EP1073624A1 - Aminomethyl-benzoic ester derivatives as tryptase inhibitors - Google Patents

Aminomethyl-benzoic ester derivatives as tryptase inhibitors

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Publication number
EP1073624A1
EP1073624A1 EP99918168A EP99918168A EP1073624A1 EP 1073624 A1 EP1073624 A1 EP 1073624A1 EP 99918168 A EP99918168 A EP 99918168A EP 99918168 A EP99918168 A EP 99918168A EP 1073624 A1 EP1073624 A1 EP 1073624A1
Authority
EP
European Patent Office
Prior art keywords
gradient
tfa
acetonitrile
water
hplc
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP99918168A
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German (de)
French (fr)
Inventor
Bohdan Waszkowycz
Sarah Elizabeth Lively
Martin James Harrison
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Tularik Ltd
Original Assignee
Proteus Molecular Design Ltd
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Publication date
Priority claimed from GBGB9808813.1A external-priority patent/GB9808813D0/en
Priority claimed from GBGB9822432.2A external-priority patent/GB9822432D0/en
Application filed by Proteus Molecular Design Ltd filed Critical Proteus Molecular Design Ltd
Publication of EP1073624A1 publication Critical patent/EP1073624A1/en
Withdrawn legal-status Critical Current

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    • C07D257/02—Heterocyclic compounds containing rings having four nitrogen atoms as the only ring hetero atoms not condensed with other rings
    • C07D257/04—Five-membered rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/21—Esters, e.g. nitroglycerine, selenocyanates
    • A61K31/215—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids
    • A61K31/235—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids having an aromatic ring attached to a carboxyl group
    • A61K31/24—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids having an aromatic ring attached to a carboxyl group having an amino or nitro group
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    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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    • A61K31/275—Nitriles; Isonitriles
    • A61K31/277—Nitriles; Isonitriles having a ring, e.g. verapamil
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/445—Non condensed piperidines, e.g. piperocaine
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/47—Quinolines; Isoquinolines
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    • C07C229/00—Compounds containing amino and carboxyl groups bound to the same carbon skeleton
    • C07C229/38—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino groups bound to acyclic carbon atoms and carboxyl groups bound to carbon atoms of six-membered aromatic rings of the same carbon skeleton
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    • C07C233/45—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by carboxyl groups
    • C07C233/46—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by carboxyl groups with the substituted hydrocarbon radical bound to the nitrogen atom of the carboxamide group by an acyclic carbon atom
    • C07C233/47—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by carboxyl groups with the substituted hydrocarbon radical bound to the nitrogen atom of the carboxamide group by an acyclic carbon atom having the carbon atom of the carboxamide group bound to a hydrogen atom or to a carbon atom of an acyclic saturated carbon skeleton
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    • C07C235/52—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups bound to carbon atoms of six-membered aromatic rings and singly-bound oxygen atoms bound to the same carbon skeleton with carbon atoms of carboxamide groups and singly-bound oxygen atoms bound to carbon atoms of the same non-condensed six-membered aromatic ring having the nitrogen atom of at least one of the carboxamide groups bound to an acyclic carbon atom of a hydrocarbon radical substituted by carboxyl groups
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    • C07C235/70—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups and doubly-bound oxygen atoms bound to the same carbon skeleton
    • C07C235/72—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups and doubly-bound oxygen atoms bound to the same carbon skeleton with the carbon atoms of the carboxamide groups bound to acyclic carbon atoms
    • C07C235/74—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups and doubly-bound oxygen atoms bound to the same carbon skeleton with the carbon atoms of the carboxamide groups bound to acyclic carbon atoms of a saturated carbon skeleton
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    • C07C235/70—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups and doubly-bound oxygen atoms bound to the same carbon skeleton
    • C07C235/72—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups and doubly-bound oxygen atoms bound to the same carbon skeleton with the carbon atoms of the carboxamide groups bound to acyclic carbon atoms
    • C07C235/76—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups and doubly-bound oxygen atoms bound to the same carbon skeleton with the carbon atoms of the carboxamide groups bound to acyclic carbon atoms of an unsaturated carbon skeleton
    • C07C235/78—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups and doubly-bound oxygen atoms bound to the same carbon skeleton with the carbon atoms of the carboxamide groups bound to acyclic carbon atoms of an unsaturated carbon skeleton the carbon skeleton containing rings
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    • C07C255/55—Carboxylic acid nitriles having cyano groups bound to carbon atoms of six-membered aromatic rings of a carbon skeleton containing cyano groups and esterified hydroxy groups bound to the carbon skeleton
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    • C07C311/00—Amides of sulfonic acids, i.e. compounds having singly-bound oxygen atoms of sulfo groups replaced by nitrogen atoms, not being part of nitro or nitroso groups
    • C07C311/22—Sulfonamides, the carbon skeleton of the acid part being further substituted by singly-bound oxygen atoms
    • C07C311/29—Sulfonamides, the carbon skeleton of the acid part being further substituted by singly-bound oxygen atoms having the sulfur atom of at least one of the sulfonamide groups bound to a carbon atom of a six-membered aromatic ring
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    • C07C311/50—Compounds containing any of the groups, X being a hetero atom, Y being any atom
    • C07C311/51—Y being a hydrogen or a carbon atom
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    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D207/00—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom
    • C07D207/02—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D207/04—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members
    • C07D207/10—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D207/16—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
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    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D211/00—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings
    • C07D211/04—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D211/06—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members
    • C07D211/36—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D211/60—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
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    • C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
    • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
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    • C07D213/63—One oxygen atom
    • C07D213/65—One oxygen atom attached in position 3 or 5
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    • C07D215/00—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems
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    • C07D215/16—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D215/20—Oxygen atoms
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    • C07D295/00—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms
    • C07D295/16—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms acylated on ring nitrogen atoms
    • C07D295/18—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms acylated on ring nitrogen atoms by radicals derived from carboxylic acids, or sulfur or nitrogen analogues thereof
    • C07D295/182—Radicals derived from carboxylic acids
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    • C07D295/16—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms acylated on ring nitrogen atoms
    • C07D295/18—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms acylated on ring nitrogen atoms by radicals derived from carboxylic acids, or sulfur or nitrogen analogues thereof
    • C07D295/182—Radicals derived from carboxylic acids
    • C07D295/192—Radicals derived from carboxylic acids from aromatic carboxylic acids

Definitions

  • the invention relates to compounds for use m the treatment of mast cell mediated diseases such as asthma and other allergic and inflammatory conditions and to pharmaceutical compositions thereof and their use m the 5 treatment of the human or animal body, and m particular to compounds which are tryptase inhibitors.
  • asthma is a complex disease frequently characterised by progressive developments of hyper- responsiveness of the trachea and bronchi as a result of - 2 - chronic inflammation reactions which irritate the epithelium lining the airway and cause pathological thickening of the underlying tissues.
  • Leukocytes and mast cells are present in the epithelium and smooth muscle tissue of the bronchi where they are activated initially by binding of specific inhaled antigens to IgE receptors. Activated mast cells release a number of preformed or primary chemical mediators of the inflammatory response in asthma as well as enzymes. Moreover, secondary mediators of inflammation are generated by enzymatic reactions of activated mast cells and a number of large molecules are released by degranulation of mast cells.
  • bronchodilator drugs which causes airways to expand.
  • the most effective bronchodilators are the (3-adrenergic agonists which mimic the actions of adrenalin. These are widely used and are simply administered to the lungs by inhalers.
  • bronchoconstrictor drugs are primarily of use in short term symptomatic relief, and do not prevent asthma attacks nor deterioration of lung function over the long term.
  • Anti -inflammatory drugs such as cromoglycate and the corticosteroids are also widely used in asthma therapy.
  • Cromoglycate has anti -inflammatory activity and has been found to be extremely safe. Although such cromolyns have minimal side effects and are currently preferred for initial preventive therapy in children, it is well known that they are of limited efficacy.
  • corticosteroids in asthma therapy was a major advance since they are very effective anti- inflammatory agents, however, steroids are very powerful, broad spectrum anti -inflammatory agents and their potency and non-specificity means that they are seriously limited by adverse side effects. Localising steroid treatment to the lungs using inhaler technology has reduced side effects but the reduced systemic exposure following inhalation still results in some undesirable effects. Hence, there is a reluctance to use steroids early in the course of the disease. There therefore still remains a need for an alternative asthma therapy which is a safe, effective, anti -inflammatory or immunomodulatory agent which can be taken to treat chronic asthma.
  • Tryptase is the major secretory protease of human mast cells and is proposed to be involved in neuropeptide processing and tissue inflammation. Tryptase is one of a large number of serine protease enzymes which play a central role in the regulation of a wide variety of physiological processes including coagulation, fibrinolysis , fertilization, development, malignancy, neuromuscular patterning and inflammation. Although a large number of serine proteases have been widely investigated, tryptase still remains relatively unexplored.
  • Mature human tryptase is a glycosylated, heparin- - 4 - associated tetramer of catalytically active subunits. Its amino-acid structure appears to have no close counterpart among the other serine proteases which have been characterised. Tryptase is stored in mast cell secretory granules and after mast cell activation, human tryptase can be measured readily in a variety of biological fluids. For example, after anaphylaxis, tryptase appears in the blood stream where it is readily detectable for several hours. Tryptase also appears in samples of nasal and lung lavage fluid from atopic subjects challenged with specific antigen.
  • Tryptase has been implicated in a variety of biological processes, including degradation of vaso- dilating and bronchorelaxing neuropeptides thereby destroying potent bronchodilatory action and modulation of bronchial responsiveness to histamine. Accordingly, mast cell tryptase may increase bronchoconstriction in asthma by destroying bronchodilating peptides. Moreover, the ability of tryptase to activate prostromelysin and procollagenase suggests that tryptase also may be involved in tissue inflammation. Accordingly, tryptase has been proposed as a potentially important mediator in the development of inflammatory response in asthma and other inflammatory diseases.
  • tryptase inhibition may be of great value in the propylaxis and treatment of a variety of mast cell mediated conditions, such as asthma, particularly in the treatment of chronic, late stage inflammatory asthma .
  • mast cell mediated conditions such as asthma
  • tryptase inhibition may be of great value in the propylaxis and treatment of a variety of mast cell mediated conditions, such as asthma, particularly in the treatment of chronic, late stage inflammatory asthma .
  • a variety of peptide based compounds are suggested as potential inhibitors of the mast cell protease tryptase.
  • a tryptase inhibitor is provided by a polypeptide obtainable from the leech hirudo medi cinalis .
  • secretory leukocyte protease inhibitor (SLPI) and active fragments thereof have been found to inhibit the proteolytic activity of - 5 - tryptase .
  • Aminomethyl-benzoic ester derivatives have previously been employed in a variety of fields. In US 5,628,803 aminomethyl-benzoic ester derivatives have been used as fuel additives. In EP 0048433, FR 2500825 and 2500826 a number of aminomethyl-benzoic ester derivatives are suggested as being useful in anti- complement compositions. The compounds are also said to have strong anti-trypsin, anti-plasmin and anti- kallikrein activity. In Japanese Abstract No. 57095908. aminomethyl-benzoic ester derivatives are disclosed as potential anti -allergic compounds and in Acta. Pharm. Nord. 3(1) 31-40 (1991) water-soluble aminoalkylbenzoate esters are suggested as prodrugs .
  • ester derivatives are generally poorly stable in biological systems, leading to a very limited duration of action in a therapeutic drug, stability to chemical and enzymatic hydrolysis can be enhanced by modification of the nature of the ester and/or by addition of stabilising chemical moieties.
  • the aminomethyl-benzoic ester derivatives of the present invention may be substituted by a range of polar, in particular acidic, moieties that may yield a large increase in plasma stability.
  • the compound of the invention will be useful not only in the treatment and prophylaxis of asthma but also of other allergic and inflammatory conditions mediated by tryptase such as allergic rhinitis, skin conditions such as eczema, atopic dermatitis and urticaria, rheumtoid arthritis, conjunctivitis and inflammatory bowel disease.
  • tryptase such as allergic rhinitis, skin conditions such as eczema, atopic dermatitis and urticaria, rheumtoid arthritis, conjunctivitis and inflammatory bowel disease.
  • the invention provides the use of a tryptase inhibitor of formula I
  • R represents hydrogen, alkyl, alkenyl, hydroxy, alkoxy, aminoalkyl, hydroxyalkyl, carboxyalkyl, alkoxyalkyl, amino, halo, cyano, nitro, thiol, alkylthio, haloalkoxy or haloalkyl;
  • Ar represents an optionally substituted carbocyclic or heterocyclic aryl group with the proviso that when Ar represents a naphthyl moiety it is not substituted by amidino or guanidine;
  • Y represents a hydrogen atom or alkyl group; or a physiologically tolerable salt thereof, e.g. a halide, phosphate, sulphate, or trifluoroacetate salt or salt with ammonium or an organic amine such as ethylamine or meglumine; in the manufacture of a medicament for use in a method of treatment of the human or non-human animal body to combat a condition responsive to said inhibitor.
  • carbocyclic aryl groups preferably contain 5 to - 7 -
  • Heterocyclic aryl groups preferably contain 5 to 10 ring atoms including 1, 2 or 3 ring heteroatoms selected from oxygen, nitrogen and sulphur.
  • Alkyl or alkenyl groups preferably contain up to 10 carbon atoms, most preferably, up to 6 carbon atoms.
  • the substituent R is preferably a short chain alkyl, e.g. Cj_ 3 alkyl such as methyl but most preferably R represents hydrogen.
  • the substituent Y is preferably a short chain alkyl, e.g. C 1-3 alkyl such as methyl but most preferably Y represents hydrogen.
  • Ar groups include optionally substituted phenyl , optionally substituted naphthyl, optionally substituted pyridyl, optionally substituted quinolyl, and optionally substituted isoquinolyl. These groups should preferably carry at least one polar, especially acidic or protected acidic substituent.
  • the substituent on the carbocyclic or heterocyclic aryl group should not be strongly basic, i.e. ArH must be less basic than benzylamine. This ensures that the compounds of the invention bind more efficiently in the tryptase active site, thus maintaining selectivity.
  • Ar represents a phenyl derivative
  • the phenyl may be substituted by one or more substituents selected from: halo, for example fluoro, chloro, bromo or iodo, methylenedioxy, -R 1 , -NR ⁇ OR 2 , C 2 _ 6 -alkenyl , -(CH 2 ) w -OR 1 ,
  • R 1 and/or R 2 are also optionally substituted with -(CH 2 ) compassion- C00R 1 , - (CH 2 ) w -CH(NHC0R 1 ) -C00R 2 , - (CH 2 ) W -CH (NR*R 2 ) -C00R 1 , - (CH 2 ) w -CONH-S0 2 -R ⁇ - (CH ⁇ -SCANHCO-R 1 , - (CH 2 ) w -tetrazole, - (CH 2 ) W -S (0) r -R 1 , or - (CH 2 ) w -P(0) 2 .
  • the substituent will comprise an electron withdrawing group and/or at least one polar moiety, most preferably an acidic or a protected form of an acidic moiety.
  • Preferred electron withdrawing groups include cyano, nitro, carboxamido, alkylsulfenyl , alkylsulfonyl , alkylaminosulfonyl , sulphonylaminoalkyl , trifluoromethyl , or halogen, and most preferably where the substituent is an electron withdrawing group the substituent will be on the 2 or 4 -position of the phenyl group .
  • the presence of at least one polar moiety provides enhanced biological stability.
  • Preferred acidic groups include alkyl or aryl carboxylic acids and esters, acyl sulphonamide, sulphonylamidocarboxyalkyl , carboxyamidosulphonylalkyl , tetrazole, sulphonic acid or phosphonic acid each bonded to the Ar ring directly or via an alkyl, sulphonamidoalkyl or carboxamidoalkyl linkage, the linkage itself being optionally substituted by small polar or apolar groups such as Cj_ 4 alkyl, NH 2 , CN, N0 2 , NHCO-alkyl, or halogen.
  • the acidic subsitutent will preferably be on the para position of the phenyl ring. In another preferred embodiment an acidic substituent is present on the ortho position of the pheny
  • Ar represents a naphthyl derivative
  • the naphthyl may be substituted by one or more substituents selected from the same group as those as listed for phenyl above.
  • substituents are cyano, nitro, carboxamido, alkyla inosulfonyl , alkylsulfonyl , alkylsulfenyl, sulphonylaminoalkyl or trifluoromethyl or as for phenyl above naphthyl may be substituted by at least one polar moiety, most preferably an acidic or a protected form of an acidic moiety, such as alkyl or aryl carboxylic acids and esters, acyl sulphonamide, sulphonylamidocarboxyalkyl , carboxyamidosulphonylalkyl , tetrazole, sulphonic acid or phosphonic acid each bonded to the Ar ring directly or via an alkyl, sulphonamido
  • aromatic heterocyclic groups include pyridyl, quinolyl, isoquinolyl, imidazolyl, indolinyl, pyrazolinyl, pyrrolidinyl, imidazolidinyl , isoindolinyl , pyrazolidinyl, furyl , pyrolyl, pyrazinyl, benzothienyl , thienyl and benzofuryl .
  • Particularly prefered heterocyclic groups are quinolyl, especially, 6 -quinolyl and pyridyl, especially 3 -pyridyl.
  • Suitable Ar groups therefore include (for convenience hydrogen atoms have been missed out) :
  • the salt is preferably a hydrochloride or other physiologically tolerated salt.
  • R and Y are as hereinbefore defined; and Z represents a phenyl group substituted by methylenedioxy, -NR x COR 2 , C 2 . 6 -alkenyl , -(CH 2 ) w -OR 1 , - (CA 6 ) -perfluoroalkyl, -(CH 2 ) W CN, -(CH 2 ) w N0 2 , -(CH 2 ) W CF 3 , -
  • R 1 and/or R 2 are also optionally substituted with -(CH 2 ) W - COOR 1 , - (CH 2 ) w -CH(NHCOR 1 ) -COOR 2 , - (CH 2 ) W -CH (NR X R 2 ) -COOR 1 , - (CH 2 ) w -CONH-S0 2 -R 1 , - (CH 2 ) w -S0 2 -NHCO-R 1 , - (CH 2 ) w -tetrazole, - (CH 2 ) w -S(0) r -R 1 , or - (CH 2 ) W -P (0) z ⁇ -R 1 ; where R 3 is an oligomer comprising 1-4 aminoacid monomers, such as the natural aminoacids
  • Z represents a naphthyl group substituted by halo, - 13 - for example fluoro, chloro, bromo or iodo, methylenedioxy, -R 1 , -NR ⁇ OR 2 , C 2 . 6 -alkenyl , -(CH 2 ) w -OR 1 ,
  • R 1 and R 2 independently represent H, C ⁇ g alkyl, C 3-7 cycloalkyl, or -(CH 2 ) W -Ph, or R 1 and R 2 are optionally connected by a bond to form a 5- 8 atom cyclic structure (eg.
