Rt = 3.68 min.
Example 816 (General procedure (L)). 1 -(4-Methylbenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000329_0003
HPLC-MS (Method B): m/z: 290 (M+1); Rt = 3.98 min.
Example 817 (General procedure (L)). 5-(2H-Tetrazol-5-yl)-1 -[4-(trifluoromethyl)benzyl]-1 H-indole
Figure imgf000329_0004
HPLC-MS (Method B): m/z: 344 (M+1); Rt = 4.18 min. Example 818 (General procedure (L)).
1 -(3-Chlorobenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000330_0001
HPLC-MS (Method B): m/z: 310 (M+1); Rt = 4.01 min.
Example 819 (General procedure (L)).
1 -(3-Methylbenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000330_0002
HPLC-MS (Method B): m/z: 290 (M+1); Rt = 3.98 min.
Example 820 (General procedure (L)).
1 -(2,4-Dichlorobenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000330_0003
HPLC-MS (Method B): m/z: 344 (M+1); Rt = 4.41 min.
Example 821 (General procedure (L)).
1 -(3-Methoxybenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000331_0001
HPLC-MS (Method B): m/z: 306 (M+1); Rt = 3.64 min.
Example 822 (General procedure (L)).
1 -(4-Fluorobenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000331_0002
HPLC-MS (Method B): m/z: 294 (M+1); Rt = 3.71 min.
Example 823 (General procedure (L)). 1 -(3-Fluorobenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000331_0003
HPLC-MS (Method B): m/z: 294 (M+1); Rt = 3.68 min.
Example 824 (General procedure (L)). 1 -(2-lodobenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000331_0004
HPLC-MS (Method B): m/z: 402 (M+1); Rt = 4.11 min. Example 825 (General procedure (L)).
1 -[(Naphthalen-2-yl)methyl]-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000332_0001
HPLC-MS (Method B): m/z: 326 (M+1); Rt = 4.18 min.
Example 826 (General procedure (L)).
1 -(3-Bromobenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000332_0002
HPLC-MS (Method B): m/z: 354 (M+1 ); Rt = 4.08 min.
Example 827 (General procedure (L)).
1 -(4-Carboxybenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000332_0003
In this preparation, a larger excess of lithium bis(trimethylsilyl)amide in tetrahydrofuran (1.0 M, 1.7 mL, 1.7 mmol) was used. HPLC-MS (Method B): m/z: 320 (M+1); Rt = 2.84 min.
Example 828 (General procedure (L)). 1 -(3-Carboxybenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000333_0001
In this preparation, a larger excess of lithium bis(trimethylsilyl)amide in tetrahydrofuran (1.0
M, 1.7 mL, 1.7 mmol) was used.
HPLC-MS (Method B): m/z: 320 (M+1); Rt = 2.91 min.
Example 829 (General procedure (L)).
1 -(2,4-Difluorobenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000333_0002
HPLC-MS (Method B): m/z: 312 (M+1); Rt = 3.78 min.
Example 830 (General procedure (L)).
1 -(3,5-Difluorobenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000333_0003
HPLC-MS (Method B): m/z: 312 (M+1); Rt = 3.78 min.
Example 831 (General procedure (L)).
1 -(3,4-Difluorobenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000334_0001
HPLC-MS (Method B): m/z: 312 (M+1); Rt = 3.81 min.
Example 832 (General procedure (L)).
1 -[4-(2-Propyl)benzyl]-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000334_0002
HPLC-MS (Method B): m/z: 318 (M+1); Rt = 4.61 min.
Example 833 (General procedure (L)). 1 -(3,5-Dimethoxybenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000334_0003
HPLC-MS (Method B): m/z: 336 (M+1); Rt = 3.68 min.
Example 834 (General procedure (L)). 1 -(2'-Cyanobiphenyl-4-ylmethyl)-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000335_0001
HPLC-MS (Method B): m/z: 377 (M+1); Rt = 4.11 min.
Example 835 (General procedure (L)). 1 -(2-Methylbenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000335_0002
HPLC-MS (Method B): m/z: 290 (M+1); Rt = 3.98 min.
Further compounds of the invention that may be prepared according to general procedure (K) and/or (L) includes:
Figure imgf000336_0001
Figure imgf000337_0001
The following compounds of the invention may be prepared eg. from 1-(4-bromobenzyl)-5- (2H-tetrazol-5-yl)-1 H-indole (example 807) or from the analogue 1-(3-bromobenzyl)-5-(2H- tetrazol-5-yl)-1 H-indole and aryl boronic acids via the Suzuki coupling reaction eg as described in Littke, Dai & Fu J. Am. Chem. Soc, 2000, 122, 4020-8 (or references cited therein), or using the methodology described in general procedure (E), optionally changing the palladium catalyst to bis(tri-ten'-butylphosphine)palladium (0).
Example 860 Example 861
Figure imgf000337_0002
Example 862 Example 863 Example 864
Figure imgf000337_0003
General procedure (M) for preparation of compounds of general formula l12:
Figure imgf000337_0004
'12 wherein T is as defined above.
The general procedure (M) is further illustrated by the following example: Example 865 (General procedure (M)). 1-Benzoyl-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000338_0001
To a solution of 5-cyanoindole (1.0 g, 7.0 mmol) in dichloromethane (8 mL) was added 4- (dimethylamino)pyridine (0.171 g, 1.4 mmol), triethylamine (1.96 mL, 1.42 g, 14 mmol) and benzoyl chloride (0.89 mL, 1.08 g, 7.7 mmol). The resulting mixture was stirred for 18 hours at room temperature. The mixture was diluted with dichloromethane (80 mL) and washed consecutively with a saturated solution of sodium hydrogencarbonate (40 mL) and brine (40 mL). The organic phase was dried with magnesium sulfate (1 hour). Filtration and concentration furnished the crude material which was purified by flash chromatography on silica gel, eluting with ethyl acetate/heptanes = 2:3. 1-Benzoyl-1H-indole-5-carbonitrile was obtained as a solid.
HPLC-MS (Method C): m/z: 247 (M+1); Rt = 4.07 min.
1-Benzoyl-1H-indole-5-carbonitrile was transformed into 1-benzoyl-5-(2H-tetrazol-5-yl)-1H- indole by the method described in example 594.
HPLC (Method C): Rt = 1.68 min.
The compound in the following example was prepared by the same procedure.
Example 866 (General procedure (M)). 1 -Benzoyl-4-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000339_0001
1-Benzoyl-1H-indole-4-carbonitrile was prepared from 4-cyanoindole according to the method described in example 865.
HPLC-MS (Method C): m/z: 247 (M+1); Rt = 4.24 min.
1-Benzoyl-4-(2H-tetrazol-5-yl)-1 H-indole was prepared from 1-benzoyl-1H-indole-4- carbonitrile according to the method described in example 594.
HPLC (Method C): Rt = 1.56 min.
Example 867 (General procedure (M)) (2-Fluoro-3-trifluoromethylphenyl)-[5-(2H-tetrazol-5-yl)-indol-1-yl]-methanone
Figure imgf000339_0002
HPLC-MS (Method B): m/z = 376 (M+1); Rt = 4.32 min.
Example 868 (General procedure (M)) (4-Methoxyphenyl)-[5-(2H-tetrazol-5-yl)-indol-1-yl]-methanone
Figure imgf000339_0003
HPLC-MS (Method B): m/z = 320 (M+1); Rt = 3.70 min. Example 869 (General procedure (M)) (3-Nitrophenyl)-[5-(2H-tetrazol-5-yl)-indol-1-yl]-methanone
Figure imgf000340_0001
HPLC-MS (Method B): m/z = 335 (M+1); Rt = 3.72 min.
Example 870 (General procedure (M)) (4-Nitrophenyl)-[5-(2H-tetrazol-5-yl)-indol-1-yl]-methanone
Figure imgf000340_0002
HPLC-MS (Method B): m/z = 335 (M+1); Rt = 3.71 min.
Example 871 (General procedure (M)) Naphthalen-2-yl-[5-(2H-tetrazol-5-yl)-indol-1-yl]-methanone
Figure imgf000340_0003
HPLC-MS (Method C): m/z = 340 (M+1); Rt = 4.25 min.
Example 872 (General procedure (M)) (2,3-Difluorophenyl)-[5-(2H-tetrazol-5-yl)-indol-1-yl]-methanone
Figure imgf000341_0001
HPLC-MS (Method B: m/z = 326 (M+1); Rt = 3.85 min.
The following known and commercially available compounds do all bind to the His B10 Zn2+ site of the insulin hexamer:
Example 873
1 -(4-Fluorophenyl)-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000341_0002
Example 874
1-Amino-3-(2H-tetrazol-5-yl)benzene
Figure imgf000341_0003
Example 875
1-Amino-4-(2H-tetrazol-5-yl)benzene
Figure imgf000341_0004
A mixture of 4-aminobenzonitrile (10 g, 84.6 mmol), sodium azide (16.5 g, 254 mmol) and ammonium chloride (13.6 g, 254 mmol) in DMF was heated at 125 °C for 16 hours. The cooled mixture was filtered and the filtrate was concentrated in vacuo. The residue was added water (200 mL) and diethyl ether (200 mL) which resulted in crystallisation. The mixture was filtered and the solid was dried in vacuo at 40 °C for 16 hours to afford 5-(4- aminophenyl)-2H-tetrazole.
1H NMR DMSO-dβ): δ = 5.7 (3H, bs), 6.69 (2H, d), 7.69 (2H, d). HPLC-MS (Method C): m/z: 162 (M+1); Rt = 0,55 min.
Example 8761 -Nitro-4-(2H-tetrazol-5-yl)benzene
Figure imgf000342_0001
Example 8771 -Bromo-4-(2H-tetrazol-5-yl)benzene
Figure imgf000342_0002
General procedure (N) for solution phase preparation of amides of general formula l13:
o ^ o
Frag — ^-OH + HN R *. Frag_ϋ_N ^ R 1 R'
1 .3 wherein Frag is any fragment carrying a carboxylic acid group, R is hydrogen, optionally substituted aryl or
Figure imgf000342_0003
Frag-C02H may be prepared eg by general procedure (D) or by other similar procedures described herein, or may be commercially available.
The procedure is further illustrated in the following example 878: Example 878 (General procedure (N))
Λ/-(4-Chlorobenzyl)-2-[3-(2,4-dioxothiazolidin-5-ylidenemethyl)-1H-indol-1-yl]acetamide
Figure imgf000343_0001
[3-(2,4-Dioxothiazolidin-5-ylidenemethyl)indol-1-yl]acetic acid (example 478, 90.7 mg, 0.3 mmol) was dissolved in NMP (1 mL) and added to a mixture of 1-ethyl-3-(3-dimethylamino- propyl)carbodiimide, hydrochloride (86.4 mg, 0.45 mmol) and 1-hydroxybenzotriazol (68.8 mg, 0.45 mmol) in NMP (1 mL). The resulting mixture was shaken at RT for 2 h. 4- Chlorobenzylamine (51 mg, 0.36 mmol) and DIPEA (46.4 mg, 0.36 mmol) in NMP (1 mL) were added to the mixture and the resulting mixture shaken at RT for 2 days. Subsequently ethyl acetate (10 mL) was added and the resulting mixture washed with 2x10 mL water followed by saturated ammonium chloride (5 mL). The organic phase was evaporated to dryness giving 75 mg (57%) of the title compound.
HPLC-MS (Method C): m/z: 426 (M+1 ); Rt. = 3.79 min.
Example 879 (General procedure (N))
Λ/-(4-Chlorobenzyl)-4-[2-chloro-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]butyramide
Figure imgf000343_0002
HPLC-MS (Method A): m/z: 465 (M+1 ); Rt = 4.35 min.
Example 880 (General procedure (N)) Λ/-(4-Chlorobenzyl)-4-[4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]butyramide
Figure imgf000343_0003
HPLC-MS (Method A): m/z: 431 (M+1 ); Rt = 3.68 min. Example 881 (General procedure (N)) 2-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]-Λ/-(4-chlorobenzyl)acetamide
Figure imgf000344_0001
HPLC-MS (Method A): m/z: 483 (M+1); Rt = 4.06 min.
Example 882 (General procedure (N)) Λ/-(4-Chlorobenzyl)-2-[3-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]acetamide
Figure imgf000344_0002
HPLC-MS (Method A): m/z: 403 (M+1); Rt = 4.03 min.
Example 883 (General procedure (N)) ιV-(4-Chlorobenzyl)-3-[4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenyl]acrylamide
Figure imgf000344_0003
HPLC-MS (Method A): m/z: 399 (M+1); Rt = 3.82.
Example 884 (General procedure (N)) Λ -(4-Chlorobenzyl)-4-[3-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]butyramide
Figure imgf000344_0004
HPLC-MS (Method A): m/z: 431 (M+1); Rt = 3.84 min.
Example 885 (General procedure (N)) 4-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]-Λ/-(4-chlorobenzyl)butyramide
Figure imgf000344_0005
HPLC-MS (Method A): m/z: 511 (M+1); Rt = 4.05 min. Example 886 (General procedure (N))
4-[2-Bromo-4-(4-oxo-2-thioxothiazolidin-5-ylidenemethyl)-phenoxy]-Λ/-(4-chlorobenzyl)- butyramide
Figure imgf000345_0001
HPLC-MS (Method A): m/z: 527 (M+1); Rt = 4.77 min.
Example 887 (General procedure (N)) Λ-(4-Chlorobenzyl)-2-[4-(2,4-dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]acetamide
Figure imgf000345_0002
HPLC-MS (Method C): m/z: 431 (M+1); Rt. = 4.03 min.
Example 888 (General procedure (N)) Λ/-(4-Chlorobenzyl)-3-[3-(2,4-dioxothiazolidin-5-ylidenemethyl)-1H-indol-1-yl]propionamide
Figure imgf000345_0003
HPLC-MS (Method C): m/z: 440 (M+1); Rt. = 3.57 min.
Example 889 (General procedure (N)) Λ/-(4-Chlorobenzyl)-4-[4-(2,4-dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]butyramide
Figure imgf000345_0004
HPLC-MS (Method C): m/z: 481 (M+1); Rt = 4.08 min. Example 890 (General procedure (N)) 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)-naphthalen-1-yloxy]-Λ/-hexylbutyramide
Figure imgf000346_0001
HPLC-MS (Method C): m/z: 441 (M+1); Rt = 4.31 min.
Example 891 (General Procedure (N))
4-({[3-(2,4-Dioxothiazolidin-5-ylidenemethyl)indole-7-carbonyl]amino}methyl)benzoic acid methyl ester
Figure imgf000346_0002
HPLC-MS (Method C): m/z: 436 (M+1); Rt.= 3.55 min.
Example 892 (General procedure (N)) Λ/-(4-Chlorobenzyl)-4-[3-(2H-tetrazol-5-yl)carbazol-9-ylmethyl]benzamide
Figure imgf000346_0003
HPLC-MS (Method C): m/z:493 (M+1); Rt = 4.19 min.
Example 893 (General procedure (N)) Λ -(4-Chlorobenzyl)-3-[3-(2H-tetrazol-5-yl)carbazol-9-ylmethyl]benzamide
Figure imgf000347_0001
HPLC-MS (Method C): m/z: 493 (M+1); Rt = 4.20 min.
Example 894 (General Procedure (N)) Λ/-(4-Chlorobenzyl)-3-methyl-4-[3-(2H-tetrazol-5-yl)-carbazol-9-ylmethyl]benzamide
Figure imgf000347_0002
HPLC-MS (Method C): m/z: 507 (M+1); Rt = 4.37min.
Example 895 (General procedure (N))
5-{2-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)-naphthalen-1-yloxy]-acetylamino}-isophthalic acid dimethyl ester
Figure imgf000347_0003
HPLC-MS (Method C): m/z = 521 (M+1); Rt. = 4.57 min.
Example 896 (General procedure (N)) 5-{2-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)-naphthalen-1-yloxy]-acetylamino}-isophthalic acid
Figure imgf000348_0001
HPLC-MS (Method C): m/z = 515 (M+23); Rt. = 3.09 min.
Example 897 (General procedure (N))
5-(3-{2-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)-naphthalen-1-yloxy]-ethyl}-ureido)- isophthalic acid monomethyl ester
Figure imgf000348_0002
HPLC-MS (Method C): m/z = 536 (M+1 ); Rt = 3,58 min.
Example 898 (General Procedure (N)). 2-{4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoylamino}succinic acid dimethyl ester
Figure imgf000348_0003
4-[3-(1H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoic acid (2.00 g, 5.41 mmol), 1- hydroxybenzotriazole (1.46 g, 10.8 mmol) and N,N-di(2-propyl)ethylamine (4.72 mL, 3.50 g, 27.1 mmol) were dissolved in dry N,N-dimethylformamide (60 mL). The mixture was cooled in an ice-water bath, and 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (1.45 g, 7.56 mmol) and (S)-aminosuccinic acid dimethyl ester hydrochloride (1.28 g, 6.48 mmol) were added. The cooling was discontinued, and the reaction mixture was stirred at room temperature for 18 hours before it was poured into hydrochloric acid (0.1 N, 600 mL). The solid was collected by filtration and washed with water (2 X 25 mL) to furnish the title compound. HPLC-MS (Method C): m/z: 513 (M+1 ); Rt = 3.65 min.
1H-NMR (DMSO-de): δ 8.90 (1 H, d), 8.86 (1H, d), 8.29 (1 H, d), 8.11 (1 H, dd), 7.87 (1H, d), 7.75 (2H, d), 7.69 (1 H, d), 7.51 (1 H, t), 7.32 (1 H, t), 7.28 (2H, d), 5.82 (2H, s), 4.79 (1 H, m), 3.61 (3H, s), 3.58 (3H, s), 2.92 (1 H, dd), 2.78 (1H, dd).
Example 899 (General Procedure (N)).
2-{4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoylamino}succinic acid
Figure imgf000349_0001
2-{4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoylamino}succinic acid dimethyl ester
(1.20 g, 2.34 mmol) was dissolved in tetrahydrofuran (30 mL). Aqueous sodium hydroxide (1 N, 14 mL) was added, and the resulting mixture was stirred at room temperature for 18 hours. The reaction mixture was poured into hydrochloric acid (0.1 N, 500 mL). The solid was collected by filtration and washed with water (2 X 25 mL) and diethyl ether (2 X 25 mL) to furnish the title compound.
HPLC-MS (Method C): m/z: 485 (M+1); Rt = 2.94 min. 1H-NMR (DMSO-de): δ 12.44 (2H, s (br)), 8.90 (1 H, d), 8.68 (1H, d), 8.29 (1 H, d), 8.11 (1H, dd), 7.87 (1 H, d), 7.75 (2H, d), 7.68 (1 H, d), 7.52 (1 H, t), 7.32 (1H, t), 7.27 (2H, d), 5.82 (2H, s), 4.70 (1 H, m), 2.81 (1H, dd), 2.65 (1 H, dd).
The compounds in the following examples were prepared in a similar fashion.
Example 900 (General procedure (N)) 2-{4-[3-(2H-Tetrazol-5-yl)-carbazol-9-ylmethyl]-benzoylamino}-succinic acid dimethyl ester
Figure imgf000350_0001
HPLC-MS (Method C): m/z = 513 (M+1 ); Rt = 3.65min.
Example 901 (General procedure (N)) 2-{4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoylamino}pentanedioic acid dimethyl ester
Figure imgf000350_0002
HPLC-MS (Method C): m/z = 527 (M+1); Rt = 3.57min.
Example 902 (General procedure (N))
(Methoxycarbonylmethyl-{4-[3-(2H-tetrazol-5-yl)-carbazol-9-ylmethyl]-benzoyl}-amino)-acetic acid methyl ester
Figure imgf000350_0003
HPLC-MS (Method C): m/z = 513 (M+1); Rt = 3,55min.
Example 903 (General procedure (N))
2-{4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoylamino}pentanedioic acid
Figure imgf000351_0001
HPLC-MS (Method C): m/z = 499 (M+1); Rt = 2.87min.
Example 904 (General procedure (N)) (Ethoxycarbonylmethyl-{4-[3-(2H-tetrazol-5-yl)-carbazol-9-ylmethyl]-benzoyl}-amino)-acetic acid ethyl ester
Figure imgf000351_0002
HPLC-MS (Method C): m/z = 541 (M+1); Rt = 3.91 min.
Example 905 (General procedure (N))
3-(3-{4-[4-(2,4-Dioxo-thiazolidin-5-ylidenemethyl)-naphthalen-1-yloxy]-butyrylamino}- propylamino)-hexanedioic acid dimethyl ester
Figure imgf000351_0003
HPLC-MS (Method C: m/z = 585 (M+1); Rt = 2,81 min.
Example 906 (General procedure (N))
3-(3-{4-[4-(2,4-Dioxo-thiazolidin-5-ylidenemethyl)-naphthalen-1-yloxy]-butyrylamino}- propylamino)-hexanedioic acid
Figure imgf000352_0001
HPLC-MS (Method C): m/z = 554 (M-3); Rt = 3,19 min.
Example 907 (General procedure (N)) (Carboxymethyl-{4-[3-(2H-tetrazol-5-yl)-carbazol-9-ylmethyl]-benzoyl}-amino)-acetic acid
Figure imgf000352_0002
HPLC-MS (Method C): m/z = 485 (M+1); Rt = 3.04 min.
Example 908 (General procedure (N)) 4-(3-{4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)-naphthalen-1 -yloxy]-butyrylamino}- propylamino)-cyclohexane-1,3-dicarboxylic acid dimethyl ester
Figure imgf000352_0003
HPLC-MS (Method C): m/z = 612 (M+1); Rt = 3,24 min.
Example 909 (General procedure (N))
2-{3-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoylamino}pentanedioic acid dimethyl ester
Figure imgf000352_0004
HPLC-MS (Method C): m/z = 527 (M+1); Rt = 3.65min. Example 910 (General procedure (N)) 2-{3-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoylamino}pentanedioic acid dimethyl ester
Figure imgf000353_0001
HPLC-MS (Method C): m/z = 527 (M+1); Rt = 3.65min.
Example 911 (General procedure (N)) 2-{3-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoylamino}pentanedioic acid dimethyl ester
Figure imgf000353_0002
HPLC-MS (Method C): m/z = 527 (M+1); Rt = 3.65min.
Example 912 (General procedure (N)) 2-{3-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoylamino}pentanedioic acid
Figure imgf000353_0003
HPLC-MS (Method C): m/z = 499 (M+1); Rt = 3.00 min.
Example 913 (General procedure (N))
(Methoxycarbonylmethyl-{3-[3-(2H-tetrazol-5-yl)carbazol-9-ylmethyl]benzoyl}amino)acetic acid methyl ester
Figure imgf000354_0001
1H-NMR (DMSO-dβ): δ 8.88 (1 H, d), 8.29 (1 H, d), 8.10 (1 H, dd), 7.85 (1H, d), 7.67 (1H, d), 7.52 (1 H, t), 7.39 (1 H, t), 7.30 (2H, m), 7.17 (2H, m), 5.79 (2H, s), 4.17 (2H, s), 4.02 (2H, s), 3.62 (3H, s), 3.49 (3H, s).
Example 914 (General procedure (N)) 2-{3-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoylamino}succinic acid dimethyl ester
Figure imgf000354_0002
HPLC-MS (Method C): m/z = 513 (M+1 ); Rt = 3.70 min.
Example 915 (General procedure (N)) 2-{3-[3-(2H-Tetrazol-5-yl)-carbazol-9-ylmethyl]-benzoylamino}-succinic acid
Figure imgf000354_0003
HPLC-MS (Method C): m/z = 485 (M+1 ); Rt = 2.96 min.
Example 916 (General procedure (N)) (Carboxymethyl-{3-[3-(2H-tetrazol-5-yl)carbazol-9-ylmethyl]benzoyl}amino)acetic acid
Figure imgf000355_0001
HPLC-MS (Method C): m/z = 485 (M+1); Rt = 2.87 min.
Example 917 (General procedure (N))
4-(4-(3-Carboxy-propylcarbamoyl)4-{4-[3-(2H-tetrazol-5-yl)carbazol-9-ylmethyl]- benzoylamino}-butyrylamino)-butyric acid
Figure imgf000355_0002
The title compound was prepared by coupling of (S)-2-{4-[3-(2H-tetrazol-5-yl)carbazol-9- ylmethyl]benzoylamino}pentanedioic acid bis-(2,5-dioxopyrrolidin-1-yl) ester (prepared from (S)-2-{4-[3-(2H-tetrazol-5-yl)carbazol-9-ylmethyl]benzoylamino}pentanedioic acid by essentially the same procedure as described for the synthesis of 4-[3-(2H-tetrazol-5-yl)carbazol-9- ylmethyl]benzoic acid 2,5-dioxopyrrolidin-1-yl ester) with 4-aminobutyric acid according to the procedure described for the preparation of 4-{4-[3-(2H-tetrazol-5-yl)carbazol-9-ylmethyl]- benzoylamino}butyric acid .
