WO2014012964A1 - New antifibrinolytic compounds - Google Patents

New antifibrinolytic compounds Download PDF

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Publication number
WO2014012964A1
WO2014012964A1 PCT/EP2013/065071 EP2013065071W WO2014012964A1 WO 2014012964 A1 WO2014012964 A1 WO 2014012964A1 EP 2013065071 W EP2013065071 W EP 2013065071W WO 2014012964 A1 WO2014012964 A1 WO 2014012964A1
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compound
formula
optionally substituted
saturated
substituents
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PCT/EP2013/065071
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French (fr)
Inventor
Josune Orbe Lopategui
Julen Oyarzabal Santamarina
José Antonio PÁRAMO FERNÁNDEZ
José Antonio RODRÍGUEZ GARCÍA
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Proyecto De Biomedicina Cima, S.L.
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Priority to US14/415,474 priority Critical patent/US9440989B2/en
Priority to JP2015522082A priority patent/JP6203257B2/en
Priority to EP13737275.1A priority patent/EP2877446B9/en
Priority to AU2013292076A priority patent/AU2013292076B2/en
Priority to ES13737275.1T priority patent/ES2603740T3/en
Priority to CA2917604A priority patent/CA2917604C/en
Publication of WO2014012964A1 publication Critical patent/WO2014012964A1/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
    • C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
    • C07D491/10—Spiro-condensed systems
    • C07D491/107—Spiro-condensed systems with only one oxygen atom as ring hetero atom in the oxygen-containing ring
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
    • A61P7/02—Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
    • A61P7/04—Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C317/00—Sulfones; Sulfoxides
    • C07C317/14—Sulfones; Sulfoxides having sulfone or sulfoxide groups bound to carbon atoms of six-membered aromatic rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C317/00—Sulfones; Sulfoxides
    • C07C317/44—Sulfones; Sulfoxides having sulfone or sulfoxide groups and carboxyl groups bound to the same carbon skeleton
    • C07C317/48—Sulfones; Sulfoxides having sulfone or sulfoxide groups and carboxyl groups bound to the same carbon skeleton the carbon skeleton being further substituted by singly-bound nitrogen atoms, not being part of nitro or nitroso groups
    • C—CHEMISTRY; METALLURGY
    • 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
    • C07D211/08—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 hydrocarbon or substituted hydrocarbon radicals directly attached to ring carbon atoms
    • C07D211/18—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 hydrocarbon or substituted hydrocarbon radicals directly attached to ring carbon atoms with substituted hydrocarbon radicals attached to ring carbon atoms
    • C07D211/34—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 hydrocarbon or substituted hydrocarbon radicals directly attached to ring carbon atoms with substituted hydrocarbon radicals attached to ring carbon atoms with hydrocarbon radicals, substituted by carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D221/00—Heterocyclic compounds containing six-membered rings having one nitrogen atom as the only ring hetero atom, not provided for by groups C07D211/00 - C07D219/00
    • C07D221/02—Heterocyclic compounds containing six-membered rings having one nitrogen atom as the only ring hetero atom, not provided for by groups C07D211/00 - C07D219/00 condensed with carbocyclic rings or ring systems
    • C07D221/20—Spiro-condensed ring systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D265/00—Heterocyclic compounds containing six-membered rings having one nitrogen atom and one oxygen atom as the only ring hetero atoms
    • C07D265/28—1,4-Oxazines; Hydrogenated 1,4-oxazines
    • C07D265/34—1,4-Oxazines; Hydrogenated 1,4-oxazines condensed with carbocyclic rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/04—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/06—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a carbon chain containing only aliphatic carbon atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D409/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms
    • C07D409/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings
    • C07D409/06—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings linked by a carbon chain containing only aliphatic carbon atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/10—Spiro-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/12—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains three hetero rings
    • C07D471/20—Spiro-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C2601/00—Systems containing only non-condensed rings
    • C07C2601/12—Systems containing only non-condensed rings with a six-membered ring
    • C07C2601/14—The ring being saturated
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C2603/00—Systems containing at least three condensed rings
    • C07C2603/93—Spiro compounds
    • C07C2603/94—Spiro compounds containing "free" spiro atoms

Definitions

  • the present invention relates to spirocyclic compounds of formula (I), to a process for their preparation, as well as to the intermediates used in this process. It also relates to pharmaceutical or veterinary compositions containing them, and to their use in medicine, in particular as antifibrinolytic and antihemorragic agents.
  • the haemostatic system is responsible for maintaining circulatory fluidity and for preventing haemorrhage in response to vascular injury.
  • Physiological hemostasis is controlled by mechanisms of coagulation and the formation of fibrin and by those favouring the degradation of fibrin (fibrinolysis).
  • Hyperfibrinolytic states caused by congenital or acquired conditions predispose to important haemorrhagic complications, often requiring transfusions and the need for re-exploration having a detrimental effect on patient outcome. Hemorhage is responsible for almost 50% of deaths occurring within 24 hours of traumatic injury and for up to 80% of
  • a major goal in surgery as well as in the treatment of major tissue damage is to avoid or minimise bleeding in order to ensure the formation of stable and solid haemostatic plugs that are not easily dissolved by fibrinolytic enzymes. Furthermore, it is of importance to ensure quick and effective formation of such plugs or clots.
  • Antifibrinolytic agents are widely used in major surgery to prevent fibrinolysis and reduce blood loss.
  • EACA epsilon- aminocaproic acid
  • TXA tranexamic acid
  • antifibrinolytics commercially available to control bleeding. These agents competitively inhibit activation of plasminogen to plasmin, an enzyme that degrades fibrin clots, fibrinogen and other plasma proteins.
  • spirocyclic compounds which comprise spirocyclic ring system containing a carbon atom (spiro atom) attached to a hydroxamic acid and a sulfonyl group, show a significant delay in the lysis time in a
  • the spirocyclic compounds of the invention also show an important reduction of the bleeding time in vivo animal models as it will be shown in detail in the examples. These characteristics of the compounds of the invention allow a rapid cessation of hemorrhage; favor an effective formation of plugs or clots; have a sustained action (persistence of the clot and prevention of hemorrhage) and aid in minimizing the adverse effects related to other antifibrinolytic/antihemorrhagic treatments having risk of thrombotic complications.
  • a first aspect of the invention relates to a compound of formula (I) (hereinafter also referred as compounds of the invention), or a
  • a and B form a spirocyclic ring system wherein the spiro atom connecting A and B is a carbon atom and wherein
  • A is a known 3- to 8-membered carbocyclic or heterocyclic monocyclic ring, saturated or partially unsaturated; or alternatively
  • A is a known 6- to 18-membered carbocyclic or heterocyclic polycyclic ring system, saturated, partially unsaturated, or partially aromatic; and B is a known 4- to 7-membered carbocyclic or heterocyclic monocyclic ring, saturated or partially unsaturated;
  • C is phenyl or a known 5- to 6-membered heteroaromatic ring
  • R1-R3 are independently selected from H, halogen, -NO 2 , -CN, R a , -OR a , -OC(Y)R a' , -OC(Y)OR a' , -OC(Y)NR R a , -OSO 2 OR a' , -NR R a , -NR C(Y)R a , -NR C(Y)OR a , -NR C(Y)NR R a , -NR S(O) 2 R a' , -NR SO 2 NR R a , -SR a' , -S(O)R a' , -S(O)OR a' , -SO 2 R a' ,-SO 2 (OR a ), -SO 2 NR R a , -SC(Y)NR R a , -C(Y)R a' ,
  • R 4 -R 7 are independently selected from halogen, -NO 2 , -CN, R c , -OR c , -NR d R c , -NR d C(Y)R c , -NR d C(Y)OR c , -NR d C(Y)NR d R c , -NR d S(O) 2 R c , -NR d SO 2 NR d R c , -SR C , -S(O)R c , -S(O)OR c , -SO 2 R c , -SO 2 R(OR c ), -SO 2 NR d R c , -SC(Y)NR d R c , -C(Y)R C , -C(Y)OR c , -C(Y)NR d R c , -C(Y)NR d OR c , and -C(O)NR
  • R a is a saturated or unsaturated (Ci-Ci 2 )alkyl optionally substituted with one or more substituents R e and/or one Cy 1 ; or alternatively R a is Cy 2 ;
  • Cy 1 and Cy 2 are independently optionally substituted with: one Cy 3 and/or one or more substituents R e , and/or one or more saturated or unsaturated (CrC 6 )alkyl groups optionally substituted with one or more substituents R e and/or one Cy 3 ; and
  • any Cy 3 is optionally substituted with one or more substituents independently selected from R e and saturated or unsaturated (CrC 6 )alkyl optionally substituted with one or more substituents R e ; each R a and R are independently H or R a ;
  • R c and each R d are independently selected from H, Cy 4 , and saturated or unsaturated (CrC 6 )alkyl optionally substituted with one or more substituents R h and/or one Cy 5 ;
  • R h independently selected from R h and saturated or unsaturated (CrC 6 )alkyl optionally substituted with one or more substituents R h ;
  • each R e is independently selected from halogen, -NO 2 , -CN, -OR f , -OC(Y)R f , -OC(Y)OR f , -OC(Y)NR 9 R f , -NR 9 R f , -NR 9 C(Y)R f , -NR 9 C(Y)OR f , -NR 9 C(Y)NR 9 R f , -NR 9 S(O) 2 R f , -NR 9 SO 2 NR 9 R f , -SR f , -S(O)R f , -S(O)OR f , -SO 2 R f , -SO 2 (OR f ), -SO 2 NR 9 R f , -SC(Y)NR 9 R f , -
  • Cy 6 is optionally substituted with: one Cy 7 , and/or one or more substituents R h , and/or one or more saturated or unsaturated (CrC 6 )alkyl groups optionally substituted with one or more substituents R h and/or one Cy 7 ; and
  • any Cy 7 is optionally substituted with one or more substituents independently selected from R h and (Ci-C 4 )alkyl optionally substituted with one or more substituents R h ; each R h is independently selected from halogen, -NO 2 , -CN, -OR', - ⁇ ( ⁇ ) ⁇ , -00(0)0 ⁇ , -OCiOJNR'R 1 , -NR'R', -NR'CiOJR 1 , -NR ⁇ OJOR 1 , -NR'C(O)NR'R', -NR ⁇ O ⁇ R 1 , -NR'SCfeNR'R 1 , -SR 1 , -S ⁇ 0)R -SOzR 1 , -SOziOR 1 ), -SCfeNR'R 1 , -C ⁇ 0)R -0(0)0 ⁇ , -CiOJNR'R 1 , and -C(O)NR'OR' each R 1 is independently H or
  • Cy 1 , Cy 2 , Cy 4 and Cy 6 are independently a C or N-attached known ring system selected from 3- to 8-membered carbocyclic or heterocyclic monocyclic ring, saturated or partially unsaturated; phenyl; 5- or 6-membered heteroaromatic ring; and 6- to 18-membered carbocyclic or heterocyclic polycyclic ring system, saturated, partially unsaturated, aromatic or partially aromatic;
  • Cy 3 , Cy 5 and Cy 7 are independently a C or N-attached known ring system selected from 3- to 8-membered carbocyclic or heterocyclic monocyclic ring, saturated or partially unsaturated; phenyl; and 5- or 6-membered
  • heteroaromatic ring wherein in the carbocyclic rings all ring members are carbon atoms; and in the heterocyclic and heteroaromatic rings one or more ring members are selected from N, O, and S; and wherein in all saturated or partially
  • Another aspect of the invention relates to a pharmaceutical or veterinary composition which comprises an effective amount of a compound of formula (I) as defined above, or a pharmaceutically or veterinary acceptable salt thereof, or any stereoisomer either of the compound of formula (I) or of its pharmaceutically or veterinary acceptable salt, together with one or more pharmaceutically or veterinary acceptable excipients or carriers.