  • ZH should be less basic than benzylamine. This ensures that the compounds of the invention bind more efficiently in the tryptase active site.
  • R is preferably a short chain alkyl, e.g. C 1-3 alkyl such as methyl but most preferably R represents hydrogen.
  • Y is preferably a short chain alkyl, e.g. C,_ 3 alkyl such as methyl but most preferably Y represents hydrogen and wherever possible Z represents a preferred substituent Ar as defined above.
  • the invention provides a tryptase inhibitor of formula (II) for use in combatting a condition responsive to said inhibitor.
  • a pharmaceutical composition comprising a compound of formula (II) , together with at least one pharmaceutically acceptable excipient .
  • the compounds of the invention may be prepared by conventional chemical synthetic routes, e.g. by ester bond formation to couple the Ar-OH compound to the aminomethylbenzoic acid derivative.
  • the readily available starting material 4- aminomethyl-benzoic acid can be utilised. Prior to esterification the amino group should be protected by any appropriate protecting group e.g. Boc, Z, Fmoc or Bpoc .
  • protecting groups e.g. Boc, Z, Fmoc or Bpoc .
  • the use of protecting groups is described in McOmie, "Protective Groups in Organic Chemistry", Plenum, 1973 and Greene, "Protective Groups in Organic Synthesis", Wiley Interscience, 1981.
  • the protected aminomethyl-benzoic acid can be simply coupled to a suitable Ar-OH derivative by conventional esterification techniques before deprotection is effected.
  • the protected aminomethyl-benzoic acid derivative can be activated by converison to its corresponding anhydride or acyl chloride, using conventional reagents, to facilitate esterification. If the aminomethyl-benzoic acid compound is to carry phenyl substituents these can be conveniently introduced prior to the esterification or protection step using conventional aromatic substitution chemistry.
  • a starting material could be employed which already carries the substituent R and the aminomethyl functionality introduced for example by reduction of a cyano group.
  • the Ar-OH alcohols are all readily available or prepared by the skilled chemist.
  • the compounds of the invention may be administered by any conventional route e.g. into the gastrointestinal - 15 - tract (e.g. rectally or orally), the nose, lungs, musculature or vasculature or transdermally .
  • the compounds may be administered by inhalation, orally, intravenously or topically to the skin or to the eye.
  • the compounds may be administered in any convenient administrative form e.g. tablets, powders, capsules, solutions, dispersions, suspensions, syrups, sprays, suppositories, gels, emulsions, patches, etc.
  • Such compositions may contain components conventional in pharmaceutical preparations, e.g.
  • the composition will be suitable for inhalation via a nebulizer or inhalable spray, e.g. a metered dose inhaler or dry powder inhaler for the treatment of lung conditions such as asthma.
  • a nebulizer or inhalable spray e.g. a metered dose inhaler or dry powder inhaler for the treatment of lung conditions such as asthma.
  • the composition will be suitable for application to the skin or mucous membranes formulated as a cream, ointment or solution. Such compositions form a further aspect of the invention.
  • the dosage of the inhibitor compound of the invention will depend upon the nature and severity of the condition being treated, the administration route and the size and species of the patient. However, in general, quantities to be administered are from 0A to 1000 mgs per day, preferably, 1 to 100 mgs per day. Conveniently, a suitable dosage, e.g 20mgs, can be admnistered by inhalation three times daily.
  • the invention provides a method of treatment of human or non-human animal body (e.g. mammalian, avian or reptilian body) to combat a condition responsive to a tryptase inhibitor, said method comprising administering to said body an effective amount of a tryptase inhibitor according to the invention.
  • human or non-human animal body e.g. mammalian, avian or reptilian body
  • the tryptase inhibitors of the invention can be administered along with other - 16 - active ingredients suitable for use in treating asthma, for example beta-adrenoceptor agonists such as salbutamol or anti-inflammatory agents such as cortiosteroids e.g. beclamethasone or cromolyns. It is envisaged that the tryptase inhibitors of the invention and the other active ingredient may act synergistically together.
  • the compounds of the invention are administered in conjunction with corticosteroids to reduce the dose of the steriod hence minimising steriod associated side effects.
  • the invention provides a pharmaceutical composition
  • a pharmaceutical composition comprising a compound of formula (I) , together with one or more anti-asthma agents together with at least one pharmaceutically acceptable excipient.
  • the compounds of the invention will be of use in combating mast cell mediated diseases such as asthma, allergic rhinitis, skin conditions such as eczema, atopic dermatitis and urticaria, rheumatoid arthritis, conjunctivitis and inflammatory bowel disease.
  • mast cell mediated diseases such as asthma, allergic rhinitis, skin conditions such as eczema, atopic dermatitis and urticaria, rheumatoid arthritis, conjunctivitis and inflammatory bowel disease.
  • a tryptase inhibitor will be primarily administered chronically as prophylaxis to prevent or diminish exacerbations of the disease. However, for some diseases a more acute relief of symptoms may also be achievable .
  • Flash column chromotography 1 was carried out using Merck silica gel Si60 (40-63mm, 230-400 mesh) .
  • H nmr (d 6 DMSO) 8.58 (3H, bs, NH 3 + ) ; 8.31-7.95 (4H, m, Ar) ; 7.85-7.30 (4H, m, Ar) ; 4.08 (2H, m, 4-CH 2 ).
  • M.S.TOF 273 (M+l) + .
  • Hplc (Jupiter5 C18, Gradient 1, water/acetonitrile/TFA) rt 12.2 min.
  • Hplc Symmetry C8 , Gradient 2, water/acetonitrile/TFA) rt 9.9 min.
  • Example 5 4' -Chlorophenyl 4- (aminomethyl) benzoate hydrochloride salt l H nmr (d 6 DMSO) 8.59 (3H, bs, NH 3 + ) ; 8.25 (2H, m, 2-H, 6- H) ; 7.80 (2H, m, 3 ' -H, 5 ' -H) ; 7.59 (2H, m, 3-H, 5-H) ; 7.40 (2H, m, 2'-H, 6 ' -H) ; 4.20 (2H, bs , 4-CH 2 ).
  • Hplc (Jupiter5 C18, Gradient 1, water/acetonitrile/TFA) rt 14.8 min.
  • Hplc (Symmetry C8 , Gradient 2, water/acetonitrile/TFA) rt 10.2 min.
  • Hplc Jupiter5 C18, Gradient 1, water/acetonitrile/TFA) rt 18.28min.
  • Hplc SymmetryC ⁇ , Gradient 2, - 20 - water/acetonitrile/TFA) rt 10.88min.
  • M.S. TOF 263 (M+l) + .
  • Hplc (Jupiter5 C18, Gradient 1, Water/acetonitrile/TFA) rt 14.71 min.
  • Hplc SymmetryC ⁇ Gradient 2, Water/acetonitrile/TFA) rt 11.29 min.
  • Hplc Jupiter5 Cl ⁇ , Gradient 1, water/acetonitrile/TFA) rt 17.64min.
  • Hplc SymmetryC ⁇ , Gradient 2, water/acetonitrile/TFA rt 10.75min.
  • Hplc (Jupiter5 Cl ⁇ , Gradient 1, water/acetonitrile/TFA) rt 17.6 min.
  • Hplc (Symmetry C ⁇ , Gradient 2, water/acetonitrile/TFA) rt 10.6 min.
  • Hplc (Jupiter5 C18, Gradient 1, Water/acetonitrile/TFA) rt 12.75 min.
  • Hplc (SymmetryC ⁇ Gradient 2, Water/acetonitrile/TFA) rt 10.00 min.
  • Hplc Jupiter5 Cl ⁇ , Gradient 1, water/acetonitrile/TFA) rt 15.57min.
  • Hplc SymmetryC ⁇ , Gradient 5, water/acetonitrile/TFA rt 8.65min
  • Hplc Jupiter5 Cl ⁇ , Gradient 1, water/acetonitrile/TFA) rt 16.82min.
  • Hplc SymmetryC ⁇ , Gradient 2, water/acetonitrile/TFA rt 10.4 ⁇ min.
  • Hplc Jupiter5 C18, Gradient 1, water/acetonitrile/TFA) rt 4.3min.
  • Hplc Symmetry C ⁇ , Gradient 3, water/acetonitrile/TFA rt 7.5 min.
  • Hplc Jupiter5 Cl ⁇ , Gradient 1, water/acetonitrile/TFA) rt 4.97min.
  • Hplc SymmetryC ⁇ , Gradient 5, water/acetonitrile/TFA rt 7.66min.
  • Hplc Jupiter5 Cl ⁇ , Gradient 1, water/acetonitrile/TFA) rt 3.77min.
  • Hplc SymmetryC ⁇ , Gradient 5, water/acetonitrile/TFA
  • Hplc Jupiter5 Cl ⁇ , Gradient 1, water/acetonitrile/TFA) rt 5.93min.
  • Hplc SymmetryC ⁇ , Gradient 5, water/acetonitrile/TFA rt 7.71min.
  • Example 55 O- [Methyl N-acetyl-L_tyrosinyl] 4- (aminomethyl) benzoate trifluoroacetate salt l n nmr (d 4 methanol) 8.22 (2H, d) ; 7.63 (2H, d) ; 7.31 (2H, d) ; 7.20 (2H, d) ; 4.72 (IH, m) ; 4.27 (2H, s) ; 3.69 (3H, S) ; 3.21 (3H, s) ; 3.21 (IH, dd) ; 3.01 (IH, dd) ; 2.92 (3H, s) .
  • H nmr (d 4 methanol) 8.28 (2H, d) ; 8.14 (2H, d) ; 7.69 (2H, d) ; 7.38 (2H, d) ; 4.27 (2H, s) .
  • Hplc Magneticellan C8, Gradient 5, Water/acetonitrile/TFA
  • LC-MS Magneticellan C18, Gradient 6, water/acetonitrile/TFA
  • Hplc Magneticellan C8, Gradient 5, Water/acetonitrile/TFA
  • LC-MS Magneticellan C18, Gradient 6, water/acetonitrile/TFA rt 2.07 min, 426 (M+l) + .
  • H nmr (d 4 methanol) 8.2 ⁇ (2H, d) ; 8.02 (2H, d) ; 7.68 (2H, d) ; 7.46 (2H, d) ; 4.27 (2H, s) .
  • Hplc Magneticellan C8, Gradient 5, Water/acetonitrile/TFA) rt ⁇ .70 min.
  • LC/MS Magneticellan Cl ⁇ Gradient 6, water/acetonitrile/TFA
  • H nmr (d 4 methanol) 8.29 (2H, d) ; 8.21 (2H, d) ; 7.96 (2H, d) ; 7.70 (2H, d) ; 7.58 (2H, d) ; 7.52 (2H, d) ; 4.29 (2H, s) ; 4.22 (2H, s) .
  • Hplc Magneticellan C8, Gradient 5, Water/acetonitrile/TFA
  • H nmr (d 4 methanol) 8.29 (2H, d) ; 7.69 (2H, d) ; 7.43 (IH, t) ; 7.24 (IH, d) ; 7.17-7.11 (2H, m) ; 4.73 (IH, dd) ; 4.29 (2H, s) ; 3.31 (IH, dd) ; 3.04 (IH, dd) ; 1.96 (3H, s) .
  • Hplc Magnellan C8, Gradient 5
  • H nmr (d 4 methanol) 8.30 (2H, d) ; 7.68 (2H, d) ; 7.51- 7.39 (2H, m) ; 7.36-7.26 (2H, m) ; 4.27 (2H, s) ; 3.82 (2H, s) ; 3.65 (2H, s) .
  • Hplc Magneticellan C8, Gradient 5, Water/acetonitrile/TFA) rt 6.42 min.
  • LC/MS Magneticellan Cl ⁇ Gradient 6, water/acetonitrile/TFA
  • H nmr (d 4 methanol) 8.29 (2H, d) ; 7.84 (IH, d) ; 7.68- 7.61 (3H, m) ; 7.39 (IH, d) ; 4.57 (IH, t); 4.27 (2H, s) ; 3.92 (IH, dd) ; 3.84 (IH, dd) .
  • Hplc Magneticellan C ⁇ ,
  • Tryptase assays were carried out at room temperature in 0.1 M phosphate buffer, 0.5 mg/ml heparin, pH 7.4 according to a method of Tapparelli et - 48 - al. (1993) J. Biol . Chem . 268., 4734-4741.
  • Purified human lung tryptase was purchased from Dr Andrew Walls, Immunopharmacology Group, Southampton General Hospital, Southampton, UK.
  • the chromogenic substrate for tryptase, S-2366 was purchased from Quadratech, Epsom, Surrey, UK.
  • K-nitroaniline was quantified by absorption at 405nm in 96 well microplates using a Dynatech MR 5000 reader (Dynex Ltd, Billingshurst , UK) .
  • K-, and Ki were calculated using SAS software.
  • a K-, value of 216 ⁇ M was determined for tryptase/S-2366.
  • Inhibitor stock solutions were prepared at 40 mM in Me 2 S0 and tested within the range lOOmM-lnM. Accuracy of K ⁇ measurements was confirmed by comparison with K values of a known inhibitor of tryptase. In agreement with published data, benzamidine inhibited tryptase with a K x value of 30 ⁇ M.
  • the tryptase pKi ' s of a number of the compounds of the invention are illustrated in Table 1 below. Also quoted are the trypsin pKi ' s of the compounds.
  • R and Y represent hydrogen in formula (I) .
  • the stability of compounds on incubation in human plasma 49 can be used to demonstrate the hydrolytic stability of the ester moiety towards the range of esterases and proteases normally present in plasma.
  • this assay is an informative measure of metabolic stability in the whole animal for compounds where ester hydrolysis is a major metabolic pathway.
  • R and Y represent hydrogen in formula (I) .

Abstract

The invention relates to the use of compounds of formula (I) wherein R represents hydrogen, alkyl, alkenyl, hydroxy, alkoxy, aminoalkyl, hydroxyalkyl, carboxyalkyl, alkoxyalkyl, amino, halo, cyano, nitro, thiol, alkylthio, haloalkoxy or haloalkyl; Ar represents an optionally substituted carbocyclic or heterocyclic aryl group with the proviso that when Ar represents a naphthyl moiety it is not substituted by amidino or guanidine; and Y represents a hydrogen atom or alkyl group; or a physiologically tolerable salt thereof; for use as tryptase inhibitors.

Description

AMINOMETHYL-BENZOIC ESTERDERIVATIVESAS TRYPTASE INHIBITORS
The invention relates to compounds for use m the treatment of mast cell mediated diseases such as asthma and other allergic and inflammatory conditions and to pharmaceutical compositions thereof and their use m the 5 treatment of the human or animal body, and m particular to compounds which are tryptase inhibitors.
Asthma, the most prevalent of all mast cell mediated conditions affects about 5% of the population m industrialised countries and there is evidence that 10 its incidence and severity are on the increase.
Furthermore, the incidence of childhood asthma is rising and there are suggestions of a link between environmental pollutants and the onset of the disease.
Initially, it was believed that 15 bronchoconstriction, i.e. the narrowing of the airways m the lungs, was the major feature of asthma. However, it is now recognised that inflammation m the lungs is an integral part of the development of the disease
The inhalation of an allergen by an asthmatic
20 generates a strong immune system response which triggers release of various inflammatory mediators, including histamme and leukotrienes from inflammatory cells. These increase the permeability of the blood vessel walls, attract inflammatory cells into the tissues and 25 contract the smooth muscle around the airways. As a result, fluid leaks from the blood and the tissues swell, further narrowing the airways. The inflammatory cells cause damage to the epithelial cells lining the airways exposing nerve endings which stimulates
30 secretion of mucous as well as augmenting the inflammation by causing the release of neurokinins .
Thus asthma is a complex disease frequently characterised by progressive developments of hyper- responsiveness of the trachea and bronchi as a result of - 2 - chronic inflammation reactions which irritate the epithelium lining the airway and cause pathological thickening of the underlying tissues.
Leukocytes and mast cells are present in the epithelium and smooth muscle tissue of the bronchi where they are activated initially by binding of specific inhaled antigens to IgE receptors. Activated mast cells release a number of preformed or primary chemical mediators of the inflammatory response in asthma as well as enzymes. Moreover, secondary mediators of inflammation are generated by enzymatic reactions of activated mast cells and a number of large molecules are released by degranulation of mast cells.
It has therefore been proposed that chemical release from mast cells properly accounts for the early bronchiolar constriction response that occurs in susceptible individuals after exposure to airborne allergens. The early asthmatic reaction is maximal at around 15 minutes after allergen exposure, recovery occurring over the ensuing 1 to 2 hours. In approximately 30% of individuals, the early asthmatic reaction is followed by a further decline in respiratory function which normally begins within a few hours and is maximal between 6 and 12 hours after exposure. This late asthmatic reaction is accompanied by a marked increase in the number of inflammatory cells infiltrating bronchiolar smooth muscle and epithelial tissues, and spilling into the airways. These cells are attracted to the site by release of mast cell derived chemotactic agents.
The most straightforward way of dealing with an asthma attack is with a bronchodilator drug which causes airways to expand. The most effective bronchodilators are the (3-adrenergic agonists which mimic the actions of adrenalin. These are widely used and are simply administered to the lungs by inhalers. However, bronchoconstrictor drugs are primarily of use in short term symptomatic relief, and do not prevent asthma attacks nor deterioration of lung function over the long term.
Anti -inflammatory drugs such as cromoglycate and the corticosteroids are also widely used in asthma therapy. Cromoglycate has anti -inflammatory activity and has been found to be extremely safe. Although such cromolyns have minimal side effects and are currently preferred for initial preventive therapy in children, it is well known that they are of limited efficacy.
The use of corticosteroids in asthma therapy was a major advance since they are very effective anti- inflammatory agents, however, steroids are very powerful, broad spectrum anti -inflammatory agents and their potency and non-specificity means that they are seriously limited by adverse side effects. Localising steroid treatment to the lungs using inhaler technology has reduced side effects but the reduced systemic exposure following inhalation still results in some undesirable effects. Hence, there is a reluctance to use steroids early in the course of the disease. There therefore still remains a need for an alternative asthma therapy which is a safe, effective, anti -inflammatory or immunomodulatory agent which can be taken to treat chronic asthma.
Tryptase is the major secretory protease of human mast cells and is proposed to be involved in neuropeptide processing and tissue inflammation. Tryptase is one of a large number of serine protease enzymes which play a central role in the regulation of a wide variety of physiological processes including coagulation, fibrinolysis , fertilization, development, malignancy, neuromuscular patterning and inflammation. Although a large number of serine proteases have been widely investigated, tryptase still remains relatively unexplored.
Mature human tryptase is a glycosylated, heparin- - 4 - associated tetramer of catalytically active subunits. Its amino-acid structure appears to have no close counterpart among the other serine proteases which have been characterised. Tryptase is stored in mast cell secretory granules and after mast cell activation, human tryptase can be measured readily in a variety of biological fluids. For example, after anaphylaxis, tryptase appears in the blood stream where it is readily detectable for several hours. Tryptase also appears in samples of nasal and lung lavage fluid from atopic subjects challenged with specific antigen.