HPLC-MS (Method C): m/z: 669 (M+1); Rt = 2.84 min.
Example 918 (General procedure (N)) [2-(2-{4-[3-(2H-Tetrazol-5-yl)-carbazol-9-ylmethyl]benzoylamino}ethoxy)ethoxy]acetic acid
Figure imgf000356_0001
HPLC-MS (Method C): m/z: 515 (M+1); Rt = 3.10 min.
Example 919 (General procedure (N))
2-{4-[3-(2H-Tetrazol-5-yl)-carbazol-9-ylmethyl]-benzoylamino}-pentanedioic acid di-tert-butyl ester
Figure imgf000356_0002
HPLC-MS (Method C): m/z = 611 (M+1 ); Rt = 4.64 min.
Example 920 (General Procedure (N)). 4-{4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoylamino}butyric Acid
Figure imgf000356_0003
HPLC-MS (Method C): m/z: 455 (M+1); Rt = 3.13 min.
Example 921 (General Procedure (N)).
[2-(2-{4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoylamino}ethoxy)ethoxy]acetic acid
Figure imgf000357_0001
The title compound was prepared by coupling of 4-[3-(2H-tetrazol-5-yl)carbazol-9- ylmethyl]benzoic acid 2,5-dioxopyrrolidin-1-yl ester with [2-(2-aminoethoxy)ethoxy]acetic acid (prepared from [2-[2-(Fmoc-amino)ethoxy]ethoxy]acetic acid by treatment with PS-Trisamine resin in DMF).
HPLC-MS (Method C): m/z: 515 (M+1); Rt = 3.10 min.
The commercially available compounds in the following examples do all bind to the HisBlO Zn2+site:
Example 922
1 -(4-Bromo-3-methylphenyl)-1 ,4-dihydrotetrazole-5-thione
Figure imgf000357_0002
Example 923
1 -(4-lodophenyl)-1 ,4-dihydrotetrazole-5-thione
Figure imgf000357_0003
Example 924 1 -(2,4,5-Trichlorophenyl)-1 H-tetrazole-5-thiol
Figure imgf000357_0004
Example 925 1-(2,6-Dimethylphenyl)-1 ,4-dihydrotetrazole-5-thione
Figure imgf000358_0001
Example 926
1 -(2,4,6-Trimethylphenyl)-1 ,4-dihydrotetrazole-5-thione
Figure imgf000358_0002
Example 927 1 -(4-Dimethylaminophenyl)-1 H-tetrazole-5-thiol
Figure imgf000358_0003
Example 928
1 -(3,4-Dichlorophenyl)-1 ,4-dihydro-1 H-tetrazole-5-thione
Figure imgf000358_0004
Example 929
1 -(4-Propylphenyl)-1 ,4-dihydro-1 H-tetrazole-5-thione
Figure imgf000358_0005
Example 930
1-(3-Chlorophenyl)-1 ,4-dihydro-1H-tetrazole-5-thione
Figure imgf000359_0001
Example 931
1 -(2-Fluorophenyl)-1 ,4-dihydro-1 H-tetrazole-5-thione
Figure imgf000359_0002
Example 932 1 -(2,4-Dichlorophenyl)-1 ,4-dihydro-1 H-tetrazole-5-thione
Figure imgf000359_0003
Example 933
1 -(4-Trifluoromethoxyphenyl)-1 ,4-dihydro-1 H-tetrazole-5-thione
Figure imgf000359_0004
Example 934 N-[4-(5-Mercaptotetrazol-1-yl)-phenyl]-acetamide
Figure imgf000359_0005
Example 935
1 -(4-Chlorophenyl)-1 ,4-dihydrotetrazole-5-thione
Figure imgf000360_0001
Example 936
1-(4-Methoxyphenyl)-1,4-dihydrotetrazole-5-thione
Figure imgf000360_0002
Example 937 1 -(3-Fluoro-4-pyrrolidin-1 -ylphenyl)-1 ,4-dihydrotetrazole-5-thione
Figure imgf000360_0003
Example 938
Λ/-[3-(5-Mercaptotetrazol-1-yl)phenyl]acetamide
Figure imgf000360_0004
Example 939 1-(4-Hydroxyphenyl)-5-mercaptotetrazole
Figure imgf000361_0001
Example 940
Figure imgf000361_0002
Preparation of 1-aryl-1,4-dihydrotetrazole-5-thiones (or the tautomeric 1-aryltetrazole-5- thiols) is described in the literature (eg. by Kauer & Sheppard, J. Org. Chem., 32, 3580-92 (1967)) and is generally performed eg. by reaction of aryl-isothiocyanates with sodium azide followed by acidification
1-Aryl-1 ,4-dihydrotetrazole-5-thiones with a carboxylic acid tethered to the aryl group may be prepared as shown in the following scheme:
Figure imgf000361_0003
Step 1 is a phenol alkylation and is very similar to steps 1 and 2 of general procedure (D) and may also be prepared similarly as described in example 481. Step 2 is a reduction of the nitro group. SnCI2, H over Pd/C and many other procedures known to those skilled in the art may be utilised.
Step 3 is formation of an arylisothiocyanate from the corresponding aniline. As reagents CS2, CSCI2, or other reagents known to those skilled in the art, may be utilised.
Step 4 is a conversion to mercaptotetrazole as described above.
Compounds of the invention include:
Example 941 Example 942
Figure imgf000362_0001
Example 943 Example 944
Figure imgf000362_0002
Example 945 Example 946
Figure imgf000362_0003
Example 947
Figure imgf000362_0004
Example 948
4-(4-Hydroxyphenyl)-1 H-[1 ,2,3]triazole-5-carbonitrile
Figure imgf000363_0001
Phenylsulphonyl acetonitrile (2.0 g, 11.04 mmol) was mixed with 4-hydroxybenzaldehyde (1.35 g, 11.04 mmol) in DMF (10 mL) and toluene (20 mL). The mixture was refluxed for 3 hours and subsequently evaporated to dryness in vacuo. The residue was treated with di- ethyl ether and toluene. The solid formed was filtered to afford 2.08 g (66%) of 2- benzenesulfonyl-3-(4-hydroxyphenyl)acrylonitrile. HPLC-MS (Method C): m/z: 286 (M+1 ); Rt. = 3.56 min.
A mixture of 2-benzenesulfonyl-3-(4-hydroxyphenyl)acrylonitrile (2.08 g, 7.3 mmol) and so- dium azide (0.47g,7.3 mmol) in DMF (50 mL) was heated at reflux temperature 2 hours. After cooling, the mixture was poured on ice. The mixture was evaporated in vacuo to almost dryness and toluene was added. After filtration, the organic phase was evaporated in vacuo. The residue was purified by silicagel chromatography eluting with a mixture of ethyl acetate and heptane (1 :2). This afforded 1.2 g (76%) of the title compound.
1 H NMR (DMSO-de): 10.2 (broad,1 H); 7.74 (d,2H); 6.99 (d,2H); 3.6-3.2 (broad,1 H). HPLC-MS (Method C) m/z: = 187 (M+1); Rt. = 1.93 min
General procedure (O) for preparation of compounds of general formula l 4:
Figure imgf000363_0002
'1
wherein
AA is as defined above,
Steps 1 and 2 are described in the literature (eg Beck & Gunther, Chem. Ber., 106, 2758-66 (1973)) Step 1 is a Knoevenagel condensation of the aldehyde AA-CHO with phenylsulfonyl- acetonitrile and step 2 is a reaction of the vinylsulfonyl compound obtained in step 1 with sodium azide. This reaction is usually performed in DMF at 90 - 110 °C.
This general procedure is further illustrated in the following example 949:
Example 949 (General Procedure (O))
[4-(5-Cyano-1H-[1 ,2,3]triazol-4-yl)phenoxy]acetic acid
Figure imgf000364_0001
Phenylsulphonylacetonitrile (0.1 g, 0.55 mmol) was mixed with 4-formylphenoxyactic acid (0.099 g, 0.55 mmol) in DMF (3 mL) and heated to 110 °C for 3 h and subsequently cooled to RT. Sodium azide (0.036 g, 0.55 mmol) was added and the resulting mixture was heated to 110 °C for 3 h and cooled to RT. The mixture was poured into water (20 mL) and centrifuged. The supernatant was discarded, ethanol (5 mL) was added and the mixture was centrifuged again. After discarding the supernatant, the residue was dried in vacuo to afford 50 mg (37%) of [4-(5-Cyano-1H-[1,2,3]triazol-4-yl)phenoxy]acetic acid.
HPLC-MS (Method C): m/z: 245 (M+1) Rt. 2.19 min.
Example 950 (General Procedure (O)) 5-(Naphthalen-1 -yl)-3H-[1 ,2,3]triazole-4-carbonitrile
Figure imgf000364_0002
HPLC-MS (Method C): m/z: 221 (M+1); Rt. 3.43 min. Example 951 (General Procedure (O)) 5-(Naphthalen-2-yl)-3H-[1 ,2,3]triazole-4-carbonitrile
Figure imgf000365_0001
HPLC-MS (Method C): m/z: 221 (M+1); Rt = 3.66 min.
Example 952 (General procedure (O))
4-[3-(5-Cyano-[1 ,2,3]triazol-4-yl)-1 ,4-dimethylcarbazol-9-ylmethyl]-benzoic acid
Figure imgf000365_0002
HPLC-MS (Method C): m/z = 422 (M+1); Rt = 3.85 min.
Preparation of intermediary aldehyde:
1 ,4 Dimethylcarbazol-3-carbaldehyde (0.68 g, 3.08 mmol) was dissolved in dry DMF (15 mL), NaH (diethyl ether washed) (0.162 g, 6.7 mol) was slowly added under nitrogen and the mixture was stirred for 1 hour at room temperature. 4-Bromomethylbenzoic acid (0.73 g, 3.4 mmol) was slowly added and the resulting slurry was heated to 40 °C for 16 hours. Water (5 mL) and hydrochloric acid (6N, 3 mL) were added. After stirring for 20 min at room temperature, the precipitate was filtered off and washed twice with acetone to afford after drying 0.38 g (34%) of 4-(3-formyl-1 ,4-dimethylcarbazol-9-ylmethyl)benzoic acid.
HPLC-MS (Method C) : m/z = 358 (M+1 ), RT. = 4.15 min.
Example 953 (General Procedure (O)) 5-(Anthracen-9-yl)-3H-[1 ,2,3]triazole-4-carbonitrile
Figure imgf000366_0001
HPLC-MS (Method C): m/z: 271 (M+1); Rt = 3.87 min.
Example 954 (General Procedure (O)) 5-(4-Methoxynaphthalen-1 -yl)-3H-[1 ,2,3]triazole-4-carbonitrile
Figure imgf000366_0002
HPLC-MS (Method C): m/z: 251 (M+1); Rt = 3.57 min.
Example 955 (General Procedure (O)) 5-(1,4-Dimethyl-9H-carbazol-3-yl)-3H-[1,2,3]triazole-4-carbonitrile
Figure imgf000366_0003
HPLC-MS (Method C): m/z: 288 (M+1); Rt = 3.67 min.
Example 956 (General procedure (O)) 5-(4'-Methoxybiphenyl-4-yl)-3H-[1 ,2,3]triazole-4-carbonitrile
Figure imgf000367_0001
HPLC-MS (Method C): m/z = 277 (M+1); Rt = 3.60 min.
Example 957 (General procedure (O)) 5-(4-Styrylphenyl)-3H-[1 ,2,3]triazole-4-carbonitrile
Figure imgf000367_0002
HPLC-MS (Method C): m/z = 273 (M+1); Rt = 4.12 min.
Example 958 (General procedure (O)) 5-(2,6-Dichloro-4-dibenzylaminophenyl)-3H-[1 ,2,3]triazole-4-carbonitrile
Figure imgf000367_0003
HPLC-MS (Method C): m/z = 434 (M+1); Rt = 4.64 min. Example 959 (General procedure (O))
5-(1 -Bromonaphthalen-2-yl)-3H-[1 ,2,3]triazole-4-carbonitrile
Figure imgf000368_0001
HPLC-MS (Method C: m/z = 300 (M+1 ); Rt. = 3.79 min.
Example 960
4-(4-Bromophenyl)-1 H-[1 ,2,3]triazole-5-carbonitrile
Figure imgf000368_0002
This compound is commercially available (MENAI).
Example 961
N-[4-(5-Cyano-1 H-[1 ,2,3]triazol-4-yl)-phenyl]-acetamide
Figure imgf000368_0003
This compound is commercially available (MENAI).
Example 962 (General procedure (O)) 5-(4'-Chlorobiphenyl-4-yl)-3H-[1 ,2,3]triazole-4-carbonitrile
Figure imgf000369_0001
HPLC-MS (Method C): m/z = 281 (M+1); Rt = 4.22 min.
The compounds in the following examples are commercially available and may be prepared using a similar methodology:
Example 963
4-(4-Trifluoromethoxyphenyl)-1 H-[1 ,2,3]triazole-5-carbonitrile
Figure imgf000369_0002
Example 964
4-Benzo[1 ,3]dioxol-5-yl-1 H-[1 ,2,3]triazole-5-carbonitrile
Figure imgf000369_0003
Example 965
4-(3-Trifluoromethylphenyl)-1 H-[1 ,2,3]triazole-5-carbonitrile
Figure imgf000370_0001
Example 966
4-Pyridin-3-yl-1 H-[1 ,2,3]triazole-5-carbonitrile
Figure imgf000370_0002
Example 967
4-(2,6-Dichlorophenyl)-1 H-[1 ,2,3]triazole-5-carbonitrile
Figure imgf000370_0003
Example 968
4-Thiophen-2-yl-1 H-[1 ,2,3]triazole-5-carbonitrile
Figure imgf000370_0004
Example 969 3,5-Dimethylisoxazole-4-carboxylic acid 4-(5-cyano-1H-[1 ,2,3]triazol-4-yl)phenyl ester
Figure imgf000371_0001
Example 970
3,3-Dimethyl-butyric acid 4-(5-cyano-1 H-[1 ,2,3]triazol-4-yl)phenyl ester
Figure imgf000371_0002
Example 971
4-Methyl-[1 ,2,3]thiadiazole-5-carboxylic acid 4-(5-cyano-1 H-[1 ,2,3]triazol-4-yl)phenyl ester
Figure imgf000371_0003
Example 972
4-Chlorobenzoic acid 4-(5-cyano-1H-[1 ,2,3]triazol-4-yl)phenyl ester
Figure imgf000371_0004
Example 973
4-(3-Phenoxyphenyl)-1 H-[1 ,2,3]triazole-5-carbonitrile
Figure imgf000372_0001
Example 974
4-(5-Bromo-2-methoxyphenyl)-1 H-[1 ,2,3]triazole-5-carbonitrile
Figure imgf000372_0002
Example 975
4-(2-Chloro-6-fluorophenyl)-1 H-[1 ,2,3]triazole-5-carbonitrile
Figure imgf000372_0003
The following cyanotriazoles are also compounds of the invention:
4-(2-Chloro-6-fluorophenyl)-1 H-[1 ,2,3]triazole-5-carbonitrile. Terephthalic acid mono[ 4-(5-cyano-1H-[1 ,2,3]triazol-4-yl)phenyl] ester. N- [4-(5-cyano-1H-[1 ,2,3]triazol-4-yl)-phenyl]terephthalamic acid 4-(4-Octyloxyphenyl)-1 H-[1 ,2,3]triazole-5-carbonitrile 4-(4-Styrylphenyl)-1 H-[1 ,2,3]triazole-5-carbonitrile. 4-(4'-Trifluoromethylbiphenyl-4-yl)-1H-[1 ,2,3]triazole-5-carbonitrile. 4-(4'-Chlorobiphenyl-4-yl)-1 H-[1 ,2,3]triazole-5-carbonitrile. 4-(4'-Methoxybiphenyl-4-yl)-1 H-[1 ,2,3]triazole-5-carbonitrile. 4-(1 -Naphthyl)-1 H-[1 ,2,3]triazole-5-carbonitrile. 4-(9-Anthranyl)-1 H-[1 ,2,3]triazole-5-carbonitrile. 4-(4-Methoxy-1 -naphthyl)- 1 H-[1 ,2,3]triazole-5-carbonitrile. 4-(4-Aminophenyl)-1 H-[1 ,2,3]triazole-5-carbonitrile. 4-(2-Naphthyl)-1 H-[1 ,2,3]triazole-5-carbonitrile.
General procedure (P) for preparation of compounds of general formula l15:
Figure imgf000373_0001
Step 3
Figure imgf000373_0002
wherein n is 1 or 3-20,
AA is as defined above,
R" is a standard carboxylic acid protecting group, such as CrCe-alkyl or benzyl and Lea is a leaving group, such as chloro, bromo, iodo, methanesulfonyloxy, toluenesulfonyloxy or the like.
This procedure is very similar to general procedure (D), steps 1 and 2 are identical.
Steps 3 and 4 are described in the literature (eg Beck & GQnther, Chem. Ber., 106, 275S-66 (1973))
Step 3 is a Knoevenagel condensation of the aldehyde obtained in step 2 with phenylsulfon- ylacetonitrile and step 4 is a reaction of the vinylsulfonyl compound obtained in step 3 with sodium azide. This reaction is usually performed in DMF at 90 - 110 °C. This General procedure (P) is further illustrated in the following two examples
Example 976 (General procedure (P))
5-[6-(5-Cyano-1 H-[1 ,2,3]triazol-4-yl)-naphthalen-2-yloxy]-pentanoic acid ethyl ester
Figure imgf000374_0001
6-Hydroxynaphthalene-2-carbaldehyde (Syncom BV. NL, 15.5 g, 90 mmol) and K2C03 (62.2 g, 450 mmol) were mixed in DMF (300mL) and stirred at room temperature for 1hour. Ethyl 5-bromovalerate (21.65 g, 103.5 mmol) was added and the mixture was stirred at room tem- perature for 16 hours. Activated carbon was added and the mixture was filtered. The filtrate was evaporated to dryness in vacuo to afford 28.4 g of crude 5-(6-formylnaphthalen-2- yloxy)pentanoic acid ethyl ester, which was used without further purification.
HPLC-MS (Method C ): m/z = 301 (M+1); Rt. = 4.39 min.
5-(6-Formylnaphthalen-2-yloxy)pentanoic acid ethyl ester (28.4 g, 94.5 mmol), phenylsulfon- ylacetonitrile (20.6 g, 113.5 mmol), and piperidine (0.94 mL) were dissolved in DMF (200 mL) and the mixture was heated at 50 °C for 16 hours. The resulting mixture was evaporated to dryness in vacuo and the residue was dried for 16 hours at 40 °C in vacuo. The solid was recrystallised from 2-propanol (800 mL) and dried again as described above. This afforded 35 g (80%) of 5-[6-(2-benzenesulfonyl-2-cyanovinyl)naphthalen-2-yloxy]pentanoic acid ethyl ester.
HPLC-MS (Method C): m/z = 486 (M+23); Rt. = 5.09 min.
5-[6-(2-Benzenesulfonyl-2-cyanovinyl)naphthalen-2-yloxy]pentanoic acid ethyl ester (35 g, 74.6 mmol) and sodium azide (4.9 g, 75.6 mmol) were dissolved in DMF (100 mL) and stirred for 16 hours at 50 °C. The mixture was evaporated to dryness in vacuo, redissolved in THF / ethanol and a small amount of precipitate was filtered off. The resulting filtrate was poured into water (2.5 L). Filtration afforded after drying 24.5 g (88%) of 5-[6-(5-cyano-1 H- [1,2,3]triazol-4-yl)naphthalen-2-yloxy]pentanoic acid ethyl ester (24.5 g, 88%).
HPLC-MS (Method C): m/z = 365 (M+1); Rt. = 4.36 min.
Example 977 (General procedure (B))
5-[6-(5-Cyano-1 H-[1 ,2,3]triazol-4-yl)-naphthalen-2-yloxy]-pentanoic acid
Figure imgf000375_0001
5-[6-(5-Cyano-1H-[1,2,3]triazol-4-yl)naphthalen-2-yloxy]pentanoicacid ethyl ester (24.5 g, 67.4 mmol) was dissolved in THF (150 mL) and mixed with sodium hydroxide (8.1 g, 202 mmol) dissolved in water (50 mL). The mixture was stirred for 2 days and the volatiles were evaporated in vacuo. The resulting aqueous solution was poured into a mixture of water (1 L) and hydrochloric acid (1N, 250 mL). The solid was isolated by filtration, dissolved in sodium hydroxide (1N, 200 mL), and the solution was washed with DCM and then ethyl acetate, the aquous layer was acidified with hydrochloric acid (12N). The precipitate was isolated by filtration, dissolved in THF / diethyl ether, the solution was treated with MgS04 and activated carbon, filtrated and evaporated in vacuo to almost dryness followed by precipitation by addition of pentane (1 L). This afforded after drying in vacuo 17.2 g ( 76%) of the title compound.
HPLC-MS (Method C): m/z = 337 (M+1); Rt. = 3.49 min.
Example 978 (General procedure (P)) 6-[6-(5-Cyano-1 H-[1 ,2,3]triazol-4-yl)naphthalen-2-yloxy]hexanoic acid
Figure imgf000376_0001
HPLC-MS (Method C): m/z = 351 (M+1); Rt = 3.68 min.
Example 979 (General procedure (P))
11 -[6-(5-Cyano-1 H-[1 ,2,3]triazol-4-yl)-naphthalen-2-yloxy]-undecanoic acid
Figure imgf000376_0002
HPLC-MS (Method C): m/z = 443 (M+23); Rt = 4.92 min.
Example 980 (General procedure (P)) 2-{3-[6-(5-Cyano-1H-[1,2,3]triazol-4-yl)-naphthalen-2-yloxy]-propyl}-malonic acid diethyl ester
Figure imgf000376_0003
HPLC-MS (Method C): m/z = 465 (M+1); Rt. = 4.95 min.
Example 981 (General procedure (P)) 2-{5-[6-(5-Cyano-1 H-[1 ,2,3]triazol-4-yl)-naphthalen-2-yloxy]-pentyl}-malonic acid diethyl ester
Figure imgf000377_0001
HPLC-MS (Method C): m/z = 465 (M+1); Rt. = 4.95 min.
Example 982 (General procedure (P))
2-{3-[6-(5-Cyano-1 H-[1 ,2,3]triazol-4-yl)-naphthalen-2-yloxy]-propyl}-malonic acid
Figure imgf000377_0002
HPLC-MS (Method C): m/z = 381 (M+1); Rt. = 3.12 min.
Example 983 (General procedure (P)) 2-{5-[6-(5-Cyano-1 H-[1 ,2,3]triazol-4-yl)-naphthalen-2-yloxy]-pentyl}-malonic acid
Figure imgf000377_0003
HPLC-MS (Method C): m/z 0 409 (M+1 ); Rt. = 3.51 min.
Example 984 (General procedure (P)) 4-[4-(5-Cyano-1 H-[1 ,2,3]triazol-4-yl)-phenoxy]butyric acid
Figure imgf000378_0001
HPLC-MS (Method C): m/z = 273 (M+1); Rt = 2.44 min.
The following compounds may be prepared according to this general procedure (P):
4-(4-(5-Cyano-1 H-[1 ,2,3]triazol-4-yl)phenoxy)butyric acid:
Figure imgf000378_0002
2-(4-(5-Cyano-1H-[1 ,2,3]triazol-4-yl)phenoxy)acetic acid:
Figure imgf000378_0003
4-(4-(5-Cyano-1 H-[1 ,2,3]triazol-4-yl)phenoxy)butyric acid ethyl ester 5-(4-(5-Cyano-1 H-[1 ,2,3]triazol-4-yl)phenoxy)pentanoic acid 8-(4-(5-Cyano-1H-[1 ,2,3]triazol-4-yl)phenoxy)octanoic acid 10-(4-(5-Cyano-1 H-[1 ,2,3]triazol-4-yl)phenoxy)decanoic acid 12-(4-(5-Cyano-1H-[1 ,2,3]triazol-4-yl)phenoxy)dodecanoic acid General procedure (R) for preparation of compounds of general formula l1 :
Figure imgf000379_0001
Figure imgf000379_0002
wherein T is as defined above and R2 and R3 are hydrogen, aryl or lower alkyl, both optionally substituted.