  • the compounds of the invention are useful as antifibrinolytic and antihemorrhagic agents. Therefore, another aspect of the invention relates to a compound of formula (I) as defined above, or a pharmaceutically or veterinary acceptable salt thereof, or any stereoisomer either of the compound of formula (I) or of its pharmaceutically or veterinary acceptable salt, for use as a medicament.
  • Another aspect of the invention relates to a compound of formula (I) as defined above, or a pharmaceutically or veterinary acceptable salt thereof, or any stereoisomer either of the compound of formula (I) or of its
  • this aspect relates to the use of a compound of formula (I) as defined above, for the manufacture of a medicament for use as antifibrinolytic and antihemorrhagic agent; and may also be formulated as a method for the treatment and/or prevention of hyperfibrinolysis and/or hemorrhages comprising administering an effective amount of the previously defined compound of formula (I), or a pharmaceutically or veterinary acceptable salt thereof, or any stereoisomer either of the compound of formula (I) or of its pharmaceutically or veterinary acceptable salt, and one or more pharmaceutically or veterinary acceptable excipients or carriers, in a subject in need thereof, including a human.
  • Another aspect of the invention relates to a compound of formula (II)
  • R is an hydroxamic acid protective group, more particularly an hydroxamic acid protective group selected from the group consisting of tetrahydro-2H-pyran-2-yloxy (THP), benzyl, 1 -naphthylmethyl and dimethyloxybenzyl (DMB).
  • THP tetrahydro-2H-pyran-2-yloxy
  • DMB dimethyloxybenzyl
  • Carbocyclic ring system refers to a known ring system wherein all the ring members are carbon atoms.
  • heterocyclic ring system refers to a known ring system wherein one or more of the ring members, preferably 1 , 2, 3, or 4 ring members, are selected from N, O, and S, where chemically possible. Unless otherwise specified, the "heterocyclic” ring system may be attached to the rest of the molecule through a C or a N atom. Both the carbocyclic and heterocyclic rings can be saturated or partially unsaturated, and may be unsubstituted or substituted as described herein, being the susbstituents placed on any available position.
  • polycyclic ring refers to a ring system which is formed by two, three or four rings which can be fused, bridged-fused, spiro-fused or can contain different types of fusion.
  • fused rings the fusion occurs through one bond which is common to two adjoining rings; in “bridged-fused” rings the fusion occurs through a sequence of atoms (bridgehead) which is common to two rings; and in “spiro-fused” rings, the fusion occurs through only one atom
  • spiro atom preferably a carbon atom, which is common to two adjoining rings (including bridged rings)
  • the polycyclic ring system can be saturated, partially unsaturated, aromatic (except in the case of ring system A) or partially aromatic; and may be unsubstituted or substituted as described herein, being the susbstituents placed on any available position.
  • the term "heteroaromatic" ring refers to a known aromatic ring system, wherein one or more of the ring members, preferably 1 , 2, 3, or 4 ring members, are selected from N, O, and S where chemically possible.
  • the heteroaromatic ring and phenyl may be unsubstituted or substituted as described herein, being the susbstituents placed on any available position.
  • one or two members of the rings are optionally C(O) and/or C(NH) and/or C[N(C C 4 )alkyl].
  • saturated or unsaturated (CrC n )alkyl refers to a saturated branched or linear hydrocarbon chain which contains from 1 to n carbon atoms. When the (C-i-Cn)alkyl is saturated it contains only single bonds. When the
  • (d-Cn)alkyl is unsaturated it contains one or two double bonds and/or one or two triple bonds.
  • the saturated or unsaturated (CrC n )alkyl may be
  • a halogen substituent means fluoro, chloro, bromo or iodo.
  • Protective group refers to a grouping of atoms that when attached to a reactive group in a molecule masks, reduces or prevents that reactivity.
  • substituted with one or more means that a group can be substituted with one or more, preferably with 1 , 2, 3 or 4 substituents, provided that this group has enough positions susceptible of being
  • a first aspect of the invention relates to compounds of formula (I) or pharmaceutically or veterinary acceptable salts thereof, or any stereoisomer either of the compound of formula (I) or of its pharmaceutically or veterinary acceptable salt.
  • compositions of formula (I) can be carried out by methods known in the art. For instance, they can be prepared from the parent compound, which contains a basic or acidic moiety, by conventional chemical methods. Generally, such salts are, for example, prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate
  • the compounds of the invention may be in crystalline form either as free solvation compounds or as solvates (e.g. hydrates) and it is intended that both forms are within the scope of the present invention.
  • Methods of solvation are generally known within the art.
  • the solvated forms with pharmaceutically or veterinary acceptable solvents such as water, ethanol and the like are equivalent to the unsolvated form for the purposes of the invention.
  • Some compounds of formula (I) can have chiral centres that can give rise to various stereoisomers.
  • the present invention relates to each of these stereoisomers and also mixtures thereof.
  • some of the compounds of the present invention can show cis/trans isomers.
  • the present invention relates to each of the geometric isomers and mixtures thereof.
  • Diastereoisomers can be separated by conventional techniques such as chromatography or fractional crystallization.
  • Optical isomers can be resolved by conventional techniques of optical resolution to give optically pure isomers. This resolution can be carried out on any chiral synthetic
  • the invention refers to a compound of formula (I) as defined above, wherein A is a known 3- to 8-membered carbocyclic or heterocyclic monocyclic ring or a known 6- to 10-membered carbocyclic or heterocyclic bicyclic ring system.
  • the invention refers to a compound of formula (I), wherein A is a monocyclic ring selected from a 3- to 6-membered carbocyclic ring, and a 5- to 6-membered heterocyclic ring.
  • the invention refers to a compound of formula (I), wherein A is a carbocyclic monocyclic ring; or a polycyclic ring system, preferably a bicyclic ring system, wherein the ring containing the spiro atom attached to the hydroxamic acid and the sulfonyl group is a carbocyclic ring.
  • the invention refers to a compound of formula (I), wherein A is selected from cyclopropane, cyclobutane, cyclopentane, cyclohexane, tetrahydrofuran, pyrrolidine, bicyclo[2.2.1 ]heptane, 2,3-dihydro- 1 H-indene, hexahydropyrrolizin-3-one, and 4-azaspiro[4.4]nonane.
  • A is selected from cyclopropane, cyclobutane, cyclopentane, cyclohexane, tetrahydrofuran, pyrrolidine, bicyclo[2.2.1 ]heptane, 2,3-dihydro- 1 H-indene, hexahydropyrrolizin-3-one, and 4-azaspiro[4.4]nonane.
  • the invention refers to a compound of formula (I), wherein A is unsubstituted, i.e. R 7 is H.
  • the invention refers to a compound of formula (I), wherein B is a 6- to 7-membered carbocyclic or heterocyclic monocyclic ring.
  • the invention refers to a compound of formula (I), wherein B is a saturated monocyclic ring, carbocyclic or heterocyclic, wherein at least one of the ring members of the heterocyclic ring is NR 4 .
  • the invention refers to a compound of formula (I), wherein B is selected from cyclohexane, piperidine, morpholine, azepane, piperazine, pyrrolidine, and azetidine.
  • B is piperidine, morpholine, azepane, pyrrolidine, and azetidine, wherein R 4 is placed on the N atom of these rings and R 5 -R 6 are H.
  • B is piperazine, wherein R 4 and R 5 are placed on the N atoms and R 6 is H.
  • the invention refers to a compound of formula (I), wherein A and B form a spirocyclic ring system selected from the group consisting of:
  • the invention refers to a compound of formula (I), wherein C is phenyl.