Tryptase has been implicated in a variety of biological processes, including degradation of vaso- dilating and bronchorelaxing neuropeptides thereby destroying potent bronchodilatory action and modulation of bronchial responsiveness to histamine. Accordingly, mast cell tryptase may increase bronchoconstriction in asthma by destroying bronchodilating peptides. Moreover, the ability of tryptase to activate prostromelysin and procollagenase suggests that tryptase also may be involved in tissue inflammation. Accordingly, tryptase has been proposed as a potentially important mediator in the development of inflammatory response in asthma and other inflammatory diseases. Accordingly, tryptase inhibition may be of great value in the propylaxis and treatment of a variety of mast cell mediated conditions, such as asthma, particularly in the treatment of chronic, late stage inflammatory asthma . In WO96/09297, 095/32945, O94/20527 and US
5,525,623 a variety of peptide based compounds are suggested as potential inhibitors of the mast cell protease tryptase. In WO95/03333 a tryptase inhibitor is provided by a polypeptide obtainable from the leech hirudo medi cinalis . In O96/08275 secretory leukocyte protease inhibitor (SLPI) and active fragments thereof have been found to inhibit the proteolytic activity of - 5 - tryptase .
However, it has now been surprisingly found that certain aminomethyl-benzoic ester derivatives are particularly effective as inhibitors of tryptase and show a surprising selectivity for tryptase over other serine proteases.
Aminomethyl-benzoic ester derivatives have previously been employed in a variety of fields. In US 5,628,803 aminomethyl-benzoic ester derivatives have been used as fuel additives. In EP 0048433, FR 2500825 and 2500826 a number of aminomethyl-benzoic ester derivatives are suggested as being useful in anti- complement compositions. The compounds are also said to have strong anti-trypsin, anti-plasmin and anti- kallikrein activity. In Japanese Abstract No. 57095908. aminomethyl-benzoic ester derivatives are disclosed as potential anti -allergic compounds and in Acta. Pharm. Nord. 3(1) 31-40 (1991) water-soluble aminoalkylbenzoate esters are suggested as prodrugs . In DE 1966174 and DE 1951061 a variety of aminomethyl-benzoic ester derivatives are suggested as having antiplasmin activity. However it has now been surprisingly found that certain aminomethyl benzoic ester derivatives are effective inhibitors of tryptase whilst showing a surprising selectivity for tryptase over other serine proteases such as Factor X, thrombin and trypsin. Moreover, certain aminomethyl-benzoic ester derivatives have a prolonged biological stability, enhancing their usefulness when administered by a systemic route, such as orally or intravenously. While ester derivatives are generally poorly stable in biological systems, leading to a very limited duration of action in a therapeutic drug, stability to chemical and enzymatic hydrolysis can be enhanced by modification of the nature of the ester and/or by addition of stabilising chemical moieties. For example, the aminomethyl-benzoic ester derivatives of the present invention may be substituted by a range of polar, in particular acidic, moieties that may yield a large increase in plasma stability.
It is envisaged that the compound of the invention will be useful not only in the treatment and prophylaxis of asthma but also of other allergic and inflammatory conditions mediated by tryptase such as allergic rhinitis, skin conditions such as eczema, atopic dermatitis and urticaria, rheumtoid arthritis, conjunctivitis and inflammatory bowel disease.
Thus, viewed from one aspect the invention provides the use of a tryptase inhibitor of formula I
( i :
wherein R represents hydrogen, alkyl, alkenyl, hydroxy, alkoxy, aminoalkyl, hydroxyalkyl, carboxyalkyl, alkoxyalkyl, amino, halo, cyano, nitro, thiol, alkylthio, haloalkoxy or haloalkyl;
Ar represents an optionally substituted carbocyclic or heterocyclic aryl group with the proviso that when Ar represents a naphthyl moiety it is not substituted by amidino or guanidine; and
Y represents a hydrogen atom or alkyl group; or a physiologically tolerable salt thereof, e.g. a halide, phosphate, sulphate, or trifluoroacetate salt or salt with ammonium or an organic amine such as ethylamine or meglumine; in the manufacture of a medicament for use in a method of treatment of the human or non-human animal body to combat a condition responsive to said inhibitor. In the compounds of the invention unless otherwise stated, carbocyclic aryl groups preferably contain 5 to - 7 -
10, more especially 5 or 6 ring atoms. Heterocyclic aryl groups preferably contain 5 to 10 ring atoms including 1, 2 or 3 ring heteroatoms selected from oxygen, nitrogen and sulphur. Alkyl or alkenyl groups preferably contain up to 10 carbon atoms, most preferably, up to 6 carbon atoms.
The substituent R is preferably a short chain alkyl, e.g. Cj_3 alkyl such as methyl but most preferably R represents hydrogen. The substituent Y is preferably a short chain alkyl, e.g. C1-3 alkyl such as methyl but most preferably Y represents hydrogen.
Representative Ar groups include optionally substituted phenyl , optionally substituted naphthyl, optionally substituted pyridyl, optionally substituted quinolyl, and optionally substituted isoquinolyl. These groups should preferably carry at least one polar, especially acidic or protected acidic substituent.
In a preferred embodiment, the substituent on the carbocyclic or heterocyclic aryl group should not be strongly basic, i.e. ArH must be less basic than benzylamine. This ensures that the compounds of the invention bind more efficiently in the tryptase active site, thus maintaining selectivity. Where Ar represents a phenyl derivative, the phenyl may be substituted by one or more substituents selected from: halo, for example fluoro, chloro, bromo or iodo, methylenedioxy, -R1, -NR^OR2, C2_6-alkenyl , -(CH2)w-OR1,
- (Ci-6) -perfluoroalkyl, -(CH2)WCN, -(CH2)wN02, -(CH2)WCF3, - (CH2)wS(0)rR1, - (CH^NR'R2, -(CH2)WCOR\ - (CH2) wC02Rx ,
- (CH2) wCONR1R2 , - (CH2) ..SC^NR^2 , - (CH2) wNHS02R: , - (CH2) STHCOR1 ,
- (CH2)wNHC02R\ -OC(=0)R1, - (CH2)w-CH(NHCOR1) -COOR2, -(CH2)W- CH(NRXR2) -COOR1, - (CH2) ^CONH-SOj-R1 , - (CH2) ..-SOj-NHCO-R1 , - (CH2)w-tetrazole, - (CH2) W-P (O) ^-R1 , - (CH2) W-C (=0) -R3 and optionally substituted aryl where R1 and R2 independently represent H, Cj__8 alkyl , C3_7 cycloalkyl , or -(CH2)W-Ph, or R1 and R2 are optionally connected by a bond to form a 5- 8 atom cyclic structure (eg. piperidine) or connected via an 0, S or N atom to form a 5-8 atom heterocyclic structure (eg. morpholine, piperazine) and where R1 and/or R2 are also optionally substituted with -(CH2)„- C00R1, - (CH2)w-CH(NHC0R1) -C00R2, - (CH2) W-CH (NR*R2) -C00R1 , - (CH2)w-CONH-S02-R\ - (CH^-SCANHCO-R1, - (CH2)w-tetrazole, - (CH2)W-S (0)r -R1, or - (CH2)w-P(0)2.3-R1; R3 is an oligomer comprising 1-4 aminoacid monomers, such as the natural aminoacids glycine, proline and serine, terminated by a free carboxylic acid, ester or amide functionality; and w=0-5 and r=0-2.
Preferably the substituent will comprise an electron withdrawing group and/or at least one polar moiety, most preferably an acidic or a protected form of an acidic moiety. Preferred electron withdrawing groups include cyano, nitro, carboxamido, alkylsulfenyl , alkylsulfonyl , alkylaminosulfonyl , sulphonylaminoalkyl , trifluoromethyl , or halogen, and most preferably where the substituent is an electron withdrawing group the substituent will be on the 2 or 4 -position of the phenyl group .
The presence of at least one polar moiety, most preferably an acidic or a protected form of an acidic moiety (e.g. an ester) provides enhanced biological stability. Preferred acidic groups include alkyl or aryl carboxylic acids and esters, acyl sulphonamide, sulphonylamidocarboxyalkyl , carboxyamidosulphonylalkyl , tetrazole, sulphonic acid or phosphonic acid each bonded to the Ar ring directly or via an alkyl, sulphonamidoalkyl or carboxamidoalkyl linkage, the linkage itself being optionally substituted by small polar or apolar groups such as Cj_4 alkyl, NH2, CN, N02, NHCO-alkyl, or halogen. The acidic subsitutent will preferably be on the para position of the phenyl ring. In another preferred embodiment an acidic substituent is present on the ortho position of the phenyl ring and a electron withdrawing substituent is present on the para position.
Where Ar represents a naphthyl derivative, the naphthyl may be substituted by one or more substituents selected from the same group as those as listed for phenyl above. Most preferred substituents are cyano, nitro, carboxamido, alkyla inosulfonyl , alkylsulfonyl , alkylsulfenyl, sulphonylaminoalkyl or trifluoromethyl or as for phenyl above naphthyl may be substituted by at least one polar moiety, most preferably an acidic or a protected form of an acidic moiety, such as alkyl or aryl carboxylic acids and esters, acyl sulphonamide, sulphonylamidocarboxyalkyl , carboxyamidosulphonylalkyl , tetrazole, sulphonic acid or phosphonic acid each bonded to the Ar ring directly or via an alkyl, sulphonamidoalkyl or carboxamidoalkyl linkage, the linkage itself being optionally substituted by small polar or apolar groups such as CA alkyl, CN, N02, NH2, NHCO-alkyl, or halogen.
Representative aromatic heterocyclic groups include pyridyl, quinolyl, isoquinolyl, imidazolyl, indolinyl, pyrazolinyl, pyrrolidinyl, imidazolidinyl , isoindolinyl , pyrazolidinyl, furyl , pyrolyl, pyrazinyl, benzothienyl , thienyl and benzofuryl . Particularly prefered heterocyclic groups are quinolyl, especially, 6 -quinolyl and pyridyl, especially 3 -pyridyl.
Optional substituents described above for one type of aryl Ar group may be present on the other types of aryl Ar groups. Suitable Ar groups therefore include (for convenience hydrogen atoms have been missed out) :
N\" "/N N-N 10 -
0
Cl A -, o
J 1 i A lA~
0
A. 0" H
A 0 o II
H y 0. J
K ^ 0 y 0
O N 0 H 50 " o
0 ^ o =f
. o o
0 ^N * n y
I' "H' It i
" If " 0 o o ^o
- A
^ 0
0 α ~CA n 0 p l' ' γ - -o
0 0 0
a
.'" ^. r« A-"
0.
0
„ 0 o 0;s'υ A
N ^0
0 a "* y 0, ,0
0 ^ A 'I . 0
^H'S-A'
A ° -^ /° 0 o
N If o O N. 11 -
Cl
Cl A -
0 ιr °
0 if rT Ϊ
0 o^A
Jx
O H ,*.. r A.0
H j 0' 0 A ,/ 0
o u^o 0
' --. y
C\ S 0
-\/
Cl
^ k
\ f 'A . /
0 A 0 *H' ^
[ D
A„
Cl o
0 H..
/
0 H'
I
I .1 'ϊ
-/ 0' H
-.. H -_A^/
<y T X o o o - 12 -
Where the compound of formula (I) is a salt, the salt is preferably a hydrochloride or other physiologically tolerated salt.
Viewed from another aspect the invention provides novel tryptase inhibitors of formula (II)
(ID
wherein R and Y are as hereinbefore defined; and Z represents a phenyl group substituted by methylenedioxy, -NRxCOR2, C2.6-alkenyl , -(CH2)w-OR1, - (CA6) -perfluoroalkyl, -(CH2)WCN, -(CH2)wN02, -(CH2)WCF3, -
(CH2)wS(0)rR1, - (CHzANR'R2, - (CH^COR1,- ( CH2 ) ^ONR'R2 , - ( CH2 ) wS02NR1R2 , - ( CH2 ) tfHSC^R1 , - ( CH2 ) wNHCOR1 , - ( CH2 ) uNHCC^R1 , -OC(=0)R1, - (CH2)w-CH(NHCOR1) -COOR2, - (CH2) W-CH (NRXR2) - COOR1, - (CH2)w-CONH-S02-R1, - (CH2) w-SO^NHCO-R1, -(CH2)W- tetrazole, - (CH2) W-P (0) 2-3-R1, - (CH2) W-C (=0) -R3 and optionally substituted aryl where R1 and R2 independently represent H, C^ alkyl , C3_7 cycloalkyl, or -(CH2)W-Ph, or R1 and R2 are optionally connected by a bond to form a 5- 8 atom cyclic structure (eg. piperidine) or connected via an 0, S or N atom to form a 5-8 atom heterocyclic structure (eg. orpholine, piperazine) and where R1 and/or R2 are also optionally substituted with -(CH2)W- COOR1, - (CH2)w-CH(NHCOR1) -COOR2, - (CH2) W-CH (NRXR2) -COOR1, - (CH2)w-CONH-S02-R1, - (CH2)w-S02-NHCO-R1, - (CH2) w-tetrazole, - (CH2)w-S(0)r -R1, or - (CH2) W-P (0) z^-R1 ; where R3 is an oligomer comprising 1-4 aminoacid monomers, such as the natural aminoacids glycine, proline and serine, terminated by a free carboxylic acid, ester or amide functionality; and w=0-5 and r=0-2; or Z represents a phenyl group subsituted in the 2- position by nitro or in the 3 -position by methoxy; or
Z represents a naphthyl group substituted by halo, - 13 - for example fluoro, chloro, bromo or iodo, methylenedioxy, -R1, -NR^OR2, C2.6-alkenyl , -(CH2)w-OR1,
- (Cj.g) -perfluoroalkyl, -(CH2)WCN, -(CH2)wN02, -(CH2)WCF3,
- (CH2)wS(0)rR1, - (CH^NR'R2, -(CH2)wCOR1, - (CH2) .CO^1 , - (CH2) AONR'R2 , - (CH2) ^O.NR'R2 , - (CH2) wNHS02R1 , - (CH2) JHCOR1 ,
- (CH2)wNHC02R\ -OC(=0)R\ - (CH2)w-CH(NHCOR1) -COOR2, -(CH2)W- CH(NR:R2) -COOR1, - (CH2) -CONH-SO^R1 , - (CH2) u-SO^NHCO-R1 , - (CH2)w-tetrazole, - (CH2) W-P (O) ^-R1 , - (CH2) W-C (=0) -R3 and optionally substituted aryl where R1 and R2 independently represent H, C^g alkyl, C3-7 cycloalkyl, or -(CH2)W-Ph, or R1 and R2 are optionally connected by a bond to form a 5- 8 atom cyclic structure (eg. piperidine) or connected via an 0, S or N atom to form a 5-8 atom heterocyclic structure (eg. morpholine, piperazine) and where R1 and/or R2 are also optionally substituted with -(CH2)W-
C00R1, - (CH2)w-CH(NHC0R1) -COOR2, - (CH2) W-CH (NRXR2) -COOR1 , - (CH2)w-CONH-S02-R1, - (CH2)w-S02-NHCO-R1, - (CH2) w-tetrazole ,
- (CH2)w-S(0)r -R1, or - (CH2)w-P(0)2_3-R1; where R3 is an oligomer comprising 1-4 aminoacid monomers, such as the natural aminoacids glycine, proline and serine, terminated by a free carboxylic acid, ester or amide functionality; and w=0-5 and r=0-2; or a heterocyclic aryl groups containing 5 to 10 ring atoms including 1, 2 or 3 ring heteroatoms selected from oxygen, nitrogen and sulphur; or a physiologically tolerable salt thereof.
In a preferred embodiment, ZH should be less basic than benzylamine. This ensures that the compounds of the invention bind more efficiently in the tryptase active site.
In the compounds of formula (II) , R is preferably a short chain alkyl, e.g. C1-3 alkyl such as methyl but most preferably R represents hydrogen. Y is preferably a short chain alkyl, e.g. C,_3 alkyl such as methyl but most preferably Y represents hydrogen and wherever possible Z represents a preferred substituent Ar as defined above. - 14 -
Viewed from a further aspect the invention provides a tryptase inhibitor of formula (II) for use in combatting a condition responsive to said inhibitor. Viewed from a yet further aspect the invention provides a pharmaceutical composition comprising a compound of formula (II) , together with at least one pharmaceutically acceptable excipient .
The compounds of the invention may be prepared by conventional chemical synthetic routes, e.g. by ester bond formation to couple the Ar-OH compound to the aminomethylbenzoic acid derivative.
The readily available starting material 4- aminomethyl-benzoic acid can be utilised. Prior to esterification the amino group should be protected by any appropriate protecting group e.g. Boc, Z, Fmoc or Bpoc . The use of protecting groups is described in McOmie, "Protective Groups in Organic Chemistry", Plenum, 1973 and Greene, "Protective Groups in Organic Synthesis", Wiley Interscience, 1981. The protected aminomethyl-benzoic acid can be simply coupled to a suitable Ar-OH derivative by conventional esterification techniques before deprotection is effected.
If necessary the protected aminomethyl-benzoic acid derivative can be activated by converison to its corresponding anhydride or acyl chloride, using conventional reagents, to facilitate esterification. If the aminomethyl-benzoic acid compound is to carry phenyl substituents these can be conveniently introduced prior to the esterification or protection step using conventional aromatic substitution chemistry.
Alternatively, a starting material could be employed which already carries the substituent R and the aminomethyl functionality introduced for example by reduction of a cyano group. The Ar-OH alcohols are all readily available or prepared by the skilled chemist.
The compounds of the invention may be administered by any conventional route e.g. into the gastrointestinal - 15 - tract (e.g. rectally or orally), the nose, lungs, musculature or vasculature or transdermally . However, it is prefered that the compounds be administered by inhalation, orally, intravenously or topically to the skin or to the eye. The compounds may be administered in any convenient administrative form e.g. tablets, powders, capsules, solutions, dispersions, suspensions, syrups, sprays, suppositories, gels, emulsions, patches, etc. Such compositions may contain components conventional in pharmaceutical preparations, e.g. diluents, carriers, pH modifiers, sweeteners, bulking agents and further active agents. Preferably, the composition will be suitable for inhalation via a nebulizer or inhalable spray, e.g. a metered dose inhaler or dry powder inhaler for the treatment of lung conditions such as asthma. For dermatological or ophthalmic indications, the composition will be suitable for application to the skin or mucous membranes formulated as a cream, ointment or solution. Such compositions form a further aspect of the invention.
The dosage of the inhibitor compound of the invention will depend upon the nature and severity of the condition being treated, the administration route and the size and species of the patient. However, in general, quantities to be administered are from 0A to 1000 mgs per day, preferably, 1 to 100 mgs per day. Conveniently, a suitable dosage, e.g 20mgs, can be admnistered by inhalation three times daily.