The general procedure (R) is further illustrated by the following example:
Example 985 (General procedure (R)) Phenyl-[3-(2H-tetrazol-5-yl)-carbazol-9-yl]-methanone
Figure imgf000379_0003
2-Chlorotritylchloride resin (100 mg, 0.114 mmol active chloride) was swelled in dichloromethane (4 mL) for 30 minutes. The solvent was drained, and a solution of 3-(2H-tetrazol-5- yl)-9H-carbazole (80 mg, 0.34 mmol) in a mixture of N,N-dimethylformamide / dichloromethane / N,N-di(2-propyl)ethylamine (5:5:1 ) (3 mL) was added. The reaction mixture was shaken at room temperature for 20 hours. The solvent was removed by filtration, and the resin was washed thoroughly with N,N-dimethylformamide (2 x 4 mL) and dichloromethane (6 x 4 mL). A solution of 4-(dimethylamino)pyridine (14 mg, 0.11 mmol) and N,N-di(2- propyl)ethylamine (0.23 mL, 171 mg, 1.32 mmol) in N,N-dimethylformamide (2 mL) was added followed by benzoyl chloride (0.13 mL, 157 mg, 1.12 mmol). The mixture was shaken for 48 hours at room temperature. The drained resin was washed consecutively with dichloromethane (2 x 4 mL), methanol (2 x 4 mL) and tetrahydrofuran (4 mL). The resin was treated for 2 hours at room temperature with a solution of dry hydrogen chloride in tetrahydrofuran / ethyl ether / ethanol = 8:1:1 (0.1 M, 3 mL). The reaction mixture was drained and concentrated. The crude product was stripped with dichloromethane (1.5 mL) three times to yield the title compound.
HPLC-MS (Method C): m/z: 340 (M+1); Rt = 3.68 min.
1H-NMR (DMSO-dβ): δ 8.91 (1 H, s), 8.34 (1 H, d), 8.05 (1H, d), 7.78 (3H, m), 7.63 (3H, m),
7.46 (2H, m), 7.33 (1 H, dd).
The compounds in the following examples were prepared in a similar fashion.
Example 986 (General procedure (R)) Phenyl-[5-(2H-tetrazol-5-yl)-indol-1-yl]-methanone
Figure imgf000380_0001
HPLC-MS (Method C): m/z: 290 (M+1 ); Rt = 3.04 min.
1H-NMR (DMSO-dβ): δ 8.46 (1H, d), 8.42 (1H, d), 8.08 (1 H, dd), 7.82 (2H, d), 7.74 (1 H, t), 7.64 (2H, t), 7.55 (1H, d), 6.93 (1H, d).
Example 987 (General procedure (R)) (2,3-Difluorophenyl)-[5-(2H-tetrazol-5-yl)-indol-1-yl]-methanone
Figure imgf000381_0001
HPLC-MS (Method B): m/z = 326 (M+1 ); Rt = 3.85 min.
Example 988 (General procedure (R)) (2-Fluoro-3-trifluoromethylphenyl)-[5-(2H-tetrazol-5-yl)-indol-1-yl]-methanone
Figure imgf000381_0002
HPLC-MS (Method B): m/z = 376 (M+1); Rt = 4.32 min.
Example 989 (General procedure (R)) (3-Nitrophenyl)-[5-(2H-tetrazol-5-yl)-indol-1 -yl]-methanone
Figure imgf000381_0003
HPLC-MS (Method B): m/z = 335 (M+1); Rt = 3.72 min.
Example 990 (General procedure (R)) (4-Nitrophenyl)-[5-(2H-tetrazol-5-yl)-indol-1-yl]-methanone
Figure imgf000381_0004
HPLC-MS (Method B): m/z = 335 (M+1); Rt = 3.71 min. Example 991 (General procedure (R)) Naphthalen-2-yl-[5-(2H-tetrazol-5-yl)-indol-1-yl]-methanone
Figure imgf000382_0001
HPLC-MS (Method C): m/z = 340 (M+1); Rt = 4.25 min.
Example 992 (General procedure (R))
Figure imgf000382_0002
HPLC-MS (Method C): m/z: 354 (M+1); Rt = 3.91 min.
Example 993 (General procedure (R))
Figure imgf000382_0003
HPLC-MS (Method C): m/z: 418 (M+1); Rt = 4.39 min.
Example 994 (General procedure (R))
Figure imgf000382_0004
HPLC-MS (Method C): m/z: 370 (M+1); Rt = 4.01 min. Example 995 (General procedure (R))
Figure imgf000383_0001
HPLC-MS (Method C): m/z: 374 (M+1); Rt = 4.28 min.
Example 996 (General procedure (R))
Figure imgf000383_0002
HPLC-MS (Method C): m/z: 416 (M+1); Rt = 4.55 min.
Example 997 (General procedure (R))
Figure imgf000383_0003
HPLC-MS (Method C): m/z: 354 (M+1); Rt = 4.22 min. Example 998 (General procedure (R))
Figure imgf000384_0001
HPLC-MS (Method C): m/z: 358 (M+1); Rt = 3.91 min.
Example 999 (General procedure (R))
Figure imgf000384_0002
HPLC-MS (Method C): m/z: 390 (M+1); Rt = 4.38 min.
Example 1000 (General procedure (R))
Figure imgf000384_0003
HPLC-MS (Method C): m/z: 418 (M+1); Rt = 4.36 min.
Example 1001 (General procedure (R))
Figure imgf000384_0004
HPLC-MS (Method C): m/z: 304 (M+1); Rt = 3.32 min. Example 1002 (General procedure (R))
Figure imgf000385_0001
HPLC-MS (Method C): m/z: 368 (M+1 ); Rt = 3.84 min.
Example 1003 (General procedure (R))
Figure imgf000385_0002
HPLC-MS (Method C): m/z: 320 (M+1 ); Rt = 3.44 min.
Example 1004 (General procedure (R))
Figure imgf000385_0003
HPLC-MS (Method C): m/z: 324 (M+1); Rt = 3.73 min. Example 1005 (General procedure (R))
Figure imgf000386_0001
HPLC-MS (Method C): m/z: 304 (M+1 ); Rt = 3.64 min.
Example 1006 (General procedure (R))
Figure imgf000386_0002
HPLC-MS (Method A): m/z: 308 (M+1); Rt = 3.61 min.
Example 1007 (General procedure (R))
Figure imgf000386_0003
HPLC-MS (Method C): m/z: 368 (M+1); Rt = 3.77 min.
Example 1008 (General procedure (R))
Figure imgf000386_0004
HPLC-MS (Method A): (sciex) m/z: 326 (M+1); Rt = 3.73 min. HPLC-MS (Method C): m/z: 326 (M+1); Rt = 3.37 min.
Example 1009 (General procedure (R))
Figure imgf000387_0001
HPLC-MS (Method C): m/z: 374 (M+1 ); Rt = 4.03 min.
General procedure (Q) for preparation of compounds of general formula l1β:
Figure imgf000387_0002
wherein PS is polymeric support, a Tentagel S RAM resin, n is 1 - 20, m is 0 - 5, and p is 0 or 1. The compounds of the invention of general formula (l2) can be prepared by means of standard peptide chemistry (General procedure H), e.g. in 0.5 mmol scale, using Fmoc strategy and HOAt or HOBT activated amino acids. The compounds prepared in the following examples according to General procedure (Q) were all isolated as the TFA salts. This procedure is further illustrated in the following:
Typically, 2 gram of Fmoc Tentagel S RAM resin (Rapp Polymere, Tubingen) with substitution 0,25 mmol/g was washed with NMP then treated with 25% piperidine in NMP for 30 min followed by wash with NMP which renders the resin ready for coupling. Step wise coupling of Fmoc-Arginine (Fmoc-Arg(Pmc)-OH), Fmoc-Glycine (Fmoc-Gly-OH) and Fmoc-4-aminobenzoic acid (Fmoc-4-Abz-OH):
To 2 mmol of Fmoc-L-Arg(Pmc)-OH (Novabiochem) was added 3,33 ml 0,6M HOAt in NMP (Perseptives) or 0,6M HOBT in NMP (Novabiochem) containing 0,2% bromphenolblue as indicator and added 330 μl of diisopropylcarbodiimide DIC (Fluka) and the solution was then added to the resin. After coupling for minimum 1 hour, or when the blue colour disappeared, the resin was washed with NMP and the Fmoc group was deprotected with 25% piperidine in NMP for 20 minutes followed by wash with NMP. This stepwise assembling of the arginine residues was repeated to give 3, 4, 5 or 6 arginines on the resin. The Fmoc-Glycine (Novabiochem) and Fmoc-4-aminobenzoic acid (Fluka and Neosystems) were coupled using the same procedure as described for Fmoc-Arg(Pmc)-OH. Coupling of A-OH, e.g. 1H-benzotriazole-5-carboxylic acid on Gly.
When A-OH, e.g. 1H-benzotriazole-5-carboxylic acid (Aldrich) was coupled on a glycine or arginine residue the coupling procedure was as described above.
Coupling of A-OH, e.g. 1H-benzotriazole-5-carboxylic acid on Abz or 4-Apac:
Due to the lower nucleophilicity of the amino group in Abz the following procedure was nec- essary. To 4 mmol of A-OH, e.g. 1 H-benzotriazole-5-carboxylic acid was added 6,66 ml of a solution of 0,6M HOAt, 0,2 mmol dimethylaminopyridine (DMAP) and 4 mmol DIC and was then added to the resin and allowed to react overnight.
Introduction of fragment 4-Apac instead of 4-Abz: 4-Nitrophenoxyacetic acid may be coupled on a glycine or arginine residue using DIC and HOBT/HOAt as described above. Subsequent reduction of the nitro group may be done using SnCI2 in NMP or DMF e.g. as described by Tumelty et al. (Tet. Lett., (1998) 7467-70>. Cleavage of the peptides from the resin. After synthesis the resin was washed extensively with diethyl ether and dried. To 1 gram of the peptidyl resin was added 25 ml of a TFA solution containing 5% thioanisole, 5% ethanol, 5% phenol and 2% triisopropylsilane and allowed to react for 2 hours. The TFA solution was filtered and concentrated with argon flow for approximately 30 minutes. Then diethylether ca. 5-7 times the residual volume of TFA was added and the peptide precipitate was extracted in 10% AcOH and washed 5 times with diethyl ether and lyophilized.
RP-HPLC analysis and purification: The crude products were analysed on RP-HPLC C18 column (4,6 x 250 mm) using one of two gradients (see table 1 and 2), temperature 25°C, wavelength 214 nm and flow rate 1 ml/min with A-buffer 0,15 % (w/w) TFA in H20 and B-
Buffer (87,5 % (w/w) MeCN, 0,13 % (w/w) TFA in H20).
The products were purified on preparative RP-HPLC C18 column (2x25 cm) using a gradient
(variable, see e.g example 1013 and similar), temperature 25°C, wavelength 214 nm and flow rate
6 ml/min with A-buffer 0,15 % (w/w) TFA in H20 and B-Buffer (87,5 % (w/w) MeCN, 0,13 % (w/w)
TFA in H20) and verified by mass spectrometry (MALDI).
Table 1 :
Figure imgf000389_0001
Table 2:
Figure imgf000389_0002
Figure imgf000390_0003
The following examples were prepared using this general procedure (O).
Example 1010 (General Procedure (Q))
Benzotriazol-5-ylcarbonyl-Gly2-Arg3-NH2 (BT-G2R3).
Figure imgf000390_0001
MS (MALDI): m/z: 746.7 g/mol; calculated: 744.2 g/mol. HPLC gradient:
Figure imgf000390_0004
Example 1011 (General Procedure (Q))
Benzotriazol-5-ylcarbonyl-Gly2-Arg4-NH2 (BT-G2R4).
Figure imgf000390_0002
MS (MALDI): m/z: 903.0 g/mol; calculated: 900.6 g/mol. HPLC gradient:
Figure imgf000391_0002
Example 1012 (General Procedure (Q))
Benzotriazol-5-ylcarbonyl-Gly2-Arg5-NH2 (BT-G2R5).
Figure imgf000391_0001
MS (MALDI): m/z: 1060.8 g/mol; calculated: 1057 g/mol. HPLC gradient
Figure imgf000391_0003
Example 1013 (General Procedure (Q)) Benzotriazol-5-ylcarbonyl-Gly2-Arg6-NH2 (BT-G2Re).
Figure imgf000392_0001
MS (MALDI): m/z: 1214.8 g/mol; calculated: 1213.4 g/mol. HPLC gradient:
Figure imgf000392_0003
Example 1014 (General Procedure (Q))
Benzotriazol-5-ylcarbonyl-4-Abz-Gly2-Arg5-NH2 (BT-4-Abz-G2R5).
Figure imgf000392_0002
MS (MALDI): m/z: 1176.7 g/mol; calculated: 1177.9 g/mol. HPLC gradient:
Figure imgf000392_0004
Figure imgf000393_0003
Example 1015 (General Procedure (Q))
Benzotriazol-5-ylcarbonyl-4-Abz-Gly-Arg5-NH2 (BT-4-Abz-GR5)
Figure imgf000393_0001
MS (MALDI): m/z: 1122 g/mol; calculated: 1120.4 g/mol. HPLC gradient:
Figure imgf000393_0004
Example 1016 (General Procedure (Q))
Benzotriazol-5-ylcarbonyl-4-Abz-Arg5-NH2 (BT-4-Abz-R5).
Figure imgf000393_0002
MS (MALDI): m/z: 1064.3 g/mol; calculated: 1063.2 g/mol. HPLC gradient:
Figure imgf000394_0003
General procedure (R) for preparation of compounds of general formula l17:
Figure imgf000394_0001
I .7 wherein X, Y, R >110U, r E, n
Figure imgf000394_0002
B2 are as defined above, p is 0 or 1 , m is 0-5 and n is 1-20.
PS is a polymeric support, e.g. TentagenS RAM resin or a Rink amide resin. This general procedure is very similar to General procedure (Q), where benzotriazole-5- carboxylic acid in the last step before cleavage from the resin is replaced with compounds optionally prepared according to general procedure (D):
Figure imgf000395_0001
Example 1017 (General Procedure (R))
4-{2-[3-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenoxy]acetylamino}benzoyl-Gly2-Arg5-NH2
Figure imgf000395_0002
Example 1018 (General Procedure (R)) 3-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenyl]acryloyl-Arg5-NH2
Figure imgf000395_0003
MS (MALDI): m/z: 1057.3 g/mol; calculated: 1055.3 g/mol.
Example 1019 (General Procedure (R)) 3-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenyl]acryloyl-Arg4-NH2
Figure imgf000396_0001
MS (MALDI): m/z: 899.1 g/mol; calculated: 901.6 g/mol.
Example 1020 (General Procedure (R))
3-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenyl]acryloyl-Arg3-NH2
Figure imgf000396_0002
MS (MALDI): m/z: 746.2 g/mol; calculated: 742.9 g/mol.
Example 1021 (General Procedure (R))
4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenoxy]butyryl-Arg5-NH2.
Figure imgf000397_0001
MS (MALDI): m/z: 1088.7 g/mol; calculated: 1087 g/mol.
Example 1022 (General Procedure (R))
4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenoxy]butyryl-Arg4-NH2.
Figure imgf000397_0002
MS (MALDI): m/z: 933.0 g/mol; calculated: 931 g/mol.
Example 1023 (General Procedure (R)) 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenoxy]butyryl-Arg3-NH2. H2
Figure imgf000397_0003
MS (MALDI): m/z: 776.9 g/mol; calculated: 774.0 g/mol.
Example 1024 (General Procedure (R))
4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]butyryl-Arg12-NH2.
Figure imgf000398_0001
MS (MALDI): m/z: 2232.9.4 g/mol; calculated: 2230.3 g/mol.
Example 1025 (General Procedure (R))
4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]butyryl-Arg8-NH2.
Figure imgf000398_0002
MS (MALDI): m/z: 1607.4 g/mol; calculated: 1605.5 g/mol.
Example 1026 (General Procedure (R))
4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]butyryl-Arg5-NH2.
Figure imgf000399_0001
MS (MALDI): m/z: 1141.9 g/mol; calculated: 1137.4 g/mol.
Example 1027 (General Procedure (R))
4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]butyryl-Arg4-NH2.
Figure imgf000399_0002
MS (MALDI): m/z: 985.4 g/mol; calculated: 981.2 g/mol.
Example 1028 (General Procedure (R)) 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]butyryl-Arg3-NH2. H2
Figure imgf000399_0003
MS (MALDI): m/z: 828.5 g/mol; calculated: 825.0 g/mol.
The following compounds were prepared according to the methodology described in general procedure (Q) and (R):
Example 1029
4-(2H-Tetrazol-5-yl)benzoyl-4-Abz-Gly2-Arg5-NH2
Figure imgf000400_0001
MS (MALDI): m/z: 1203.8 g/mol; calculated: 1203.8 g/mol.
Example 1030
4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoyl-Arg5-NH2
Figure imgf000400_0002
MS (MALDI): m/z: 1152.5 g/mol; calculated: 1149.3 g/mol.
Example 1031
4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoyl-Arg8-NH2
Figure imgf000401_0001
MS (MALDI): m/z: 1621.0 g/mol; calculated: 1617.5 g/mol.
Example 1032
4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoyl-Arg12-NH2
Figure imgf000401_0002
MS (MALDI): m/z: 2247.9 g/mol; calculated: 2242.3 g/mol.
General procedure (S) for preparation of compounds of general formula l18:
H-(
H-(Gly)p-
Figure imgf000402_0001
CGr-Lnk-(Gly)p-(Lys)-(Arg)-N-PS
H
CGr-Lnk-(Gly)p-(Lys)-(Arg)-NH2 <
'« Wherein PS is polymeric support, a Rink Amide AM resin, n is 0 - 20, m is 0 - 20, with the proviso that n + m <20, wherein (Gly)p is defined as in a broader sense as Frg1 above, p is 0 - 5, and furthermore the Frg2 (the Lys and Arg residues) may comprise one or more neutral amino acids independently selected from the group consisting of Gly, Ala, Thr, and Ser, and wherein CGr-Lnk-OH, CGr, and Lnk is as described above.
The compounds of the invention of general formula (l18) can be prepared by means of standard peptide chemistry (as e.g. in General Procedure Q or R), e.g. in 7 mmol scale, using Fmoc strategy and HOAt or HOBt activated amino acids. The compounds prepared in the following examples according to General Procedure (S) were all isolated as the TFA salts. This procedure is further illustrated in the following:
Typically, 10 gram (7 mmol) of Rink Amide AM resin (Novabiochem 200-400 mesh) with substitution 0,70 mmol/g was treated with NMP by shaking for 3 h or more -then treated with NMP/Piperidine/DBU (80/20/2) for (30 - 60 min) x 2 followed by wash with NMP x 6 which renders the resin ready for coupling. Step wise coupling of Fmoc-Arginine (Fmoc-Arg(Pbf)-OH).
21 mmol (13.6 g) of Fmoc-L-Arg(Pbf)-OH (MultiSyn Tech Gmbh., Germany) was dissolved in NMP (80 mL) together with HOBt-hydrate (21 mmol, 3.21 g). DIC (21 mmol, 3.27 mL) was added to the solution and the mixture kept for 2-5 min before it was added to the resin which was shaken for a minimum 3 h, the resin was washed with NMP (80 mL). After each coupling step a capping step was performed using 20 x molar excess of Acetic acid / HOBt/DIC (8.4 g/21.4 g/ 21 mL) in NMP (80 mL) followed by washing with NMP (80 mL) x 4 and the coupling was checked with TNBS (no red colour) The Fmoc group was deprotected with NMP/Piperidine/DBU (80/20/2) for (30 - 60 min) x 2 followed by wash with NMP. Then another Fmoc-L-Arg(Pbf)-OH group was coupled as described above. This stepwise assembling of the arginine residues was repeated to give the wanted number of arginines on the resin. When more than 6 residues were added double couplings were performed on each coupling step, one coupling carried out for 3 h or more- the other overnight. When Lysine or Glycine were part of the synthesised molecules the same procedure as described above was used changing Fmoc-L-Arg(Pbf)-OH to Fmoc-L-Lys(Boc)-OH or Fmoc- Gly-OH, respectively.
Coupling of Ligand (CGr-Lnk-OH) to the polymer supported amino acid chain. The attachment of the ligand was carried out after deprotection of Fmoc as described above followed by coupling with the ligand containing a carboxylic acid using HOAt /DIC for activation using the same ratio as described above.
Double couplings and capping as described above were performed.
Cleavage from the resin.
The resin was washed with DCM (80 mL) x 4 followed by diethyl ether (80 mL ) x 4.
Subsequently the resin was dried well and then added to a 20 fold excess of a mixture of TFA / Thioanisol / Ethanol (90/5/5) and the mixture was stirred at RT overnight, evaporated to almost dryness and the residue was poured into a 20 fold excess of cold diethyl ether. The mixture was stirred for 30 min, filtered and the precipitate was washed with ether twice and dried. The dried compound was then dissolved in water and freeze dried, and the freeze dried material was then purified by HPLC. RP-HPLC purification: The crude products were purified on RP-HPLC Kromasil RP18 10θA 50x350 mm, flow lOOmL/min; gradient: 0.0-2.0 min: 20% acetonotrile,0.1 % TFA; 2.0 17.0 min: 20 % acetonitrile, ,0.1 % TFA to 28 % acetonitrile, ,0.1 % TFA; 17.0-25.0 100 % acetonitrile, 0.1% TFA. Alternatively other HPLC systems were used. The identity was verified by mass spectrometry (MALDI-TOF).
The following examples were prepared using this general procedure
Example 1033 (General Procedure (S)) 5-[6-(5-Cyano-1H-[1,2,3]triazol-4-yl)naphthalen-2-yloxy]pentanoyl-Arg12-NH2
Figure imgf000404_0001
MS (MALDI-TOF): m/z: 2210 g/mol; calculated: 2209.2 g/mol.
Example 1034 (General Procedure (S)) 4'-[5-(2H-Tetrazol-5-yl)indol-1-ylmethyl]biphenyl-4-carbonyl-Arg12-NH2
Figure imgf000404_0002
MS (MALDI-TOF): m/z: 2268 g/mol; calculated: 2269 g/mol.
Example 1035 (General Procedure (S)) 4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoyl-Arg14-NH2
Figure imgf000404_0003
MS (MALDI-TOF): m/z: 2553 g/mol; calculated: 2554 g/mol.
Example 1036 (General Procedure (S)) 4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoyl-Lys2-Arg12-NH2
Figure imgf000405_0001
MS (MALDI-TOF): m/z: 2498 g/mol; calculated: 2499 g/mol
Example 1037 (General Procedure (S)) 4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoyl-Gly4-Arg8-NH2
Figure imgf000405_0002
MS (MALDI-TOF): m/z: 1845 g/mol; calculated: 1846 g/mol.
Example 1038 (General Procedure (S)) 3-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoyl-Arg12- NH2
Figure imgf000405_0003
MS (MALDI-TOF): m/z: 2242 g/mol; calculated: 2243 g/mol. Example 1039 (General Procedure (S)) 4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoyl-Gly2-Arg12-
Figure imgf000406_0001
MS (MALDI-TOF): m/z: 2497.5 g/mol; calculated: 2496 g/mol.
Example 1040 (General Procedure (S)) 4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoyl-Argιβ-
NH2
Figure imgf000406_0002
MS (MALDI-TOF): m/z: 2873 g/mol; calculated: 2864 g/mol.
Example 1041 (General Procedure (S))
4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoyl-Lys12-
NH2
Figure imgf000406_0003
MS (MALDI-TOF): m/z: 1905 g/mol; calculated: 1904 g/mol. Example 1042 (General Procedure (S)) 4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoyl-Arg6-
Figure imgf000407_0001
MS (MALDI-TOF): m/z: 1303 g/mol; calculated: 1304 g/mol.
Example 1043 (General Procedure (S)) 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]butyryl-Arg6-
Figure imgf000407_0002
MS (MALDI-TOF): m/z: 1293 g/mol; calculated: 1292 g/mol.