  • C is phenyl substituted with R 1 at the orto, meta or para position, and R 2 and R 3 are independently selected from H, halogen, R a , -OR a' ,and -NR R a ; wherein R a , R a' and R are
  • C is independently selected from H and -(Ci-C 4 )alkyl optionally substituted with one or more halogen atoms.
  • C is phenyl substituted with R-i at the para position and R 2 and R 3 are H.
  • C is phenyl substituted with R 1 at the orto position and R 2 and R 3 are H.
  • C is phenyl substituted with R 1 at the meta position and R 2 and R 3 are H.
  • C is phenyl substituted with R 1 at the meta position, and R 2 at the para position, R 3 being H; or alternatively, C is phenyl substituted with R 1 at the para position, and R 2 at the meta position, R 3 being H; wherein R 2 is selected from H, halogen, R a , -OR a' ,and -NR R a ; and R a , R a and R are independently selected from H and -(Ci-C 4 )alkyl optionally substituted with one or more halogen atoms.
  • the invention refers to a compound of formula (I), wherein in R (relating to Ri-R 3 ), Cy 1 and Cy 2 are independently optionally substituted with one or more substituents selected from R e and saturated or unsaturated (CrC 6 )alkyl optionally substituted with one or more substituents
  • R h independently selected from R h and saturated or unsaturated (CrC 6 )alkyl optionally substituted as previously defined, more particularly with one or more substituents R h .
  • the invention refers to a compound of formula (I), wherein in R (relating to Ri-R 3 ), R is H and saturated or unsaturated (Ci-Ci 2 )alkyl optionally substituted with one or more substituents R e , more particularly wherein in R e , R f and each R 9 are independently selected from H and saturated or unsaturated (CrC 6 )alkyl optionally substituted with one or more fluorine atoms.
  • the invention refers to a compound of formula (I), wherein in R R 3 , R e is selected from halogen, -NO 2 , -CN, -OR f , -OC(O)R f , -OC(O)OR f , -OC(O)NR 9 R f , -NR 9 R f , -NR 9 C(O)R f , -NR 9 C(O)OR f ,
  • the invention refers to a compound of formula (I), wherein in Ri-R 3 , R f and each R 9 are independently selected from H and saturated or unsaturated (CrC 6 )alkyl optionally substituted with one or more fluorine atoms.
  • the invention refers to a compound of formula (I), wherein in Ri-R 3 , Cy 1 and Cy 2 are independently optionally substituted with one or more substituents selected from R e and saturated or unsaturated (CrC 6 )alkyl optionally substituted as previously defined; and Cy 6 is optionally substituted with one or more substituents independently selected from R h and saturated or unsaturated (CrC 6 )alkyl optionally substituted as previously defined.
  • the invention refers to a compound of formula (I), wherein R R 3 are independently selected from H, halogen, -NO 2 , -CN, R a , -OR 3' , -OC(O)R a' , -OC(O)OR a' , -OC(O)NR R a' , -NR R a , -NR C(O)R a ,
  • the invention refers to a compound of formula (I), wherein in R 4 -R 7 , R h is selected from halogen, -NO 2 , -CN, -OR', - ⁇ ( ⁇ ) ⁇ , -00(0)0 ⁇ , -OCiOJNR'R 1 , -N R 1 , -NR'CiO) ⁇ , -NRtyOJOR 1 , -NR'C(O)NR'R', -NR ⁇ O), ⁇ , -S , -S(O)R, -SO 2 R, -SO 2 NRR, -C(O)R, -C(O)OR, and
  • the invention refers to a compound of formula (I), wherein R 4 -R 7 are independently selected from halogen, -NO 2 , -CN, R c , -OR c , -NR d R c , -NR d C(O)R c , -NR d C(O)OR c , -NR d C(O)NR d R c , -NR d S(O) 2 R c , -SR C , -S(O)R c , -SO 2 R c , -SO 2 NR d R c , -C(O)R c , -C(O)OR c , and -C(O)NR d R c .
  • the invention refers to a compound of formula (I), wherein R 2 and R 3 are independently selected from H, halogen, R a , -OR a ,and -NR R a ; and R 5 -R 7 are independently selected from H, halogen, R c , -OR c , and -NR d R c , wherein R a , R a' , R , R c and R d are independently selected from H and -(Ci-C 4 )alkyl optionally substituted with one or more fluorine atoms.
  • the invention refers to a compound of formula (I), wherein is selected from H, halogen, -NO 2 , -CN, R a , -OR 3' , -OR 3' ,
  • R 4 is selected from halogen, -NO 2 , -CN, R c , -OR c , -NR d R c , -NR d C(O)R c , -NR d C(O)OR c , -NR d C(O)OR 3' ;
  • the present invention also relates to the combination of any of the specific embodiments defined above for any of the variables A, B, C, and R 1 -R 7 .
  • the compound of formula (I) is selected from the group consisting of:
  • R is an hydroxamic acid protective group, more particularly an hydroxamic acid protective group selected from the group consisting of tetrahydro-2H-pyran-2-yloxy (THP), benzyl,
  • the first conversion can be carried out in the presence of an activating agent such as 1 -ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride
  • EDC.HCI Hydroxybenzotriazole
  • HOBt Hydroxybenzotriazole
  • a base such as N-methylmorpholine (NMM)
  • NMM N-methylmorpholine
  • suitable solvent such as dichloromethane, chloroform or dimethylformamide
  • hydroxy protective groups include those where the hydroxy group is either acylated or alkylated such as benzyl, and trityl ethers as well as alkyl ethers, tetrahydropyranyl ethers, trialkylsilyl ethers and allyl ethers.
  • the hydroxamic acid protective group is tetrahydro-2H-pyran-2-yloxy (THP), benzyl,
  • DMB dimethyloxybenzyl
  • (CrC 6 )alkyl benzyl, p-methoxyphenyl, trimethylsilyl, or [2-(Trimethylsilyl)- ethoxy]methyl (SEM).
  • the carboxy protective group is (CrC 6 )alkyl
  • the deprotection is carried out in basic medium, for example with LiOH in a suitable solvent such as tetrahydrofuran-methanol.
  • a compound of formula (lla) can be reacted with a compound of formula H-Q-alk (VIII) in the presence of a base, such as NaH, optionally in a suitable solvent or without solvent, and preferably heating. After removal of the protective group in the resulting compound of formula (lie), compound of fomula (lb) is obtained.
  • a base such as NaH
  • a compound of formula (lla) can be reacted with a compound of formula H-NCy 2 R a (X), optionally in a suitable solvent or without solvent, and preferably heating. After removal of the protective group in the resulting compound of formula (lie), compound of formula (Id) is obtained.
  • R 1 is -NR C(Y)R a , -NR C(Y)OR a ,
  • -NR C(Y)NR R a , -NR S(O) 2 R a' , -NR SO 2 NR R a' may be obtained from a compound of formula (llg), which can be prepared as shown in the following scheme:
  • compound of formula (Ila) is reacted with benzylamine, preferably heating to give a compound of formula (llf) which can be
  • a compound of formula (llg) hydrogenated in the presence of Pd/C to give a compound of formula (llg).
  • This compound can be reacted for example with an acyl halide in the presence of a base, such as triethylamine, in a suitable solvent, such as tetrahydrofuran, at a temperature comprised from room temperature to the reflux temperature of the solvent, to give an amide; or with an isocyanate to give a urea.
  • a base such as triethylamine
  • a suitable solvent such as tetrahydrofuran
  • compound of formula (Ila) is reacted with benzyl alcohol in the presence of a base, such as NaH, in a suitable solvent, such as
  • This compound can be reacted for example with an acyl halide in the presence of a base, such as triethylamine, in a suitable solvent, preferably at room temperature to give an ester.
  • a base such as triethylamine
  • the reactions described above can be carried out in a different order.
  • the reactions carried out on compounds of formula (II) i.e. on the protected hydroxamic acid
  • compounds of formula (III) i.e. the carboxylic ester
  • the present invention also relates to a pharmaceutical or veterinary composition
  • a pharmaceutical or veterinary composition comprising an effective amount of a compound of formula (I) as defined above, or a pharmaceutically or veterinary acceptable salt thereof, or any stereoisomer either of the compound of formula (I) or of its
  • an effective amount refers to the amount of a compound that, when administered, is sufficient to prevent development of, or alleviate to some extent, one or more of the symptoms of the disease which is addressed.
  • the specific dose of the compound of the invention to obtain a therapeutic benefit may vary depending on the particular circumstances of the individual patient including, among others, the size, weight, age and sex of the patient, the nature and stage of the disease, the aggressiveness of the disease, and the route of administration. For example, a dose of from about 0.01 to about 300 mg/kg may be used.
  • pharmaceutically or veterinary acceptable excipients or carriers refers to pharmaceutically or veterinary acceptable materials, compositions or vehicles. Each component must be pharmaceutically or veterinary acceptable in the sense of being compatible with the other ingredients of the pharmaceutical or veterinary composition. It must also be suitable for use in contact with the tissue or organ of humans and animals without excessive toxicity, irritation, allergic response, immunogenicity or other problems or complications commensurate with a reasonable benefit/risk ratio.
  • the election of the pharmaceutical or veterinary formulation will depend upon the nature of the active compound and its route of administration. Any route of administration may be used, for example oral, parenteral and topical administration.
  • the pharmaceutical or veterinary composition may be formulated for oral administration and may contain one or more physiologically
  • compatible carriers or excipients in solid or liquid form.
  • These preparations may contain conventional ingredients such as binding agents, fillers, lubricants, and acceptable wetting agents.
  • the pharmaceutical or veterinary composition may be formulated for parenteral administration in combination with conventional injectable liquid carriers, such as water or suitable alcohols.