Thus, viewed from a further aspect the invention provides a method of treatment of human or non-human animal body (e.g. mammalian, avian or reptilian body) to combat a condition responsive to a tryptase inhibitor, said method comprising administering to said body an effective amount of a tryptase inhibitor according to the invention.
In a preferred embodiment the tryptase inhibitors of the invention can be administered along with other - 16 - active ingredients suitable for use in treating asthma, for example beta-adrenoceptor agonists such as salbutamol or anti-inflammatory agents such as cortiosteroids e.g. beclamethasone or cromolyns. It is envisaged that the tryptase inhibitors of the invention and the other active ingredient may act synergistically together. In a preferred embodiment the compounds of the invention are administered in conjunction with corticosteroids to reduce the dose of the steriod hence minimising steriod associated side effects.
Thus, viewed from a yet further aspect the invention provides a pharmaceutical composition comprising a compound of formula (I) , together with one or more anti-asthma agents together with at least one pharmaceutically acceptable excipient.
It is envisaged that the compounds of the invention will be of use in combating mast cell mediated diseases such as asthma, allergic rhinitis, skin conditions such as eczema, atopic dermatitis and urticaria, rheumatoid arthritis, conjunctivitis and inflammatory bowel disease.
Although not wanting to be bound by any theory, it is expected that a tryptase inhibitor will be primarily administered chronically as prophylaxis to prevent or diminish exacerbations of the disease. However, for some diseases a more acute relief of symptoms may also be achievable .
For all the above conditions the compounds of the invention could be used alone or along side existing treatments and therapies.
The invention will now be described with reference to the following non-limiting examples.
Experimental
Abbreviations used follow IUPAC-IUB nomenclature. Additional abbreviations are Hplc, high-performance - 17 - liquid chromotography; MALDI-TOF, matrix assisted laser desorbtion ionisation - time of flight; rt , retention time; nmr, nuclear magnetic resonance. Alcohols and 4- aminomethylbenzoic acid were purchased from Aldrich
(Gillingham, UK) , Lancaster (Morecombe, UK) , Avocado
(Heysham, UK) or Ubichem.
Purification: Flash column chromotography1 was carried out using Merck silica gel Si60 (40-63mm, 230-400 mesh) .
Analysis: Proton nuclear magnetic resonance (1H) spectra were recorded on Bruker DPX300 (300MHz) . Analytical Hplc was on a Shimadzu LC6 gradient system equipped with an autosampler. Eluant A consisted of aqueous TFA (0.1%) and eluant B 90% MeCN in aqueous TFA (0.1%) with gradient elution (Gradient 1, Omin 20%B then 20%B to 100%B over 15min then 100%B for 5min; Gradient 2, Omin 20%B then 20%B to 100%B over 11 min; Gradient 3, Omin 50%B then 50%B to 100%B over 14min) . Columns used were Jupiter 5 C18 column (2.1x150mm, 5μm particle size) and SymmetryShield RP8 column (2.1x50mm, 3.5μm particle size) . Purified products were further analysed by MALDI- TOF.
Example 1
Preparation of 2' -Naphthyl 4- (aminomethyl)benzoate trifluoroacetate salt t-Butyloxycarbonylaminomethyl benzoic acid2 (500mg, 2.0mmol) dissolved in dichloromethane (5ml) was treated with 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (420mg, 2.2mmol). After stirring for 10 minutes 2-naphthol (316mg, 2.2mmol) and a catalytic ammount of N,N-dimethylaminopyridine were added. The resulting solution was stirred overnight at room temperature. After dilution with dichloromethane (20ml) the mixture was washed with saturated aqueous citric acid (25ml) , saturated aqueous sodium bicarbonate (25ml) and water (25ml) , then dried over magnesium sulphate and concentrated under reduced pressure. The crude product was purifed by flash column chromotography (10% - 30% ethyl acetate/hexane) to yield the 2 ' -naphthyl -4- (t- 5 butyloxycarbonylaminomethyl) benzoate as a thick oil (620mg, 82% yield) .
The deprotection was effected by dissolution in a mixture of trifluoroacetate (5ml) and dichloromethane (5ml) . After stirring for 1 hour the mixture was 10 concentrated under reduced pressure. Trituration with diethyl ether afforded 2 ' -naphthyl 4-
(aminomethyl) benzoate trifluoracetic acid salt (580mg, 75% yield) .
15. XH nmr (d4 methanol) 8.20 (2H, d, J=8.2Hz, 2-H, 6-H) ;
8.05-7.60 (3H, m, Ar) ; 7.58 (2H, d, J=8.2Hz, 3-H, 5-H) ;
7.45-7.20 (4H, m, Ar) ; 4.18 (2H, s, 4-CH2). M.S.TOF 278
(M+l)+'Hplc (Jupiter5 C18, Gradient 1, water/acetonitrile/TFA) rt 16.6 min. Hplc (Symmetry C8 , 20 Gradient 2, water/acetonitrile/TFA) rt 10.5 min.
Further examples were synthesised in an analagous manner using alcohols either commercially available or prepared using standard literature procedures. In several cases 25 the deprotection step was effected using hydrogen chloride dissolved in diethyl ether to afford the products as hydrochloride salts.
Example 2 30 1' -Naphthyl 4- (aminomethyl) benzoate hydrochloride salt
XH nmr (d4 methanol) 8.41 (2H, d, J=8.4Hz, 2-H, 6-H) ; 7.99 (1H, d, J=7.5Hz, Ar) ; 7.88 (2H, d, J=8.0Hz, Ar) ; 7.75 (2H, d, J=8.4Hz, 3-H, 5-H); 7.57 (3H, m, Ar) ; 7.40 (1H, m, Ar) ; 4.29 (2H, s, 4-CH2). M.S.TOF 278 (M+l) + . 5 Hplc (Jupiter5 C18, Gradient 1, water/acetonitrile/TFA) rt 16.03min. Hplc (SymmetryC8, Gradient 2, water/acetonitrile/TFA) rt 10.43min. - 19 -
Example 3
Phenyl 4- (aminomethyl) benzoate hydrochloride salt λE nmr (d4 methanol) 8.27 (2H, d, J=8.3Hz, 2-H, 6-H) ; 7.68 (2H, d, J=8.3Hz, 3-H, 5-H); 7.52-7.20 (5H, m, Ph) ; 4.26 (2H, s, 4-CH2) . M.S.TOF 228 (M+l) + . Hplc (Jupiter5 C18, Gradient 1, water/acetonitrile/TFA) rt 11.4 min. Hplc (Symmtry C8, Gradient 2, water/acetonitrile/TFA) rt 9.6 min.
Example 4
4' -Nitrophenyl 4- (aminomethyl) benzoate hydrochloride salt
:H nmr (d6 DMSO) 8.58 (3H, bs, NH3 +) ; 8.31-7.95 (4H, m, Ar) ; 7.85-7.30 (4H, m, Ar) ; 4.08 (2H, m, 4-CH2). M.S.TOF 273 (M+l)+. Hplc (Jupiter5 C18, Gradient 1, water/acetonitrile/TFA) rt 12.2 min. Hplc (Symmetry C8 , Gradient 2, water/acetonitrile/TFA) rt 9.9 min.
Example 5 4' -Chlorophenyl 4- (aminomethyl) benzoate hydrochloride salt lH nmr (d6 DMSO) 8.59 (3H, bs, NH3 +) ; 8.25 (2H, m, 2-H, 6- H) ; 7.80 (2H, m, 3 ' -H, 5 ' -H) ; 7.59 (2H, m, 3-H, 5-H) ; 7.40 (2H, m, 2'-H, 6 ' -H) ; 4.20 (2H, bs , 4-CH2). M.S.TOF 262.5 (M+l)+. Hplc (Jupiter5 C18, Gradient 1, water/acetonitrile/TFA) rt 14.8 min. Hplc (Symmetry C8 , Gradient 2, water/acetonitrile/TFA) rt 10.2 min.
Example 6 1' -Bromo-2' -naphthyl 4- (aminomethyl) benzoate trifluoroacetate salt
'H nmr (d4 methanol) 8.27 (2H, d, J=8.4Hz, 2-H,6-H); 8.20 (1H, d, J=8.5Hz, Ar) ; 7.92 (2H, m, Ar) ; 7.61 (2H, d, J=8.4Hz, 3-H, 5-H) ; 7.54 (2H, m, Ar) ; 7.38 (1H, d, J=8.9Hz, Ar) ; 4.18 (2H, s, 4-CH2). M.S.TOF 357 (M+l) + .
Hplc (Jupiter5 C18, Gradient 1, water/acetonitrile/TFA) rt 18.28min. Hplc (SymmetryCδ, Gradient 2, - 20 - water/acetonitrile/TFA) rt 10.88min.
Example 7
4 ' -Methoxyphenyl 4- (aminomethyl) benzoate hydrochloride salt
:H nmr (d4 methanol) 8.26 (2H, d, J=8.3Hz, 2-H, 6-H) ;
7.66 (2H, d, J=8.3Hz, 3-H, 5-H); 7.16 (2H, d, J=9.1Hz, 2'-H, 6'-H); 7.00 (2H, d, J=9.1Hz, 3 ' -H, 5 ' -H) ; 4.25(2H, s, 4-CH2); 3.83 (3H, s, OMe) . M.S.TOF 258 (M+l)+. Hplc (Jupiter5 C18, Gradient 1, water/acetonitrile/TFA) rt 11.84min. Hplc (SymmetryCδ, Gradient 5, water/acetonitrile/TFA) rt 9.00min.
Example 8 6 ' -Bromo- 2 ' -naphthyl 4- (aminomethyl) benzoate hydrochloride salt
XH nmr (d4 methanol) 8.15 (2H, d, J= 8.4Hz, 2-H, 6-H) ; 7.99 (IH, d, J=1.9Hz, 5'H); 7.78 (IH, d, J=9.0Hz, 4 ' -H) ;
7.67 (IH, d, J=8.8Hz, 8 ' -H) ; 7.61 (IH, d, J=2.2Hz, 1'- H) ; 7.52 (2H, d, J=8.4Hz, 3-H, 5-H) ; 7.48 (IH, dd,
J=8.8, 1.9Hz, 7'-H); 7.29 (IH, dd, J= 9.0, 2.2Hz, 3 ' -H) ; 4.11 (2H, s, 4-CH2) . M.S. TOF 357 (M+l) + . Hplc (Jupiter5 C18, Gradient 1, Water/acetonitrile/TFA) rt 19.8 min. Hplc (SymmetryCδ Gradient 2, Water/acetonitrile/TFA) rt 10.93 min.
Example 9
7 ' -Methoxy-2 ' -naphthyl 4 - (aminomethyl) benzoate hydrochloride salt :H nmr (d4 methanol) 8.32 (2H, d, J=8.3Hz, 2-H, 6-H) ;
7.87 (IH, d, J=8.8Hz, 4 ' -H) ; 7.82 (IH, d, J=9.0Hz, 5'- H) ; 7.69 (2H, d, J=8.3Hz, 3-H, 5-H); 7.65 (IH, d, J=2.2Hz, l'-H); 7.28 (IH, d, J=2.4Hz, 8 ' -H) ; 7.21 (IH, dd, J=8.8, 2.2Hz, 3 ' -H) ; 7.17 (IH, dd, J=9.0, 2.4, 6'- H) ; 4.28 (2H, s, 4-CH2); 3.93 (3H, s, 7 ' -OMe) . M.S. TOF 308 (M+l)+. Hplc (Jupiter5 C18, Gradient 1, Water/acetonitrile/TFA) rt 16.57 min. Hplc (SymmetryCδ - 21 - Gradient 2, Water/acetonitrile/TFA) rt 10.63 min.
Example 10
4 ' -Cyanopheny1 4- (aminomethyl) benzoate hydrochloride salt
:H nmr (d4 methanol) 8.29 (2H, d, J=8.3Hz, '2-H, 6-H) ; 7.87 (2H, d, J= 11.2Hz, 3'-H, 5 ' -H) ; 7.69 (2H, d, J=8.3Hz, 3-H, 5-H) ; 7.50 (2H, d, J=11.2Hz, 2 ' -H, 6 ' -H) ; 4.2δ (2H, s, 4-CH2) ; M.S. TOF 253 (M+l) + . Hplc (Jupiter5 Clδ, Gradient 1, Water/acetonitrile/TFA) rt 9.99 min.
Hplc (SymmetryCδ Gradient 2, Water/acetonitrile/TFA) rt 9.38 min.
Example 11 3 ' -Chlorophenyl 4- (aminomethyl) benzoate hydrochloride salt
:H nmr (d4 methanol) δ.27 (2H, d, J=δ.3Hz, 2-H, 6-H) ; 7.67 (2H, d, J=δ.3Hz, 3-H, 5-H); 7.50-7.44 (IH, dd, J=δ.5, δ.2Hz, 5'-H); 7.38-7.33 (2H, m, Ar) ; 7.25-7.21 (IH, m, Ar) ; 4.27 (2H, s, 4-CH2). M.S. TOF 263 (M+l)+. Hplc (Jupiter5 C18, Gradient 1, Water/acetonitrile/TFA) rt 14.71 min. Hplc (SymmetryCδ Gradient 2, Water/acetonitrile/TFA) rt 11.29 min.
Example 12
6 ' -Methoxycarbonyl-2 ' -napthy1 4- (aminomethyl) benzoate trifluoroacetate salt
:H nmr (d4 methanol) 8.60 (IH, s, 5 ' -H) ; 8.45 (2H, d, J=δ.3Hz, 2-H, 6-H) ; δ.23 (2H, m, 7'-H, δ ' -H) ; δ.10 (IH, d, J=δ.7Hz, 3'-H); 7.97 (IH, d, J=2.2Hz, 1 ' -H) ; 7.δl
(2H, d, J=δ.3Hz, 3-H, 5-H) ; 7.62 (IH, dd, J=δ.7, 2.2Hz, 4 ' -H) ; 4.40 (2H, s, 4-CH2); 4. lδ (3H, s, 6 ' -C02Me) . M.S.TOF 36 (M+l) + . Hplc (Jupiter5 Clδ, Gradient 1, water/acetonitrile/TFA) rt 16.1 min. Hplc (Symmetry Cδ , Gradient 2, water/acetonitrile/TFA) rt 10.5 min.
Example 13 - 22 -
3' -Methoxycarbonyl-2' -naphthyl 4- (aminomethyl) benzoate trifluoroacetate salt lH nmr (d4 methanol) δ.70 (IH, s, 1 ' -H) ; 8.31 (2H, d, J=δ.4Hz, 2-H, 6-H) ; δ.09 (IH, d, J=δ.2Hz, 8 ' -H) ; 7.97 (IH, d, J=7.9Hz, 5'-H); 7.78 (IH, s, 4 ' -H) ; 7.76-7.60 (4H, m, Ar) ; 4.28 (2H, s, 4-CH2); 3.80 (3H, s, 3 ' -C02Me) . M.S.TOF 336 (M+l)+. Hplc (Jupiter5 C18, Gradient 1, water/acetonitrile/TFA) rt 16.4 min. Hplc (Symmetry Cδ , Gradient 2, water/acetonitrile/TFA) rt 10.4 min.
Example 14
1' -Methoxycarbonyl-2' -naphthyl 4- (aminomethyl) benzoate trifluoroacetate salt
XH nmr (d4 methanol) δ . Oδ (2H, d, J=7.9Hz, 2-H, 6-H) ; δ.02 (IH, d, J=9.0Hz, 3 ' -H) ; 7.95- 7.δ5 (2H, m, 5 ' -H, δ'-H); 7.60 (2H, d, J=7.9Hz, 3-H, 5-H); 7.50 (2H, m, 6'- H, 7'-H); 7.3δ (IH, d, J=9.0Hz, 4 ' -H) ; 4.25 (2H, s, 4- CH2) ; 3.7δ (3H, s, 1 ' -C02Me) . M.S.TOF 336 (M+l) + . Hplc (Jupiter5 Clδ, Gradient 1, water/acetonitrile/TFA) rt 16.6 min. Hplc (Symmetry Cδ , Gradient 2, water/acetonitrile/TFA) rt 10.4 min.
Example 15
5',6',7' , 8' -Tetrahydro-2-naphthyl 4- (aminomethyl) benzoate trifluoroacetate salt
:H nmr (d4 methanol) 6.13 (2H, d, J=δ.4Hz, 2-H, 6-H) ; 7.54 (2H, d, J=8.4Hz, 3-H, 5-H) ; 7.01 (IH, d, J=9.0Hz, Ar) ; 6.61 (2H, m, Ar) ; 4.12 (2H, s, 4-CH2) ; 2.70 (4H, m, 5'-H, δ'-H); 1.73 (4H, m, 6'-H, 7 ' -H) . M.S.TOF 262 (M+l)+. Hplc (Jupiter5 Clδ, Gradient 1, water/acetonitrile/TFA) rt 17.64min. Hplc (SymmetryCδ, Gradient 2, water/acetonitrile/TFA) rt 10.75min.
Example 16 6' -Quinolinyl 4- (aminomethyl) benzoate dihydrochloride salt
:H nmr (d4 methanol) 9.46 (2H, m, Ar) ; δ.61 (IH, d, - 23 -
J=9.3Hz, Ar) ; 8.54 (3H, m, Ar) ; 8.37 (2H, m, Ar) ; 7.93 (2H, d, J=8.3Hz, 3-H, 5-H); 4.46 (2H, s, 4-CH2). M.S.TOF 279 (M+l)+. Hplc (Jupiter5 Clδ, Gradient 1, water/acetonitrile/TFA) rt 7.07min. Hplc (SymmetryCδ, Gradient 5, water/acetonitrile/TFA) rt 3.44min.
Example 17
3' -iso-Propylphenyl 4- (aminomethyl) benzoate trifluoroacetate salt XH nmr (d4 methanol) 8.16 (2H, d, J=8.4Hz, 2-H, 6-H) ;
7.57 (2H, d, J=8.4Hz, 3-H, 5-H) ; 7.27 (IH, t, J=7.8Hz, 5'-H); 7.09 (IH, d, J=7.8Hz, 6 ' -H) ; 7.00 (IH, m, 2 ' -H) ; 6.93 (IH, m, 4 ' -H) ; 4.17 (2H, s, 4-CH2); 2.88 (IH, sep, J=6.9Hz, 3'-CH); 1.08 (6H, d, J=6.9Hz, 3 ' -CMe2) . M.S.TOF 270 (M+l) + . Hplc (Jupiter5 Clδ, Gradient 1, water/acetonitrile/TFA) rt 17.6 min. Hplc (Symmetry Cδ , Gradient 2, water/acetonitrile/TFA) rt 10.6 min.
Example 18 4'-Biphenyl 4- (aminomethyl) benzoate trifluoroacetate salt
XH nmr (d4 methanol) 8.1δ (2H, d, J=8.3Hz, 2-H, 6-H) ; 7.61 (2H, d, J=8.3Hz, 3-H, 5-H) ; 7.57-7.17 (9H, m, Ar) ; 4.16 (2H, s, 4-CH2) . M.S.TOF 304 (M+l) + . Hplc (Jupiter5 Clδ, Gradient 1, water/acetonitrile/TFA) rt lδ .9 min. Hplc (Symmetry C8, Gradient 2, water/acetonitrile/TFA) rt 10.8 min.