Other preferred compounds of the invention that may be prepared according to general procedures (Q), (R) and / or general procedure (S) include:
Building block from example 469:
4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]butyryl-Arg10-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]butyryl-Arg9-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]butyryl-Arg7-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]butyryl-Arg11-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]butyryl-Arg13-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]butyryl-Arg14-NH2 4-[4-(2,4-Dioxothiazolidin-5-yIidenemethyl)naphthalen- -yloxyjbutyry l-Arg15-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen- -yloxy]butyry l-Arg16-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen- -yloxyjbutyry l-Arg17-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen- -yloxyjbutyry l-Arg18-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen- -yloxyjbutyry l-Arg19-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen- -yloxyjbutyry I-Arg20-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen- -yloxyjbutyry l-Lys6-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen- -yloxyjbutyry l-Lys5-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen- -yloxyjbutyryl l-Lys4-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen- -yloxyjbutyryl l-Lys3-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen- -yloxyjbutyry l-Lys7-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen- -yloxyjbutyryl l-Lys8-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen- -yloxyjbutyry l-Lys9-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen- -yloxyjbutyry! l-Lys10-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen- -yloxyjbutyry l-LySl 1-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen- -yloxyjbutyry l-Lys12-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen- -yloxyjbutyry l-Lys13-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen- -yloxyjbutyry l-Lys14-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen- -yloxyjbutyry l-Lys15-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen- -yloxyjbutyryl l-Lys16-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen- -yloxyjbutyryl l-Lys17-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen- -yloxyjbutyryl l-Lys18-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen- -yloxyjbutyryl l-Lys19-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen- -yloxyjbutyryl l-Lys2o-NH2
Building block from example 470:
5-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen- -yloxy]pentanoyl-Arg6-NH2
5-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen- -yloxy]pentanoyl-Arg5-NH2
5-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen- -yloxy]pentanoyl-Arg4-NH2
5-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen- -yloxy]pentanoyl-Arg3-NH2
Building block from page 232:
6-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen- -yloxy]hexanoyl-Arg3-NH2
6-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen- -yloxy] hexa noyl-Arg4-N H2
6-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen- -yloxyJhexanoyl-Arg5-NH2
Building block from page 232:
7-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen -1 -yloxy]heptanoyl-Arg3-NH2 7-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxyJheptanoyl-Arg4-NH2
7-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]heptanoyl-Arg5-NH2
Building block from page 232:
8-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]octanoyl-Arg3-NH2
8-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]octanoyl-Arg4-NH2
8-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]octanoyl-Arg5-NH2
Building block from page 232:
10-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]decanoyl-Arg3-NH2
10-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]decanoyl-Arg4-NH2
10-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]decanoyl-Arg5-NH2
Building block from page 232:
11-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]undecanoyl-Arg3-NH2
11-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]undecanoyl-Arg4-NH2
11 -[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]undecanoyl-Arg5-NH2
Building block from page 232:
12-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxyJdodecanoyl-Arg3-NH2
12-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]dodecanoyl-Arg4-NH2
12-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxyJdodecanoyl-Arg5-NH2
Building block from page 233:
15-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]pentadecanoyl-Arg3-NH2
15-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxyJpentadecanoyl-Arg4-NH2
15-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]pentadecanoyl-Arg5-NH2
Building block from example 283:
4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1-yloxy]butyryl-Arg10-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1-yloxyJbutyryl-Argg-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1-yloxy]butyryl-Arg8-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1-yloxy]butyryl-Arg7-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1-yloxyJbutyryl-Arg6-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1-yloxy]butyryl-Arg5-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1-yloxy]butyryl-Arg4-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1-yloxyJbutyryl-Arg3-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1-yloxy]butyryl-Arg11-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1-yloxy]butyryl-Arg12-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1-yloxy]butyryl-Arg13-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1-yloxy]butyryl-Arg14-NH2 4-[4-(2,4-Dioxothiazolidin-5-ylmethy )naphthalen 1 -yloxyjbutyry l-Arg15-NH2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl )naphthalen 1 -yloxyjbutyry l-Arg16-NH2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl )naphthalen 1 -yloxyjbutyry l-Arg17-NH2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl )naphthalen 1 -yloxyjbutyry l-Arg18-NH2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl )naphthalen- 1 -yloxyjbutyry l-Arg19-NH2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl )naphthalen- 1 -yloxyjbutyry l-Arg20-NH2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl )naphthalen 1 -yloxyjbutyryl l-Lys6-NH2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl )naphthalen- 1 -yloxyjbutyry l-Lys5-NH2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl )naphthalen- 1 -yloxyjbutyry l-Lys4-NH2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl )naphthalen 1 -yloxyjbutyry] l-Lys3-NH2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl )naphthalen- 1 -yloxyjbutyryl l-Lys7-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen 1 -yloxyjbutyryl l-Lys8-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylmethyl )naphthalen 1 -yloxyjbutyry! l-Lys9-NH2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl )naphthalen- 1 -yloxyjbutyryl l-Lys10-NH2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl )naphthalen 1 -yloxyjbutyry l-Lysι NH2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl )naphthalen 1 -yloxyjbutyryl l-Lysι2-NH2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl )naphthalen 1 -yloxyjbutyry l-Lysι3-NH2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl )naphthalen 1 -yloxyjbutyry l-Lys14-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen- 1 -yloxyjbutyryl l-Lys15-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen 1 -yloxyjbutyryl l-Lys16-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen 1 -yloxyjbutyry l-Lys17-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen- 1 -yloxyjbutyryl l-Lys18-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen 1 -yloxyjbutyryl l-Lys19-NH2
4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen 1 -yloxyjbutyryl l-Lys2o-NH2
Building block from example 476:
2-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]acetyl-Arg6-NH2
2-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalene-1-yloxy]acetyl-Arg5-NH2
2-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalene-1-yloxy]acetyl-Arg4-NH2
2-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalene-1-yloxy]acetyl-Arg3-NH2
Building block from example 480:
2-{5-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)benzylidene]-4-oxo-2-thioxothiazolidin-3- yl}acetyl-Arge-NH2
2-{5-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)benzylideneJ-4-oxo-2-thioxothiazolidin-3- yl}acetyl-Arg5-NH2
2-{5-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)benzylidene]-4-oxo-2-thioxothiazolidin-3- yl}acetyl-Arg4-NH2
2-{5-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)benzylidene]-4-oxo-2-thioxothiazolidin-3- yl}acetyl-Arg3-NH2
4-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]butyryl-Arg6-NH2
4-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]butyryl-Arg5-NH2
4-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]butyryl-Arg4-NH2
4-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]butyryl-Arg3-NH2
15-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]pentadecanoyl-Arg6-NH2
15-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]pentadecanoyl-Arg5-NH2
15-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]pentadecanoyl-Arg4-NH2
15-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]pentadecanoyl-Arg3-NH2
5-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]pentanoyl-Arg6-
NH2
5-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]pentanoyl-Arg5-
NH2
5-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]pentanoyl-Arg4-
NH2
5-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]pentanoyl-Arg3-
NH2
Building block from example 462:
3-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenyl]acryloyl-Arg6-NH2
Building block from example 473:
2-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]acetyl-Arg6-NH2
2-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]acetyl-Arg5-NH2
2-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]acetyl-Arg4-NH2
2-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]acetyl-Arg3-NH2
8-[6-(2,4-Dioxothiazolidin-5-yl denemethyl)naphthalen-2-yloxy]octanoyl-Arg6-NH2 8-[6-(2,4-Dioxothiazolidin-5-yl denemethyl)naphthalen-2-yloxyJoctanoyl-Arg5-NH2 8-[6-(2,4-Dioxothiazolidin-5-yl denemethyl)naphthalen-2-yloxy]octanoyl-Arg4-NH2 8-[6-(2,4-Dioxothiazolidin-5-yl denemethyl)naphthalen-2-yloxy]octanoyl-Arg3-NH2 6-[6-(2,4-Dioxothiazolidin-5-yl denemethyl)naphthalen-2-yloxy]hexanoyl-Arg6-NH2 6-[6-(2,4-Dioxothiazolidin-5-yl denemethyl)naphthalen-2-yloxy]hexanoyl-Arg5-NH2 6-[6-(2,4-Dioxothiazolidin-5-yl denemethyl)naphthalen-2-yloxy]hexanoyl-Arg4-NH2 6-[6-(2,4-Dioxothiazolidin-5-yl denemethyl)naphthalen-2-yloxy]hexanoyl-Arg3-NH2
Building block from example 466: 4-[2-Chloro-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]butyryl-Arg6-NH2
4-[2-Chloro-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]butyryl-Arg5-NH2
4-[2-Chloro-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]butyryl-Arg4-NH2
4-[2-Chloro-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]butyryl-Arg3-NH2
Building block from example 460:
4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenoxy]butyryl-Arg6-NH2
Building block from example 467:
4-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]butyryl-Arg6-NH2
4-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]butyryl-Arg5-NH2
4-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]butyryl-Arg4-NH2
4-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]butyryl-Arg3-NH2
11-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]undecanoyl-Arg6-NH2
11-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]undecanoyl-Arg5-NH2
11-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]undecanoyl-Arg4-NH2
11-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]undecanoyl-Arg3-NH2
4-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]butyryl-Arg6-NH2
4-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxyJbutyryl-Arg5-NH2
4-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]butyryl-Arg4-NH2
4-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxyJbutyryl-Arg3-NH2
Building block from example 464:
4-(2,4-Dioxothiazolidin-5-ylidenemethyl)benzoyl-Arg6-NH2
4-(2,4-Dioxothiazolidin-5-ylidenemethyl)benzoyl-Arg5-NH2
4-(2,4-Dioxothiazolidin-5-ylidenemethyl)benzoyl-Arg4-NH2
4-(2,4-Dioxothiazolidin-5-ylidenemethyl)benzoyl-Arg3-NH2
Building block from example 463:
2-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenoxyJacetyl-Arg6-NH2
2-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenoxy]acetyl-Arg5-NH2
2-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenoxy]acetyl-Arg4-NH2
2-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenoxy]acetyl-Arg3-NH2
Building block from example 461 :
2-[3-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenoxy]acetyl-Arg6-NH2
2-[3-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenoxy]acetyl-Arg5-NH2
2-[3-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenoxy]acetyl-Arg4-NH2
2-[3-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenoxy]acetyl-Arg3-NH2
Building block from example 474: 4-[3-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenoxy]butyryl-Arg6-NH2 4-[3-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenoxy]butyryl-Arg5-NH2 4-[3-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenoxy]butyryl-Arg4-NH2 4-[3-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenoxy]butyryl-Arg3-NH2 Building block from example 468:
4-[2-Bromo-4-(4-oxo-2-thioxothiazolidin-5-ylidenemethyl)phenoxy]butyryl-Arg6-NH2 4-[2-Bromo-4-(4-oxo-2-thioxothiazolidin-5-ylidenemethyl)phenoxy]butyryl-Arg5-NH2 4-[2-Bromo-4-(4-oxo-2-thioxothiazolidin-5-ylidenemethyl)phenoxy]butyryl-Arg4-NH2 4-[2-Bromo-4-(4-oxo-2-thioxothiazolidin-5-ylidenemethyl)phenoxy]butyryl-Arg3-NH2 Building block from example 738:
4-[3-(2H-Tetrazol-5-yl l)carbazol 9-ylmethyl]benzoyl l-Arg4-NH2 4-[3-(2H-Tetrazol-5-y i)carbazol 9-ylmethyl]benzoy l-Arg3-NH2 4-[3-(2H-Tetrazol-5-yl l)carbazol 9-ylmethyl]benzoyl l-Arg7-NH2 4-[3-(2H-Tetrazol-5-y l)carbazol 9-ylmethyrjbenzoyl l-Arg9-NH2 4-[3-(2H-Tetrazol-5-yl l)carbazol 9-ylmethyl]benzoyl l-Arg10-NH2 4-[3-(2H-Tetrazol-5-y l)carbazol 9-ylmethyl]benzoyl l-Argn-NH2 4-[3-(2H-Tetrazol-5-yl l)carbazol 9-ylmethyl]benzoyl l-Arg13-NH2 4-[3-(2H-Tetrazol-5-yl l)carbazol 9-ylmethyl] benzoyl l-Arg15-NH2 4-[3-(2H-Tetrazol-5-yl l)carbazol 9-ylmethyl]benzoy l-Arg17-NH2 4-[3-(2H-Tetrazol-5-yl l)carbazol •9-ylmethyl]benzoy l-Arg18-NH2 4-[3-(2H-Tetrazol-5-yl l)carbazol 9-ylmethyl]benzoy l-Arg19-NH2 4-[3-(2H-Tetrazol-5-yl l)carbazol 9-ylmethyl]benzoy l-Arg20-NH2 4-[3-(2H-Tetrazol-5-yl l)carbazol 9-ylmethyl]benzoy l-Lys6-NH2 4-[3-(2H-Tetrazol-5-yl l)carbazol •9-ylmethyl]benzoy l-Lys4-NH2 4-[3-(2H-Tetrazol-5-yl l)carbazol 9-ylmethyl]benzoyl l-Lys3-NH2 4-[3-(2H-Tetrazol-5-yl l)carbazol ■9-ylmethyl]benzoy l-Lys7-NH2 4-[3-(2H-Tetrazol-5-yl l)carbazol •9-ylmethyl]benzoy l-Lys8-NH2 4-[3-(2H-Tetrazol-5-yl l)carbazol 9-ylmethyl]benzoy l-Lys9-NH2 4-[3-(2H-Tetrazol-5-yl l)carbazol ■9-ylmethyl]benzoy l-LySιo-NH2 4-[3-(2H-Tetrazol-5-yl l)carbazol ■9-ylmethyl]benzoy I-Lysn-NH, 4-[3-(2H-Tetrazol-5-yl l)carbazol ■9-ylmethyl]benzoyl l-Lys13-NH2 4-[3-(2H-Tetrazol-5-yl l)carbazol ■9-ylmethyl]benzoy l-Lys14-NH2 4-[3-(2H-Tetrazol-5-yl l)carbazol •9-ylmethyl]benzoy l-Lys15-NH2 4-[3-(2H-Tetrazol-5-yl l)carbazol •9-ylmethyl]benzoyl l-Lys16-NH2 4-[3-(2H-Tetrazol-5-yl l)carbazol •9-ylmethyl]benzoyl l-Lys17-NH2 4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoyl-Lys18-NH2 4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoyl-Lys19-NH2 4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoyl-Lys2o-NH2 Building block from page 322:
4,-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]biphenyl-4-carbonyl-Arg6-NH2 4'-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]biphenyl-4-carbonyl-Arg5-NH2 4'-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]biphenyl-4-carbonyl-Arg4-NH2 4'-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]biphenyl-4-carbonyl-Arg3-NH2 Building block from example 743:
3-[3-(2H-Tetrazol-5-yl)carbazo! l-9-ylmethyl]benzoyl-Arg6-NH2 3-[3-(2H-Tetrazol-5-yl)carbazol l-9-ylmethyl]benzoyl-Arg5-NH2 3-[3-(2H-Tetrazol-5-yl)carbazol l-9-ylmethyl]benzoyl-Arg4-NH2 3-[3-(2H-Tetrazol-5-yl)carbazol l-9-ylmethyl]benzoyl-Arg3-NH2 3-[3-(2H-Tetrazol-5-yl)carbazol l-9-ylmethyl]benzoyl-Arg7-NH2 3-[3-(2H-Tetrazol-5-yl)carbazol l-9-ylmethyl]benzoyl-Arg8-NH2 3-[3-(2H-Tetrazol-5-yl)carbazol l-9-ylmethyl] benzoyl-Arg9-N H2 3-[3-(2H-Tetrazol-5-yl)carbazol l-9-y I methyl] benzoy l-Arg i 0-N H2 3-[3-(2H-Tetrazol-5-yl)carbazo l-9-ylmethyl]benzoyl-Argn-NH2 3-[3-(2H-Tetrazol-5-yl)carbazo l-9-y I methyl] benzoy l-Arg 12-N H2 3-[3-(2H-Tetrazol-5-yl)carbazo l-9-ylmethyl]benzoyl-Arg13-NH2 3-[3-(2H-Tetrazol-5-yl)carbazo l-9-yl methyl] benzoy l-Arg , 4-N H2 3-[3-(2H-Tetrazol-5-yl)carbazo l-9-ylmethyl]benzoyl-Arg15-NH2 3-[3-(2H-Tetrazol-5-yl)carbazo l-9-ylmethyl]benzoyl-Arg16-NH2
Example 1044
Equilibrium Solubility. For pH-solubility profiles, a 0.6 mM insulin stock solution containing 0.3 mM Zn2+, 30 mM phenol, 1.6% glycerol and Zn2+ -binding ligand as required were prepared and the pH was adjusted to the desired value corresponding to the alkaline endpoint of the pH-solubility profile. From these stock solutions samples were withdrawn, the pH adjusted to the desired value in the pH 3-8 range, and samples were incubated at 23 C for 24 hours. After centrifugation (20,000 g in 20 min at 23 °C) of each sample, pH was measured and the solubility was determined by quantification of insulin contents in the supernatant by SEC HPLC analysis The effect of various concentration of the ligand 4-[3-(2H-tetrazol-5-yl)carbazol-9- ylmethyl]benzoyl-Arg12-NH2 (Example 1032) on the pH-dependence of Gly*21, Asp628 insulin solubility is illustrated in Figure 1.
Example 1045
The effect of a high concentration of the ligand 4-[3-(2H-tetrazol-5-yl)carbazol-9- ylmethyl]benzoyl-Arg12-NH2 (Example 1032)on the pH-dependence of Gly*21 insulin solubility is illustrated in Figure 2. The solubility was determined as in example 1044. Solution conditions: 0.6 mM human insulin, 0.3 mM mM Zn2+, 30 mM phenol, 1.6% glycerol, 23 °C.
Example 1046
The slow release (prolonged action) properties of certain formulations of the present invention was characterized by the disappearance rate from the subcutaneous depot following subcutaneous injections in pigs. T5o% is the time when 50% of the A14 Tyr(125l) insulin has disappeared from the site of injection as measured with an external v-counter (Ribel et al., The Pig as a Model for Subcutaneous Absorption in Man. In: M. Serrano-Rtios and P.J. Le- febre (Eds): Diabetes (1985) Proceedings of the 12th congress of the International Diabetes Federation, Madrid, Spain, 1985 (Excerpta Medica, Amsterdam (1986), 891-896). The com- position of a series of protracted formulations is given in the table below together with the Tso% values. The disappearance curves are illustrated in Figure 3 a-c, e-f. For comparison, the T500- for the corresponding insulin preparations formulated without the ligands would be about 2 hours. The disappearance curve for B29-Nε-myristoyl-des(B30) human insulin (Figure 3 d) is 11 hours.
The induction of slow release by addition of exogenous ligands of the invention affords further advantages in terms of versatility regarding the choice of insulin species and release patterns. Consequently, human or mutant insulins such as Gly*21, Gly^Asp828 may be formulated as slow- or dual-release preparations by adding variable amounts of His810 Zn2+-site ligand. This is illustrated below for Gly*21, Asp628 human insulin and Gly*21 human insulin employing different Zn2+-site ligands. As shown in the table below and in Figure 3 panels a-c and e-f, addition of ligand produces a slow release preparation with Tso% in the range of 5 to 16 hours.
Figure imgf000416_0001
Figure 3 shows the disappearance from the subcutaneous depot (pig model) of insulin preparations. Curves a)-c) are Gly*21, AspB28 human insulin formulated with an excess concentration compared to Zn2+ of a) 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1- yloxy]butyryl-Arg3-NH2, b) 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1- yloxy]butyryl-Arg5-NH2 and c) 4-(2H-Tetrazol-5-yl)benzoyl-Abz-Gly2-Arg5-NH2. Curve d) is B29-Nε-myristoyl-des(B30) human insulin. Curves e) and f) are Gly*21 human insulin formulated with two different excess concentrations compared to Zn2+ of 4-[3-(2H-Tetrazol-5- yl)carbazol-9-ylmethyl]benzoyl-Arg12-NH2. ANALYTICAL METHODS
Assays to quantify the binding affinity of ligands to the metal site of the insulin R6 hexamers:
4H3N-assay:
The binding affinity of ligands to the metal site of insulin Re hexamers are measured in a UV/vis based displacement assay. The UV/vis spectrum of 3-hydroxy-4-nitro benzoic acid (4H3N) which is a known ligand for the metal site of insulin Re shows a shift in absorption maximum upon displacement from the metal site to the solution (Huang et al., 1997, Biochemistry 36, 9878-9888). Titration of a ligand to a solution of insulin Re hexamers with 4H3N mounted in the metal site allows the binding affinity of these ligands to be determined following the reduction of absorption at 444 nm. A stock solution with the following composition 0.2 mM human insulin, 0.067 mM Zn-acetate, 40 mM phenol, 0.101 mM 4H3N is prepared in a 10mL quantum as described below. Buffer is always 50mM tris buffer adjusted to pH=8.0 with NaOH/CI04 ".
1000 μL of 2.0mM human insulin in buffer 66.7 μL of 10mM Zn-acetate in buffer 800 μL of 500mM phenol in H20 201 μL of 4H3N in H20 7.93 ml buffer
The ligand is dissolved in DMSO to a concentration of 20 mM.
The ligand solution is titrated to a cuvette containing 2 mL stock solution and after each addition the UV/vis spectrum is measured. The titration points are listed in Table 3 below. Table 3
Figure imgf000418_0002
The UV/vis spectra resulting from a titration of the compound 3-hydroxy-2-naphthoic acid is shown in Figure 4. Inserted in the upper right corner is the absorbance at 444nm vs. the concentration of ligand.
The following equation is fitted to these datapoints to determine the two parameters KD(obs), the observed dissociation constant, and absmax the absorbance at maximal ligand concentration.
abs ([ligandjfree) = (absmax * [ligand]free)/ (KD(obs) + [ligand]freΘ)
The observed dissociation constant is recalculated to obtain the apparent dissociation constant
KD(app) = KD(obs) / ( 1+[4H3N]/K4H3N )
The value of
Figure imgf000418_0001
μM is taken from Huang et al., 1997, Biochemistry 36, 9878-9888.
TZD-assay:
The binding affinity of ligands to the metal site of insulin R6 hexamers are measured in a fluo- rescense based displacement assay. The fluorescence of 5-(4- dimethylaminobenzylidene)thiazolidine-2,4-dione (TZD) which is a ligand for the metal site of insulin R6 is quenched upon displacement from the metal site to the solution. Titration of a ligand to a stock solution of insulin Re hexamers with this compound mounted in the metal site allows the binding affinity of these ligands to be determined measuring the fluorescence at 455nm upon excitation at 41 Onm.
Preparation
Stock solution: 0.02 mM human insulin, 0.007 mM Zn-acetate, 40 mM phenol, 0.01 mM TZD in 50mM tris buffer adjusted to pH=8.0 with NaOH/CI04 '. The ligand is dissolved in DMSO to a concentration of 5 mM and added in aliquots to the stock solution to final concentrations of 0-250 μM.
Measurements
Fluorescence measurements were carried out on a Perkin Elmer Spectrofluorometer LS50B.The main absorption band was excited at 410 nm and emission was detected at 455 nm. The resolution was 10 nm and 2.5 nm for excitation and emission, respectively.
The fluorescence spectra resulting from a titration of the compound 5-(4- dimethylaminobenzylidene)thiazolidine-2,4-dione (TZD) is shown in Figure 5. Inserted in the upper right corner is the fluorescence at 455 nm upon exitation at 410 nM vs. the concentration of ligand.
Data analysis
This equation is fitted to the datapoints ΔF(455nm)) = ΔFmax * [ligand]freβ/( KD(app) * ( 1 +[TZD]/KTZD )+ [ligand]free))
K (app) is the apparent dissociation constant and Fmax is the fluorescence at maximal ligand concentration. The value of KT2D is measured separately to 230 nM
Two different fitting-procedures can be used. One in which both parameters, KD(app) and Fmax, are adjusted to best fit the data and a second in which the value of Fmax is fixed (Fmax=1 ) and only KD(app) is adjusted. The given data are from the second fitting procedure. The Solver module of Microsoft Excel can be used to generate the fits from the datapoints.

Claims

1. A pharmaceutical preparation comprising
4. Acid-stabilised insulin
5. Zinc ions
6. A zinc-binding ligand of the following general formula (I)
CGr-Lnk-Frg1-Frg2-X (I)
wherein:
CGr is a chemical group which reversibly binds to a His Zn site of an insulin hexamer;
Lnk is a linker selected from • a valence bond • a chemical group GB of the formula -B1-B2-C(0)-, -B1-B2-S02-, -B1-B2-CH2-, or -B1-
B2-NH-; wherein B1 is a valence bond, -0-, -S-, or -NR6B-,
B2 is a valence bond, d-C18-alkylene, C2-C18-alkenylene, C2-Cι8-alkynylene, arylene, heteroarylene, -d-C18-alkyl-aryl-, -C2-C18-alkenyl-aryl-, -C2-C18-alkynyl-aryl-, -C(=O)- C C18-alkyl-C(=0)-, -C(=0)-C1-C18-alkenyl-C(=0)-, -C(=0)-C1-C18-alkyl-0-C1-C18- alkyl-C(=0)-, -C(=0)- C1-C18-alkyl-S-C1-C18-alkyl-C(=0)-l -C(=0)-C1-C18-alkyl-NRβ-C1-
C18-alkyl-C(=0)-, -C(=0)-aryl-C(=0)-, -C(=0)-heteroaryl-C(=0)-; wherein the alkylene, alkenylene, and alkynylene moieties are optionally substituted by -CN, -CF3, -OCF3, -OR6B, or -NR6BR7B and the arylene and heteroarylene moieties are optionally substituted by halogen, -C(0)OR6B, -C(0)H, OCOR6B, -S02, -CN, -CF3, -OCF3, -N02, -OR6B, -NR6BR7B, d-C18-alkyl, or d-C18-alkanoyl;
R6Band R7B are independently H, d-C4-alkyl;
Frg1 is a fragment consisting of 0 to 5 neutral α- or β-amino acids
Frg2 is a fragment comprising 1 to 20 positively charged groups independently selected from amino or guanidino groups; and
X is -OH, -NH2 or a diamino group, or a salt thereof with a pharmaceutically acceptable acid or base, or any optical isomer or mixture of optical isomers, including a racemic mixture, or any tautomeric forms.