  • conventional pharmaceutical or veterinary excipients for injection such as stabilizing agents, solubilizing agents, and buffers, may be included in such compositions.
  • compositions may be injected intramuscularly, intraperitoneal ⁇ , or intravenously.
  • the pharmaceutical or veterinary composition may be formulated for topical administration.
  • Formulations include creams, lotions, gels, powders, solutions and patches wherein the compound is dispersed or dissolved in suitable excipients.
  • the topical compositions of the invention may be administered by means of a carrier material, which can be a solid support.
  • a topical composition comprising a carrier material, which can be a solid support.
  • solid supports include intelligent textiles, dressings, coatings, sponges, band-aids, sanitary pads, compresses, plasters, etc.
  • the manufacture of such compositions can be obtained by conventional methods, for example, by mixing the
  • compositions may be in any form, including, among others, tablets, pellets, capsules, aqueous or oily solutions,
  • suspensions, emulsions, or dry powdered forms suitable for reconstitution with water or other suitable liquid medium before use, for immediate or retarded release can readily be determined by those skilled in the art according to the type of formulation being prepared.
  • the compounds of the present invention are useful as antihemorrhagic and antifibrinolytic agents and can be used in a broad range of therapeutic applications.
  • antifibrinolytic agents in addition to reducing postoperative hemorrhage, can be an alternative to blood transfusion and other hemoderivatives for example in heart, liver and orthopedic surgery, and also in the setting of oncologic surgery in organs rich in fibrinolysis activators (prostate, uterus).
  • antifibrinolytic agents can reduce all- cause mortality and death due to bleeding.
  • the antifibrinolytic agents of the invention can also be used to control bleeding in trombolytic therapy, e.g.
  • the antifibrinolytic agents of the invention are useful in the treatment of local hemorrhages, e.g. after teeth extraction, in particular in patients with congenital coagulopathies, such as hemophilia, or patients with diabetes; in the treatment of menorrhage in women associated with congenital or acquired coagulopathies, as well as in post-partum haemorrhage, and in the treatment of hemorrhages of
  • the HPLC measurement was performed using Gilson 281 from 233 pump (binary), an autosampler, and a UV detector. The fractions was detected by
  • the MS detector was configured with an electrospray ionization source.
  • the source temperature was maintained at 300-350 oC.
  • Reverse phase HPLC was carried out on Luna C18 (100x30 mm; 4um).
  • Solvent A water with 0.075% trifluoroacetic acid
  • Solvent B acetonitrile with 0.075% trifluoroacetic acid.
  • Gradient At room temperature, 20% of B to 40% of B within 6 min at 25 mL/min; then 40% B at 25 mL/min over 2 min, UV detector.
  • PE Petroleum ether
  • Q is O or S
  • Q' is Q or SO 2
  • Cy is phenyl or a 5- to 6- membered heteroaryl and can be optionally subtituted.
  • R is a hydrocarbon chain which can be optionally subtituted.
  • R is a hydrocarbon chain optionally substituted, a carbocyclic or heteroaliphatic ring optionally substituted, a phenyl or 5- to 6- membered heteroaryl optionally substituted.
  • B is boronic acids, boronate esters or trifluoroborate salts and R is phenyl or a 5- to 6- membered heteroaryl or a 3- to 7- heterocyclic or carbocyclic aliphatic ring or a hydrocarbon chain which can be optionally substituted.
  • R-10a 4-bromobenzenesulfonyl fluoride
  • R and R' are hydrogen, X is a halogen and R" is phenyl or a 5- to 6- membered heteroaryl or a 3- to 7- heterocyclic or carbocyclic aliphatic ring or a hydrocarbon chain which can be optionally substituted.
  • Q is O or NH
  • R is a 3- to 7- heterocyclic or carbocyclic aliphatic ring or a hydrocarbon chain which can be optionally substituted and R' is halogen or alcoxy.
  • Q is O or S and Cy is phenyl or a 5- to 6- membered heteroaryl and can be optionally substituted.
  • Q is O or NH
  • R is hydrogen
  • X is a halogen
  • R' is phenyl or a 5- to 6- membered heteroaryl or a 3- to 7- heterocyclic or carbocyclic aliphatic ring or a hydrocarbon chain which can be optionally substituted.
  • Synthesized compounds are obtained as racemic mixtures.
  • Corresponding isomers are purified by supercritical fluid chromatography (SFC) to obtain two enantiomers from each racemic compound.
  • Thromboelastometry is a viscoelastometric method for haemostasis testing in whole blood.
  • TEM® measures the interactions of coagulation factors, inhibitors and cellular components during the phases of clotting and
  • tPA Tissue plasminogen activator
  • CaCI 2 start-tern reagent
  • CMs tested compounds
  • Table 3 shows the results in human blood as effective concentration to delay lysis time by 50% (EC 50 LT); where, EC 50 LT ⁇ 25 ⁇ (+), 1 0 ⁇ ⁇ EC 50 LT ⁇ 25 ⁇ (++), 1 ⁇ ⁇ ECSO LT ⁇ 1 0 ⁇ (+++), EC 50LT ⁇ 1 ⁇ (++++) at all the assayed concentrations (1 000-0.2 ⁇ ).
  • Example EC50LT Example EC50LT
  • Table 4 shows the results in mice blood as effective concentration to delay lysis time by 50% (EC 50 LT); where, EC 50 LT ⁇ 10 ⁇ (+),1 ⁇ ⁇ EC 50 LT ⁇ 0 ⁇ (++),1 nM ⁇ EC50LT ⁇ 1 ⁇ (+++) and EC 50LT ⁇ 1 nM (++++) for all the assayed concentrations (1000-0.2 ⁇ ).
  • Antifibrinolvtic effect in vivo tail bleeding assay
  • Hyperfibrinolytic bleeding model consisted in injection of 0.5 mg/kg tPA into the ocular plexus to prolong bleeding time due to excessive fibrinolysis.
  • tPA 0.5 mg/kg tPA
  • polyurethane catheter (Microcannula 72-9030, Harvard Apparatus) for agents administration.
  • the catheter was connected to a syringe pump (AL-1000, WPI) for the infusion of 200 ⁇ _ (10% bolus, 90% perfusion during 40 minutes) of tested agents.
  • tPA 0.5 mg/kg
  • saline or the different compounds was infused through the femoral catheter to ensure systemic distribution of all the agents.
  • Reference compounds, TXA and Aprotinin were administered at 300 and 10 mg/Kg respectively; however, all compounds of the invention were administered at 1 mg/Kg. Five minutes later, 5 mm of tail tip were removed using a scalpel blade and the tail tip bathed in 1 mL of sterile saline at 37 °C.
  • the time of bleeding was defined as the interval between initial transections and the visual cessation of bleeding, that was measured up to 30 minutes. A value of 30 min was assigned to those animals bleeding longer than the observation period.
  • Table 6 shows the results reporting bleeding time (BT); where BT > 20 minutes (+),10 minutes ⁇ BT ⁇ 20 minutes (++), 5 minutes ⁇
  • tested compounds of the invention show a very significant reduction of the bleeding time when compared to the control or TXA. In all the cases the dose of tested compounds were lower than TXA or Aprotinin doses.

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Abstract

It relates to spirocyclic compounds of formula (I),or pharmaceutically or veterinary acceptable salts thereof, or any stereoisomers either of the compounds of formula (I) or of their pharmaceutically or veterinary acceptable salts, wherein A and B form a spirocyclic ring system wherein the spiro atom connecting A and B is a carbonatom and wherein A is a known 3- to 8-membered carbocyclic or heterocyclic monocyclic ring or a known 6- to 18-membered carbocyclic or heterocyclic polycyclic ring system; B is a known 4- to 7-membered carbocyclic or heterocyclic monocyclic ring; C is phenyl or a known 5- to 6-membered heteroaromatic ring; and R1-R7 are as defined herein. It also relates to a process for their preparation, as well as to the intermediates used in this process; to pharmaceutical or veterinary compositions containing them, and to their use in medicine, in particular as antifibrinolytic and antihemorragic agents.

Description

New antifibrinolvtic compounds
The present invention relates to spirocyclic compounds of formula (I), to a process for their preparation, as well as to the intermediates used in this process. It also relates to pharmaceutical or veterinary compositions containing them, and to their use in medicine, in particular as antifibrinolytic and antihemorragic agents.
BACKGROUND ART
The haemostatic system is responsible for maintaining circulatory fluidity and for preventing haemorrhage in response to vascular injury. Physiological hemostasis is controlled by mechanisms of coagulation and the formation of fibrin and by those favouring the degradation of fibrin (fibrinolysis).
Hyperfibrinolytic states caused by congenital or acquired conditions predispose to important haemorrhagic complications, often requiring transfusions and the need for re-exploration having a detrimental effect on patient outcome. Hemorhage is responsible for almost 50% of deaths occurring within 24 hours of traumatic injury and for up to 80% of
intraoperative trauma mortality. In western countries about one third of in- hospital deaths due to trauma is caused by abnormal blood loss which is an important contributory factor for other causes of death, particularly multi- organ failure, requiring massive blood transfusion. Failure to start appropriate early management in bleeding trauma patients is a leading cause of preventable death from trauma. Post-partum hemorrhage (PPH) is another leading cause of death in the developing world, accounting for 25% of maternal deaths, and rose in the the developed world from 1 .5% in 1999 to 4.1 % in 2009. The risk of haemorrhage can also be important in
cardiovascular patients on anti-coagulant therapy. Pharmacological approaches are an important part of multimodal therapy aiming to reducing bleeding and transfusion in order to reverse specific defects associated with such states; among them, the role of fibrinolysis inhibitors is growing. It is well known that subjects who bleed excessively in association with surgery or major trauma and need blood transfusions develop more complications than those who do not experience any bleeding. However, moderate bleeding requiring the administration of human blood products may lead to complications associated with the risk of transferring human viruses. Extensive bleeding requiring massive blood transfusions may lead to the development of multiple organ failure including lung or kidney function.