Example 19 4' - (t-Butyl) phenyl 4- (aminomethyl) benzoate trifluoroacetate salt lH nmr (d4 methanol) 8.15 (2H, d, J=8.4Hz, 2-H, 6-H) ; 7.55 (2H, d, J=8.4Hz, 3-H, 5-H) ; 7.38 (2H, d, J=6.8Hz, 2'-H, 6'-H); 7.08 (2H, d, J=8.6Hz, 3 ' -H, 5 * -H) ; 4.18 (2H, s, 4-CH2) ; 1.26 (9H, s, 4 ' -sBu) . M.S.TOF 284 (M+l) + . Hplc (Jupiter5 Clδ, Gradient 1, water/acetonitrile/TFA) rt 19.5 min. Hplc (Symmetry Cδ, Gradient 2, - 24 - water/acetonitrile/TFA) rt 10.9 min.
Example 20
7' -Hydroxy-2' -naphthyl 4- (aminomethyl) benzoate hydrochloride salt
:H nmr (d4 methanol) 8.32 (2H, d, J=δ.3Hz, 2-H, 6-H) ; 7.64 (IH, d, J= δ.9Hz, 4 ' -H) ; 7.78 (IH, d, J=8.7Hz, 5'- H) ; 7.69 (2H, d, J=8.3Hz, 3-H, 5-H); 7.51 (IH, d, J=2.2Hz, l'-H); 7.16-7.08 (3H, m, 3 ' -H, 6 ' -H, 8 ' -H) ; 4.28 (2H, s, 4-CH2) . M.S. TOF 294 (M+l) + . Hplc (Jupiter5 C18, Gradient 1, Water/acetonitrile/TFA) rt 12.75 min. Hplc (SymmetryCδ Gradient 2, Water/acetonitrile/TFA) rt 10.00 min.
Example 21
4' -iso-Propylphenyl 4- (aminomethyl) benzoate hydrochloride salt lK nmr (d4 methanol) 8.26 (2H, d, J=8.4Hz, 2-H, 6-H) ; 7.67 (2H, d, J=8.4Hz, 3-H, 5-H); 7.34 (2H, d, J=8.6Hz, 2'-H, 6'-H); 7.15 (2H, d, J=8.6Hz, 3 ' -H, 5 ■ -H) ; 4.26 (2H, s, 4-CH2) ; 2.98 (IH, sep, J= 6.8Hz, 4 ' -CH) ; 1.30 (6H, d, J=6.δHz, 4'-CMe2). M.S. TOF 270 (M+l) + . Hplc (Jupiter5 Clδ, Gradient 1, Water/acetonitrile/TFA) rt 18.27 min. Hplc (SymmetryCδ Gradient 2, Water/acetonitrile/TFA) rt 10.70 min.
Example 22
4' -Benzylphenyl 4- (aminomethyl) benzoate hydrochloride salt XH nmr (d4 methanol) 8.3δ (2H, d, J=8.3Hz, 2-H, 6-H) ;
7.76 (2H, d, J=δ.3Hz, 3-H, 5-H); 7.45-7.25 (9H, m, Ar) ;
4.38 (2H, s, 4-CH2) ; 4.15 (2H, s, 4 ' -CH2) . M.S. TOF 318 (M+l)+. Hplc (Jupiter5 Clδ, Gradient 1,
Water/acetonitrile/TFA) rt 19.80 min. Hplc (SymmetryCδ Gradient 2, Water/acetonitrile/TFA) rt 10.96 min.
Example 23 - 25 -
3 ' -Methoxyphenyl 4- (aminomethyl) benzoate hydrochloride salt H nmr (d4 methanol) 8.11 (2H, d, J=8.3 Hz, 2-H, 6-H) ;
7.51 (2H, d, J=8.3 Hz, 3-H, 5-H); 7.24-7.18 (IH, m, 5'- H) ; 6.76-6.65 (3H, m, 2 ' -H, 4 ' -H, 6 ' -H) ; 4.11 (2H, s, 4-
CH2) ; 3.68 (3H, s, 3 ' -OMe) . M.S. TOF 258 (M+l) + . Hplc
(Jupiter5 C18, Gradient 1, Water/acetonitrile/TFA) rt
12.56 min. Hplc (SymmetryCδ Gradient 2,
Water/acetonitrile/TFA) rt 9.69 min.
Example 24
8' -Methoxycarbonyl-2' -naphthyl 4- (aminomethyl) benzoate trifluoroacetate salt
XH nmr (d4 methanol) 8.88 (IH, d, J=9.4Hz, 4 ' -H) ; 8.19 (2H, d, J=8.4Hz, 2-H, 6-H); 8.10 (IH, d, J=8.3Hz, Ar) ;
8.10 (IH, dd, J=7.3, 1.1Hz, Ar) ; 8.02 (IH, d, J=8.3Hz,
Ar) ; 7.72 (IH, d, J=2.4Hz, 1 ' -H) ; 7.57 (2H, d, J=8.4Hz,
3-H, 5-H) ; 7.49 (IH, m, Ar) ; 7.42 (IH, dd, J=9.4, 2,4Hz,
3 ' -H) ; 4.16 (2H, s, 4-CH2); 3.90 (3H, s, 5 ' -C02Me) . M.S.TOF 336 (M+l)+. Hplc (Jupiter5 Clδ, Gradient 1, water/acetonitrile/TFA) rt 16.6 min. Hplc (Symmetry Cδ ,
Gradient 2, water/acetonitrile/TFA) rt 10.5 min.
Example 25 1' -Amino-2' -naphthyl 4- (aminomethyl) benzoate dihydrochloride salt
:H nmr (d6 DMSO) δ.59 (3H, bs, 1 ' -NH) ; 8.47 (IH, d, J=δ.lHz, Ar) ; 8.32 (2H, d, J=8.2Hz, 2-H, 6-H) ; 6.15 (IH, d, J=8.2Hz, Ar) ; 8.01 (2H, s, Ar) ; 7.77 (3H, d, J=8.2Hz, Ar) ; 7.64 (IH, t, J=7.1Hz, Ar) ; 4.15 (2H, s, 4-CH2). M.S.TOF 294 (M+2)+. Hplc (Jupiter5 Clδ, Gradient 1, water/acetonitrile/TFA) rt 15.57min. Hplc (SymmetryCδ, Gradient 5, water/acetonitrile/TFA) rt 8.65min
Example 26
7' -Quinolinyl 4- (aminomethyl) benzoate dihydrochloride salt - 26 - lE nmr (d4 methanol) 9.15 (IH, d, J=4.1Hz, Ar) ; 8.98 (IH, d, J=8.2Hz, Ar) ; 8.36 (3H, d, J=8.3Hz, Ar) ; 8.18 (IH, d, J=1.5Hz, Ar) ; 7.94 (IH, dd, J=8.3, 5.0Hz, Ar) ; 7.85 (IH, m) ; 7.74 (2H, d, J=8.2Hz, 3-H, 5-H) ; 4.29 (2H, s, 4-CH2). M.S.TOF 282 (M+2)+. Hplc (Jupiter5 C18, Gradient 1, water/acetonitrile/TFA) rt 2.15min. Hplc (SymmetryCδ, Gradient 5, water/acetonitrile/TFA) rt 4.76min.
Example 27 3' - (t-Butyl) phenyl 4- (aminomethyl) benzoate hydrochloride salt
Η nmr (d4 methanol) 8.17 (2H, d, J=8.3Hz, 2-H, 6-H) ; 7.56 (2H, d, J=8.3Hz, 3-H, 5-H, ) ; 7.27 (2H, m, Ar) ; 7.14
(IH, m, Ar) ; 6.93 (IH, m, Ar) ; 4.16 (2H, s, 4-CH2); 1.24 (9H, s, 3 ' -lBu) . M.S.TOF 284 (M+l) + . Hplc (Jupiter5 Clδ, Gradient 1, water/acetonitrile/TFA) rt lδ.78min. Hplc
(SymmetryCδ, Gradient 2, water/acetonitrile/TFA) rt 10.δ5min.
Example 28
3'-Biphenyl 4- (aminomethyl) benzoate hydrochloride salt
Η nmr (d4 methanol) 8.31 (2H, d, J=8.4Hz, 2-H, 6-H) ; 8.12 (2H, d, J=7.8Hz, Ar) ; 7.74-7.23 (7H, m, Ar) ; 7.01 (2H, d, J=7.7Hz, Ar) ; 4.2δ (2H, s, 4-CH2). M.S.TOF 304 (M+l)+. Hplc (Jupiter5 Clδ, Gradient 1, water/acetonitrile/TFA) rt 18.57min. Hplc (SymmetryC8, Gradient 2, water/acetonitrile/TFA) rt 10.90min.
Example 29 4' -Phenoxyphenyl 4- (aminomethyl) benzoate hydrochloride salt
'H nmr (d4 methanol) 7.93 (2H, d, J=8.3Hz, 2-H, 6-H) ; 7.58 (2H, d, J=9.0Hz, Ar) ; 7.51 (2H, d, J=8.3Hz, 3-H, 5- H) ; 7.25 (2H, , Ar) ; 7.01 (IH, m, Ar) ; 6.91 (4H, m, Ar) ; 4.11 (2H, s, 4-CH2). M.S.TOF 320 (M+l)+. Hplc
(Jupiter5 Clδ, Gradient 1, water/acetonitrile/TFA) rt 16.55min. Hplc (SymmetryCδ, Gradient 2, - 27 - water/acetonitrile/TFA) rt 10.55min.
Example 30
3 '-Phenoxyphenyl 4- (aminomethyl) benzoate hydrochloride salt lH nmr (d4 methanol) 7.90 (2H, d, J=δ.2Hz, 2-H, 6-H) ; 7.50 (2H, d, J=8.2Hz, 3-H, 5-H); 7.4-7.2 (5H, m, Ar) ; 7.03 (IH, t, J=7.4Hz, Ar) ; 6.93 (2H, d, J=7.7Hz, Ar) ; 6.68 (IH, m, Ar) ; 4.11 (2H, s, 4-CH2). M.S.TOF 320 (M+l)+. Hplc (Jupiter5 Clδ, Gradient 1, water/acetonitrile/TFA) rt 16.82min. Hplc (SymmetryCδ, Gradient 2, water/acetonitrile/TFA) rt 10.4δmin.
Example 31 5' -Amino-2' -naphthyl 4- (aminomethyl) benzoate ditrifluoracetic acid salt
XE nmr (d4 methanol) 8.21 (2H, d, J=8.3Hz, 2-H, 6-H) ; 7.9δ (IH, d, J=9.2Hz, Ar) ; 7.71 (IH, d, J=2.3Hz, 1 ' -H) ; 7.63 (IH, d, J=8.4Hz, Ar) ; 7.56 (2H, d, J=8.3Hz, 3-H, 5- H) ; 7.40 (2H, m, Ar) ; 7.21 (IH, m, Ar) ; 4.15 (2H, s, 4- CH2) . M.S.TOF 293 (M+l) + . Hplc (Jupiter5 C18, Gradient 1, water/acetonitrile/TFA) rt 4.3min. Hplc (Symmetry Cδ, Gradient 3, water/acetonitrile/TFA) rt 7.5 min.
Example 32
8' -Amino-2' -naphthyl 4- (aminomethyl) benzoate ditrifluoroacetate salt H nmr (d4 methanol) 8.20 (2H, d, J=8.4Hz, 2-H, 6-H) ; 7.62 (IH, d, J=9.0Hz, Ar) ; 7.76 (IH, d, J=2.2Hz, 1 ' -H) ; 7.58 (2H, d, J=8.4Hz, 3-H, 5-H) ; 7.42 (IH, d, J=8.3Hz,
Ar) ; 7.27 (2H, m, Ar) ; 6.98 (IH, m, Ar) ; 4.15 (2H, s, 4- CH2) . M.S.TOF 292 (M+l) + . Hplc (Jupiter5 C18, Gradient 1, water/acetonitrile/TFA) rt 8.4 min. Hplc (Symmetry C8 , Gradient 3, water/acetonitrile/TFA) rt 7.4 min.
Example 33
5' -Quinolinyl 4- (aminomethyl) benzoate ditrifluoroacetate - 26 - salt lH nmr (d4 methanol) 6.97 (IH, d, J=4.3Hz, Ar) ; δ.59 (IH, d, J=8.5Hz, Ar) ; 8.30 (2H, d, J=8.1Hz, 2-H, 6-H) ; 8.01
(IH, d, J=8.6Hz, Ar) ; 7.91 (IH, t, J=8.1Hz, Ar) ; 7.70- 7.60 (2H, m, Ar) ; 7.63 (2H, d, J=8.1Hz, 3-H, 5-H) ; 4.20
(2H, s, 4-CH2) . M.S. TOF 279 (M+l) + . HPLC (Jupiter5 Clδ, water/acetonitrile/TFA, gradient 2) rt 9.δ3 min.
Example 34 8' -Quinolinyl 4- (aminomethyl) benzoate dihydrochloride salt l¥L nmr (d4 methanol) 9.12 (IH, dd, J=1.4, δ.4Hz, Ar) ; 9.06 (IH, dd, J=1.4, 5.2Hz, Ar) ; 6.33 (2H, d, J=8.4Hz, 2-H, 6-H) ; 8.19 (IH, dd, J=1.7, 7.9Hz , Ar) ; 8.03-7.92 (3H, m, Ar) ; 7.65 (2H, d, J=8.4Hz, 3-H, 5-H) ; 4.21 (2H, s, 4-CH2) . M.S. TOF 279 (M+l) + . HPLC (Jupiter5 Clδ, water/acetonitrile/TFA, gradient 2) rt 11.95 min.
Example 35 5' -Isoquinolinyl 4- (aminomethyl) benzoate dihydrochloride salt H nmr (d4 methanol) 9.84 (IH, s, 1 ' -H) ; 8.69 (IH, d, J=6.6Hz, Ar) ; 8.58 (2H, m, Ar) ; 8.47 (2H, d, 8.4Hz, 2-H, 6-H) ; 8.29 (IH, dd, J=6.7, 7.8Hz, Ar) ; 8.20 (IH, d, J=7.9Hz, 4'-H); 7.81 (2H, d, J=8.4Hz, 2-H, 5-H) ; 4.21
(2H, s, 4-CH2) . M.S. TOF 279 (M+l) + . HPLC (Jupiter5 C16, water/acetonitrile/TFA, gradient 2) rt 9.79 min.
Example 36 4' -Quinolinyl 4- (aminomethyl) benzoate dihydrochloride salt lH nmr (d4 methanol) 9.22 (IH, d, J=6.4Hz, 2 ' -H) ; 8.50 (IH, d, J=8.4Hz, Ar) ; 8.37 (2H, d, J=8.3Hz, 2-H, 6-H) ; 8.25 (IH, d, J=6.3Hz, 3 ' -H) ; 8.22-8.17 (2H, m, Ar) ; 8.00-7.94 (IH, m, Ar) ; 7.70 (2H, d, J=8.3Hz, 3-H, 5-H) ,
4.23 (2H, s, 4-CH2) . M.S. TOF 279 (M+l) + . HPLC (Jupiter5 Clδ, water/acetonitrile/TFA, gradient 2) rt 9.45 min. - 29 -
Example 37
3' -Pyridyl 4- (aminomethyl) benzoate dihydrochloride salt
XH nmr (d4 methanol) 8.98 (IH, d, J=2.2Hz, 2 ' -H) ; 8.72 (IH, d, J=5.4Hz, 4 ' -H) ; 8.53-8.49 (IH, m, 6 ' -H) ; 8.25 (2H, d, J=8.5Hz, 2-H, 6-H) ; 8.07 (IH, dd, J=5.6 , 8.7Hz , 5 ' -H) ; 7.62 (2H, d, J=8.5Hz, 3-H, 5-H); 4.1δ (2H, s, 4- CH2) . M.S. TOF 229 (M+l) + . HPLC (Jupiter5 Clδ, water/acetonitrile/TFA, gradient 2) rt 7.26 min.
Example 38
3 ' - (N-Morpholino) carbonylphenyl 4- (aminomethyl) benzoate hydrochloride salt
:H nmr (d4 methanol) 6.17 (2H, d, J=δ.2Hz, 2-H, 6-H) ;
7.5δ (2H, d, J=8.2Hz, 3-H, 5-H); 7.48 (IH, m, Ar) ; 7.30 (IH, d, J=1.5Hz, Ar) ; 7.27 (2H, s, Ar) ; 4.13 (2H, s, 4-
CH2) ; 3.7-3.3 (8H, m, 3 ' -N (CH2CH2) 2) . M.S.TOF 341 (M+l) + .
Hplc (Jupiter5 Clδ, Gradient 1, water/acetonitrile/TFA) rt 4.97min. Hplc (SymmetryCδ, Gradient 5, water/acetonitrile/TFA) rt 7.66min.
Example 39
4' - (N-Morpholino) carbonylphenyl 4- (aminomethyl) benzoate hydrochloride salt
Η nmr (d4 methanol) 6.05 (2H, d, J=8.3Hz, 2-H, 6-H) ; 7.49 (2H, d, J=8.3Hz, 3-H, 5-H) ; 7.36 (2H, d, J=δ.5Hz,
3'-H, 5'-H); 7.17 (2H, d, J=8.5Hz, 2 ' -H, 6 ' -H) ; 4.06 (2H, s, 4-CH2) ; 3.6-3.2 (8H, m, 4 ' -N (CH2CH2) 2) . M.S.TOF
341 (M+l)+. Hplc (Jupiter5 Clδ, Gradient 1, water/acetonitrile/TFA) rt 3.77min. Hplc (SymmetryCδ, Gradient 5, water/acetonitrile/TFA) rt 7.69min.
Example 40
2 ' - (N-Morpholino) carbonylphenyl 4- (aminomethyl) benzoate hydrochloride salt lU nmr (d4 methanol) δ.06 (2H, d, J=8.3Hz, 2-H, 6-H) ;
7.50 (2H, d, J=8.3 Hz, 3-H, 5-H); 7.41 (IH, m, Ar) ; 7.25 (3H, m, Ar) ; 4.07 (2H, s, 4-CH2) ; 3.6-3.1 (δH, m, 2'- - 30 -
N(CH2CH2)2). M.S.TOF 341 (M+l) + . Hplc (Jupiter5 C18, Gradient 1, water/acetonitrile/TFA) rt 5.66min. Hplc
(SymmetryCδ, Gradient 5, water/acetonitrile/TFA) rt 7. δOmin.