2. A pharmaceutical preparation according to claim 1 wherein CGr is a chemical structure selected from the group consisting of carboxylates, dithiocarboxylates, phenolates, thiophe- nolates, alkylthiolates, sulfonamides, imidazoles, triazoles, 4-cyano-1 ,2,3-triazoles, benzimi- dazoles, benzotriazoles, purines, thiazolidinediones, tetrazoles, 5-mercaptotetrazoles, rhodanines, N-hydroxyazoles, hydantoines, thiohydantoines, barbiturates, naphthoic acids and salicylic acids.
3. A pharmaceutical preparation according to claim 2 wherein CGr is a chemical structure selected from the group consisting of benzotriazoles, 3-hydroxy 2-napthoic acids, salicylic acids, tetrazoles, thiazolidinediones, 5-mercaptotetrazoles, or 4-cyano-1 ,2,3-triazoles.
4. A pharmaceutical composition according to any one of the claims 1 to 3 wherein CGr is
Figure imgf000421_0001
wherein
X is =0, =S or =NH
Y is -S-, -O- or -NH-
R1, R1* and R4 are independently selected from hydrogen or d-C6-alkyl, R2 and R2* are hydrogen or d-Cβ-alkyl or aryl, R1 and R2 may optionally be combined to form a double bond, R1* and R2* may optionally be combined to form a double bond, R3, R3* and R5 are independently selected from hydrogen, halogen, aryl optionally substi- tuted with one or more substituents independently selected from R1β, d-Cβ-alkyl, or -C(0)NR11R12,
A, A1 and B are independently selected from d-C6-alkyl, aryl, aryl-d-C6-alkyl, -NR11-aryl, aryl-C2-C6-alkenyl or heteroaryl, wherein the alkyl or alkenyl is optionally substituted with one or more substituents independently selected from R6 and the aryl or heteroaryl is optionally substituted with up to four substituents R7, R8, R9, and R10, A and R3 may be connected through one or two valence bonds, B and R5 may be connected through one or two valence bonds,
R6 is independently selected from halogen, -CN, -CF3, -OCF3, aryl, -COOH and -NH2, R7, R8, R9 and R10 are independently selected from
• hydrogen, halogen, -CN, -CH2CN, -CHF2, -CF3, -OCF3, -OCHF2, -OCH2CF3, -OCF2CHF2, -S(0)2CF3, -OS(0)2CF3, -SCF3, -N02, -OR11, -NR11R12, -SR11, -NR11S(0)2R12, -S(0)2NR11R12, -S(0)NR11R12, -S(0)R11, -S(0)2R11, -OS(0)2 R11, -C(0)NR11R12, -OC(0)NR11R12, -NR11C(0)R12, -CH2C(0)NR11R12, -Od-C6-alkyl-C(0)NR11R12, -CH2OR11, -CH2OC(0)R11, -CH2NR11R12, -OC(0)R11,
-Od-C15-alkyl-C(0)OR11, -Od-C6-alkyl-OR11, -Sd-C6-alkyl-C(0)OR11 -C2-C6-alkenyl-C(=0)OR11, -NR11-C(=0)-d-C6-alkyl-C(=0)OR11,
-NR11-C(=O)-d-Cβ-alkenyl-C(=O)OR11 , -C(0)OR11, C(0)R11, or -C2-C6-alkenyl- C(=0)R11, =0, or -C2-Ce-alkenyl-C(=0)-NR11R12,
• CrCe-alkyl, C2-C6-alkenyl or C2-C6-alkynyl, each of which may optionally be substituted with one or more substituents independently selected from R13,
• aryl, aryloxy, aryloxycarbonyl, aroyl, arylsulfanyl, aryl-d-C6-alkoxy, aryl-d-Ce-alkyl, aryl-C2-C6-alkenyl, aroyl-C2-Cβ-alkenyl, aryl-C2-Cβ-alkynyl, heteroaryl, heteroaryl-d-
Cβ-alkyl, heteroaryl-C2-C6-alkenyl, heteroaryl-C2-C6-alkynyl, or C3-C6 cycloalkyl,
of which each cyclic moiety may optionally be substituted with one or more substituents independently selected from R14,
R11 and R 2 are independently selected from hydrogen, OH, d-C20-alkyl, aryl-d-C6-alkyl or aryl, wherein the alkyl groups may optionally be substituted with one or more substituents independently selected from R 5, and the aryl groups may optionally be substituted one or more substituents independently selected from R16; R11 and R12 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
R13 is independently selected from halogen, -CN, -CF3, -OCF3, -OR11, -C(0)OR11 , -NR11R12, and -C(0)NR11R12, R14 is independently selected from halogen, -C(0)OR11, -CH2C(0)OR11, -CH2OR11, -CN, - CF3, -OCF3, -N02, -OR11, -NR11R12, -NR11C(0)R11, -S(0)2R11, aryl and d-C6-alkyl,
R15 is independently selected from halogen, -CN, -CF3, =0, -OCF3, -Od-C6-alkyl, -C(0)Od- Cβ-alkyl, -COOH and -NH2,
R16 is independently selected from halogen, -C(O)Od-C6-alkyl, -COOH, -CN, -CF3, -OCF3, - N02, -OH, -OCi-Ce-alkyl, -NH2, C(=0) or d-Cβ-alkyl, or any enantiomer, diastereomer, in- eluding a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base.
5. A pharmaceutical composition according to claim 4 wherein X is =0 or =S.
6. A pharmaceutical composition according to claim 5 wherein X is =0.
7. A pharmaceutical composition according to claim 5 wherein X is =S.
8. A pharmaceutical composition according to any one of the claims 4 to 7 wherein Y is -O- or -S-.
9. A pharmaceutical composition according to claim 8 wherein Y is -0-.
10. A pharmaceutical composition according to claim 8 wherein Y is -NH-.
11. A pharmaceutical composition according to claim 8 wherein Y is -S-.
12. A pharmaceutical composition according to any one of the claims 4 to 11 wherein A is aryl optionally substituted with up to four substituents, R7, R8, R9, and R10 which may be the same or different.
13. A pharmaceutical composition according to claim 12 wherein A is selected from ArG1 optionally substituted with up to four substituents, R7, R8, R9, and R10 which may be the same or different.
14. A pharmaceutical composition according to claim 13 wherein A is phenyl or naphtyl optionally substituted with up to four substituents, R7, R8, R9, and R10 which may be the same or different.
15. A pharmaceutical composition according to claim 14 wherein A is
Figure imgf000423_0001
16. A pharmaceutical composition according to claim 14 wherein A is phenyl.
17. A pharmaceutical composition according to any one of the claims 4 to 11 wherein A is heteroaryl optionally substituted with up to four substituents, R7, R8, R9, and R10 which may be the same or different.
18. A pharmaceutical composition according to claim 17 wherein A is selected from Het1 optionally substituted with up to four substituents, R7, R8, R9, and R10 which may be the same or different.
19. A pharmaceutical composition according to claim 18 wherein A is selected from Het2 optionally substituted with up to four substituents, R7, R8, R9, and R10 which may be the same or different.
20. A pharmaceutical composition according to claim 19 wherein A is selected from Het3 optionally substituted with up to four substituents, R7, R8, R9, and R10 which may be the same or different.
21. A pharmaceutical composition according to claim 20 wherein A is selected from the group consisting of indolyl, benzofuranyl, quinolyl, furyl, thienyl, or pyrrolyl, wherein each heteroaryl may optionally substituted with up to four substituents, R7, R8, R9, and R10 which may be the same or different.
22. A pharmaceutical composition according to claim 20 wherein A is benzofuranyl optionally substituted with up to four substituents R7, R8, R9, and R10 which may be the same or differ- ent.
23. A pharmaceutical composition according to claim 22 wherein A is
Figure imgf000424_0001
24. A pharmaceutical composition according to claim 20 wherein A is carbazolyl optionally substituted with up to four substituents R7, R8, R9, and R10 which may be the same or differ- ent.
25. A pharmaceutical composition according to claim 24 wherein A is
26. A pharmaceutical composition according to claim 20 wherein A is quinolyl optionally substituted with up to four substituents R7, R8, R9, and R10 which may be the same or different.
27. A pharmaceutical composition according to claim 26 wherein A is
Figure imgf000425_0001
28. A pharmaceutical composition according to claim 20 wherein A is indolyl optionally substituted with up to four substituents R7, R8, R9, and R10 which may be the same or different.
29. A pharmaceutical composition according to claim 28 wherein A is
Figure imgf000425_0002
30. A pharmaceutical composition according to any one of the claims 4 to 29 wherein R1 is hydrogen.
31. A pharmaceutical composition according to any one of the claims 4 to 30 wherein R2 is hydrogen.
32. A pharmaceutical composition according to any one of the claims 4 to 29 wherein R1 and R2 are combined to form a double bond.
33. A pharmaceutical composition according to any one of the claims 4 to 32 wherein R3 is d-Cβ-alkyl, halogen, or C(0)NR16R17.
34. A pharmaceutical composition according to claim 33 wherein R3 is d-Cβ-alkyl or C(0)NR16R17.
35. A pharmaceutical composition according to claim 34 wherein R3 is methyl.
36. A pharmaceutical composition according to any one of the claims 4 to 11 wherein B is phenyl optionally substituted with up to four substituents, R7, R8, R9, and R10 which may be the same or different.
37. A pharmaceutical composition according to any one of the claims 4 to 11 or 36 wherein
R4 is hydrogen.
38. A pharmaceutical composition according to any one of the claims 4 to 11 or 36 to 37 wherein R5 is hydrogen.
39. A pharmaceutical composition according to any one of the claims 4 to 38 wherein R6 is aryl.
40. A pharmaceutical composition according to claim 39 wherein R6 is phenyl.
41. A pharmaceutical composition according to any one of the claims 4 to 40 wherein R7, R8, R9 and R10 are independently selected from
• hydrogen, halogen, -N02, -OR11, -NR11R12, -SR11, -NR11S(0)2R12, -S(0)2NR11R12, -S(0)NR11R12, -S(0)R11, -S(0)2R11, -OS(0)2 R11, -NR11C(0)R12, -CH2OR11, - CH2OC(0)R11, -CH2NR11R12, -OC(0)R11, -Od-C6-alkyl-C(0)OR11, -Od-C6- alkyl-C(0)NR11R12, -Od-C6-alkyl-OR11, -Sd-C6-alkyl-C(0)OR11, -C2-C6-alkenyl-
C(=0)OR11, -C(0)OR11, or -C2-C6-alkenyl-C(=0)R11,
• d-Ce-alkyl, C2-C6-alkenyl or C2-C6-alkynyl, which may each optionally be substituted with one or more substituents independently selected from R13
• aryl, aryloxy, aroyl, arylsulfanyl, aryl-d-Cβ-alkoxy, aryl-d-C6-alkyl, aryl-C2- Cβ-alkenyl, aroyl-C2-C6-alkenyl, aryl-C2-Cβ-alkynyl, heteroaryl, heteroaryl-d-Ce-alkyl, wherein each of the cyclic moieties optionally may be substituted with one or more substituents independently selected from R14.
42. A pharmaceutical composition according to claim 41 wherein R7, R8, R9 and R10 are independently selected from
• hydrogen, halogen, -N02, -OR11, -NR11R12, -SR11, -S(0)2R11, -OS(0)2 R11, - CH2OC(0)R11, -OC(0)R11, -Od-C6-alkyl-C(0)OR11, -Od-C6-alkyl-OR11, -SC,-C6- alkyl-C(0)OR11, -C(0)OR11, or -C2-C6-alkenyl-C(=0)R11,
• d-C6-alkyl or Ci-Cβ-alkenyl which may each optionally be substituted with one or more substituents independently selected from R13
•aryl, aryloxy, aroyl, aryl-d-C6-alkoxy, aryl-d-Cβ-alkyl, heteroaryl,
of which each of the cyclic moieties optionally may be substituted with one or more substituents independently selected from R14.
43. A pharmaceutical composition according to claim 42 wherein R7, R8, R9 and R10 are inde- pendently selected from • hydrogen, halogen, -N02, -OR11, -NR11R12, -SR11, -S(0)2R11, -OS(0)2 R11, - CH2OC(0)R11, -OC(0)R11, -Od-C6-alkyl-C(0)OR11, -Od-Ce-alkyl-OR11, -SCι-C6- alkyl-C(0)OR11, -C(0)OR11, or -C2-C6-alkenyl-C(=0)R11,
• d-Cβ-alkyl or d-C6- which may each optionally be substituted with one or more substituents independently selected from R13
• aryl, aryloxy, aroyl, aryl-d-Ce-alkoxy, aryl-d-C6-alkyl, heteroaryl,
of which each of the cyclic moieties optionally may be substituted with one or more substituents independently selected from R14.
44. A pharmaceutical composition according to claim 43 wherein R7, R8, R9 and R10 are independently selected from . hydrogen, halogen, -OR11, -Od-C6-alkyl-C(0)OR11, or -C(0)OR11,
• d-Cβ-alkyl which may each optionally be substituted with one or more substituents independently selected from R13
•aryl, aryloxy, aryl-d-Cβ-alkoxy,
of which each of the cyclic moieties optionally may be substituted with one or more substituents independently selected from R14.
45. A pharmaceutical composition according to claim 44 wherein R7, R8, R9 and R10 are independently selected from
• hydrogen, halogen, -OR11, -Od-C6-alkyl-C(0)OR11, or -C(0)OR11,
• CrCe-alkyl which may each optionally be substituted with one or more substituents independently selected from R13
• ArG1 , ArGloxy, ArG1-d-C6-alkoxy,
of which each of the cyclic moieties optionally may be substituted with one or more substituents independently selected from R14.
46. A pharmaceutical composition according to claim 45 wherein R7, R8, R9 and R10 are independently selected from
• hydrogen, halogen, -OR11, -Od-C6-alkyl-C(0)OR11, or -C(0)OR11,
•d-Cβ-alkyl which may optionally be substituted with one or more substituents independently selected from R13
• phenyl, phenyloxy, phenyl-d-C6-alkoxy, wherein each of the cyclic moieties optionally may be substituted with one or more substituents independently selected from R14.
47. A pharmaceutical composition according to any one of the claims 4 to 46 wherein R11 and R12 are independently selected from hydrogen, d-C2o-alkyl, aryl or aryl-d-Ce-alkyl, wherein the alkyl groups may optionally be substituted with one or more substituents independently selected from R15, and the aryl groups may optionally be substituted one or more substitu- ents independently selected from R16; R11 and R12 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds.
48. A pharmaceutical composition according to claim 47 wherein R1 and R12 are independ- ently selected from hydrogen, Cι-C20-alkyl, aryl or aryl-d-Ce-alkyl, wherein the alkyl groups may optionally be substituted with one or more substituents independently selected from R15, and the aryl groups may optionally be substituted one or more substituents independently selected from R16.
49. A pharmaceutical composition according to claim 48 wherein R11 and R12 are independ- ently selected from phenyl or phenyl-d-C6-alkyl.
50. A pharmaceutical composition according to claim 48 wherein one or both of R11 and R12 are methyl.
51. A pharmaceutical composition according to any one of the claims 4 to 50 wherein R13 is independently selected from halogen, CF3, OR11 or NR11R12.
52. A pharmaceutical composition according to claim 51 wherein R13 is independently selected from halogen or OR11.
53. A pharmaceutical composition according to claim 52 wherein R13 is OR11.
54. A pharmaceutical composition according to any one of the claims 4 to 53 wherein R14 is independently selected from halogen, -C(0)OR11, -CN, -CF3, -OR11, S(0)2R11, and d- Cβ-alkyl.
55. A pharmaceutical composition according to claim 54 wherein R14 is independently selected from halogen, -C(0)OR11, or -OR11.
56. A pharmaceutical composition according to any one of the claims 4 to 55 wherein R15 is independently selected from halogen, -CN, -CF3, -C(0)OCrC6-alkyl,and -COOH.
57. A pharmaceutical composition according to claim 56 wherein R15 is independently selected from halogen or -C(0)OCrC6-alkyl.
58. A pharmaceutical composition according to any one of the claims 4 to 57 wherein R16 is independently selected from halogen, -C(0)OCrCβ-alkyl, -COOH, -N02, -Od-Cβ-alkyl, -NH2, C(=0) or d-Ce-alkyl.
59. A pharmaceutical composition according to claim 58 wherein R16 is independently selected from halogen, -C(0)OCrC6-alkyl, -COOH, -N02, or d-C6-alkyl.
60. A pharmaceutical composition according to any one of the claims 1 to 3 wherein CGr is
Figure imgf000429_0001
wherein
R19 is hydrogen or d-Cβ-alkyl, R20 is hydrogen or d-Cβ-alkyl,
D, D1 and F are a valence bond, d-C6-alkylene or d-Cβ-alkenylene optionally substituted with one or more substituents independently selected from R72,
R72 is independently selected from hydroxy, d-Cβ-alkyl, or aryl,
E is d-Cβ-alkyl, aryl or heteroaryl, wherein the aryl or heteroaryl is optionally substituted with up to three substituents R21, R22 and R23,
G and G1 are d-Ce-alkyl, aryl or heteroaryl, wherein the aryl or heteroaryl is optionally substituted with up to three substituents R24, R25 and R26,
R17, R18, R21, R22, R23, R24, R25 and R26 are independently selected from • hydrogen, halogen, -CN, -CH2CN, -CHF2, -CF3, -OCF3, -OCHF2, -OCH2CF3, -OCF2CHF2, -S(0)2CF3, -SCF3, -N02, =0, -OR27, -NR27R28, -SR27, -NR27S(0)2R28, -S(0)2NR27R28, -S(0)NR27R28, -S(0)R27, -S(0)2R27, -C(0)NR27R28, -OC(0)NR27R28, -NR27C(0)R28, -NR27C(0)OR28, -CH2C(0)NR27R28, -OCH2C(0)NR27R28, -CH20R27, -CH2NR27R28, -OC(0)R27, -OCrC6-alkyl-C(0)OR27, -SCrC6-alkyl-C(0)OR27, -C2-C6- alkenyl-C(=0)OR27, -NR27-C(=0)-d-C6-alkyl-C(=0)OR27, -NR27-C(=0)-d-C6- alkenyl-C(=0)OR27, -C(=0)NR27-d-Cβ-alkyl-C(=0)OR27, -d-C6-alkyl-C(=0)OR27,or -C(0)OR27,
• d-Ce-alkyl, C2-C6-alkenyl or C2-C6-alkynyl,
which may optionally be substituted with one or more substituents independently selected from R29,
• aryl, aryloxy, aryloxycarbonyl, aroyl, aryl-d-C6-alkoxy, aryl-d-Ce-alkyl, aryl-C2- Cβ-alkenyl, aryl-C2-C6-alkynyl, heteroaryl, heteroaryl-d-Cβ-alkyl, heteroaryl-C2-C6- alkenyl or heteroaryl-C2-C6-alkynyl,
of which the cyclic moieties optionally may be substituted with one or more substituents selected from R30,
R27 and R28 are independently selected from hydrogen, d-Cβ-alkyl, aryl-d-Ce-alkyl or aryl, or R27 and R28 when attached to the same nitrogen atom together with the said nitrogen atom may form a 3 to 8 membered heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
R29 is independently selected from halogen, -CN, -CF3, -OCF3, -OR27, and -NR27R28,
R30 is independently selected from halogen, -C(0)OR27, -CN, -CF3, -OCF3, -N02, -OR27, -NR27R28 and d-Ce-alkyl, or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base.
61. A pharmaceutical composition according to claim 60 wherein D is a valence bond.
62. A pharmaceutical composition according to claim 60 wherein D is CrC6-alkylene optionally substituted with one or more hydroxy, d-Cβ-alkyl, or aryl.
63. A pharmaceutical composition according to any one of the claims 60 to 62 wherein E is aryl or heteroaryl, wherein the aryl or heteroaryl is optionally substituted with up to three sub- stituents independently selected from R21, R22 and R23.
64. A pharmaceutical composition according to claim 63 wherein E is aryl optionally substituted with up to three substituents independently selected from R21, R22 and R23.
65. A pharmaceutical composition according to claim 64 wherein E is selected from ArG1 and optionally substituted with up to three substituents independently selected from R21, R22 and R23.
66. A pharmaceutical composition according to claim 65 wherein E is phenyl optionally substituted with up to three substituents independently selected from R21, R22 and R23.
67. A pharmaceutical composition according to claim 66 wherein CGr is
Figure imgf000431_0001
68. A pharmaceutical composition according to any one of the claims 60 to 67 wherein R21, R22 and R23 are independently selected from
• hydrogen, halogen, -CHF2, -CF3, -OCF3, -OCHF2, -OCH2CF3, -OCF2CHF2, -SCF3, - N02, -OR27, -NR27R28, -SR27, -C(0)NR27R28, -OC(0)NR27R28, -NR27C(0)R28, -NR 7C(0)OR28, -CH2C(0)NR27R28, -OCH2C(0)NR27R28, -CH2OR27, -CH2NR27R28,
-OC(0)R27, -OCrC6-alkyl-C(0)OR27, -Sd-C6-alkyl-C(0)OR27, -C2-C6-alkenyl- C(=0)OR27, -NR27-C(=0)-CrC6-alkyl-C(=0)OR27, -NR27-C(=0)-d-C6- alkenyl-C(=0)OR27-, -C(=0)NR27-CrC6-alkyl-C(=0)OR27, -CrC6-alkyl-C(=0)OR27, or -C(0)OR27,
• d-Cβ-alkyl, C2-Ce-alkenyl or C2-Ce-alkynyl,
which may optionally be substituted with one or more substituents independently selected from R29 • aryl, aryloxy, aryloxycarbonyl, aroyl, aryl-d -Ce-alkoxy, aryl-d-Cβ-alkyl, aryl-C2- C6-alkenyl, aryl-C2-C6-alkynyl, heteroaryl, heteroaryl-CrC6-alkyl, heteroaryl-C2-C6- alkenyl or heteroaryl-C2-C6-alkynyl,
of which the cyclic moieties optionally may be substituted with one or more substituents selected from R30.
69. A pharmaceutical composition according to claim 68 wherein R21, R22 and R23 are independently selected from
. hydrogen, halogen, -OCF3, -OR27, -NR27R28, -SR27, -NR27C(0)R28, -NR27C(0)OR28,
-OC(0)R27, -OCrC6-alkyl-C(0)OR27, -SCrC6-alkyl-C(0)OR27, -C2-C6-alkenyl- C(=0)OR27, -C(=0)NR27-d-Cβ-alkyl-C(=0)OR27, -CrC6-alkyl-C(=0)OR27, or -C(0)OR27,
• d-Cβ-alkyl optionally substituted with one or more substituents independently selected from R29
• aryl, aryloxy, aroyl, aryl-d-Cβ-alkoxy, aryl-d-Cβ-alkyl, heteroaryl, heteroaryl-d-Cβ- alkyl,
of which the cyclic moieties optionally may be substituted with one or more substituents selected from R30.
70. A pharmaceutical composition according to claim 69 wherein R21, R22 and R23 are independently selected from
• hydrogen, halogen, -OCF3, -OR27, -NR27R28, -SR27, -NR27C(0)R28, -NR27C(0)OR28, -OC(0)R27, -OCrC6-alkyl-C(0)OR27, -SCrC6-alkyl-C(0)OR27, -C2-C6-alkenyl- C(=0)OR27, -C(=0)NR27-d-Cβ-alkyl-C(=0)OR27, -CrC6-alkyl-C(=0)OR27, or -C(0)OR27,
• methyl, ethyl propyl optionally substituted with one or more substituents independently selected from R29
• aryl, aryloxy, aroyl, aryl-d-Cβ-alkoxy, aryl-d-Ce-alkyl, heteroaryl, heteroaryl-d-Cβ- alkyl of which the cyclic moieties optionally may be substituted with one or more substituents selected from R30.