Therefore, a major goal in surgery as well as in the treatment of major tissue damage is to avoid or minimise bleeding in order to ensure the formation of stable and solid haemostatic plugs that are not easily dissolved by fibrinolytic enzymes. Furthermore, it is of importance to ensure quick and effective formation of such plugs or clots.
Antifibrinolytic agents are widely used in major surgery to prevent fibrinolysis and reduce blood loss. Currently two synthetic lysine analogs, epsilon- aminocaproic acid (EACA) and tranexamic acid (TXA), are the only
antifibrinolytics commercially available to control bleeding. These agents competitively inhibit activation of plasminogen to plasmin, an enzyme that degrades fibrin clots, fibrinogen and other plasma proteins.
However, there are some concerns with these currently available
antifibrinolytic agents due to the potential risk of thrombotic complications. There is still a need for improved treatment of subjects experiencing bleeding episodes, including subjects where the bleeding episodes are due to surgery, trauma, or other forms of tissue damage, as well as in clinical scenarios characterized by excessive fibrinolysis. SUMMARY OF THE INVENTION
Inventors have conceived and produced found a novel series of spirocyclic compounds that shows good antifibrinolytic and antihemorrhagic properties. In particular, the spirocyclic compounds which comprise spirocyclic ring system containing a carbon atom (spiro atom) attached to a hydroxamic acid and a sulfonyl group, show a significant delay in the lysis time in a
thromboelastometry assay. In addition, the spirocyclic compounds of the invention also show an important reduction of the bleeding time in vivo animal models as it will be shown in detail in the examples. These characteristics of the compounds of the invention allow a rapid cessation of hemorrhage; favor an effective formation of plugs or clots; have a sustained action (persistence of the clot and prevention of hemorrhage) and aid in minimizing the adverse effects related to other antifibrinolytic/antihemorrhagic treatments having risk of thrombotic complications.
Therefore, a first aspect of the invention relates to a compound of formula (I) (hereinafter also referred as compounds of the invention), or a
pharmaceutically or veterinary acceptable salt thereof, or any stereoisomer either of the compound of formula (I) or of its pharmaceutically or veterinary acceptable salt
Figure imgf000004_0001
(I)
wherein
A and B form a spirocyclic ring system wherein the spiro atom connecting A and B is a carbon atom and wherein
A is a known 3- to 8-membered carbocyclic or heterocyclic monocyclic ring, saturated or partially unsaturated; or alternatively
A is a known 6- to 18-membered carbocyclic or heterocyclic polycyclic ring system, saturated, partially unsaturated, or partially aromatic; and B is a known 4- to 7-membered carbocyclic or heterocyclic monocyclic ring, saturated or partially unsaturated;
C is phenyl or a known 5- to 6-membered heteroaromatic ring;
R1-R3 are independently selected from H, halogen, -NO2, -CN, Ra, -ORa, -OC(Y)Ra', -OC(Y)ORa', -OC(Y)NR Ra, -OSO2ORa', -NR Ra, -NR C(Y)Ra, -NR C(Y)ORa, -NR C(Y)NR Ra, -NR S(O)2Ra', -NR SO2NR Ra, -SRa', -S(O)Ra', -S(O)ORa', -SO2Ra',-SO2(ORa), -SO2NR Ra, -SC(Y)NR Ra, -C(Y)Ra',
-C(Y)ORa', -C(Y)NR Ra, -C(Y)NR ORa, and -C(O)NR SO2Ra';
R4-R7 are independently selected from halogen, -NO2, -CN, Rc, -ORc, -NRdRc, -NRdC(Y)Rc, -NRdC(Y)ORc, -NRdC(Y)NRdRc, -NRdS(O)2Rc, -NRdSO2NRdRc, -SRC, -S(O)Rc, -S(O)ORc, -SO2Rc, -SO2R(ORc), -SO2NRdRc, -SC(Y)NRdRc, -C(Y)RC, -C(Y)ORc, -C(Y)NRdRc, -C(Y)NRdORc, and -C(O)NRdSO2Rc
Ra is a saturated or unsaturated (Ci-Ci2)alkyl optionally substituted with one or more substituents Re and/or one Cy1; or alternatively Ra is Cy2;
wherein Cy1 and Cy2 are independently optionally substituted with: one Cy3 and/or one or more substituents Re, and/or one or more saturated or unsaturated (CrC6)alkyl groups optionally substituted with one or more substituents Re and/or one Cy3; and
wherein any Cy3 is optionally substituted with one or more substituents independently selected from Re and saturated or unsaturated (CrC6)alkyl optionally substituted with one or more substituents Re; each Ra and R are independently H or Ra;
Rc and each Rd are independently selected from H, Cy4, and saturated or unsaturated (CrC6)alkyl optionally substituted with one or more substituents Rh and/or one Cy5;
wherein Cy4 is optionally substituted with one or more substituents
independently selected from Rh and saturated or unsaturated (CrC6)alkyl optionally substituted with one or more substituents Rh; and
wherein Cy5 is optionally substituted with one or more substituents
independently selected from Rh and saturated or unsaturated (CrC6)alkyl optionally substituted with one or more substituents Rh; each Re is independently selected from halogen, -NO2, -CN, -ORf, -OC(Y)Rf, -OC(Y)ORf, -OC(Y)NR9Rf, -NR9Rf, -NR9C(Y)Rf, -NR9C(Y)ORf, -NR9C(Y)NR9Rf, -NR9S(O)2Rf, -NR9SO2NR9Rf, -SRf, -S(O)Rf, -S(O)ORf, -SO2Rf, -SO2(ORf), -SO2NR9Rf, -SC(Y)NR9Rf, -C(Y)Rf, -C(Y)ORf, -C(Y)NR9Rf, -C(Y)NR9ORf, and -C(O)NR9SO2Rf. Rf and each R9 are independently selected from H, Cy6, and saturated or unsaturated (CrC6)alkyl optionally substituted with one or more substituents Rh and/or one Cy7;
wherein Cy6 is optionally substituted with: one Cy7, and/or one or more substituents Rh, and/or one or more saturated or unsaturated (CrC6)alkyl groups optionally substituted with one or more substituents Rh and/or one Cy7; and
wherein any Cy7 is optionally substituted with one or more substituents independently selected from Rh and (Ci-C4)alkyl optionally substituted with one or more substituents Rh; each Rh is independently selected from halogen, -NO2, -CN, -OR', -ΟΟ(Ο)^, -00(0)0^, -OCiOJNR'R1, -NR'R', -NR'CiOJR1, -NR^OJOR1, -NR'C(O)NR'R', -NR^O^R1, -NR'SCfeNR'R1, -SR1, -S{0)R -SOzR1, -SOziOR1), -SCfeNR'R1, -C{0)R -0(0)0^, -CiOJNR'R1, and -C(O)NR'OR' each R1 is independently H or -(Ci-C4)alkyl optionally substituted with one or more halogen atoms; Y is O, S, or NR';
Cy1, Cy2, Cy4 and Cy6 are independently a C or N-attached known ring system selected from 3- to 8-membered carbocyclic or heterocyclic monocyclic ring, saturated or partially unsaturated; phenyl; 5- or 6-membered heteroaromatic ring; and 6- to 18-membered carbocyclic or heterocyclic polycyclic ring system, saturated, partially unsaturated, aromatic or partially aromatic;
Cy3, Cy5 and Cy7 are independently a C or N-attached known ring system selected from 3- to 8-membered carbocyclic or heterocyclic monocyclic ring, saturated or partially unsaturated; phenyl; and 5- or 6-membered
heteroaromatic ring; wherein in the carbocyclic rings all ring members are carbon atoms; and in the heterocyclic and heteroaromatic rings one or more ring members are selected from N, O, and S; and wherein in all saturated or partially
unsaturated rings one or two members of the rings are optionally C(O) and/or C(NH) and/or C[N(C C4)alkyl]. Another aspect of the invention relates to a pharmaceutical or veterinary composition which comprises an effective amount of a compound of formula (I) as defined above, or a pharmaceutically or veterinary acceptable salt thereof, or any stereoisomer either of the compound of formula (I) or of its pharmaceutically or veterinary acceptable salt, together with one or more pharmaceutically or veterinary acceptable excipients or carriers.
As previously described, the compounds of the invention are useful as antifibrinolytic and antihemorrhagic agents. Therefore, another aspect of the invention relates to a compound of formula (I) as defined above, or a pharmaceutically or veterinary acceptable salt thereof, or any stereoisomer either of the compound of formula (I) or of its pharmaceutically or veterinary acceptable salt, for use as a medicament.
Another aspect of the invention relates to a compound of formula (I) as defined above, or a pharmaceutically or veterinary acceptable salt thereof, or any stereoisomer either of the compound of formula (I) or of its
pharmaceutically or veterinary acceptable salt, for use as antifibrinolytic and antihemorrhagic agent. Thus, this aspect relates to the use of a compound of formula (I) as defined above, for the manufacture of a medicament for use as antifibrinolytic and antihemorrhagic agent; and may also be formulated as a method for the treatment and/or prevention of hyperfibrinolysis and/or hemorrhages comprising administering an effective amount of the previously defined compound of formula (I), or a pharmaceutically or veterinary acceptable salt thereof, or any stereoisomer either of the compound of formula (I) or of its pharmaceutically or veterinary acceptable salt, and one or more pharmaceutically or veterinary acceptable excipients or carriers, in a subject in need thereof, including a human.