Example 41
3' - ( (N-Morpholino) carbonylmethyl) phenyl 4-
(aminomethyl) benzoate hydrochloride salt Η nmr (d< methanol) 8.50 (2H, d, J=8.3Hz, 2-H, 6-H) ; 7.92 (2H, d, J=δ.3Hz, 3-H, 5-H); 7.67 (IH, m, Ar) ; 7.48
(IH, d, J=7.6Hz, Ar) ; 7.40 (2H, d, J=8.0Hz, Ar) ; 4.50
(2H, S, 3'-CH2); 4.09 (2H, s, 4-CH2); 3.9-3.7 (8H, m, 3'- N(CH2CH2)2). M.S.TOF 355 (M+l) + . Hplc (Jupiter5 Clδ, Gradient 1, water/acetonitrile/TFA) rt 6.54min. Hplc (SymmetryCδ, Gradient 5, water/acetonitrile/TFA) rt 7.76min.
Example 42
4' - ( (N-Morpholino) carbonylmethyl) phenyl 4- (aminomethyl) benzoate hydrochloride salt
Η nmr (d4 methanol) 8.15 (2H, d, J=8.3Hz, 2-H, 6-H) ; 7.55 (2H, d, J=8.3Hz, 3-H, 5-H); 7.26 (2H, d, J=8.6Hz, Ar) ; 7.11 (2H, d, J=8.6Hz, Ar) ; 4.14 (2H, s, 4-CH2); 3.71 (2H, s, 4'-CH2); 3.7-3.3 (8H, m, 4 ' -N (CH2CH2) 2) . M.S.TOF 355 (M+l)+. Hplc (Jupiter5 Clδ, Gradient 1, water/acetonitrile/TFA) rt 5.93min. Hplc (SymmetryCδ, Gradient 5, water/acetonitrile/TFA) rt 7.71min.
Example 43 2' - ( (N-Morpholino) carbonylmethyl) phenyl 4-
(aminomethyl) benzoate hydrochloride salt lE nmr (d4 methanol) 6.21 (2H, s, Ar) ; 7.67 (2H, s, Ar) ; 7.35 (2H, m, Ar) ; 7.26 (2H, m, Ar) ; 4.21 (2H, s, 4-CH2); 3.75 (2H, s, 2'-CH2); 3.6-3.3 (8H, m, 2 ' -N (CH2CH2) 2) . M.S.TOF 355 (M+l)+. Hplc (Jupiter5 C18, Gradient 1, water/acetonitrile/TFA) rt 5.6δmin. Hplc (SymmetryCδ, Gradient 5, water/acetonitrile/TFA) rt δ.58min. - 31 -
Example 44
2'-Biphenyl 4- (aminomethyl) benzoate trifluoroacetate salt XH nmr (d4 methanol) 8.06 (2H, d, J=8.3Hz, 2-H, 6-H) ;
7.57 (2H, d, J=8.3Hz, 3-H, 5-H); 7.51-7.39 (5H, m, Ar) ; 7.33-7.21 (4H, m, Ar) ; 4.21 (2H, s, 4-CH2); M.S. TOF 304 (M+l)+. Hplc (Jupiter5 Clδ, Gradient 1,
Water/acetonitrile/TFA) rt 16.04 min. Hplc (SymmetryCδ Gradient 2, Water/acetonitrile/TFA) rt 10.54 min.
Example 45
2' - (t-Butyl) phenyl 4- (aminomethyl) benzoate trifluoroacetate salt XH nmr (d4 methanol) 8.30 (2H, d, J=8.4Hz, 2-H, 6-H) ;
7.70 (2H, d, J=δ.4Hz, 3-H, 5-H) ; 7.53-7.46 (IH, m, 6'- H) ; 7.33- 7.21 (2H, m, 5 ' -H, 4 ' -H) ; 7.05-7.11 (IH, m, 3 ' -H) ; 4.2δ (2H, s, 4-CH2); 1.36 (9H, s, 2 ' -lBu) ; M.S. TOF 284 (M+l) + . Hplc (Jupiter5 Clδ, Gradient 1, Water/acetonitrile/ TFA) rt 16.64 min. Hplc (SymmetryC8 Gradient 2, Water/acetonitrile/TFA) rt 10.69 min.
Example 46
4 ' - (2 -Phenylethenyl) phenyl 4- (aminomethyl) benzoate trifluoroacetate salt
2H nmr (d4 methanol) 8.16 (2H, d, J=8.3Hz, 2-H, 6-H) ; 7.57-7.07 (13H, m, Ar, CH=CH) ; 4.15 (2H, s, 4-CH2). M.S. TOF 330 (M+l) + . Hplc (Jupiter5 Clδ, Gradient δ, Water/acetonitrile/TFA) rt 13.47 min. Hplc (SymmetryCδ Gradient 2, Water/acetonitrile/TFA) rt 11.31 min.
Example 47
4' - (2-Phenylethyl) phenyl 4- (aminomethyl) benzoate trifluoroacetate salt :H nmr (d4 methanol) 8.14 (2H, d, J=8.3Hz, 2-H, 6-H) ;
7.57 (2H, d, J=8.3Hz, 3-H, 5-H); 7.16-6.97 (9H, m, Ar) ; 4.15 (2H, s, 4-CH2) ; 2.84 (4H, s, 4 ' -CH2CH2) . M.S. TOF - 32 -
332 (M+l)+. Hplc (Jupiter5 C18, Gradient 8, Water/acetonitrile/TFA) rt 13.47 min. Hplc (SymmetryCδ Gradient 2, Water/acetonitrile/TFA) rt 11.25 min.
Example 48
2' -Nitrophenyl 4- (aminomethyl) benzoate hydrochloride salt λE nmr (d4 methanol) 8.16 (2H, d, J=8.1, 2-H, 6-H) ; δ.09 (IH, dd, J=1.0, 8.1Hz, 3 ' -H) ; 7.73 (IH, t, J=7.2Hz, 5'- H) ; 7.59 (2H, d, J=8.1Hz, 3-H, 5-H); 7.46 (IH, t,
J=7.8Hz, 4 ' -H) ; 7.41 (IH, d, J=8.0Hz, 6 ' -H) ; 4.17 (2H, s, 4-CH2) . M.S. TOF 273 (M+l) + . HPLC (Jupiter5 C18, water/acetonitrile/TFA, gradient 2) rt
Example 49
2 ' -Cyanopheny1 4- (aminomethyl) benzoate hydrochloride salt
:H nmr (d4 methanol) 8.21 (2H, d, J=8.4Hz, 2-H, 6-H) ; 7.77-7.67 (2H, m, Ar) ; 7.59 (2H, d, J=8.4Hz, 3H, 5H) ; 7.46-7.36 (2H, m, Ar) ; 4.17 (2H, s, 4-CH2). M.S. TOF 253 (M+l)+.
Example 50
2 ' -Methylphenyl 4- (aminomethyl) benzoate hydrochloride salt H nmr (d4 methanol) 8.17 (2H, d, J=8.2Hz, 2-H, 6-H) ; 7.59 (2H, d, J=6.2Hz, 3-H, 5-H) ; 7.23-7.01 (4H, m, Ar) ; 4.16 (2H, s, 4-CH2) . M.S. TOF 242 (M+l) +
The following examples were prepared in a similar manner. Suitably protected alcohols were bought or synthesised by conventional means and aminomethylbenzoic acid was purchased and used as previously described.
Analysis: Proton nuclear magnetic resonance (XH) spectra were recorded on Bruker DPX300 (300MHz) . Analytical Hplc was on a Shimadzu LC6 gradient system equipped with an - 33 - autosampler. Eluant A consisted of aqueous TFA (0.1%) and eluant B 90% MeCN in aqueous TFA (0.1%) with gradient elution (Gradient 4, Omin 20%B then 20%B to 53.3%B over 15min the 53.3%B for lmin then 100%B for lmin; Gradient 5, Omin 20%B then 20%B to 100%B over
15min then 100%B for 5min; Gradient 6, Omin 20%B then 20%B to 100%B over 5 min then 100%B for lmin) . Columns used were Magellan Cδ column (2.1 x 150mm, 5μm particle size) and Magellan Clδ column (4.6 x 30mm, 3μm particle size) . Purified products were further analysed by LC-MS (PESCIEX Single Quadropole API-150EX) .
Example 51
4- (Methoxycarbonylmethyl) phenyl 4- (aminomethyl) benzoate trifluoroacetate salt lE nmr (d4 methanol) 8.25 (2H, d) ; 7.65 (2H, d) ; 7.38 (2H, d) ; 7.20 (2H, d) ; 4.29 (2H, s);3.72 (5H,s) M.S.TOF 300 (M+l)+, Hplc (Magellan C8, Gradient 4, water/acetonitrile/TFA) rt 12.06min, LC-MS (Magellan C18, Gradient 6, water/acetonitrile/TFA) rt 2.27min, 300 (M+l)+.
Example 52
4- [ (2 -Acetylamino-2-carboxy) ethyl] phenyl 4- (aminomethyl) benzoate trifluoroacetate salt λE nmr (d4 methanol) δ.00 (2H, d) ; 7.43 (2H, d) ; 6.98 (2H, d) ; 4.48 (IH, m) ; 4.0 (2H, s) ; 3.01 (IH, dd) ; 2.78 (IH, dd) ; 1.72 (3H, s) . Hplc (magellan C8, Gradient 5, water/acetonitrile/TFA) rt 9.4min. LC-MS (magellan Clδ, Gradient 6, water/acetonitrile/TFA) rt 1.69min, 357 (M+l)+.
Example 53
4- [ (N-Carboxymethyl) carboxamido] phenyl 4- (aminomethyl) benzoate trifluoroacetate salt H nmr (d4 methanol) 6.31 (2H, d) ; 8.02 (2H, d) ; 7.71 (2H, d) ; 7.42 (2H, d) ; 4.29 (2H, s) ; 4.16 (2H, s) . Hplc - 34 -
(Magellan C8 , Gradient 5, Water/acetonitrile/TFA) rt 7.23 min. LC-MS (Magellan Clδ, Gradient 6, water/acetonitrile/TFA) rt 1.45 min, 329 (M+l)+.
Example 54
4- (Carboxymethyl) phenyl 4- (aminomethyl) benzoate trifluoroacetate salt λU nmr (d4 methanol) 8.27 (2H, d) ; 7.68 (2H, d) ; 7.39 (2H, d) ; 7.20 (2H, d) ; 4.24 (2H, s);3.69 (2H, s) , Hplc (Magellan C8 , Gradient 4, water/acetonitrile/TFA) rt 9.94min, LC-MS (Magellan C18, Gradient 6, water/acetonitrile/TFA) rtl.δOmin, 266 (M+l)+.
Example 55 O- [Methyl N-acetyl-L_tyrosinyl] 4- (aminomethyl) benzoate trifluoroacetate salt ln nmr (d4 methanol) 8.22 (2H, d) ; 7.63 (2H, d) ; 7.31 (2H, d) ; 7.20 (2H, d) ; 4.72 (IH, m) ; 4.27 (2H, s) ; 3.69 (3H, S) ; 3.21 (3H, s) ; 3.21 (IH, dd) ; 3.01 (IH, dd) ; 2.92 (3H, s) . Hplc (magellan Cδ , gradient 5, water/acetonitrile/TFA) rt δ.3min. LC-MS (magellan Clδ, Gradient 5, water/acetonitrile/TFA) rt 1.98 min, 371 (M+l)+.
Example 56
4- [N- [ (Methoxycarbonyl) methyl] carboxamido] phenyl 4- (aminomethyl) benzoate trifluoroacetate salt
XH nmr (d4 methanol) 8.30 (2H, d) ; 7.95 (2H, d) ; 7.32 (2H, d) ; 4.28 (2H, s) ; 4.12 (2H, s) ; 3.71 (3H, s) . Hplc (magellan Cδ , Gradient 5, water/acetonitrile/TFA) rt 9.10min. LC-MS (magellan Clδ, Gradient 5, water/acetonitrile/TFA) rt 1.65min, 343 (M+l)+.
Example 57 4- [4-Carboxypiperidin-l-oyl] phenyl 4-
(aminomethyl) benzoate trifluoroacetate salt
:H nmr (d4 methanol) 8.10 (2H, d) ; 7.39 (2H, d) ; 7.32 - 35 -
(2H, d) ; 7.18 (2H, d) ; 4.32 (IH, m) ; 4.12 (2H, s) ; 3.65 (IH, m) ; 3.05 (2H, m) ; 2.60 (IH, m) ; 2.82 (2H, m) ; 1.50 (2H, ) . Hplc (magellan C8 , Gradient 5, water/acetonitrile/TFA) rt 8.4min. LC-MS (magellan Clδ, Gradient 5, water/acetonitrile/TFA) rt 1.69min, 3δ3 (M+l)+.
Example 58
4- (2 -Carboxypyrrolidin-1-oyl) phenyl 4- (aminomethyl) benzoate trifluoroacetate salt
:H nmr (d4 methanol) 6.15 (2H, d) ; 7.56 (4H, m) ; 7.29 (2H, d) ; 4.53 (IH, m) ; 4.15 (2H, s) ; 3.70-3.50 (2H, m) ; 2.30 (IH, m) ; 2.10-1.60 (3H, m) , Hplc (Magellan Cδ , Gradient 4, water/acetonitrile/TFA) rt 9.34min, LC-MS (Magellan C18, Gradient 6, water/acetonitrile/TFA) rt 1.73min, 369 (M+l)+.
Example 59
4- [4- (Ethoxycarbonyl)piperidin-l-oyl] phenyl 4- (aminomethyl) benzoate trifluoroacetate salt
A nmr (d4 methanol) 8.27 (2H, d) ; 7.62 (2H, d) ; 7.52 (2H, d) ; 7.39 (2H, d) ; 4.48 (IH, s) ; 4.29 (2H, s) ; 4.15 (2H, ; 3.77 (IH, m) ; 3.22 (2H, m) ; 2.73 (IH, m) ; 2.05 (2H, m) ; 1.70 (2H, m) ; 1.30 (3H, t). Hplc (magellan C8, Gradient 5, water/acetonitrile/TFA) rt 7.29 min. LC-MS (magellan Clδ, Gradient 5, water/acetonitrile/TFA) rt 2.41min, 411 (M+l)+.
Example 60 3- (4-Carboxypiperidin-1-oyl) phenyl 4-
(aminomethyl) benzoate trifluoroacetate salt H nmr (d4 methanol) 6.32 (2H, d) ; 7.69 (2H, d) ; 7.62 (IH, t) ; 7.43-7.36 (3H, m) ; 4.56-4.45 (IH, m) ; 4.29 (2H, s) ; 3.86-3.74 (IH, m) , 3.36-3.09 (2H, m) ; 2.75-2.64 (IH, m) ; 2.16-1.89 (2H, m) ; 1.82-1.96 (2H, m) . Hplc
(Magellan Cδ , Gradient 5, Water/acetonitrile/TFA) rt δ.03 min. LC-MS (Magellan Clδ, Gradient 6, - 36 - water/acetonitrile/TFA) rt 1.68 min, 3δ3 (M+l)+.
Example 61
2- (4-Carboxypiperidin-l-oyl) phenyl 4- (aminomethyl) benzoate trifluoroacetate salt lR nmr (d4 methanol) 8.23 (2H, d) ; 7.67 (2H, d) ; 7.62 (IH, m) ; 7.48-7.37 (3H, m) ; 4.45-4.30 (IH, m) ; 4.26 (2H, s) ; 3.67-3.57 (IH, m) , 3.25-3.08 (IH, m) ; 2.99-2.87 (IH, m) ; 2.62-2.51 (IH, m) ; 1.95-1.81 (2H, m) ; 1.72-1.26 (2H, m) . Hplc (Magellan C8, Gradient 5,
Water/acetonitrile/TFA) rt 8.03 min. LC-MS (Magellan C18, Gradient 6, water/acetonitrile/TFA) rt 1.80 min, 383 (M+l) + .
Example 62
4- [2- (N- (l-Carboxy-2-hydroxy) ethyl) carboxamido- pyrrolidin-1-oyl] phenyl 4- (aminomethyl) benzoate trifluoroacetate salt H nmr (d4 methanol) 8.15 (2H, d) ; 7.64 (2H, d) ; 7.57 (2H, d) ; 7.24(2H, d) ; 4.60 (IH, m) ; 4.48 (IH, m) ; 4.17
(2H, s) ;3.90-3.20 (4H, ) ; 2.28 (IH, m) ; 2.10-1.60 (3H, m) , Hplc (Magellan C8, Gradient 4, water/acetonitrile/TFA) rt 8.17min, LC-MS (Magellan C18,
Gradient 6, water/acetonitrile/TFA) rt 1.35min, 456 (M+l)+.
Example 63
4- [ (4-Ethoxycarbonylmethyl)piperazin-l-oylphenyl] 4-
(aminomethyl) benzoate triflouroacetate salt lE nmr (d4 methanol) 8.31 (2H, d) ; 7.83 (2H, d) ; 7.79
(2H, d) ; 7.57 (2H, d) ; 4.50 (2H, q) ; 4.42 (2H, s) ; 4.30 (2H, s) ; 4.10 (4H, bs) ; 3.58 (4H, bs) ; 1.50 (3H, t) . Hplc (magellan C8, Gradient 5, water/acetonitrile/TFA) rt 7.37min. LC-MS (magellan C18, Gradient 5, water/acetonitrile/TFA) rt 1.40min, 426 (M+l)+.
Example 64 - 37 -
4- [4- (Phenylsulfonylcarboxamido)piperidin-l-oyl] phenyl 4- (aminomethyl) benzoate trif luoroacetate salt lE nmr (d4 methanol) 8.30 (2H, d) ; 8.05 (2H, d) ; 7.76- 7.60 (5H, m) ; 7.54 (2H, d) ; 7.39 (2H, d) , 4.60-4.49 (IH, m) ; 4.29 (2H, s) ; 3.87-3.74 (IH, m) , 3.26-2.95 (2H, m) ; 2.64-2.53 (IH, m) ; 1.95-1.70 (2H, m) ; 1.68-1.53 (2H, m) . Hplc (Magellan C8 , Gradient 5, Water/acetonitrile/TFA) rt 9.17 min. LC-MS (Magellan C18, Gradient 6, water/acetonitrile/TFA) rt 2.29 min, 522 (M+l)+.
Example 65
4-Carboxyphenyl 4- (aminomethyl) benzoate trif luoroacetate salt
:H nmr (d4 methanol) 8.28 (2H, d) ; 8.14 (2H, d) ; 7.69 (2H, d) ; 7.38 (2H, d) ; 4.27 (2H, s) . Hplc (Magellan C8, Gradient 5, Water/acetonitrile/TFA) rt 7.81 min. LC-MS (Magellan C18, Gradient 6, water/acetonitrile/TFA) rt 2.27 min, 272 (M+l)+.
Example 66
2- (2 -Carboxypyrrolidin-1-oyl) phenyl 4-
(aminomethyl) benzoate trifluoroacetate salt lE nmr (d4 methanol) 8.25 (2H, d) ; 7.70-7.30 (6H, m) ; 4.40 (IH, m) ; 4.25 (2H, s) ; 3.50 (2H, m) ; 2.40-1.80 (4H, m) , Hplc (Magellan C8 , Gradient 4, water/acetonitrile/TFA) rt 8.76min, LC-MS (Magellan Clδ, Gradient 6, water/acetonitrile/TFA) rt 1.83min, 369 (M+l)+.