71. A pharmaceutical composition according to claim 70 wherein R21, R22 and R23 are independently selected from
• hydrogen, halogen, -OCF3, -OR27, -NR 7R28, -SR27, -NR27C(0)R28, -NR27C(0)OR28, -OC(0)R27, -OCrC6-alkyl-C(0)OR27, -Sd-C6-alkyl-C(0)OR27, -C2-C6-alkenyl- C(=0)OR27, -C(=0)NR27-CrC6-alkyl-C(=0)OR27, -CrC6-alkyl-C(=0)OR27, or -C(0)OR27,
• methyl, ethyl propyl optionally substituted with one or more substituents independently selected from R29
• ArG1, ArG1-0-, ArG1-C(0)-, ArG1-d-Ce-alkoxy, ArG1-d-C6-alkyl, Het3, Het3-d- Cβ-alkyl of which the cyclic moieties optionally may be substituted with one or more substituents selected from R30.
72. A pharmaceutical composition according to claim 71 wherein R21, R22 and R23 are independently selected from
• hydrogen, halogen, -OCF3, -OR27, -NR27R28, -SR27, -NR27C(0)R28, -NR27C(0)OR28, -OC(0)R27, -OCrC6-alkyl-C(0)OR27, -SCrC6-alkyl-C(0)OR27, -C2-C6-alkenyl- C(=0)OR27, -C(=0)NR27-CrC6-alkyl-C(=0)OR27, -CrC6-alkyl-C(=0)OR27, or -C(0)OR27,
• CrCe-alkyl optionally substituted with one or more substituents independently selected from R29
• phenyl, phenyloxy, phenyl-d-Cβ-alkoxy, phenyl-d-Ce-alkyl, of which the cyclic moieties optionally may be substituted with one or more substituents selected from R30.
73. A pharmaceutical composition according to any one of the claims 60 to 72 wherein R19 is hydrogen or methyl.
74. A pharmaceutical composition according to claim 73 wherein R19 is hydrogen.
75. A pharmaceutical composition according to any one of the claims 60 to 74 wherein R27 is Hydrogen, d-C6-alkyl or aryl.
76. A pharmaceutical composition according to claim 75 wherein R27 is hydrogen or CrC6- alkyl.
77. A pharmaceutical composition according to any one of the claims 60 to 76 wherein R28 is hydrogen or d-Ce-alkyl.
78. A pharmaceutical composition according to claim 60 wherein F is a valence bond.
79. A pharmaceutical composition according to claim 60 wherein F is d-C6-alkylene optionally substituted with one or more hydroxy, d-C6-alkyl, or aryl.
80. A pharmaceutical composition according to any one of the claims 60 or 78 to 79 wherein G is CrCe-alkyl or aryl, wherein the aryl is optionally substituted with up to three substituents R24, R25 and R26.
81. A pharmaceutical composition according to any one of the claims 60 or 78 to 79 wherein G is d-Cβ-alkyl or ArG1 , wherein the aryl is optionally substituted with up to three substitu- ents R24, R25 and R26.
82. A pharmaceutical composition according to claim 80 wherein G is d-Cβ-alkyl.
83. A pharmaceutical composition according to claim 82 wherein G is phenyl optionally substituted with up to three substituents R24, R25 and R26.
84. A pharmaceutical composition according to any one of the claims 60 to 83 wherein R24, R25 and R26 are independently selected from
• hydrogen, halogen, -CHF2, -CF3, -OCF3, -OCHF2, -OCH2CF3, -OCF2CHF2, -SCF3, - N02, -OR27, -NR27R28, -SR27, -C(0)NR27R28, -OC(0)NR27R28, -NR27C(O)R28, -NR27C(O)OR28, -CH2C(0)NR27R28, -OCH2C(0)NR27R28, -CH2OR27, -CH2NR27R28, -OC(0)R27, -OCι-C6-alkyl-C(0)OR27, -SCrC6-alkyl-C(0)OR27, -C2-C6-alkenyl-
C(=0)OR27, -NR27-C(=0)-CrC6-alkyl-C(=0)OR27, -NR27-C(=0)-d-C6- alkenyl-C(=0)OR27-, -C(=0)NR27-CrC6-alkyl-C(=0)OR27, -d-Cβ-alkyl-C(=0)OR27, or -C(0)OR27,
• d-Ce-alkyl, C2-C6-alkenyl or C2-C6-alkynyl,
which may optionally be substituted with one or more substituents independently selected from R29 • aryl, aryloxy, aryloxycarbonyl, aroyl, aryl-d-Cβ-alkoxy, aryl-CrC6-alkyl, aryl-C2- Cβ-alkenyl, aryl-C2-C6-alkynyl, heteroaryl, heteroaryl-d-Cβ-alkyl, heteroaryl-C2-Ce- alkenyl or heteroaryl-C2-C6-alkynyl,
of which the cyclic moieties optionally may be substituted with one or more substituents selected from R30.
85. A pharmaceutical composition according to claim 84 wherein R24, R25 and R26 are independently selected from
• hydrogen, halogen, -OCF3, -OR27, -NR27R28, -SR27, -NR27C(0)R28, -NR27C(0)OR28,
-OC(0)R27, -OCrC6-alkyl-C(0)OR27, -Sd-C6-alkyl-C(0)OR27, -C2-C6-alkenyl- C(=0)OR27, -C(=0)NR27-d-Cβ-alkyl-C(=0)OR27, -CrC6-alkyl-C(=0)OR27, or -C(0)OR27,
• d-Cβ-alkyl, C2-C6-alkenyl or C2-C6-alkynyl,
which may optionally be substituted with one or more substituents independently selected from R29
'aryl, aryloxy, aryloxycarbonyl, aroyl, aryl-CrC6-alkoxy, aryl-d-Ce-alkyl, aryl-C2-
Cβ-alkenyl, aryl-C2-C6-alkynyl, heteroaryl, heteroaryl-CrC6-alkyl, heteroaryl-C2-Ce- alkenyl or heteroaryl-C2-C6-alkynyl,
of which the cyclic moieties optionally may be substituted with one or more substitu- ents selected from R30.
86. A pharmaceutical composition according to claim 85 wherein R24, R25 and R26 are independently selected from
• hydrogen, halogen, -OCF3, -OR27, -NR27R28, -SR27, -NR27C(0)R28, -NR27C(0)OR28, -OC(0)R27, -OCrC6-alkyl-C(0)OR27, -Sd-C6-alkyl-C(0)OR27, -C2-C6-alkenyl-
C(=0)OR27, -C(=0)NR27-d-Ce-alkyl-C(=0)OR27, -CrC6-alkyl-C(=0)OR27, or -C(0)OR27,
• CrCe-alkyl optionally substituted with one or more substituents independently se- lected from R29 • aryl, aryloxy, aroyl, aryl-d-Ce-alkoxy, aryl-d-Cβ-alkyl, heteroaryl, heteroaryl-d-Ce- alkyl,
of which the cyclic moieties optionally may be substituted with one or more substituents selected from R30.
87. A pharmaceutical composition according to claim 86 wherein R21, R22 and R23 are independently selected from
. hydrogen, halogen, -OCF3, -OR27, -NR27R28, -SR27, -NR27C(0)R28, -NR27C(0)OR28,
-OC(0)R27, -Od-C6-alkyl-C(0)OR27, -Sd-Cβ-alkyl-C(0)OR27, -C2-C6-alkenyl- C(=0)OR27, -C(=0)NR27-d-C6-alkyl-C(=0)OR27, -d-C6-alkyl-C(=0)OR27, or -C(0)OR27,
. methyl, ethyl propyl optionally substituted with one or more substituents independently selected from R29
• ArG1 , ArG1-0-, ArG1-C(0)-, ArG1-d-C6-alkoxy, ArG1 -d-Ce-alkyl, Het3, Het3-d- Ce-alkyl of which the cyclic moieties optionally may be substituted with one or more substituents selected from R30.
88. A pharmaceutical composition according to claim 87 wherein R21, R22 and R23 are independently selected from
. hydrogen, halogen, -OCF3, -OR27, -NR27R28, -SR27, -NR27C(0)R28, -NR27C(0)OR28,
-OC(0)R27, -OCrC6-alkyl-C(0)OR27, -Sd-C6-alkyl-C(0)OR27, -C2-C6-alkenyl- C(=0)OR27, -C(=0)NR27-Cι-C6-alkyl-C(=0)OR27, -d-C6-alkyl-C(=0)OR27, or -C(0)OR27,
• methyl, ethyl propyl optionally substituted with one or more substituents independently selected from R29
• ArG1 , ArGI-O-, ArG1-C(0)-, ArG1 -d-Ce-alkoxy, ArG1 -d-Ce-alkyl, Het3, Het3-d- Cβ-alkyl of which the cyclic moieties optionally may be substituted with one or more substituents selected from R30.
89. A pharmaceutical composition according to claim 88 wherein R21, R22 and R23 are independently selected from
• hydrogen, halogen, -OCF3, -OR27, -NR27R28, -SR27, -NR27C(0)R28, -NR27C(0)OR28, -OC(0)R27, -Od-C6-alkyl-C(0)OR27, -SCrC6-alkyl-C(0)OR27, -C2-C6-alkenyl- C(=0)OR27, -C(=0)NR27-d-C6-alkyl-C(=0)OR27, -CrC6-alkyl-C(=0)OR27, or -C(0)OR27, • methyl, ethyl propyl optionally substituted with one or more substituents independently selected from R29
• ArG1 , ArG1-0-, ArG1 -d-Ce-alkoxy, ArG1-CrC6-alkyl, of which the cyclic moieties optionally may be substituted with one or more substituents se- lected from R30.
90. A pharmaceutical composition according to any one of the claims 60 or 78 to 89 wherein R20 is hydrogen or methyl.
91. A pharmaceutical composition according to claim 90 wherein R20 is hydrogen.
92. A pharmaceutical composition according to any one of the claims 60 or 78 to 91 wherein R27 is hydrogen, d-C6-alkyl or aryl.
93. A pharmaceutical composition according to claim 92 wherein R27 is hydrogen or CrCe- alkyl or ArG1.
94. A pharmaceutical composition according to claim 93 wherein R27 is hydrogen or CrC6- alkyl.
95. A pharmaceutical composition according to any one of the claims 60 or 78 to 93 wherein R28 is hydrogen or d-C6-alkyl.
96. A pharmaceutical composition according to claim 60 wherein R17 and R18 are independently selected from
. hydrogen, halogen, -CN, -CF3, -OCF3, -N02, -OR27, -NR27R28, -SR27, -S(O)R27,
-S(O)2R27, -C(O)NR27R28, -CH2OR27, -OC(O)R27, -OCrC6-alkyl-C(0)OR27, -SCrC6- alkyl-C(0)OR27, or -C(0)OR27,
• CrCe-alkyl, C2-C6-alkenyl or C2-C6-alkynyl, optionally substituted with one or more substituents independently selected from R29 • aryl, aryloxy, aroyl, aryl-d-Cβ-alkoxy, aryl-d-Ce-alkyl, heteroaryl, heteroaryl-d-Cβ- alkyl,
of which the cyclic moieties optionally may be substituted with one or more substituents selected from R30.
97. A pharmaceutical composition according to claim 96 wherein R17 and R18 are independently selected from
• hydrogen, halogen, -CN, -CF3, -N02, -OR27, -NR27R28, or -C(0)OR27,
• d-Cβ-alkyl optionally substituted with one or more substituents independently selected from R29
»aryl, aryloxy, aroyl, aryl-d-Ce-alkoxy, aryl-d-Ce-alkyl, heteroaryl, heteroaryl-d-Ce- alkyl,
of which the cyclic moieties optionally may be substituted with one or more substituents selected from R30.
98. A pharmaceutical composition according to claim 97 wherein R17 and R18 are independently selected from
• hydrogen, halogen, -CN, -CF3, -N02, -OR27, -NR27R28, or -C(0)OR27
• methyl, ethyl propyl optionally substituted with one or more substituents independently selected from R29 •aryl, aryloxy, aroyl, aryl-d-Cβ-alkoxy, aryl-CrC6-alkyl, heteroaryl, heteroaryl-C-ι-C6- alkyl of which the cyclic moieties optionally may be substituted with one or more substituents selected from R30.
99. A pharmaceutical composition according to claim 98 wherein R17 and R18 are independ- ently selected from
• hydrogen, halogen, -CN, -CF3, -N02, -OR27, -NR27R28, or -C(0)OR27
• methyl, ethyl propyl optionally substituted with one or more substituents independently selected from R29
• ArG1 , ArG1-0-, ArG1-C(0)-, ArG1 -d-Ce-alkoxy, ArG1-CrC6-alkyl, Het3, Het3-d- Cβ-alkyl of which the cyclic moieties optionally may be substituted with one or more substituents selected from R30.
100. A pharmaceutical composition according to claim 99 wherein R17 and R18 are independently selected from
• hydrogen, halogen, -CN, -CF3, -N02, -OR27, -NR27R28, or -C(0)OR27
• d-Cβ-alkyl optionally substituted with one or more substituents independently selected from R29
• phenyl, phenyloxy, phenyl-CrC6-alkoxy, phenyl-d-Cβ-alkyl, of which the cyclic moieties optionally may be substituted with one or more substituents selected from R30.
101. A pharmaceutical composition according to any one of the claims 60 to 100 wherein R27 is hydrogen or d-C6-alkyl.
102. A pharmaceutical composition according to claim 101 wherein R27 is hydrogen, methyl or ethyl.
103. A pharmaceutical composition according to any one of the claims 60 to 102 wherein R28 is hydrogen or d-C6-alkyl.
104. A pharmaceutical composition according to claim 103 wherein R28 is hydrogen, methyl or ethyl.
105. A pharmaceutical composition according to any one of the claims 60 to 104 wherein R72 is -OH or phenyl.
106. A pharmaceutical composition according to claim 60 wherein CGr is
Figure imgf000439_0001
107. A pharmaceutical composition according to any one of the claims 1 to 3 wherein CGr is of the form H-l-J-
wherein H is
Figure imgf000439_0002
wherein the phenyl, naphthalene or benzocarbazole rings are optionally substituted with one or more substituents independently selected from R31
I is selected from
• a valence bond,
• -CH2N(R32)- or -S02N(R33)-,
Figure imgf000440_0001
is S(0)2 or CH2, Z2 is -NH-, -O-or -S-, and n is 1 or 2,
J is 0 • d-Cβ-alkyl, C2-C6-alkenyl or C2-Cβ-alkynyl, which may each optionally be substituted with one or more substituents selected from R34,
• Aryl, aryloxy, aryl-oxycarbonyl-, aroyl, aryl-d-Cβ-alkoxy-, aryl-d-Cβ-alkyl-, aryi-C2- Cβ-alkenyl-, aryl-C2-C6-alkynyl-, heteroaryl, heteroaryl-d-Cβ-alkyl-, heteroaryl-C2-C6- alkenyl- or heteroaryl-C2-C6-alkynyl-, wherein the cyclic moieties are optionally substi- 5 tuted with one or more substituents selected from R37,
• hydrogen,
R31 is independently selected from hydrogen, halogen, -CN, -CH2CN, -CHF2, -CF3, -OCF3, -OCHF2, -OCH2CF3, -OCF2CHF2, -S(0)2CF3, -SCF3, -N02, -OR35, -C(0)R35, -NR35R36, -SR35, 0 -NR35S(0)2R36, -S(0)2NR35R36, -S(0)NR35R36, -S(0)R35, -S(0)2R35, -C(0)NR35R36, -OC(0)NR35R36, -NR35C(0)R36, -CH2C(0)NR35R36, -OCH2C(0)NR35R36, -CH2OR35, -CH2NR35R36, -OC(O)R35, -OCrC6-alkyl-C(0)OR35, -Sd-Cβ-alkyl-C(0)OR35 -C2-C6-alkenyl- C(=O)OR35, -NR35-C(=0)-d-C6-alkyl-C(=0)OR35, -NR35-C(=0)-CrC6-alkenyl-C(=0)OR35-, d-Ce-alkyl, d-C6-alkanoyl or -C(0)OR35, 5
Rr and R are independently selected from hydrogen, d-C6-alkyl or CrC6-alkanoyl,
R34 is independently selected from halogen, -CN, -CF3, -OCF3, -OR35, and -NR35R36,
i0 R35 and R36 are independently selected from hydrogen, d-Ce-alkyl, aryl-d-Ce-alkyl or aryl, or R35 and R36 when attached to the same nitrogen atom together with the said nitrogen atom may form a 3 to 8 membered heterocyclic ring optionally containing one or two further het- eroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
R37 is independently selected from halogen, -C(0)OR35, -C(0)H, -CN, -CF3, -OCF3, -NO2, - OR35, -NR35R36, d-Cβ-alkyl or d-C6-alkanoyl,
or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base.
108. A pharmaceutical composition according to claim 107 wherein CGr is of the form H-l-J, wherein H is
Figure imgf000441_0001
wherein the phenyl, naphthalene or benzocarbazole rings are optionally substituted with one or more substituents independently selected from R3 ,
I is selected from • a valence bond,
• -CH2N(R32)- or -S02N(R33)-,
Figure imgf000441_0002
wherein Z1 is S(0)2 or CH2, Z2 is N.-O-or -S-, and n is 1 or 2,
J is • d-Cβ-alkyl, C2-C6-alkenyl or C2-C6-alkynyl, which may each optionally be substituted with one or more substituents selected from R34,
• Aryl, aryloxy, aryl-oxycarbonyl-, aroyl, aryl-d-C6-alkoxy-, aryl-d-Cβ-alkyl-, aryl-C2- Ce-alkenyl-, aryl-C2-C6-alkynyl-, heteroaryl, heteroaryl-d-Cβ-alkyl-, heteroaryl-C2-C6- alkenyl- or heteroaryl-C2-C6-alkynyl-, wherein the cyclic moieties are optionally substi- tuted with one or more substituents selected from R37,
• hydrogen, R31 is independently selected from hydrogen, halogen, -CN, -CH2CN, -CHF2, -CF3, -OCF3, -OCHF2, -OCH2CF3, -OCF2CHF2, -S(0)2CF3, -SCF3, -N02, -OR35, -C(0)R35, -NR35R36, -SR35, -NR35S(0)2R36, -S(0)2NR35R36, -S(0)NR35R36, -S(0)R35, -S(0)2R35, -C(0)NR35R36, -OC(0)NR35R36, -NR35C(0)R36, -CH2C(0)NR35R36, -OCH2C(0)NR35R3β, -CH2OR35, -CH2NR35R36, -OC(O)R35, -OC1-C6-alkyl-C(O)OR35, -Sd-C6-alkyl-C(0)OR35 -C2-C6-alkenyl- C(=0)OR35, -NR35-C(=0)-C1-Ce-alkyl-C(=0)OR35, -NR35-C(=0)-CrC6-alkenyl-C(=0)OR35-, CrCe-alkyl, d-Cβ-alkanoyl or -C(0)OR35,
R32 and R33 are independently selected from hydrogen, d-C6-alkyl or d-Ce-alkanoyl,
R^ is independently selected from halogen, -CN, -CF3, -OCF3, -OR35, and -NR35R36,
R and R36 are independently selected from hydrogen, d-Cβ-alkyl, aryl-d-C6-alkyl or aryl, or R35 and R36 when attached to the same nitrogen atom together with the said nitrogen atom may form a 3 to 8 membered heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
R37 is independently selected from halogen, -C(0)OR35, -C(0)H, -CN, -CF3, -OCF3, -NO2, - OR35, -NR35R36, d-Cβ-alkyl or d-Cβ-alkanoyl,
or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base, With the proviso that R31 and J cannot both be hydrogen.
109. A pharmaceutical composition according to any one of the claims 107 or 108 wherein H is
Figure imgf000442_0001
110. A pharmaceutical composition according to claim 109 wherein H is
Figure imgf000443_0001
111. A pharmaceutical composition according to claim 109 wherein H is
Figure imgf000443_0002
112. A pharmaceutical composition according to any one of the claims 107 to 111 wherein I is a valence bond, -CH2N(R32)-, or -SO2N(R33)-.
113. A pharmaceutical composition according to claim 112 wherein I is a valence bond.
114. A pharmaceutical composition according to any one of the claims 107 to 113 wherein J is
• hydrogen, • d-Cβ-alkyl, C2-C6-alkenyl or C2-C6-alkynyl, which may optionally be substituted with one or more substituents selected from halogen, -CN, -CF3, -OCF3, -OR35, and -NR35R36,
• aryl, or heteroaryl, wherein the cyclic moieties are optionally substituted with one or more substituents independently selected from R37.
115. A pharmaceutical composition according to claim 114 wherein J is
• hydrogen,
• aryl or heteroaryl, wherein the cyclic moieties are optionally substituted with one or more substituents independently selected from R37.
116. A pharmaceutical composition according to claim 114 wherein J is • hydrogen,
• ArG1 or Het3, wherein the cyclic moieties are optionally substituted with one or more substituents independently selected from R37.
117. A pharmaceutical composition according to claim 116 wherein J is
• hydrogen, • phenyl or naphthyl optionally substituted with one or more substituents independently selected from R37.
118. A pharmaceutical composition according to claim 117 wherein J is hydrogen.
119. A pharmaceutical composition according to any one of the claims 107 to 118 wherein R32 and R33 are independently selected from hydrogen or d-C6-alkyl.
120. A pharmaceutical composition according to any one of the claims 107 to 119 wherein R34 is hydrogen, halogen, -CN, -CF3, -OCF3, -SCF3, -N02, -OR35, -C(0)R35, -NR35R3β, -SR35, -C(0)NR35R36, -OC(0)NR35R36, -NR35C(0)R3β, -OC(0)R35, -Od-C6-alkyl-C(0)OR35, -Sd-C6- alkyl-C(0)OR35 or -C(0)OR35.
121. A pharmaceutical composition according to claim 120 wherein R34 is hydrogen, halogen, -CF3, -N02, -OR35, -NR35R36, -SR35, -NR35C(0)R36, or -C^OR35.
122. A pharmaceutical composition according to claim 121 wherein R34 is hydrogen, halogen, -CF3, -N02, -OR35, -NR35R36, or -NR35C(0)R36.
123. A pharmaceutical composition according to claim 122 wherein R34 is hydrogen, halogen, or -OR35.
124. A pharmaceutical composition according to any one of the claims 107 to 123 wherein R35 and R36 are independently selected from hydrogen, d-Cβ-alkyl, or aryl.
125. A pharmaceutical composition according to claim 124 wherein R35 and R36 are independently selected from hydrogen or d-Cβ-alkyl.
126. A pharmaceutical composition according to any one of the claims 107 to 125 wherein R37 is halogen, -C(0)OR35, -CN, -CF3, -OR35, -NR35R36, d-C6-alkyl or d-C6-alkanoyl.
127. A pharmaceutical composition according to claim 126 wherein R37 is halogen, - C(0)OR35, -OR35, -NR35R36, d-C6-alkyl or d-Ce-alkanoyl.