Processes for the preparation of compounds of formula (I) are also part of the invention as well as intermediates used in these processes. Accordingly, another aspect of the invention relates to a compound of formula (III)
Figure imgf000008_0001
(III) wherein A, B, C, R1-R7 are as defined above, and R' is H or a carboxy protective group, more particularly a carboxy protective group selected from the group consisting of (CrC6)alkyl, benzyl, p-methoxyphenyl, trimethylsilyl and [2-(Trimethylsilyl)ethoxy]methyl (SEM), with the proviso that compound (III) is other than 7-methoxycarbonyl-7-phenylsulphonyl-2-oxaspiro[2.4]- heptane and (2S*, 4R*)-2-Phenylsulfonyl-4-iodomethyl-6, 1 1 -dioxaspiro[4.6]- undecane-2-carboxylic acid methyl ester.
Another aspect of the invention relates to a compound of formula (II)
Figure imgf000008_0002
(II) wherein A, B, C, R1-R7 are as defined above, and R is an hydroxamic acid protective group, more particularly an hydroxamic acid protective group selected from the group consisting of tetrahydro-2H-pyran-2-yloxy (THP), benzyl, 1 -naphthylmethyl and dimethyloxybenzyl (DMB). DETAILED DESCRIPTION OF THE INVENTION
All terms as used herein in this application, unless otherwise stated, shall be understood in their ordinary meaning as known in the art. Other more specific definitions for certain terms as used in the present application are as set forth below and are intended to apply uniformly through-out the specification and claims unless an otherwise expressly set out definition provides a broader definition. For the purposes o the present invention, in the spirocyclic ring system formed by the ring system A and ring system B, the spiro atom connecting A and B is a carbon atom.
The term "carbocyclic" ring system refers to a known ring system wherein all the ring members are carbon atoms. The term "heterocyclic" ring system refers to a known ring system wherein one or more of the ring members, preferably 1 , 2, 3, or 4 ring members, are selected from N, O, and S, where chemically possible. Unless otherwise specified, the "heterocyclic" ring system may be attached to the rest of the molecule through a C or a N atom. Both the carbocyclic and heterocyclic rings can be saturated or partially unsaturated, and may be unsubstituted or substituted as described herein, being the susbstituents placed on any available position.
According to the present invention, the term "polycyclic" ring refers to a ring system which is formed by two, three or four rings which can be fused, bridged-fused, spiro-fused or can contain different types of fusion. For the purposes of the present invention, in "fused" rings the fusion occurs through one bond which is common to two adjoining rings; in "bridged-fused" rings the fusion occurs through a sequence of atoms (bridgehead) which is common to two rings; and in "spiro-fused" rings, the fusion occurs through only one atom
(spiro atom), preferably a carbon atom, which is common to two adjoining rings (including bridged rings)
The polycyclic ring system can be saturated, partially unsaturated, aromatic (except in the case of ring system A) or partially aromatic; and may be unsubstituted or substituted as described herein, being the susbstituents placed on any available position. The term "heteroaromatic" ring refers to a known aromatic ring system, wherein one or more of the ring members, preferably 1 , 2, 3, or 4 ring members, are selected from N, O, and S where chemically possible. The heteroaromatic ring and phenyl may be unsubstituted or substituted as described herein, being the susbstituents placed on any available position.
The term "known" ring system as used herein refers to a ring system which is chemically feasible and is known in the art and so intends to exclude those ring systems that are not chemically possible.
For the purposes of the present invention, in all saturated or partially unsaturated rings, one or two members of the rings are optionally C(O) and/or C(NH) and/or C[N(C C4)alkyl].
The term saturated or unsaturated (CrCn)alkyl refers to a saturated branched or linear hydrocarbon chain which contains from 1 to n carbon atoms. When the (C-i-Cn)alkyl is saturated it contains only single bonds. When the
(d-Cn)alkyl is unsaturated it contains one or two double bonds and/or one or two triple bonds. The saturated or unsaturated (CrCn)alkyl may be
substituted or unsubstituted as described herein.
A halogen substituent means fluoro, chloro, bromo or iodo. "Protective group" (PG) refers to a grouping of atoms that when attached to a reactive group in a molecule masks, reduces or prevents that reactivity.
The expression "substituted with one or more" means that a group can be substituted with one or more, preferably with 1 , 2, 3 or 4 substituents, provided that this group has enough positions susceptible of being
substituted.
For the purposes of the invention, room temperature is 20-25 °C. As mentioned above, a first aspect of the invention relates to compounds of formula (I) or pharmaceutically or veterinary acceptable salts thereof, or any stereoisomer either of the compound of formula (I) or of its pharmaceutically or veterinary acceptable salt. There is no limitation on the type of salt that can be used, provided that these are pharmaceutically or veterinary acceptable when they are used for therapeutic purposes. The term "pharmaceutically or veterinary acceptable salts", embraces salts commonly used to form alkali metal salts and to form addition salts of free acids or free bases.
The preparation of pharmaceutically or veterinary acceptable salts of the compounds of formula (I) can be carried out by methods known in the art. For instance, they can be prepared from the parent compound, which contains a basic or acidic moiety, by conventional chemical methods. Generally, such salts are, for example, prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate
pharmaceutically or veterinary acceptable base or acid in water or in an organic solvent or in a mixture of them. The compounds of formula (I) and their salts may differ in some physical properties but they are equivalent for the purposes of the present invention.
The compounds of the invention may be in crystalline form either as free solvation compounds or as solvates (e.g. hydrates) and it is intended that both forms are within the scope of the present invention. Methods of solvation are generally known within the art. In general, the solvated forms with pharmaceutically or veterinary acceptable solvents such as water, ethanol and the like are equivalent to the unsolvated form for the purposes of the invention.
Some compounds of formula (I) can have chiral centres that can give rise to various stereoisomers. The present invention relates to each of these stereoisomers and also mixtures thereof. Moreover, some of the compounds of the present invention can show cis/trans isomers. The present invention relates to each of the geometric isomers and mixtures thereof.
Diastereoisomers can be separated by conventional techniques such as chromatography or fractional crystallization. Optical isomers can be resolved by conventional techniques of optical resolution to give optically pure isomers. This resolution can be carried out on any chiral synthetic
intermediate or on products of general formula (I). Optically pure isomers can also be individually obtained using enantiospecific synthesis. In one embodiment, the invention refers to a compound of formula (I) as defined above, wherein A is a known 3- to 8-membered carbocyclic or heterocyclic monocyclic ring or a known 6- to 10-membered carbocyclic or heterocyclic bicyclic ring system.
In another embodiment, the invention refers to a compound of formula (I), wherein A is a monocyclic ring selected from a 3- to 6-membered carbocyclic ring, and a 5- to 6-membered heterocyclic ring.
In another embodiment, the invention refers to a compound of formula (I), wherein A is a carbocyclic monocyclic ring; or a polycyclic ring system, preferably a bicyclic ring system, wherein the ring containing the spiro atom attached to the hydroxamic acid and the sulfonyl group is a carbocyclic ring.
In another embodiment, the invention refers to a compound of formula (I), wherein A is selected from cyclopropane, cyclobutane, cyclopentane, cyclohexane, tetrahydrofuran, pyrrolidine, bicyclo[2.2.1 ]heptane, 2,3-dihydro- 1 H-indene, hexahydropyrrolizin-3-one, and 4-azaspiro[4.4]nonane.
In another embodiment, the invention refers to a compound of formula (I), wherein A is unsubstituted, i.e. R7 is H.
In another embodiment, the invention refers to a compound of formula (I), wherein B is a 6- to 7-membered carbocyclic or heterocyclic monocyclic ring.
In another embodiment, the invention refers to a compound of formula (I), wherein B is a saturated monocyclic ring, carbocyclic or heterocyclic, wherein at least one of the ring members of the heterocyclic ring is NR4.
In another embodiment, the invention refers to a compound of formula (I), wherein B is selected from cyclohexane, piperidine, morpholine, azepane, piperazine, pyrrolidine, and azetidine. In another embodiment, B is piperidine, morpholine, azepane, pyrrolidine, and azetidine, wherein R4 is placed on the N atom of these rings and R5-R6 are H. In another embodiment, B is piperazine, wherein R4 and R5 are placed on the N atoms and R6 is H. In another embodiment, the invention refers to a compound of formula (I), wherein A and B form a spirocyclic ring system selected from the group consisting of:
Figure imgf000013_0001
In another embodiment, the invention refers to a compound of formula (I), wherein C is phenyl. In another embodiment, C is phenyl substituted with R1 at the orto, meta or para position, and R2 and R3 are independently selected from H, halogen, Ra, -ORa',and -NR Ra; wherein Ra, Ra'and R are
independently selected from H and -(Ci-C4)alkyl optionally substituted with one or more halogen atoms. In another embodiment, C is phenyl substituted with R-i at the para position and R2 and R3 are H. In another embodiment, C is phenyl substituted with R1 at the orto position and R2 and R3 are H. In another embodiment, C is phenyl substituted with R1 at the meta position and R2 and R3 are H. In another embodiment, C is phenyl substituted with R1 at the meta position, and R2 at the para position, R3 being H; or alternatively, C is phenyl substituted with R1 at the para position, and R2 at the meta position, R3 being H; wherein R2 is selected from H, halogen, Ra, -ORa',and -NR Ra; and Ra, Raand R are independently selected from H and -(Ci-C4)alkyl optionally substituted with one or more halogen atoms.
In another embodiment, the invention refers to a compound of formula (I), wherein in R (relating to Ri-R3), Cy1 and Cy2 are independently optionally substituted with one or more substituents selected from Re and saturated or unsaturated (CrC6)alkyl optionally substituted with one or more substituents
Re; and Cy6 is optionally substituted with one or more substituents
independently selected from Rh and saturated or unsaturated (CrC6)alkyl optionally substituted as previously defined, more particularly with one or more substituents Rh.