Example 67
3- (2 -Carboxypyrrolidin-1-oyl) phenyl 4-
(aminomethyl) benzoate trifluoroacetate salt H nmr (d4 methanol) 8.40 (2H, d) ; 7.80(2H, d) ; 7.70-7.40 (4H, m) ; 4.80-4.50 (IH, m) ; 4.39 (2H, s) ; 3.60 (2H, m) ; 2.50 (IH, m) ; 2.15 (3H, m) , Hplc (Magellan Cδ , Gradient 4, water/acetonitrile/TFA) rt 7.6δmin, LC-MS (Magellan Clδ, Gradient 6, water/acetonitrile/TFA) rt 1.93min, 369 - 38 - (M+ l ) \
Example 68
4- (N-Phenylsulfonylcarboxamido) phenyl 4- (aminomethyl) benzoate trifluoroacetate salt
XH nmr (d4 methanol) 8.27 (2H, d) ; 8.12 (2H, d) ; 8.00 (2H, d) ; 7.71-7.58 (5H, m) ; 7.37 (2H, d) ; 4.29 (2H, s) . Hplc (Magellan Cδ , Gradient 5, Water/acetonitrile/TFA) rt 9.74 min. LC-MS (Magellan Clδ, Gradient 6, water/acetonitrile/TFA) rt 2.61 min, 411 (M+l)+.
Example 69
4- (Tetrazolylmethyl) phenyl 4- (aminomethyl) benzoate trifluoroacetate salt XH nmr (d4 methanol) 8.27 (2H, d) ; 7.68 (2H, d) ; 7.49
(2H, d) ; 7.30 (2H, d) ; 4.27 (2H, s);3.97 (2H, s) M.S.TOF 310 (M+l)+, Hplc (Magellan C8, Gradient 4, water/acetonitrile/TFA) rt 10.15min, Hplc (Magellan Clδ, Gradient 6, water/acetonitrile/TFA) rt 2.12min.
Example 70
4- (N-Methanesulfonylcarboxamido) phenyl 4-
(aminomethyl) benzoate trifluoroacetate salt λE nmr (d4 methanol) δ.25 (2H, d) ; 8.01 (2H, d) ; 7.68 (2H, d) ; 7.43 (2H, d) ; 4.28 (2H, s) ; 3.39 (3H, s) . Hplc (Magellan C8 , Gradient 5, Water/acetonitrile/TFA) rt 7.68 min. LC-MS (Magellan C18, Gradient 6, water/acetonitrile/TFA) rt 1.74 min, 349 (M+l)+.
Example 71
4- [ (2 -Amino-2 -carboxy) ethyl] phenyl 4-
(aminomethyl) benzoate ditrifluoroacetate salt lE nmr (d4 methanol) 8.28 (2H, d) ; 7.70 (2H, d) ; 7.46 (2H, d) ; 7.29 (2H, d) ; 4.29 (2H, s) ; 4.19 (IH, dd) ; 3.41 (IH, dd) ; 3.20 (IH, dd) . Hplc (Magellan C8 , Gradient 5, Water/acetonitrile/TFA) rt 6.77 min. LC-MS (Magellan C18, Gradient 6, water/acetonitrile/TFA) rt 1.35 min, - 39 - 315 (M+l) + .
Example 72
[4- (4-Carboxypiperidin-l-oyl) -2 -nitro] phenyl 4- (aminomethyl) benzoate trifluoroacetate salt
*H nmr (d4 methanol) 8.33-8.28 (3H, m) ; 7.91 (IH, dd) ; 7.73 (2H, d) ; 7.66 (IH, d) ; 4.57-4.38 (IH, m) ; 4.31 (2H, s) ; 3.83-3.70 (IH, m) , 3.36-3.13 (2H, m) ; 2.78-2.66 (IH, m) ; 2.17-1.92 (2H, m) ; 1.87-1.69 (2H, m) . Hplc (Magellan C8 , Gradient 5, Water/acetonitrile/TFA) rt 8.49 min. LC-MS (Magellan C18, Gradient 6, water/acetonitrile/TFA) rt 1.83 min, 428 (M+l)+.
Example 73 [2- (4-Carboxypiperidin-l-oyl) -4-nitro] phenyl 4- (aminomethyl) benzoate trifluoroacetate salt
:H nmr (d„ methanol) 8.48 (IH, dd) ; 8.39 (IH, br s) ; 8.26 (2H, d) ; 7.77-7.67 (3H, m) ; 4.51-4.36 (IH, m) ; 4.30 (2H, s) ; 3.68-3.59 (IH, m) , 3.31-3.18 (IH, m) ; 3.02 (IH, td) ; 2.68-2.58 (IH, m) ; 2.00-1.90 (2H, m) ; 1.78-1.30 (2H, m) . Hplc (Magellan C8, Gradient 5, Water/acetonitrile/TFA) rt 9.18 min. LC-MS (Magellan C18, Gradient 6, water/acetonitrile/TFA) rt 2.07 min, 426 (M+l)+.
Example 74
4-Sulphonamidophenyl-4- (aminomethyl) benzoate trifluoroacetate salt
:H nmr (d4 methanol) 8.2δ (2H, d) ; 8.02 (2H, d) ; 7.68 (2H, d) ; 7.46 (2H, d) ; 4.27 (2H, s) . Hplc (Magellan C8, Gradient 5, Water/acetonitrile/TFA) rt δ.70 min. LC/MS (Magellan Clδ Gradient 6, water/acetonitrile/TFA) rt 1.17 min, 307 (M+l)+.
Example 75 (4-Carboxymethyl-2-nitro)phenyl-4- (aminomethyl) benzoate trifluoroacetate salt
:H nmr (d4 methanol) H nmr (d4 methanol) 6.29 (2H, d) ; - 40 -
8.18 (IH, d) ; 7.78 (IH, dd) ; 7.71 (2H, d) ; 7.49 (IH, d) ; 4.30 (2H, s) ; 3.85 (2H, s) . Hplc (Magellan Cδ, Gradient 5, Water/acetonitrile/TFA) rt 9.25 min. LC/MS (Magellan Clδ Gradient 6, water/acetonitrile/TFA) rt 1.25 min, 331 (M+l)+.
Example 76
4- [2- [ (Pyrrolidin-2-oyl) amino-2-carboxy] ethyl] phenyl-4-
(aminomethyl) benzoate trifluoroacetate salt XH nmr (d4 methanol) δ.25 (2H, d) ; 7.67 (2H, d) ; 7.38
(2H, d) ; 7.19 (2H, d) ; 4.87-4.72 (2H, m) ; 4.27-4.21 (3H, m) ; 3.46-3.31 (2H, m) ; 3.07 (IH, dd) ; 2.50-2.35 (IH, m) ; 2.16-1.97 (3H, m) . Hplc (Magellan C8 , Gradient 5, Water/acetonitrile/TFA) rt 7.88 min. LC/MS (Magellan C18 Gradient 6, water/acetonitrile/TFA) rt 1.00 min, 412 (M+l)+.
Example 77
4- [ (4-Carboxypiperidin-l-oyl)methyl]phenyl-4- (aminomethyl) benzoate trifluoroacetate salt lE nmr (d4 methanol) 8.27 (2H, d) ; 7.68 (2H, d) ; 7.38 (2H, d) ; 7.24 (2H, d) ; 4.40 (IH, d) ; 4.27 (2H, s) ; 4.01 (IH, d) ; 3.86 (2H, s) ; 3.23 (IH, t); 2.93 (IH, t) ; 2.67- 2.54 (IH, m) ; 2.00-1.87 (2H, m) ; 1.65-1.45 (2H, m) . Hplc (Symmetry Rpδ, Gradient 8, Water/acetonitrile/TFA) rt 5.04 min. LC/MS (Magellan Clδ Gradient 6, water/acetonitrile/TFA) rt 1.32 min, 397 (M+l)+.
Example 78 [ (4-Carboxymethylpiperazin-l-oyl)methyl] phenyl-4- (aminomethyl) benzoate ditrifluoroacetate salt lE nmr (d4 methanol) 8.25 (2H, d) ; 7.67 (2H, d) ; 7.37 (2H, d) ; 7.23 (2H, d) ; 4.26 (2H, s) ; 4.13 (2H, s) ; 4.03- 3.86 (6H, m) ; 3.47-3.34 (4H, m) . Hplc (Symmetry Rpδ, Gradient δ, Water/acetonitrile/TFA) rt 3.90 min. LC/MS (Magellan Clδ Gradient 6, water/acetonitrile/TFA) rt 1.07 min, 412 (M+l)\ - 41 -
Example 79
4- [ (N-Carboxymethyl) acetamido] phenyl-4 -
(aminomethyl) benzoate trifluoroacetate salt :H nmr (d4 methanol) 8.27 (2H, d) ; 7.66 (2H, d) ; 7.44
(2H, d) ; 7.21 (2H, d) ; 4.26 (2H, s) ; 3.95 (2H, s) ; 3.6 (2H, s) . Hplc (Symmetry Rpδ , Gradient 8,
Water/acetonitrile/TFA) rt 4.42 min. LC/MS (Magellan C18 Gradient 6, water/acetonitrile/TFA) rt 1.17 min, 343 (M+l)+.
Example 80
4- [ (2-Carboxypyrrolidin-l-oyl)methyl] phenyl-4-
(aminomethyl) benzoate trifluoroacetate salt lE nmr (d4 methanol) 8.26 (2H, d) ; 7.66 (2H, d) ; 7.40
(2H, d) ; 7.21 (2H, d) ; 4.48 (IH, dd) ; 4.26 (2H, s) ; 3.82 (2H, s) ; 3.73-3.59 (2H, m) ; 2.35-1.98 (4H, m) . Hplc (Symmetry Rp8, Gradient 8, Water/acetonitrile/TFA) rt 4.97 min. LC/MS (Magellan C18 Gradient 6, water/acetonitrile/TFA) rt 1.35 min, 383 (M+l)+.
Example 81
4- [4- (Carboxymethyl)piperazin-l-oyl] phenyl-4-
(aminomethyl) benzoate ditrifluoroacetate salt XH nmr (d4 methanol) 8.28 (2H, d) ; 7.69 (2H, d) ; 7.64
(2H, d) ; 7.42 (2H, d) ; 4.27 (2H, s) ; 4.19 (2H, s) ; 4.08- 3.90 (4H, m) ; 3.56-3.44 (4H, m) . Hplc (Magellan C8, Gradient 5, Water/acetonitrile/TFA) rt 9.74 min. LC/MS (Magellan Clδ Gradient 6, water/acetonitrile/TFA) rt O.δO min, 398 (M+l)+.
Example 82
4- (Carboxymethyl-aminosulphonyl) phenyl-4-
(aminomethyl) benzoate trifluoroacetate salt H nmr (d4 methanol) 8.30 (2H, d) ; 8.01 (2H, d) ; 7.71
(2H, d) ; 7.50 (2H, d) ; 4.31 (2H, s) ; 3.79 (2H, s) . Hplc (Magellan C8 , Gradient 5, Water/acetonitrile/TFA) rt - 42 -
9.04 min. LC/MS (Magellan Clδ Gradient 6, water/acetonitrile/TFA) rt 1.20 min, 365 (M+l)+.
Example 83 4- (N-Benzoylaminosulphonyl)phenyl-4-
(aminomethyl) benzoate trifluoroacetate salt H nmr (d4 methanol) 8.33 (2H, d) ; 8.27 (2H, d) ; 7.89 (2H, d) ; 7.73-7.62 (3H, m) ; 7.59-7.50 (4H, m) ; 4.30 (2H, s) . Hplc (Magellan C8 , Gradient 5, Water/acetonitrile/TFA) rt 13.76 min. LC/MS (Magellan
C18 Gradient 6, water/acetonitrile/TFA) rt 1.83 min, 411 (M+l)+.
Example 84 4- [4- (Aminomethyl) benzoylaminosulphonyl] phenyl-4- (aminomethyl) benzoate ditrifluoroacetate salt
:H nmr (d4 methanol) 8.29 (2H, d) ; 8.21 (2H, d) ; 7.96 (2H, d) ; 7.70 (2H, d) ; 7.58 (2H, d) ; 7.52 (2H, d) ; 4.29 (2H, s) ; 4.22 (2H, s) . Hplc (Magellan C8, Gradient 5, Water/acetonitrile/TFA) rt 10.44 min. LC/MS (Magellan
Clδ Gradient 6, water/acetonitrile/TFA) rt 1.12 min, 440 (M+l)+.
Example 85 4- (Tetrazol-5-yl)phenyl-4- (aminomethyl) benzoate trifluoroacetate salt
XH nmr (d„ methanol) 8.24 (2H, d) ; 8.0δ (2H, d) ; 7.61 (2H, d) ; 7.32 (2H, d) ; 4.21 (2H, s) . Hplc (Magellan C8 , Gradient 5, Water/acetonitrile/TFA) rt 9.81 min. LC/MS (Magellan C18 Gradient 6, water/acetonitrile/TFA) rt 1.17 min, 296 (M+l)+.
Example 86
4- [N- (Tetrazol-5-ylmethyl) carboxamido] phenyl-4- (aminomethyl)benzoate trifluoroacetate salt
:H nmr (d4 methanol) 6.31 (2H, d) ; 8.06 (2H, d) ; 7.70 (2H, d) ; 7.43 (2H, d) ; 4.98 (2H, s) , 4.30 (2H, s) . Hplc - 43 -
(Magellan Cδ , Gradient 5, Water/acetonitrile/TFA) rt 8.64 min. LC/MS (Magellan Clδ Gradient 6, water/acetonitrile/TFA) rt 1.05 min, 353 (M+l)+.
Example 87
3- [ (2-Carboxy-2-acetylamino) ethyl] phenyl-4- (aminomethyl) benzoate trifluoroacetate salt
:H nmr (d4 methanol) 8.29 (2H, d) ; 7.69 (2H, d) ; 7.43 (IH, t) ; 7.24 (IH, d) ; 7.17-7.11 (2H, m) ; 4.73 (IH, dd) ; 4.29 (2H, s) ; 3.31 (IH, dd) ; 3.04 (IH, dd) ; 1.96 (3H, s) . Hplc (Magellan C8, Gradient 5,
Water/acetonitrile/TFA) rt 8.75 min. LC/MS (Magellan Clδ Gradient 6, water/acetonitrile/TFA) rt 1.13 min, 357 (M+l)+.
Example 88
2 [ (2 -Carboxy-2 -acetylamino) ethyl] phenyl-4-
(aminomethyl) benzoate trifluoroacetate salt lE nmr (d4 methanol) 8.34 (2H, d) ; 7.68 (2H, d) ; 7.40- 7.34 (2H, m) ; 7.30-7.20 (2H, m) ; 4.77 (IH, dd) ; 4.27 (2H, s) ; 3.25 (IH, dd) ; 2.90 (IH, dd) ; 1.88 (3H, s) . Hplc (Magellan Cδ , Gradient 5, Water/acetonitrile/TFA) rt 9.26 min. LC/MS (Magellan Clδ Gradient 6, water/acetonitrile/TFA) rt 1.27 min, 357 (M+l)+.
Example 89
2- [ (Carboxymethyl) acetamido] phenyl-4-
(aminomethyl) benzoate trifluoroacetate salt
:H nmr (d4 methanol) 8.30 (2H, d) ; 7.68 (2H, d) ; 7.51- 7.39 (2H, m) ; 7.36-7.26 (2H, m) ; 4.27 (2H, s) ; 3.82 (2H, s) ; 3.65 (2H, s) . Hplc (Magellan C8, Gradient 5, Water/acetonitrile/TFA) rt 6.42 min. LC/MS (Magellan Clδ Gradient 6, water/acetonitrile/TFA) rt 1.20 min, 343 (M+l)+.
Example 90
2- [ [ (l-Carboxy-2-hydroxy) ethyl] acetamido] phenyl-4- - 44 -
(aminomethyl) benzoate trifluoroacetate salt lE nmr (d4 methanol) 8.32 (2H, d) ; 7.68 (2H, d) ; 7.52- 7.39 (2H, m) ; 7.37-7.26 (2H, m) ; 4.43 (IH, t); 4.28 (2H, s) ; 3.86 (IH, dd) ; 3.74 (IH, dd) ; 3.68 (2H, s) . Hplc (Magellan Cδ , Gradient 5, Water/acetonitrile/TFA) rt δ.69 min. LC/MS (Magellan Clδ Gradient 6, water/acetonitrile/TFA) rt 1.15 min, 373 (M+l)+.
Example 91 2- [ (alpha-Carboxy)benzylacetamido] phenyl-4- (aminomethyl) benzoate trifluoroacetate salt
:H nmr (d4 methanol) 6.25 (2H, d) ; 7.62 (2H, d) ; 7.48-
7.26 (9H, m) ; 5.33 (IH, s) ; 4.27 (2H, s); 3.67 (2H, s) . Hplc (Magellan C8, Gradient 5, Water/acetonitrile/TFA) rt 10.11 min. LC/MS (Magellan Clδ Gradient 6, water/acetonitrile/TFA) rt 1.53 min, 419 (M+l)+.
Example 92
2- (Carboxymethylamido) phenyl-4- (aminomethyl) benzoate trifluoroacetate salt lE nmr (d4 methanol) 8.30 (2H, d) ; 7.78 (IH, d) ; 7.71- 7.62 (3H, m) ; 7.46 (IH, t) ; 7.37 (IH, d) ; 4.28 (2H, s) ; 4.00 (2H, s) . Hplc (Magellan C8, Gradient 5, Water/acetonitrile/TFA) rt 7.93 min. LC/MS (Magellan C18 Gradient 6, water/acetonitrile/TFA) rt 1.25 min, 329 (M+l) + .
Example 93
2- [ (1-Carboxy-2 -hydroxy) ethylamido] phenyl-4- (aminomethyl) benzoate trifluoroacetate salt lE nmr (d4 methanol) δ.29 (2H, d) ; 7.83 (IH, d) ; 7.67- 7.61 (3H, m) ; 7.46 (IH, t) ; 7.36 (IH, d) ; 4.58 (IH, t) ;
4.27 (2H, s) ; 3.92 (IH, dd) ; 3,83 (IH, dd) . Hplc (Magellan C8, Gradient 5, Water/acetonitrile/TFA) rt 8.44 min. LC/MS (Magellan C18 Gradient 6, water/acetonitrile/TFA) rt 1.03 min, 359 (M+l)+. - 45 -
Example 94
2- [ (1-Carboxy-2 -phenyl) ethylamido] phenyl-4-
(aminomethyl) benzoate trifluoroacetate salt
:H nmr (d4 methanol) 8.19 (2H, d) ; 7.66-7.57 (4H, m) ; 7.44-7.30 (2H, m) ; 7.28-7.17 (5H, m) ; 4.78 (IH, dd) ;
4.28 (2H, s) ; 3.27 (IH, dd) ; 3.04 (IH, dd) . Hplc (Magellan C8, Gradient 5, Water/acetonitrile/TFA) rt 9.70 min. LC/MS (Magellan C18 Gradient 6, water/acetonitrile/TFA) rt 1.72 min, 419 (M+l)+.