128. A pharmaceutical composition according to claim 127 wherein R37 is halogen, - C(0)OR35 or -OR35.
129. A pharmaceutical composition according to any one of the claims 1 to 3 wherein CGr is
Figure imgf000444_0001
wherein K is a valence bond, d-Cβ-alkylene, -NH-C(=0)-U-, -d-Cβ-alkyl-S-, -d-Cβ-alkyl-O-, -C(=0)-, or -C(=0)-NH-, wherein any C C6-alkyl moiety is optionally substituted with R38,
U is a valence bond, d-C6-alkenylene, -CrC6-alkyl-0- or CrC6-alkylene wherein any d- Cβ-alkyl moiety is optionally substituted with d-C6-alkyl,
R38 is d-Cβ-alkyl, aryl, wherein the alkyl or aryl moieties are optionally substituted with one or more substituents independently selected from R39, R39 is independently selected from halogen, cyano, nitro, amino,
M is a valence bond, arylene or heteroarylene, wherein the aryl or heteroaryl moieties are optionally substituted with one or more substituents independently selected from R40,
R40 is selected from
• hydrogen, halogen, -CN, -CH2CN, -CHF2, -CF3, -OCF3, -OCHF2, -OCH2CF3, -OCF2CHF2, -S(0)2CF3, -OS(0)2CF3, -SCF3, -N02, -OR41, -NR41R42, -SR41, -NR41S(0)2R42, -S(0)2NR41R42, -S(0)NR41R42, -S(0)R41, -S(0)2R41, -OS(0)2 R41,
-C(0)NR41R42, -OC(0)NR41R42, -NR41C(0)R42, -CH2C(0)NR41R42, -Od-C6- alkyl-C(0)NR41R42, -CH2OR41, -CH2OC(0)R41, -CH2NR41R42, -OC(0)R41, -Od-C6- alkyl-C(0)OR41, -OCrC6-alkyl-OR41, -S-d-Ce-alkyl-C(0)OR41, -C2-C6-alkenyl- C(=0)OR41, -NR41-C(=0)-d-C6-alkyl-C(=0)OR41, -NR 1-C(=0)-d-C6- alkenyl-C(=0)OR41 , -C(0)OR41, -C2-C6-alkenyl-C(=0)R41, =0, -NH-C(=0)-0-d-
Ce-alkyl, or -NH-C(=0)-C(=0)-0-CrC6-alkyl,
• d-Cβ-alkyl, C2-C6-alkenyl or C2-C6-alkynyl, which may each optionally be substituted with one or more substituents selected from R43,
• aryl, aryloxy, aryloxycarbonyl, aroyl, arylsulfanyl, aryl-d-Ce-alkoxy, aryl-d-Ce-alkyl, aryl-C2-C6-alkenyl, aroyl-C2-C6-alkenyl, aryl-C2-C6-alkynyl, heteroaryl, heteroaryl-d- Ce-alkyl, heteroaryl-C2-C6-alkenyl or heteroaryl-C2-C6-alkynyl, wherein the cyclic moieties optionally may be substituted with one or more substituents selected from R44,
R41 and R42 are independently selected from hydrogen, -OH, d-Ce-alkyl, d-C6-alkenyl, aryl- d-Ce-alkyl or aryl, wherein the alkyl moieties may optionally be substituted with one or more substituents independently selected from R45, and the aryl moieties may optionally be substi- tuted with one or more substituents independently selected from R46; R41 and R42 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds, R43 is independently selected from halogen, -CN, -CF3, -OCF3, -OR41, and -NR41R42
R44 is independently selected from halogen, -C(0)OR41, -CH2C(0)OR41, -CH2OR41, -CN, - CF3, -OCF3, -N02, -OR41, -NR41R42 and d-Ce-alkyl, R45 is independently selected from halogen, -CN, -CF3, -OCF3, -0-CrCβ-alkyl, -C(0)-O-d- C6-alkyl, -COOH and -NH2,
R46 is independently selected from halogen, -C(0)OCrC6-alkyl, -COOH, -CN, -CF3, -OCF3, - NO2, -OH, -Od-Cβ-alkyl, -NH2, C(=0) or d-Ce-alkyl,
Q is a valence bond, d-Ce-alkylene, -d-C6-alkyl-0-, -d-Ce-alkyl-NH-, -NH-d-Cβ-alkyl, -NH-C(=0)-, -C(=0)-NH-, -0-d-Cβ-alkyl, -C(=0)-, or -d-C6-alkyl-C(=0)-N(R47)- wherein the alkyl moieties are optionally substituted with one or more substituents independently selected from R48,
R47 and R48 are independently selected from hydrogen, d-Cβ-alkyl, aryl optionally substituted with one or more R49,
R49 is independently selected from halogen and -COOH,
T is
• hydrogen,
• d-Cβ-alkyl, C2-C6-alkenyl , C2-C6-alkynyl, d-Cβ-alkyloxy-carbonyl, wherein the alkyl, alkenyl and alkynyl moieties are optionally substituted with one or more substituents independently selected from R50,
• aryl, aryloxy, aryloxy-carbonyl, aryl-d-Cβ-alkyl, aroyl, aryl-d-Cβ-alkoxy, aryl-C2- Cβ-alkenyl, aryl-C2-C6-alkyny-, heteroaryl, heteroaryl-d-Cβ-alkyl, heteroaryl-C2- Cβ-alkenyl, heteroaryl-C2-Cβ-alkynyl,
wherein any alkyl, alkenyl , alkynyl, aryl and heteroaryl moiety is optionally substituted with one or more substituents independently selected from R50,
R50 is CrCe-alkyl, d-Ce-alkoxy, aryl, aryloxy, aryl-d-Ce-alkoxy, -C(=0)-NH-CrC6-alkyl-aryl,
-C(=O)-NR50*-CrC6-alkyl, -C(=0)-NH-(CH2CH20)mCrC6-alkyl-COOH, heteroaryl, het- eroaryl-d-Cβ-alkoxy, -d-Ce-alkyl-COOH, -O-d-Ce-alkyl-COOH, -S(0)2R51, -C2-C6-alkenyl-COOH, -OR51, -N02, halogen, -COOH, -CF3, -CN, =0, -N(R51R52), wherein m is 1 , 2, 3 or 4, and wherein the aryl or heteroaryl moieties are optionally substituted with one or more R53, and the alkyl moieties are optionally substituted with one or more R50B. R50A and R50B are independently selected from -C(0)Od-C6-alkyl, -COOH, -d-C6-alkyl- C(O)OCrC6-alkyl, -d-Cβ-alkyl-COOH, or d-C6-alkyl,
R51 and R52 are independently selected from hydrogen and d-Cβ-alkyl,
R53 is independently selected from d-Cβ-alkyl, d-Cβ-alkoxy, -CrC6-alkyl-COOH, -C2-
Cβ-alkenyl-COOH, -OR51, -N02, halogen, -COOH, -CF3, -CN, or -N(R51R52),
or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base.
130. A pharmaceutical composition according to claim 129 wherein K is a valence bond, d-
Cβ-alkylene, -NH-C(=0)-U-, -d-Cβ-alkyl-S-, -d-Ce-alkyl-O-, or -C(=0)-, wherein any d-
Cβ-alkyl moiety is optionally substituted with R38.
131. A pharmaceutical composition according to claim 130 wherein K is a valence bond, d-
Ce-alkylene, -NH-C(=0)-U-, -d-Cβ-alkyl-S-, or -d-Ce-alkyl-O, wherein any C C6-alkyl moiety is optionally substituted with R38.
132. A pharmaceutical composition according to claim 131 wherein K is a valence bond, d- Cβ-alkylene, or -NH-C(=0)-U, wherein any d-C6-alkyl moiety is optionally substituted with R38.
133. A pharmaceutical composition according to claim 132 wherein K is a valence bond or d-Cβ-alkylene, wherein any d-C6-alkyl moiety is optionally substituted with R38.
134. A pharmaceutical composition according to claim 132 wherein K is a valence bond or -NH-C(=0)-U.
135. A pharmaceutical composition according to claim 133 wherein K is a valence bond.
136. A pharmaceutical composition according to any one of the claims 129 to 135 wherein U is a valence bond or -d-Cβ-alkyl-O-.
137. A pharmaceutical composition according to claim 136 wherein U is a valence bond.
138. A pharmaceutical composition according to any one of the claims 129 to 137 wherein M is arylene or heteroarylene, wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from R40.
139. A pharmaceutical composition according to claim 138 wherein M is ArG1 or Het1 , wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from R40.
140. A pharmaceutical composition according to claim 139 wherein M is ArG1 or Het2, wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from R40.
141. A pharmaceutical composition according to claim 140 wherein M is ArG1 or Het3, wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from R40.
142. A pharmaceutical composition according to claim 141 wherein M is phenylene optionally substituted with one or more substituents independently selected from R40.
143. A pharmaceutical composition according to claim 141 wherein M is indolylene optionally substituted with one or more substituents independently selected from R40.
144. A pharmaceutical composition according to claim 143 wherein M is
Figure imgf000448_0001
145. A pharmaceutical composition according to claim 141 wherein M is carbazolylene optionally substituted with one or more substituents independently selected from R40.
146. A pharmaceutical composition according to claim 145 wherein M is
Figure imgf000448_0002
147. A pharmaceutical composition according to any one of the claims 129 to 146 wherein R40 is selected from
• hydrogen, halogen, -CN, -CF3, -OCF3, -N02, -OR41, -NR41R42, -SR41, -S(0)2R41, -NR41C(0)R42, -OCι-C6-alkyl-C(0)NR41R42, -C2-C6-alkenyl-C(=0)OR41, -C(0)OR41,
=O, -NH-C(=0)-0-d-C6-alkyl, or -NH-C(=0)-C(=0)-0-d-C6-alkyl,
d-Cβ-alkyl or C2-C6- alkenyl which may each optionally be substituted with one or more substituents independently selected from R43,
• aryl, aryloxy, aryl-d-Cβ-alkoxy, aryl-d-Ce-alkyl, aryl-C2-C6-alkenyl, heteroaryl, het- eroaryl-d-Cβ-alkyl, or heteroaryl-C2-C6-alkenyl, wherein the cyclic moieties optionally may be substituted with one or more substituents selected from R44.
148. A pharmaceutical composition according to claim 147 wherein R40 is selected from • hydrogen, halogen, -CN, -CF3, -OCF3, -N02, -OR41, -NR41R42, -SR41, -S(0)2R41, -NR41C(0)R42, -OCrC6-alkyl-C(0)NR41R42, -C2-C6-alkenyl-C(=0)OR41, -C(0)OR41, =O, -NH-C(=0)-0-d-Cβ-alkyl, or -NH-C(=0)-C(=0)-0-CrC6-alkyl,
d-Cβ-alkyl or C2-C6- alkenyl which may each optionally be substituted with one or more substituents independently selected from R43,
• ArG1 , ArG1-0-, ArG1-CrC6-alkoxy, ArGI-d-Cβ-alkyl, ArG1-C2-C6-alkenyl, Het3, Het3-CrC6-alkyl, or Het3-C2-C6-alkenyl, wherein the cyclic moieties optionally may be substituted with one or more substituents selected from R44.
149. A pharmaceutical composition according to claim 148 wherein R40 is selected from
• hydrogen, halogen, -CF3, -N02, -OR41, -NR41R42, -C(0)OR41, =0, or -NR41C(0)R42,
• d-Ce-alkyl,
• ArG1.
150. A pharmaceutical composition according to claim 149 wherein R40 is hydrogen.
151. A pharmaceutical composition according to claim 149 wherein R40 is selected from
• halogen, -N02, -OR41, -NR41R42, -C(0)OR41, or -NR41C(0)R42,
• methyl,
• phenyl.
152. A pharmaceutical composition according to any one of the claims 129 to 151 wherein R41 and R42 are independently selected from hydrogen, d-Ce-alkyl, or aryl, wherein the aryl moieties may optionally be substituted with halogen or -COOH.
153. A pharmaceutical composition according to claim 152 wherein R41 and R42 are independently selected from hydrogen, methyl, ethyl, or phenyl, wherein the phenyl moieties may optionally be substituted with halogen or -COOH.
154. A pharmaceutical composition according to any one of the claims 129 to 153 wherein Q is a valence bond, d-Cβ-alkylene, -CrC6-alkyl-0-, -d-C6-alkyl-NH-, -NH-d-C8-alkyl, -NH-C(=0)-, -C(=0)-NH-, -O-d-Cβ-alkyl, -C(=0)-, or -d-C6-alkyl-C(=0)-N(R47)- wherein the alkyl moieties are optionally substituted with one or more substituents independently selected from R48.
155. A pharmaceutical composition according to claim 154 wherein Q is a valence bond, -CH2-, -CH2-CH2-, -CH2-0-, -CH2-CH2-0-, -CH2-NH-, -CH2-CH2-NH-, -NH-CH2-, -NH-CH2-CH2-, -NH-C(=0)-, -C(=0)-NH-, -0-CH2-, -0-CH2-CH2-, or -C(=0)-.
156. A pharmaceutical composition according to any one of the claims 129 to 155 wherein R47 and R48 are independently selected from hydrogen, methyl and phenyl.
157. A pharmaceutical composition according to any one of the claims 129 to 156 wherein T is
• hydrogen,
• CrCe-alkyl optionally substituted with one or more substituents independently se- lected from R50,
• aryl, aryl-CrC6-alkyl, heteroaryl, wherein the alkyl, aryl and heteroaryl moieties are optionally substituted with one or more substituents independently selected from R50.
158. A pharmaceutical composition according to claim 157 wherein T is
• hydrogen, "CrCe-alkyl optionally substituted with one or more substituents independently selected from R50,
• ArG1 , ArGI-d-Cβ-alkyl, Het3, wherein the alkyl, aryl and heteroaryl moieties are optionally substituted with one or more substituents independently selected from R50.
159. A pharmaceutical composition according to claim 158 wherein T is • hydrogen,
• d-Cβ-alkyl, optionally substituted with one or more substituents independently selected from R50,
• phenyl, phenyl-CrC6-alkyl, wherein the alkyl and phenyl moieties are optionally substituted with one or more substituents independently selected from R50.
160. A pharmaceutical composition according to claim 159 wherein T is phenyl substituted with R50.
161. A pharmaceutical composition according to any one of the claims 129 to 160 wherein R50 is d-Cβ-alkyl, d-Cβ-alkoxy, aryl, aryloxy, aryl-d-Ce-alkoxy, -C(=0)-NH-CrC6-alkyl-aryl, -C(=O)-NR50*-C1-Cβ-alkyl, -C(=0)-NH-(CH2CH20)mCrCβ-alkyl-COOH, heteroaryl, -d- Ce-alkyl-COOH, -0-CrC6-alkyl-COOH, -S(0)2R51, -C2-C6-alkenyl-COOH, -OR51, -N02, halogen, -COOH, -CF3, -CN, =0, -N(R51R52), wherein the aryl or heteroaryl moieties are optionally substituted with one or more R53.
162. A pharmaceutical composition according to claim 161 wherein R50 is d-Ce-alkyl, d- Cβ-alkoxy, aryl, aryloxy, -C(=O)-NR50*-CrC6-alkyl, -C(=0)-NH-(CH2CH20)mCrC6-alkyl- COOH, aryl-d-Ce-alkoxy , -OR51, -N02, halogen, -COOH, -CF3, wherein any aryl moiety is optionally substituted with one or more R53.
163. A pharmaceutical composition according to claim 162 wherein R50 is d-Cβ-alkyl, aryloxy, -C(=O)-NR50*-C1-C6-alkyl, -C(=0)-NH-(CH2CH20)mCrC6-alkyl-COOH, aryl-C
C6-alkoxy, -OR51, halogen, -COOH, -CF3, wherein any aryl moiety is optionally substituted with one or more R53.
164. A pharmaceutical composition according to claim 163 wherein R50 is d-Cβ-alkyl, ArG1-0-, -C(=O)-NR50A-CrC6-alkyl, -C(=0)-NH-(CH2CH20)mCrC6-alkyl-COOH, ArG1-d- Ce-alkoxy , -OR51, halogen, -COOH, -CF3, wherein any aryl moiety is optionally substituted with one or more R53.
165. A pharmaceutical composition according to claim 164 wherein R50 is -C(=O)-NR50*CH2, -C(=0)-NH-(CH2CH20)2CH2l-COOH, or -C(=O)-NR50*CH2CH2.
166. A pharmaceutical composition according to claim 164 wherein R50 is phenyl, methyl or ethyl.
167. A pharmaceutical composition according to claim 166 wherein R50 is methyl or ethyl.
168. A pharmaceutical composition according to any one of the claims 129 to 167 wherein m is 1 or 2.
169. A pharmaceutical composition according to any one of the claims 129 to 168 wherein R51 is methyl.
170. A pharmaceutical composition according to any one of the claims 129 to 169 wherein R53 is d-Cβ-alkyl, d-Ce-alkoxy, -OR51, halogen.or -CF3.
171. A pharmaceutical composition according to any one of the claims 129 to 170 wherein R50* is -C(0)OCH3, -C(0)OCH2CH3 -COOH, -CH2C(0)OCH3, -CH2C(0)OCH2CH3, -CH2CH2C(0)OCH3, -CH2CH2C(0)OCH2CH3, -CH2COOH, methyl, or ethyl.
172. A pharmaceutical composition according to any one of the claims 129 to 171 wherein R50B is -C(0)OCH3, -C(0)OCH2CH3 -COOH, -CH2C(0)OCH3, -CH2C(0)OCH2CH3,
-CH2CH2C(0)OCH3, -CH2CH2C(0)OCH2CH3, -CH2COOH, methyl, or ethyl.
173. A pharmaceutical composition according to any one of the claims 1 to 3 wherein CGr is
Figure imgf000451_0001
wherein V is d-Cβ-alkyl, aryl, heteroaryl, aryl-Ci-β-alkyl- or aryl-C2-β-alkenyl-, wherein the alkyl or alkenyl is optionally substituted with one or more substituents independently selected from R54, and the aryl or heteroaryl is optionally substituted with one or more substituents independently selected from R55,
R54 is independently selected from halogen, -CN, -CF3, -OCF3, aryl, -COOH and -NH2, R55 is independently selected from • hydrogen, halogen, -CN, -CH2CN, -CHF2, -CF3, -OCF3, -OCHF2, -OCH2CF3, -OCF2CHF2, -S(0)2CF3, -OS(0)2CF3, -SCF3, -N02, -OR56, -NR R57, -SR56, -NR56S(0)2R57, -S(0)2NR56R57, -SfOJNR R57, -S(0)R56, -S(0)2R56, -OS(0)2 R56, -C(0)NR56R57, -OC(0)NR56R57, -NR56C(0)R57, -CH2C(0)NR56R57, -Od-C6- alkyl-C(0)NR56R57, -CHzOR56, -CH2OC(0)R56, -CH2NR56R57, -OC(0)R56, -Od-C8- alkyl-C(0)OR56, -Od-Ce-alkyl-OR56, -Sd-Cβ-alkyl-C(0)OR56, -C2-C6-alkenyl- C(=0)OR56, -NR56-C(=0)-CrC6-alkyl-C(=0)OR56, -NR56-C(=0)-d-C6- alkenyl-C(=0)OR56 , -C(0)OR56, or -C2-C6-alkenyl-C(=0)R56,
• d-Ce-alkyl, C2-C6-alkenyl or C2-C6-alkynyl,
which may optionally be substituted with one or more substituents selected from R58,
• aryl, aryloxy, aryloxycarbonyl, aroyl, arylsulfanyl, aryl-d-Ce-afkoxy, aryl-d-Cβ-alkyl, aryl-C2-C6-alkenyl, aroyl-C2-C6-alkenyl, aryl-C2-Ce-alkynyl, heteroaryl, heteroaryl-d- Ce-alkyl, heteroaryl-C2-C6-alkenyl or heteroaryl-C2-C6-alkynyl,
of which the cyclic moieties optionally may be substituted with one or more substituents selected from R59,
R56 and R57 are independently selected from hydrogen, OH, CF3, d-C12-alkyl, aryl-d-Ce- alkyl, -C(=0)-CrC6-alkyl or aryl, wherein the alkyl groups may optionally be substituted with one or more substituents independently selected from R60, and the aryl groups may optionally be substituted with one or more substituents independently selected from R61; R56 and R57 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
R58 is independently selected from halogen, -CN, -CF3, -OCF3, -OR56, and -NR∞R57,
R59 is independently selected from halogen, -C(0)OR56, -CH2C(0)OR56, -CHzOR56, -CN, - CF3, -OCF3, -N02, -OR56, -NR∞R57 and d-Cβ-alkyl, R60 is independently selected from halogen, -CN, -CF3, -OCF3, -OCrC6-alkyl, -C(0)OCrC6- alkyl, -C(=0)-R62, -COOH and -NH2,
R61 is independently selected from halogen, -C(0)OCrC6-alkyl, -COOH, -CN, -CF3, -OCF3, - N02, -OH, -Od-Ce-alkyl, -NH2, C(=0) or d-Cβ-alkyl,
R62 is CrCe-alkyl, aryl optionally substituted with one or more substituents independently selected from halogen, or heteroaryl optionally substituted with one or more d-C6-alkyl independently, or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base.
174. A pharmaceutical composition according to claim 173 wherein V is aryl, heteroaryl, or aryl-d-e-alkyl-, wherein the alkyl is optionally substituted with one or more substituents independently selected R54, and the aryl or heteroaryl is optionally substituted with one or more substituents independently selected from R55.
175. A pharmaceutical composition according to claim 174 wherein V is aryl, Het1 , or aryl-d- e-alkyl-, wherein the alkyl is optionally substituted with one or more substituents independently selected from R54, and the aryl or heteroaryl moiety is optionally substituted with one or more substituents independently selected from R55.
176. A pharmaceutical composition according to claim 175 wherein V is aryl, Het2, or aryl-d- e-alkyl-, wherein the alkyl is optionally substituted with one or more substituents independently selected from R54, and the aryl or heteroaryl moiety is optionally substituted with one or more substituents independently selected from R55.
177. A pharmaceutical composition according to claim 176 wherein V is aryl, Het3, or aryl-d. e-alkyl-, wherein the alkyl is optionally substituted with one or more substituents independently selected from R54, and the aryl or heteroaryl moiety is optionally substituted with one or more substituents independently selected from R55.
178. A pharmaceutical composition according to claim 177 wherein V is aryl optionally substituted with one or more substituents independently selected from R55.
179. A pharmaceutical composition according to claim 178 wherein V is ArG1 optionally substituted with one or more substituents independently selected from R55.
180. A pharmaceutical composition according to claim 179 wherein V is phenyl, naphthyl or anthranyl optionally substituted with one or more substituents independently selected from R55.
181. A pharmaceutical composition according to claim 180 wherein V is phenyl optionally substituted with one or more substituents independently selected from R55.
182. A pharmaceutical composition according to any one of the claims 173 to 181 wherein R55 is independently selected from .halogen, d-Cβ-alkyl, -CN, -OCF3 ,-CF3, -N02, -OR56, -NR56R57, -NR56C(0)R57
-SR56, -Od-C8-alkyl-C(0)OR56, or -dOJOR56,
• CrCe-alkyl optionally substituted with one or more substituents independently selected from R58
• aryl, aryl-d-Cβ-alkyl, heteroaryl, or heteroaryl-d-Ce-alkyl of which the cyclic moieties optionally may be substituted with one or more substituents independently selected from R59.
183. A pharmaceutical composition according to claim 182 wherein R55 is independently selected from
• halogen, d-Ce-alkyl, -CN, -OCF3 ,-CF3, -N02, -OR56, -NR∞R57, -NR56C(O)R57 -SR56, -Od-Cs-alkyl-dOJOR56, or -C(0)OR56
• d-C6-alkyl optionally substituted with one or more substituents independently selected from R58
• ArG1, ArG1-d-Ce-alkyl, Het3, or Het3-d-C6-alkyl of which the cyclic moieties optionally may be substituted with one or more substitu- ents independently selected from R59.
184. A pharmaceutical composition according to claim 183 wherein R55 is independently selected from halogen, -OR56, -NR∞R57, -C(0)OR56, -OC1-C8-alkyl-C(0)OR56, -NR56C(0)R57 or d-Ce-alkyl.
185. A pharmaceutical composition according to claim 184 wherein R55 is independently se- lected from halogen, -OR56, -NR56R57, -C(0)OR56, -Od-C8-alkyl-C(0)OR56, -NR56C(O)R57, methyl or ethyl.
186. A pharmaceutical composition according to any one of the claims 173 to 185 wherein R56 and R57 are independently selected from hydrogen, CF3, d-C12-alkyl, or -C(=O)-CrC6-alkyl; R56 and R57 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom.
187. A pharmaceutical composition according to claim 186 wherein R56 and R57 are independently selected from hydrogen or d-C12-alkyl, R56 and R57 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom.
188. A pharmaceutical composition according to claim 187 wherein R56 and R57 are inde- pendently selected from hydrogen or methyl, ethyl, propyl butyl, R56 and R57 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom.