In a more particular embodiment, the invention refers to a compound of formula (I), wherein in R (relating to Ri-R3), R is H and saturated or unsaturated (Ci-Ci2)alkyl optionally substituted with one or more substituents Re, more particularly wherein in Re, Rf and each R9 are independently selected from H and saturated or unsaturated (CrC6)alkyl optionally substituted with one or more fluorine atoms.
In another embodiment, the invention refers to a compound of formula (I), wherein in R R3, Re is selected from halogen, -NO2, -CN, -ORf, -OC(O)Rf, -OC(O)ORf, -OC(O)NR9Rf, -NR9Rf, -NR9C(O)Rf, -NR9C(O)ORf,
-NR9C(O)NR9Rf, -NR9S(O)2Rf, -SRf, -S(O)Rf, -SO2Rf, -SO2NR9Rf, -C(O)Rf, -C(O)ORf, -C(O)NR9Rf, and -C(O)NR9ORf. In another embodiment, the invention refers to a compound of formula (I), wherein in Ri-R3, Rf and each R9 are independently selected from H and saturated or unsaturated (CrC6)alkyl optionally substituted with one or more fluorine atoms.
In another embodiment, the invention refers to a compound of formula (I), wherein in Ri-R3, Cy1 and Cy2 are independently optionally substituted with one or more substituents selected from Re and saturated or unsaturated (CrC6)alkyl optionally substituted as previously defined; and Cy6 is optionally substituted with one or more substituents independently selected from Rh and saturated or unsaturated (CrC6)alkyl optionally substituted as previously defined. In another embodiment, the invention refers to a compound of formula (I), wherein R R3 are independently selected from H, halogen, -NO2, -CN, Ra, -OR3', -OC(O)Ra', -OC(O)ORa', -OC(O)NR Ra', -NR Ra, -NR C(O)Ra,
-NR C(O)ORa', -NR C(O)NR Ra, -NR S(O)2Ra', -SRa', -S(O)Ra', -SO2Ra', -SO2NR Ra, -C(O)Ra', -C(O)ORa', -C(O)NR Ra, and -C(O)NR ORa'.
In another embodiment, the invention refers to a compound of formula (I), wherein in R4-R7, Rh is selected from halogen, -NO2, -CN, -OR', -ΟΟ(Ο)^, -00(0)0^, -OCiOJNR'R1, -N R1, -NR'CiO)^, -NRtyOJOR1, -NR'C(O)NR'R', -NR^O),^, -S , -S(O)R, -SO2R, -SO2NRR, -C(O)R, -C(O)OR, and
-CiOJNR^.
In another embodiment, the invention refers to a compound of formula (I), wherein R4-R7 are independently selected from halogen, -NO2, -CN, Rc, -ORc, -NRdRc, -NRdC(O)Rc, -NRdC(O)ORc, -NRdC(O)NRdRc, -NRdS(O)2Rc, -SRC, -S(O)Rc, -SO2Rc, -SO2NRdRc, -C(O)Rc, -C(O)ORc, and -C(O)NRdRc.
In another embodiment, the invention refers to a compound of formula (I), wherein R2 and R3 are independently selected from H, halogen, Ra, -ORa,and -NR Ra; and R5-R7 are independently selected from H, halogen, Rc, -ORc, and -NRdRc, wherein Ra, Ra', R , Rc and Rd are independently selected from H and -(Ci-C4)alkyl optionally substituted with one or more fluorine atoms. In another embodiment, the invention refers to a compound of formula (I), wherein is selected from H, halogen, -NO2, -CN, Ra, -OR3', -OR3',
-OC(O)R3', -OC(O)OR3', -OC(O)NR R3', -NR R3', -NR C(O)R3', -NR C(O)OR3', -NR C(O)NR R3', -NR S(O)2R3', -SR3', -S(O)R3', -SO2R3', -SO2NR R3', -C(O)R3', -C(O)OR3', -C(O)NR R3', and -C(O)NR OR3'; R4 is selected from halogen, -NO2, -CN, Rc, -ORc, -NRdRc, -NRdC(O)Rc, -NRdC(O)ORc, -NRdC(O)NRdRc, -NRdS(O)2Rc, -SRC, -S(O)Rc, -SO2Rc, -SO2NRdRc, -C(O)Rc, -C(O)ORc, and -C(O)NRdRc; and R2, R3, and R5-R7 are independently selected from H, halogen, -(C C4)alkyl, -OH, -O[(C C4)alkyl], -NH2, -NH[(C C4)alkyl],
-N[(Ci-C4)alkyl]2, wherein each (Ci-C4)alkyl is independently optionally substituted with one or more fluorine atoms.
The present invention also relates to the combination of any of the specific embodiments defined above for any of the variables A, B, C, and R1-R7.
In another embodiment of the invention, the compound of formula (I) is selected from the group consisting of:
Figure imgf000016_0001
1 -04 1 -05 1 -06
Figure imgf000017_0001
-19 1-20 1-21
Figure imgf000018_0001
Figure imgf000019_0001
Figure imgf000020_0001
-55 1-56
Figure imgf000021_0001
Figure imgf000022_0001
-91 1-92 1-93
Figure imgf000023_0001
-106 1-107 1-108
Figure imgf000024_0001
-124 1-125 1-126
Figure imgf000025_0001
Isomer B*
Isomer A* Isomer B*
1 -136 1 -137 1 -138
*Regarding these compounds, no attempt has been made to identify the absolute configurations. However, in the examples it is clearly indicated which of the two isomers is concerned in relative terms by differentiating
unambiguously between their physical and/or spectroscopic properties.
Generally, compounds of formula (I) as defined above may be obtained by reacting a compound of formula (III)
Figure imgf000025_0002
with a hydroxylamine of formula RO-NH2 (IV), wherein A, B, C, R1-R7 are as previously defined; R' is H; and R is an hydroxamic acid protective group, more particularly an hydroxamic acid protective group selected from the group consisting of tetrahydro-2H-pyran-2-yloxy (THP), benzyl,
1 -naphthylmethyl and dimethyloxybenzyl (DMB), to give a compound of formula (II)
Figure imgf000026_0001
(II) and subsequently removing the protective group of the hydroxamic acid to give a compound of formula (I).
The first conversion can be carried out in the presence of an activating agent such as 1 -ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride
(EDC.HCI) and Hydroxybenzotriazole (HOBt), preferably in the presence of a base, such as N-methylmorpholine (NMM), in a suitable solvent, such as dichloromethane, chloroform or dimethylformamide, at a temperature comprised from room temperature to the temperature of the boiling point of the solvent, preferably at room temperature.
The removal of the protective group of the hydroxamic acid is carried out by standard methods well-known in the art as described for example in T. W. Green and P. G. M. Wuts, Protective Groups in Organic Chemistry (Wiley, 3rd ed. 1999, Chapter 2, pp. 17-200). Representative hydroxy protective groups include those where the hydroxy group is either acylated or alkylated such as benzyl, and trityl ethers as well as alkyl ethers, tetrahydropyranyl ethers, trialkylsilyl ethers and allyl ethers. For example, the hydroxamic acid protective group is tetrahydro-2H-pyran-2-yloxy (THP), benzyl,
1 -naphthylmethyl or dimethyloxybenzyl (DMB). When the hydroxamic acid protective group is THP, the deprotection is carried out in acidic medium, for example with HCI, in a suitable solvent such as dioxane.
Intermediate compounds of formula (II), wherein A, B, C, R1-R7 and R are as previously defined, also form part of the invention.
Compounds of formula (III) wherein R' is H can be obtained by removing the protective group of a compound of formula (III) wherein R' is a carboxy protective group by standard methods well-known in the art as described for example in T. W. Green and P. G. M. Wuts, Protective Groups in Organic Chemistry (Wiley, 3rd ed. 1999, Chapter 5, pp. 369-451 ). Representative carboxy protective groups include alkyl, aryl or benzyl esters, sylil esters, amides or hydrazides. For example, the carboxy protective group is
(CrC6)alkyl, benzyl, p-methoxyphenyl, trimethylsilyl, or [2-(Trimethylsilyl)- ethoxy]methyl (SEM). When the carboxy protective group is (CrC6)alkyl, the deprotection is carried out in basic medium, for example with LiOH in a suitable solvent such as tetrahydrofuran-methanol.
Intermediate compounds of formula (III) wherein A, B, C, R1-R7 are as previously defined and R' is H or a carboxy protective group, more particularly a carboxy protective group selected from the group consisting of (CrC6)alkyl, benzyl, p-methoxyphenyl, trimethylsilyl and [2-(Trimethylsilyl)ethoxy]methyl (SEM) form also part of the invention, with the proviso that compound (III) is other than 7-methoxycarbonyl-7-phenylsulphonyl-2-oxaspiro[2.4]heptane and (2S*, 4R*)-2-Phenylsulfonyl-4-iodomethyl-6,1 1 -dioxaspiro[4.6]undecane-2- carboxylic acid methyl ester, form also part of the invention. These
compounds are cited in D. Bouyssi et al., "Rearrangement of
oxaspiroheptanes to cyclohexanones mediated by lithium iodide", Synlett 2000, vol. 5, pp. 749-751 and Osamu Kitagawa et al., "Stereoselective Iodine Atom Transfer [3 + 2] Cycloaddition Reaction with Alkenes Using
Unsymmetrical Allylated Active Methine Radicals", The Journal of Organic Chemistry 2004, vol. 69, pp. 2607-2610, respectively, which describe only a synthetic process without describing any therapeutical application.