Example 95
2- [N- (1(R) -Carboxy-5-aminopenyl) carboxamido] phenyl-4-
(aminomethyl) benzoate trifluoroacetate salt
:H nmr (d4 methanol) 8.29 (2H, d) ; 7.78 (IH, d) ; 7.71- 7.62 (3H, m) ; 7.47 (IH, t) ; 7.36 (IH, d) ; 4.51 (IH, dd) ;
4.29 (2H, s) ; 2.92-2.83 (2H, m) ; 2.00-1.85 (IH, m) ; 1.85-1.56 (3H, m) ; 1.56-1.39 (2H, m) . Hplc (Symmetry Rp8, Gradient 7, Water/acetonitrile/TFA) rt 5.65 min. LC/MS (Magellan C18 Gradient 6, water/acetonitrile/TFA) rt 1.05 min, 400 (M+l)+.
Example 96
2- [N- (MS) -Carboxy-5-aminopenyl) carboxamido] phenyl-4-
(aminomethyl) benzoate trifluoroacetate salt XH nmr (d4 methanol) 8.28 (2H, d) ; 7.80-7.73 (IH, m) ;
7.73-7.59 (3H, m) ; 7.46 (IH, t) ; 7.35 (IH, d) ; 4.51 (IH, dd) ; 4.29 (2H, s) ; 2.90-2.79 (2H, m) ; 2.00-1.86 (IH, m) ; 1.86-1.57 (3H, m) ; 1.53-1.39 (2H, m) . Hplc (Symmetry Rpδ, Gradient 7, Water/acetonitrile/TFA) rt 4.37 min. LC/MS (Magellan Clδ Gradient 6, water/acetonitrile/TFA) rt 1.06 min, 400 (M+l)+.
Example 97
[4-Chloro-2- (carboxymethyl) amido] phenyl-4- (aminomethyl) benzoate trifluoroacetate salt lE nmr (d4 methanol) 8.2δ (2H, d) ; 7.79 (IH, d) ; 7.69- 7.62 (3H, m) ; 7.39 (IH, d) ; 4.28 (2H, s) ; 3.99 (2H, s) . - 46 -
Hplc (Magellan C8 , Gradient 5, Water/acetonitrile/TFA) rt 9.45 min. LC/MS (Magellan C18 Gradient 6, water/acetonitrile/TFA) rt 1.28 min, 363 (M+l)+.
Example 98
[4-Chloro-2- (l-carboxy-2-hydroxy) ethylamido] phenyl-4- (aminomethyl) benzoate trifluoroacetate salt
:H nmr (d4 methanol) 8.29 (2H, d) ; 7.84 (IH, d) ; 7.68- 7.61 (3H, m) ; 7.39 (IH, d) ; 4.57 (IH, t); 4.27 (2H, s) ; 3.92 (IH, dd) ; 3.84 (IH, dd) . Hplc (Magellan Cδ ,
Gradient 5, Water/acetonitrile/TFA) rt 8.65 min. LC/MS (Magellan Clδ Gradient 6, water/acetonitrile/TFA) rt 1.30 min, 393 (M+l)+.
Example 99
[4-Chloro-2- [ (1-carboxy-2 -phenyl) ethylamido] ]phenyl-4- (aminomethyl) benzoate trifluoroacetate salt
XH nmr (d4 methanol) 8.14 (2H, d) ; 7.63-7.51 (4H, m) ; 7.32 (IH, d) ; 7.26-7.15 (5H, m) ; 4.73 (IH, dd) ; 4.25 (2H, s) ; 3.25 (IH, dd) ; 3.00 (IH, dd) . Hplc (Magellan C8, Gradient 5, Water/acetonitrile/TFA) rt 10.59 min. LC/MS (Magellan Clδ Gradient 6, water/acetonitrile/TFA) rt 2.75 min, 453 (M+l)+.
Example 100
[4-Chloro-2- [ (alpha-carboxy)benzylamido] ]phenyl-4- (aminomethyl) benzoate trifluoroacetate salt H nmr (d4 methanol) 8.18 (2H, d) ; 7.77 (IH, d) ; 7.66- 7.60 (3H, m) ; 7.39-7.29 (6H, d) ; 5.52 (IH, s) ; 4.29 (2H, s) . Hplc (Magellan Cδ, Gradient 5,
Water/acetonitrile/TFA) rt 10.45 min. LC/MS (Magellan Clδ Gradient 6, water/acetonitrile/TFA) rt 1.77 min, 439 (M+l)+.
Example 101
2- (Tetrazol-5-yl)phenyl-4- (aminomethyl) benzoate trifluoroacetate salt - 47 - lE nmr (d4 methanol) 8.27 (2H, d) ; 8.00 (IH, d) ; 7.75 (IH, t) ; 7.67 (2H, d) ; 7.62-7.50 (2H, m) ; 4.29 (2H, s) . Hplc (Magellan Cδ , Gradient 5, Water/acetonitrile/TFA) rt 10.26 min. LC/MS (Magellan Clδ Gradient 6, water/acetonitrile/TFA) rt 1.37 min, 296 (M+l)+.
Example 102
Phenyl 4- (aminomethyl) -2 -methylbenzoate hydrochloride salt λE nmr (d4 methanol) δ.25 (IH, d) ; 7.54-7.47 (4H, m) ;
7.38-7.32 (IH, m) ; 7.30-7.25 (2H, m) ; 4.24 (2H, s) ; 2.74 (3H, s) . M.S. TOF 242 (M+l)+. Hplc (Jupiter5 C18, Gradient 1, Water/acetonitrile/TFA) rt 9.69 min. Hplc (SymmetryCδ Gradient 2, Water/acetonitrile/TFA) rt 9.91 min.
Example 103
Phenyl 4- (aminomethyl) -3 -methylbenzoate hydrochloride salt XH nmr (d4 methanol) 8.17-7.75 (2H, m) ; 7.62 (IH, d) ;
7.51 (2H, t) ; 7.35 (IH, t) ; 7.27 (2H, d) ; 4.32 (2H, s) ; 2.57 (3H, s) . M.S. TOF 242 (M+l)+. Hplc (Jupiter5 Clδ, Gradient 1, Water/acetonitrile/TFA) rt 9.40 min. Hplc (SymmetryCδ Gradient 2, Water/acetonitrile/TFA) rt 9.82 min.
References
1. Still, W.C.; Kahn, M. ; Mitra, A. J. Org. Chem 1978, 43 , 2923 2. Smith, J. , Liras, J.L., Schneider, S.E., Anslyn, E.V. J" Org Chem 1996, 61 , 8811
Protocol for Tryptase Inhibition Assay
Tryptase assays were carried out at room temperature in 0.1 M phosphate buffer, 0.5 mg/ml heparin, pH 7.4 according to a method of Tapparelli et - 48 - al. (1993) J. Biol . Chem . 268., 4734-4741. Purified human lung tryptase was purchased from Dr Andrew Walls, Immunopharmacology Group, Southampton General Hospital, Southampton, UK. The chromogenic substrate for tryptase, S-2366, was purchased from Quadratech, Epsom, Surrey, UK. Product (p-nitroaniline) was quantified by absorption at 405nm in 96 well microplates using a Dynatech MR 5000 reader (Dynex Ltd, Billingshurst , UK) . K-, and Ki were calculated using SAS software. A K-, value of 216μM was determined for tryptase/S-2366. Inhibitor stock solutions were prepared at 40 mM in Me2S0 and tested within the range lOOmM-lnM. Accuracy of Kλ measurements was confirmed by comparison with K values of a known inhibitor of tryptase. In agreement with published data, benzamidine inhibited tryptase with a Kx value of 30μM.
The tryptase pKi ' s of a number of the compounds of the invention are illustrated in Table 1 below. Also quoted are the trypsin pKi ' s of the compounds.
Table 1
E . No . Ar pKi tryptase p^ trypsin
10 4 -Cyanophenyl 9.22 7.99
16 6 -quinolyl 8.33 7.46
24 6 ' -carboxymethyl- 8.17 7.42 2 -naphthyl
In all cases R and Y represent hydrogen in formula (I) .
These results clearly illustrate the efficacy of the compounds of the invention as well as their selectivity for tryptase over the serine protease trypsin.
Protocol for assessment of plasma stability,
The stability of compounds on incubation in human plasma 49 can be used to demonstrate the hydrolytic stability of the ester moiety towards the range of esterases and proteases normally present in plasma. As such, this assay is an informative measure of metabolic stability in the whole animal for compounds where ester hydrolysis is a major metabolic pathway.
In general, 10 uM samples of compound were incubated at 37 C in 3 ml human plasma. Aliquots were removed at intervals over a 4 hour incubation, extracted by solid phase techniques and analysed by HPLC.
The half-life of hydrolysis of some representative esters are illustrated in Table 2 below.
Table 2
Ex . No . Ar tl/2 of hydrolysis on plasma incubation
10 4 -cyanophenyl < 5 minutes
57 4- [4 -Carboxy piperidin- 107 min 1-oyl] phenyl
52 4- [ (2-Acetylamino-2- 22 hours carboxy) ethyl] phenyl
In all cases R and Y represent hydrogen in formula (I) .
These results indicate that the stability to hydrolysis by plasma enzymes is greatly enhanced for some compounds, which may therefore be preferred for therapeutic appliction, particularly for systemic adminstration.

Claims

- 50 - Claims
1. Use of a tryptase inhibitor of formula I
( i ;
wherein R represents hydrogen, alkyl, alkenyl , hydroxy, alkoxy, aminoalkyl, hydroxyalkyl, carboxyalkyl, alkoxyalkyl, amino, halo, cyano, nitro, thiol, alkylthio, haloalkoxy or haloalkyl; Ar represents an optionally substituted carbocyclic or heterocyclic aryl group with the proviso that when Ar represents a naphthyl moiety it is not substituted by amidino or guanidine; and
Y represents a hydrogen atom or alkyl group; or a physiologically tolerable salt thereof; in the manufacture of a medicament for use in a method of treatment of the human or non-human animal body to combat a condition responsive to said inhibitor.
2. Use as claimed in claim 1 wherein Y represents hydrogen.
3. Use as claimed in either one of claims 1 or 2 wherein R represents hydrogen.
4. Use as claimed in any one of claims 1 to 3 wherein Ar represents optionally subsituted phenyl, naphthyl, pyridyl, quinolyl or isoquinolyl .
5. Use as claimed in claim 4 wherein Ar represents a phenyl or naphthyl group substituted by one or more substituents selected from: halo, methylenedioxy, -R1, - 51 -
-NRxCOR2, C2_6-alkenyl, -(CH2)w-0R1, - (Cx_6) -perfluoroalkyl , -(CH2)WCN, -(CH2)wN02, -(CH2)WCF3, - (CH2) WS (0) rRx ,
- (CH2) WNR:R2 , - (CH2) COR1 , - (CH2) wCOjR1 , - (CH2) wCONRxR2 ,
- (CH2) wS02 R1R2 , - (CH2) NHSO^1 , - (CH2) ^HCOR1 , - (CH2) wNHC02Rx , -OC(=0)R1, - (CH2)w-CH(NHCOR1) -COOR2, - (CH2) W-CH (NRXR2) -
COOR1, - (CH2)w-CONH-S02-R1, - (CH2) u-SO^NHCO-R1 , -(CH2)W- tetrazole, - (CH2) W-P (0) 2-3-R1, - (CH2) W-C (=0) -R3 and optionally substituted aryl where R1 and R2 independently represent H, C1_8 alkyl , C3_7 cycloalkyl, or -(CH2)W-Ph, or R1 and R2 are optionally connected by a bond to form a 5- 8 atom cyclic structure or connected via an O, S or N atom to form a 5-8 atom heterocyclic structure and where R1 and/or R2 are also optionally substituted with -(CH2)W- C00R1, - (CH2)w-CH(NHC0R1) -COOR2, - (CH2) W-CH (NR:R2) -COOR1, - (CH2)w-CONH-S02-R1, - (CH2) ^SO^NHCO-R1 , - (CH2) w-tetrazole ,
- (CH2)w-S(0)r -R1, or - (CH2)w-P(0)2.3-R1; R3 is an oligomer comprising 1-4 aminoacid monomers, terminated by a free carboxylic acid, ester or amide functionality; and w=0-5 and r=0-2.
6. Use as claimed in claim 5 wherein Ar is substituted by an electron-withdrawing group and/or a protected or unprotected acidic group.
7. Use as claimed in claim 6 wherein said electron withdrawing group is selected from cyano, nitro, carboxamido, alkylsulfenyl , alkylsulfonyl , alkylaminosulfonyl, trifluoromethyl or halogen.
8. Use as claimed in claim 6 or 7 wherein said acidic group is selected from alkyl or aryl carboxylic acids and esters, acyl sulphonamide , sulphonylamidocarboxyalkyl , carboxyamidosulphonylalkyl , tetrazole, sulphonic acid or phosphonic acid each bonded to the Ar ring directly or via an alkyl, sulphonamidoalkyl or carboxamidoalkyl linkage, the linkage itself being optionally substituted by C^ - 52 - alkyl, CN, N02, NH2, NHCO-alkyl, or halogen.
9. Use as claimed in claim ╬┤ wherein if Ar is a phenyl derivative then said acidic substituent is present on the para position of the phenyl ring.
10. Use as claimed in claim 6 wherein if Ar is phenyl said acidic substituent is present on the ortho position and said electron withdrawing substituent is present on the para position of the phenyl ring.
11. A compound of formula (II)
R
OZ
K A o (II)
wherein R represents hydrogen, alkyl, alkenyl, hydroxy, alkoxy, aminoalkyl, hydroxyalkyl, carboxyalkyl, alkoxyalkyl, amino, halo, cyano, nitro, thiol, alkylthio, haloalkoxy or haloalkyl;
Y represents a hydrogen atom or alkyl group; ; and Z represents a phenyl group substituted by methylenedioxy, -NR^OR2, C2.6-alkenyl , -(CH2)w-0R1,
- (C!_6) -perfluoroalkyl , -(CH2)WCN, -(CH2)wN02, -(CH2)WCF3, - (CH2)wS(0)rR1, - (CH^NR'R2, - (CH2)wCOR\- (CH2) ^ONR'R2,
- (CH2) uSOsNR^2, - (CH2) wNHS02R1 , - (CH2) tfHCOR1 , - (CH2) wNHC02R1, -OC(=0)R1, - (CH2)w-CH(NHCOR1) -COOR2, - (CH2) W-CH (NR^2) -
COOR1, - (CH2)w-CONH-S02-R1, - (CH2) u-SO^NHCO-R1 , -(CH2)W- tetrazole, - (CH2) W-P (O) ^-R1, - (CH2) W-C (=0) -R3 and optionally substituted aryl where R1 and R2 independently represent H, C^g alkyl , C3.7 cycloalkyl, or -(CH2)W-Ph, or R1 and R2 are optionally connected by a bond to form a 5- ╬┤ atom cyclic structure (eg. piperidine) or connected via an O, S or N atom to form a 5-╬┤ atom heterocyclic - 53 - structure (eg. morpholine, piperazine) and where R1 and/or R2 are also optionally substituted with -(CH2)W- COOR1, - (CH2)w-CH(NHCOR1) -COOR2, - (CH2) W-CH (NR:R2) -COOR1 , - (CH2)w-CONH-S02-R1, - (CH^-SO^NHCO-R1, - (CH2) w-tetrazole , - (CH2)W-S (0)r -R1, or - (CH2) W-P (0) 2_3-Rx ; where R3 is an oligo er comprising 1-4 aminoacid monomers, such as the natural aminoacids glycine, proline and serine, terminated by a free carboxylic acid, ester or amide functionality; and w=0-5 and r=0-2; or Z represents a phenyl group subsituted in the 2- position by nitro or in the 3 -position by methoxy; or
Z represents a naphthyl group substituted by halo, methylenedioxy, -R1, -NR^OR2, C2.6-alkenyl , -(CH2)w-OR1,
- (Ci.g) -perfluoroalkyl, -(CH2)WCN, -(CH2)wN02, -(CH2)WCF3, -(CH2)wS(0)rR1, -(CH2)WNR1R2, -(CH2)wCOR\ - (CH2) ^R1,
- (CH2) ^ONR'R2 , - (CH2) wS02NR1R2 , - (CH2) wNHS02R1 , - (CH2) NHCOR1 ,
- (CH^NHCO^1, -0C(=0)R1, - (CH2) W-CH (NHCOR1) -COOR2, -(CH2)W- CH(NR1R2) -COOR1, - (CH2) u-CONH-SO^R1 , - (CH2) u-SO^NHCO-R1 , - (CH2)w-tetrazole, - (CH2) W-P (O) ^-R1 , - (CH2) W-C (=0) -R3 and optionally substituted aryl where R1 and R2 independently represent H, C^g alkyl, C3.7 cycloalkyl, or -(CH2)W-Ph, or R1 and R2 are optionally connected by a bond to form a 5- 8 atom cyclic structure or connected via an O, S or N atom to form a 5-8 atom heterocyclic structure and where R1 and/or R2 are also optionally substituted with -(CH2)W- COOR1, - (CH2)W-CH (NHCOR1) -COOR2, - (CH2) W-CH (NR:R2) -COOR1 , - (CH2)w-CONH-S02-R1, - (CH2)w-S02-NHCO-R1, - (CH2) w-tetrazole,
- (CH2)w-S(0)r -R1, or - (CH2)w-P(0)2.3-R1; where R3 is an oligomer comprising 1-4 aminoacid monomers, terminated by a free carboxylic acid, ester or amide functionality; and w=0-5 and r=0-2; or a heterocyclic aryl groups containing 5 to 10 ring atoms including 1, 2 or 3 ring heteroatoms selected from oxygen, nitrogen and sulphur; or a physiologically tolerable salt thereof.
12. A compound of formula (II) as claimed in claim 11 - 54 - for use in combatting a condition responsive to said inhibitor.
13. A pharmaceutical composition comprising a compound of formula (II) as claimed in claim 11, together with at least one pharmaceutically acceptable excipient.
14. A pharmaceutical composition comprising a compound of formula (I) as claimed in claims 1 to 10, together with one or more anti-asthma agents and at least one pharmaceutically acceptable excipient.
15. A method of treatment of human or non-human animal body to combat a condition responsive to a tryptase inhibitor, said method comprising administering to said body an effective amount of a tryptase inhibitor as claimed in any one of claims 1 to 10.
EP99918168A 1998-04-24 1999-04-23 Aminomethyl-benzoic ester derivatives as tryptase inhibitors Withdrawn EP1073624A1 (en)

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