189. A pharmaceutical composition according to any one of the claims 1 to 3 wherein CGr is
Figure imgf000455_0001
wherein AA is d-Ce-alkyl, aryl, heteroaryl, aryl-d-β-alkyl- or aryl-C2-β-alkenyl-, wherein the alkyl or alkenyl is optionally substituted with one or more substituents independently selected from R63, and the aryl or heteroaryl is optionally substituted with one or more substituents independently selected from R64,
R63 is independently selected from halogen, -CN, -CF3, -OCF3, aryl, -COOH and -NH2l
R64 is independently selected from
• hydrogen, halogen, -CN, -CH2CN, -CHF2, -CF3, -OCF3, -OCHF2, -OCH2CF3, -OCF2CHF2, -S(0)2CF3, -OS(0)2CF3, -SCF3, -N02, -OR65, -NR65R66, -SR65, -NR65S(0)2R66, -S(0)2NR65R66, -S(0)NR65R66, -S(0)R65, -S(0)2R65, -OS(0)2 R65, -C(0)NR65R66, -OC(0)NR65R66, -NR65C(0)R66, -CH2C(0)NR65R66, -OCrC6- alkyl-C(0)NR65R66, -CH2OR65, -CH2OC(0)R65, -CH2NR65R66, -OC(0)R65, -OCrC6- alkyl-C(0)OR65, -Od-Ce-alkyl-OR65, -SCrC6-alkyl-C(0)OR65, -C2-C6-alkenyl- C(=0)OR65, -NR65-C(=0)-d-C6-alkyl-C(=0)OR65, -NR65-C(=0)-CrC6- alkenyl-C(=0)OR65 , -C(0)OR65, or -C2-C6-alkenyl-C(=0)R65,
•CrCβ-alkyl, C2-C6-alkenyl or C2-C6-alkynyl, each of which may optionally be substituted with one or more substituents selected from R67,
• aryl, aryloxy, aryloxycarbonyl, aroyl, arylsulfanyl, aryl-d-Cβ-alkoxy, aryl-d-Ce-alkyl, aryl-C2-C6-alkenyl, aroyl-C2-C6-alkenyl, aryl-C2-C6-alkynyl, heteroaryl, heteroaryl-d- Ce-alkyl, heteroaryl-C2-C6-alkenyl or heteroaryl-C2-C6-alkynyl, of which the cyclic moieties optionally may be substituted with one or more substituents selected from R68,
R65 and R66 are independently selected from hydrogen, OH, CF3, Cι-C12-alkyl, aryl-d-Cβ- alkyl, -C(=0)-R69, aryl or heteroaryl, wherein the alkyl groups may optionally be substituted with one or more substituents selected from R70, and the aryl and heteroaryl groups may optionally be substituted with one or more substituents independently selected from R71; R65 and R66 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
R67 is independently selected from halogen, -CN, -CF3, -OCF3, -OR65, and -NR65R66,
R68 is independently selected from halogen, -C(0)OR65, -CH2C(0)OR65, -CH2OR65, -CN, - CF3, -OCF3, -N02, -OR65, -NR65R66 and d-Ce-alkyl,
R69 is independently selected from d-Ce-alkyl, aryl optionally substituted with one or more halogen, or heteroaryl optionally substituted with one or more d-Ce-alkyl,
R70 is independently selected from halogen, -CN, -CF3, -OCF3, -OCrC6-alkyl, -C(0)Od-C6- alkyl, -COOH and -NH2,
R71 is independently selected from halogen, -C(0)OCrCβ-alkyl, -COOH, -CN, -CF3, -OCF3, - NO2, -OH, -Od-Ce-alkyl, -NH2, C(=0) or d-Cβ-alkyl,
or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base.
190. A pharmaceutical composition according to claim 189 wherein AA is aryl, heteroaryl or aryl-Ci-e-alkyl-, wherein the alkyl is optionally substituted with one or more R63, and the aryl or heteroaryl is optionally substituted with one or more substituents independently selected from R64.
191. A pharmaceutical composition according to claim 190 wherein AA is aryl or heteroaryl optionally substituted with one or more substituents independently selected from R64.
192. A pharmaceutical composition according to claim 191 wherein AA is ArG1 or Het1 optionally substituted with one or more substituents independently selected from R64.
193. A pharmaceutical composition according to claim 192 wherein AA is ArG1 or Het2 optionally substituted with one or more substituents independently selected from R64.
194. A pharmaceutical composition according to claim 193 wherein AA is ArG1 or Het3 optionally substituted with one or more substituents independently selected from R64.
195. A pharmaceutical composition according to claim 194 wherein AA is phenyl, naphtyl, anthryl, carbazolyl, thienyl, pyridyl, or benzodioxyl optionally substituted with one or more substituents independently selected from R64.
196. A pharmaceutical composition according to claim 195 wherein AA is phenyl or naphtyl optionally substituted with one or more substituents independently selected from R64.
197. A pharmaceutical composition according to any one of the claims 189 to 196 wherein R64 is independently selected from hydrogen, halogen, -CF3, -OCF3, -OR65, -NR65R66, d-Ce- alkyl , -OC(0)R65, -OCrC6-alkyl-C(0)OR65, aryl-C2-C6-alkenyl, aryloxy or aryl, wherein d- Ce-alkyl is optionally substituted with one or more substituents independently selected from R67, and the cyclic moieties optionally are substituted with one or more substituents independently selected from R68.
198. A pharmaceutical composition according to claim 197 wherein R64 is independently selected from halogen, -CF3, -OCF3, -OR65, -NR65R66, methyl, ethyl, propyl, -OC(0)R65, -OCH2-C(0)OR65, -OCH2-CH2-C(0)OR65, phenoxy optionally substituted with one or more substituents independently selected from R68.
199. A pharmaceutical composition according to any one of the claims 189 to 198 wherein R65 and R66 are independently selected from hydrogen, CF3, d-C12-alkyl, aryl, or heteroaryl optionally substituted with one or more substituents independently selected from R71.
200. A pharmaceutical composition according to claim 199 wherein R65 and R66 are independently hydrogen, d-C12-alkyl, aryl, or heteroaryl optionally substituted with one or more substituents independently selected from R71.
201. A pharmaceutical composition according to claim 200 wherein R65 and R66 are independently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, ArG1 or Het1 optionally substituted with one or more substituents independently selected from R71.
202. A pharmaceutical composition according to claim 201 wherein R65 and R66 are independently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, ArG1 or Het2 optionally substituted with one or more substituents independently selected from R71.
203. A pharmaceutical composition according to claim 202 wherein R65 and R66 are independently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, ArG1 or Het3 optionally substituted with one or more substituents independently selected from R71.
204. A pharmaceutical composition according to claim 203 wherein R65 and R66 are inde- pendently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, phenyl, naphtyl, thiadiazolyl optionally substituted with one or more R71 independently; or isoxazoiyl optionally substituted with one or more substituents independently selected from R71.
205. A pharmaceutical composition according to any one of the claims 189 to 204 wherein R71 is halogen or CrCβ-alkyl.
206. A pharmaceutical composition according to claim 205 wherein R71 is halogen or methyl.
207. A pharmaceutical preparation according to any one of the claims 1 to 205 wherein Frg1 consists of 0 to 5 neutral amino acids independently selected from the group consisting of Gly, Ala, Thr, and Ser.
208. A pharmaceutical preparation according to claim 207 wherein Frg1 consists of 0 to 5 Gly.
209. A pharmaceutical preparation according to claim 208 wherein Frg 1 consists of 0 Gly.
210. A pharmaceutical preparation according to claim 208 wherein Frg1 consists of 1 Gly.
211. A pharmaceutical preparation according to claim 208 wherein Frg1 consists of 2 Gly.
212. A pharmaceutical preparation according to claim 208 wherein Frg 1 consists of 3 Gly.
213. A pharmaceutical preparation according to claim 208 wherein Frg1 consists of 4 Gly.
214. A pharmaceutical preparation according to claim 208 wherein Frg1 consists of 5 Gly.
215. A pharmaceutical preparation according to any one of the claims 1 to 214 wherein GB is of the formula B1-B2-C(0)-, B1-B2-S02- or B1-B2-CH2-, wherein B1 and B2 are as defined in claim 1.
216. A pharmaceutical preparation according to any one of the claims 1 to 214 wherein GB is of the formula B1-B2-C(0)-, B1-B2-S02- or B1-B2-NH-, wherein B1 and B2 are as defined in claim 1.
217. A pharmaceutical preparation according to any one of the claims 1 to 214 wherein GB is of the formula B1-B2-C(0)-, B1-B2-CH2- or B1-B2-NH-, wherein B1 and B2 are as defined in claim 1.
218. A pharmaceutical preparation according to any one of the claims 1 to 214 wherein GB is of the formula B1-B2-CH2-, B1-B2-S02- or B1-B2-NH-, wherein B1 and B2 are as defined in claim 1.
219. A pharmaceutical preparation according to any one of the claims 215 or 216 wherein GB is of the formula B1-B2-C(0)- or B1-B2-S02-, wherein B1 and B2 are as defined in claim 1.
220. A pharmaceutical preparation according to any one of he claims 215 or 217 wherein GE is of the formula B1-B2-C(0)- or B1-B2-CH2-, wherein B1 and B2 are as defined in claim 1.
221. A pharmaceutical preparation according to any one of he claims 216 or 217 wherein GE is of the formula B1-B2-C(0)- or B1-B2-NH-, wherein B1 and B2 are as defined in claim 1
222. A pharmaceutical preparation according to any one of he claims 215 or 218 wherein G is of the formula B1-B2-CH2- or B1-B2-S02- , wherein B1 and B2 are as defined in claim 1.
223. A pharmaceutical preparation according to any one of he claims 216 or 218 wherein GB is of the formula B1-B2-NH- or B1-B2-S02- , wherein B1 and B2 are as defined in claim 1
224. A pharmaceutical preparation according to any one of he claims 217 or 218 wherein GB is of the formula B1-B2-CH2- or B1-B2-NH- , wherein B1 and B2 are as defined in claim 1.
225. A pharmaceutical preparation according to any one of he claims 219, 220, or 221 wherein GB is of the formula B1-B2-C(0)-.
226. A pharmaceutical preparation according to any one of he claims 220, 222 or 224 wherein GB is of the formula B1-B2-CH2-.
227. A pharmaceutical preparation according to any one of he claims 220, 222 or 223 wherein GB is of the formula B1-B2-S02-.
228. A pharmaceutical preparation according to any one of he claims 221, 223 or 224 wherein GB is of the formula B1-B2-NH-.
229. A pharmaceutical preparation according to any one of he claims 1 to 228 wherein B1 is a valence bond, -0-, or -S-.
230. A pharmaceutical preparation according to any one of he claims 1 to 228 wherein B1 is a valence bond, -0-, or -N(R6B)-.
231. A pharmaceutical preparation according to any one of he claims 1 to 228 wherein B1 is a valence bond, -S-, or -N(R6B)-.
232. A pharmaceutical preparation according to any one of he claims 1 to 228 wherein B1 is -0-, -S- or -N(R6B)-.
233. A pharmaceutical preparation according to any one of he claims 229 or 230 wherein B1 is a valence bond or -0-.
234. A pharmaceutical preparation according to any one of he claims 229 or 231 wherein B1 is a valence bond or -S-.
235. A pharmaceutical preparation according to any one of he claims 230 or 231 wherein B1 is a valence bond or -N(R6B)-.
236. A pharmaceutical preparation according to any one of he claims 229 or 232 wherein B1 is -O-or -S-.
237. A pharmaceutical preparation according to any one of the claims 230 or 232 wherein B1 is -O-or -N(R6B)-.
238. A pharmaceutical preparation according to any one of the claims 231 or 232 wherein B1 is -S-or -N(R6B)-.
239. A pharmaceutical preparation according to any one of the claims 233, 234 or 235 wherein B1 is a valence bond.
240. A pharmaceutical preparation according to any one of the claims 233, 236 or 237 wherein B1 is -O-.
241. A pharmaceutical preparation according to any one of the claims 234, 236 or 238 wherein B1 is -S-.
242. A pharmaceutical preparation according to any one of the claims 235, 237 or 238 wherein B1 is -N(R6B)-.
243. A pharmaceutical preparation according to any one of the claims 1 to 242 wherein B2 is a valence bond, CrCι8-alkylene, C2-C18-alkenylene, C2-C18-alkynylene, arylene, heteroary- lene, -d-C18-alkyl-aryl-, -C(=O)-d-C18-alkyl-C(=O)-, -C(=O)-CrC18-alkyl-O-CrC18-alkyl- C(=0)-, -C(=0)-CrCi8-alkyl-S-Ci-C18-alkyl-C(=0)-, -C(=0)-CrCi8-alkyl-NR6-CrC18-alkyl- C(=0)-; and the alkylene and arylene moieties are optionally substituted as defined in claim 1.
244. A pharmaceutical preparation according to claim 243 wherein B2 is a valence bond, d- C18-alkylene, C2-C18-alkenylene, C2-C18-alkynylene, arylene, heteroarylene, -d-C18-alkyl-aryl-
, -C(=0)-CrC18-alkyl-C(=0)-, -C(=0)-C1-C18-alkyl-0-C1-C18-alkyl-C(=0)-, and the alkylene and arylene moieties are optionally substituted as defined in claim 1.
245. A pharmaceutical preparation according to claim 244 wherein B2 is a valence bond, d- C18-alkylene, C2-C18-alkenylene, C2-C18-alkynylene, arylene, heteroarylene, -d-Cia-alkyl-aryl- , -C(=0)-d-C18-alkyl-C(=0)-, and the alkylene and arylene moieties are optionally substituted as defined in claim 1.
246. A pharmaceutical preparation according to claim 245 wherein B2 is a valence bond, d- Ciβ-alkylene, arylene, heteroarylene, -d-Cι8-alkyl-aryl-, -C(=0)-d-C18-alkyl-C(=0)-, and the alkylene and arylene moieties are optionally substituted as defined in claim 1.
247. A pharmaceutical preparation according to claim 246 wherein B2 is a valence bond, d- dβ-alkylene, arylene, heteroarylene, -CrC18-alkyl-aryl-, and the alkylene and arylene moieties are optionally substituted as defined in claim 1.
248. A pharmaceutical preparation according to claim 247 wherein B2 is a valence bond, d- Cι8-alkylene, arylene, -d-C18-alkyl-aryl-, and the alkylene and arylene moieties are optionally substituted as defined in claim 1.
249. A pharmaceutical preparation according to claim 248 wherein B2 is a valence bond or -CrCι8-alkylene, and the alkylene moieties are optionally substituted as defined in claim 1.
250. A pharmaceutical preparation according to any one of the claims 1 to 249 whereiin Frg2 comprises 1 to 16 positively charged groups.
251. A pharmaceutical preparation according to claim 250 wherein Frg2 comprises 1 to 12 positively charged groups.
252. A pharmaceutical preparation according to claim 251 wherein Frg2 comprises 1 to 10 positively charged groups.
253. A pharmaceutical preparation according to any one of the claims 1 to 249 wherein Frg2 comprises 10 to 20 positively charged groups.
254. A pharmaceutical preparation according to claim 253 wherein Frg2 comprises 12 to 20 positively charged groups.
255. A pharmaceutical preparation according to claim 254 wherein Frg2 comprises 16 to 20 positively charged groups.
256. A pharmaceutical preparation according to any one of the claims 250 to 255 wherein the positively charged groups of Frg2 are basic amino acids independently selected from the group consisting of Lys and Arg and D-isomers of these.
257. A pharmaceutical preparation according to claim 256 wherein the basic amino acids are all Arg.
258. A pharmaceutical preparation according to any one of the claims 250 to 257, wherein Frg2 comprises one or more neutral amino acids independently selected from the group consisting of Gly, Ala, Thr, and Ser.
259. A pharmaceutical preparation according to claim 258, wherein Frg2 comprises one or more Gly.
260. A pharmaceutical preparation according to any one of the claims 1 to 259 wherein X is - OH or -NH2.
261. A pharmaceutical preparation according to claim 260 wherein X is -NH2.
262. A pharmaceutical preparation according to any one of the claims 1 to 261 which further comprises at least 3 phenolic molecules per putative insulin hexamer.
263. A pharmaceutical preparation according to any one of the claims 1 to 262 wherein the acid-stabilised insulin is an analogue of human insulin wherein A21 is Ala, Gin, Glu, Gly, His, He, Leu, Met, Phe, Ser, Thr, Trp, Tyr, Val, and hSer.
264. A pharmaceutical preparation according to claim 263 wherein the acid-stabilised insulin is an analogue of human insulin wherein A21 is Ala, Gly, He, Leu, Phe, Ser, Thr, Val, and hSer.
265. A pharmaceutical preparation according to claim 264 wherein the acid-stabilised insulin is an analogue of human insulin wherein A21 is Ala or Gly.
266. A pharmaceutical preparation according to claim 265 wherein the acid-stabilised insulin is an analogue of human insulin wherein A21 is Gly.
267. A pharmaceutical preparation according to any one of the claims 263 to 266 wherein the acid-stabilised insulin is an analogue of human insulin further modified by exchange or deletion of one or more amino acid residues according to the following: B3 is selected from Thr, Ser, Lys or Ala A18 is Gln B28 is Lys, Asp or Glu B29 is Pro or Glu B9 is Glu or Asp BIO is Glu B25 is deleted B30 is deleted.
268. A pharmaceutical preparation according to claim 267 wherein the acid-stabilised insulin is an analogue of human insulin further modified by exchange of B28 to Lys or Asp.
269. A pharmaceutical preparation according to claim 268 wherein the acid-stabilised insulin is an analogue of human insulin further modified by exchange of B28 to Asp.
270. A pharmaceutical preparation according to claim 268 wherein the acid-stabilised insulin is an analogue of human insulin further modified by exchange of B28 to Lys.
271. A pharmaceutical preparation according any one of the claims 263 to 270 wherein the acid-stabilised insulin is an analogue of human insulin further modified by exchange of B29 to Pro.
272. A pharmaceutical preparation according any one of the claims 263 to 271 wherein the acid-stabilised insulin is an analogue of human insulin further modified by exchange of B3 to Lys or Ala.
273. A pharmaceutical preparation according any one of the claims 263 to 272 wherein the acid-stabilised insulin is an analogue of human insulin further modified by exchange of A18 to Gin.
274. A pharmaceutical preparation according any one of the claims 263 to 273 wherein the acid-stabilised insulin is an analogue of human insulin further modified by deletion of B25.
275. A pharmaceutical preparation according any one of the claims 263 to 274 wherein the acid-stabilised insulin is an analogue of human insulin further modified by deletion of B30.
276. A pharmaceutical preparation according to claim 263 wherein the acid-stabilised insulin is selected from the group
A21G
A21G, B28K, B29P A21G, B28D
A21G, B28E
A21G, B3K, B29E
A21G, desB27
A21G, B9E A21G, B9D
A21G, B10ε
A21G, desB25
A21G, desB30
A21G, B28K, B29P, desB30 A21G, B28D, desB30
A21G, B28E, desB30
A21G, B3K, B29E, desB30
A21G, desB27, desB30
A21G, B9E, desB30 A21G, B9D, desB30
A21G, B10E, desB30
A21G, desB25, desB30.
277. A pharmaceutical preparation according to any one of the claims 1 to 276 wherein zinc ions are present in an amount corresponding to 10 to 40 μg Zn/100 U insulin.
278. A pharmaceutical preparation according to claim 277 wherein zinc ions are present in an amount corresponding to 10 to 26 μg Zn 100 U insulin.
279. A pharmaceutical preparation according to any one of the claims 1 to 278 wherein the ratio between insulin and the zinc-binding ligand according to any one of the claims 1 to 261 is in the range from 99:1 to 1 :99.
280. A pharmaceutical preparation according to claim 279 wherein the ratio between insulin and the zinc-binding ligand according to any one of the claims 1 to 261 is in the range from 95:5 to 5:95.
281. A pharmaceutical preparation according to claim 280 wherein the ratio between between insulin and the zinc-binding ligand according to any one of the claims 1 to 261 is in the range from 80:20 to 20:80.
282. A pharmaceutical preparation according to claim 281 wherein the ratio between between insulin and the zinc-binding ligand according to any one of the claims 1 to 261 is in the range from 70:30 to 30:70.
283. A pharmaceutical preparation according to any one of the claims 1 to 282 wherein the concentration of insulin is 60 to 3000 nmol/ml.
284. A pharmaceutical preparation according to claim 283 wherein the concentration of insulin is 240 to 1200 nmol/ml.
285. A pharmaceutical preparation according to claim 284 wherein the concentration of insulin is about 600 nmol/ml.
286. A method of preparing a zinc-binding ligand according to claim 1 comprising the steps of
• Identifying starter compounds that binds to the R-state HisB10-Zn2+ site
• optionally attaching a fragment consisting of 0 to 5 neutral α- or β-amino acids
• attaching a fragment comprising 1 to 20 positively charged groups independently se- lected from amino or guanidino groups.
287. Method of prolonging the action of an acid-stabilised insulin preparation which comprises adding a zinc-binding ligand according to any of claims 1 to 261 to the acid-stabilised insulin preparation.
288. A method of treating type 1 or type 2 diabetes comprising administering to a patient in need thereof a theraputically effective amount of a pharmaceutical preparation according to any one of the claims 1 to 282.
289. Use of a preparation according to any one of the claims 1 to 282 for the preparation of a medicament for treatment of type 1 or type 2 diabetes.
PCT/DK2004/000158 2003-03-11 2004-03-11 Pharmaceutical preparations comprising acid-stabilised insulin WO2004080480A1 (en)

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WO2007104786A1 (en) 2006-03-15 2007-09-20 Novo Nordisk A/S Mixtures of amylin and insulin
JP2007291079A (en) * 2006-03-27 2007-11-08 Otsuka Pharmaceut Co Ltd Pharmaceutical composed of carbostyril compound
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EP2282736A2 (en) * 2008-04-30 2011-02-16 Wellstat Therapeutics Corporation Tetrazole compounds for reducing uric acid
US8148541B2 (en) * 2006-01-09 2012-04-03 Korea Research Institute Of Bioscience And Biotechnology Rhodanine derivatives, a process for the preparation thereof and pharmaceutical composition containing the same
US8404645B2 (en) 2007-09-11 2013-03-26 Novo Nordisk As Stable formulations of amylin and its analogues
WO2013066134A2 (en) * 2011-11-04 2013-05-10 한국생명공학연구원 Novel cinnamyl-rhodanine derivatives and pharmaceutical composition comprising same as active ingredients
WO2013096093A1 (en) * 2011-12-21 2013-06-27 Merck Sharp & Dohme Corp. Compounds as dgat-1 inhibitors
WO2013124245A1 (en) 2012-02-21 2013-08-29 Bayer Intellectual Property Gmbh Herbicidally active 4-nitro-substituted n-(tetrazol-5-yl)-, n-(triazol-5-yl)-, and n-(1,3,4-oxadiazol-2-yl)aryl carboxylic acid amides
WO2014118355A1 (en) 2013-02-04 2014-08-07 Sanofi Stabilized pharmaceutical formulations of insulin analogues and/or insulin derivatives
US9061067B2 (en) 2004-07-08 2015-06-23 Novo Nordisk A/S Polypeptide protracting tags
US9364519B2 (en) 2011-09-01 2016-06-14 Sanofi-Aventis Deutschland Gmbh Pharmaceutical composition for use in the treatment of a neurodegenerative disease
US9408893B2 (en) 2011-08-29 2016-08-09 Sanofi-Aventis Deutschland Gmbh Pharmaceutical combination for use in glycemic control in diabetes type 2 patients
US9499521B2 (en) 2014-12-11 2016-11-22 President And Fellows Of Harvard College Inhibitors of cellular necrosis and related methods
US9526764B2 (en) 2008-10-17 2016-12-27 Sanofi-Aventis Deutschland Gmbh Combination of an insulin and a GLP-1-agonist
US9707176B2 (en) 2009-11-13 2017-07-18 Sanofi-Aventis Deutschland Gmbh Pharmaceutical composition comprising a GLP-1 agonist and methionine
US9821032B2 (en) 2011-05-13 2017-11-21 Sanofi-Aventis Deutschland Gmbh Pharmaceutical combination for improving glycemic control as add-on therapy to basal insulin
US9839692B2 (en) 2014-01-09 2017-12-12 Sanofi Stabilized pharmaceutical formulations of insulin analogues and/or insulin derivatives
US9895423B2 (en) 2014-01-09 2018-02-20 Sanofi Stabilized pharmaceutical formulations of insulin aspart
US9895424B2 (en) 2014-01-09 2018-02-20 Sanofi Stabilized pharmaceutical formulations of insulin analogues and/or insulin derivatives
US9950039B2 (en) 2014-12-12 2018-04-24 Sanofi-Aventis Deutschland Gmbh Insulin glargine/lixisenatide fixed ratio formulation
US9981013B2 (en) 2010-08-30 2018-05-29 Sanofi-Aventis Deutschland Gmbh Use of AVE0010 for the treatment of diabetes mellitus type 2
CN108164594A (en) * 2017-12-08 2018-06-15 珠海冀百康生物科技有限公司 The recovery method that a kind of insulin precurosor for removing 30 amino acids residue of B chains precipitates
US10029011B2 (en) 2009-11-13 2018-07-24 Sanofi-Aventis Deutschland Gmbh Pharmaceutical composition comprising a GLP-1 agonist, an insulin and methionine
US10159713B2 (en) 2015-03-18 2018-12-25 Sanofi-Aventis Deutschland Gmbh Treatment of type 2 diabetes mellitus patients
US10434147B2 (en) 2015-03-13 2019-10-08 Sanofi-Aventis Deutschland Gmbh Treatment type 2 diabetes mellitus patients
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US7521465B2 (en) 2003-01-17 2009-04-21 Bexel Pharmaceuticals, Inc. Diphenyl ether derivatives
US7087576B2 (en) 2003-10-07 2006-08-08 Bexel Pharmaceuticals, Inc. Dipeptide phenyl ethers
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US9061067B2 (en) 2004-07-08 2015-06-23 Novo Nordisk A/S Polypeptide protracting tags
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US8148541B2 (en) * 2006-01-09 2012-04-03 Korea Research Institute Of Bioscience And Biotechnology Rhodanine derivatives, a process for the preparation thereof and pharmaceutical composition containing the same
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