Given that the chemical structure of intermediate compounds of formula (II) and formula (III) coincides largely with the chemical structure of the final compounds of formula (I), the particular embodiments defined above for the compounds of formula (I) regarding the variables A, B, C, and R1-R7 are also particular embodiments of the intermediate compounds of formula (II) and formula (III).
Compounds of formula (III) can be generally obtained by cou
compound of formula (V) with a compound of formula (VI):
Figure imgf000028_0001
(V) (VI) (III) wherein A, B, C, R1-R7 are as previously defined and R' is carboxy protective group. This conversion is carried out in the presence of a base such as lithium diisopropylamide (LDA), in a suitable solvent such as tetrahydrofuran, and at a suitable temperature, preferably -78 °C.
Additionally, compounds of formula (I), wherein R1 is Q-Cy2, wherein Q is O, S, SO or SO2 and Cy2 is as defined above (i.e. compound of formula (la)) may be obtained in general as shown in the following scheme:
Figure imgf000028_0002
(la) (Mb) (l la)
Scheme 1 A compound of formula (V), wherein A, B, R4-R7 are as previously defined and R' is carboxy protective group, is reacted as described above with a compound of formula (Via) wherein C, R2 and R3 are as previously defined, to give a compound of formula (Ilia), which is deprotected to give the carboxylic acid of formula (1Mb) and subsequently reacted with a compound of formula (IV) as already described to give compound (lla).
Compound of formula (lla) is reacted with a compound of formula H-Q'-Cy2 (VII), wherein Q' is O or S, and Cy2 is as defined above, to give a compound of formula (Mb). This reaction is carried out in the presence of a base, such as cesium carbonate, optionally in a suitable solvent or without solvent, and preferably heating. Further, a compound of formula (Mb) wherein Q is S can be oxidized to a compound of formula (Mb) wherein Q is SO or SO2 in the presence of an oxidizing agent, such as m-chloroperbenzoic acid, in a suitable solvent, such as dichlorormethane, and preferably at room temperature. Deprotection of a compound of formula (Mb) yields a compound of formula (la) as defined above.
Additionally, compounds of formula (I), wherein R1 is Q-alk, Q is O or S, and alk is saturated or unsaturated -(Ci-Ci2)alkyl optionally substituted (i.e.
compound of formula (lb)) may be obtained in general as shown in the following scheme:
Figure imgf000029_0001
Scheme 2
Thus, a compound of formula (lla) can be reacted with a compound of formula H-Q-alk (VIII) in the presence of a base, such as NaH, optionally in a suitable solvent or without solvent, and preferably heating. After removal of the protective group in the resulting compound of formula (lie), compound of fomula (lb) is obtained.
Compounds of formula (I), wherein R1 is -NR Ra , R is saturated or unsaturated -(CrCi2)alkyl optionally substituted and Ra is H or saturated or unsaturated -(CrCi2)alkyl optionally substituted (i.e. compound of formula lc)) may be obtained in general as shown in the following scheme:
Figure imgf000030_0001
Scheme 3 Thus, a compound of formula (lla) can be reacted with a compound of formula H-NR Ra (IX), optionally in a suitable solvent or without solvent, and preferably heating. After removal of the protective group in the resulting compound of formula (lid), compound of formula (lc) is obtained.
Compounds of formula (I), wherein R1 is -NCy2Ra and Cy2 is an optionally substituted aromatic, heteroaromatic or aliphatic ring (i.e. compound of formula (Id)) may be obtained in general as shown in the following scheme:
Figure imgf000030_0002
Scheme 4
Thus, a compound of formula (lla) can be reacted with a compound of formula H-NCy2Ra (X), optionally in a suitable solvent or without solvent, and preferably heating. After removal of the protective group in the resulting compound of formula (lie), compound of formula (Id) is obtained. Compounds of formula (I), wherein R1 is -NR C(Y)Ra, -NR C(Y)ORa,
-NR C(Y)NR Ra, -NR S(O)2Ra', -NR SO2NR Ra' (i.e. compound of formula (le)) may be obtained from a compound of formula (llg), which can be prepared as shown in the following scheme:
Figure imgf000031_0001
Scheme 5
In a first step, compound of formula (Ila) is reacted with benzylamine, preferably heating to give a compound of formula (llf) which can be
hydrogenated in the presence of Pd/C to give a compound of formula (llg). This compound can be reacted for example with an acyl halide in the presence of a base, such as triethylamine, in a suitable solvent, such as tetrahydrofuran, at a temperature comprised from room temperature to the reflux temperature of the solvent, to give an amide; or with an isocyanate to give a urea.
Compounds of formula (I), wherein R1 is -OC(Y)Ra (i.e. compound of formula (If)) may be obtained from a compound of formula (Mi), which can be prepared as shown in the following scheme:
Figure imgf000031_0002
(Ila) (llh) (Mi)
Scheme 6
In a first step, compound of formula (Ila) is reacted with benzyl alcohol in the presence of a base, such as NaH, in a suitable solvent, such as
tetrahydrofuran, to give a compound of formula (llh) which can be
hydrogenated in the presence of Pd/C to give a compound of formula (Mi). This compound can be reacted for example with an acyl halide in the presence of a base, such as triethylamine, in a suitable solvent, preferably at room temperature to give an ester.
Compounds of formula (I), wherein R1 is Ra (i.e. compound of formula (lg)) may be obtained from a compound of formula (Ilk), which can be prepared as shown in the following scheme:
Figure imgf000032_0001
(llg) (llj) (|lk)
Scheme 7 In a first step, compound of formula (llg) is reacted with potassium iodide and NaNO2 in a suitable solvent, such as acetonitrile and then an acid, such as concentrated HCI, is added to give a compound of formula (llj). This compound can be reacted for example with boronic acid derivatives in the presence of a palladium catalyst (Suzuki coupling) to give a compound of formula (Ilk).
The same type of reactions shown above for can be aplied to R2-R7.
Alternatively, the reactions described above can be carried out in a different order. Thus, the reactions carried out on compounds of formula (II) (i.e. on the protected hydroxamic acid) can be carried out on compounds of formula (III) (i.e. the carboxylic ester), and can be deprotected to give the corresponding carboxylic acids, which can be converted to the compounds of formula (I) as described above.
The compounds of formulas (V) to (XI) are commercially available or can be obtained by conventional synthetic processes.
The present invention also relates to a pharmaceutical or veterinary composition comprising an effective amount of a compound of formula (I) as defined above, or a pharmaceutically or veterinary acceptable salt thereof, or any stereoisomer either of the compound of formula (I) or of its
pharmaceutically or veterinary acceptable salt, together with pharmaceutically or veterinary acceptable excipients or carriers. The expression "effective amount" as used herein, refers to the amount of a compound that, when administered, is sufficient to prevent development of, or alleviate to some extent, one or more of the symptoms of the disease which is addressed. The specific dose of the compound of the invention to obtain a therapeutic benefit may vary depending on the particular circumstances of the individual patient including, among others, the size, weight, age and sex of the patient, the nature and stage of the disease, the aggressiveness of the disease, and the route of administration. For example, a dose of from about 0.01 to about 300 mg/kg may be used. The expression "pharmaceutically or veterinary acceptable excipients or carriers" refers to pharmaceutically or veterinary acceptable materials, compositions or vehicles. Each component must be pharmaceutically or veterinary acceptable in the sense of being compatible with the other ingredients of the pharmaceutical or veterinary composition. It must also be suitable for use in contact with the tissue or organ of humans and animals without excessive toxicity, irritation, allergic response, immunogenicity or other problems or complications commensurate with a reasonable benefit/risk ratio. The election of the pharmaceutical or veterinary formulation will depend upon the nature of the active compound and its route of administration. Any route of administration may be used, for example oral, parenteral and topical administration. For example, the pharmaceutical or veterinary composition may be formulated for oral administration and may contain one or more physiologically
compatible carriers or excipients, in solid or liquid form. These preparations may contain conventional ingredients such as binding agents, fillers, lubricants, and acceptable wetting agents.
The pharmaceutical or veterinary composition may be formulated for parenteral administration in combination with conventional injectable liquid carriers, such as water or suitable alcohols. Conventional pharmaceutical or veterinary excipients for injection, such as stabilizing agents, solubilizing agents, and buffers, may be included in such compositions. These
pharmaceutical or veterinary compositions may be injected intramuscularly, intraperitoneal^, or intravenously.
The pharmaceutical or veterinary composition may be formulated for topical administration. Formulations include creams, lotions, gels, powders, solutions and patches wherein the compound is dispersed or dissolved in suitable excipients. The topical compositions of the invention may be administered by means of a carrier material, which can be a solid support. Thus, it also forms part of the invention a topical composition comprising a carrier material, which can be a solid support. Illustrative, non-limiting examples of solid supports include intelligent textiles, dressings, coatings, sponges, band-aids, sanitary pads, compresses, plasters, etc. The manufacture of such compositions can be obtained by conventional methods, for example, by mixing the
combinations of the invention and the material carrier.
The pharmaceutical or veterinary compositions may be in any form, including, among others, tablets, pellets, capsules, aqueous or oily solutions,
suspensions, emulsions, or dry powdered forms suitable for reconstitution with water or other suitable liquid medium before use, for immediate or retarded release. The appropriate excipients and/or carriers, and their amounts, can readily be determined by those skilled in the art according to the type of formulation being prepared.
The compounds of the present invention are useful as antihemorrhagic and antifibrinolytic agents and can be used in a broad range of therapeutic applications. In surgery, antifibrinolytic agents, in addition to reducing postoperative hemorrhage, can be an alternative to blood transfusion and other hemoderivatives for example in heart, liver and orthopedic surgery, and also in the setting of oncologic surgery in organs rich in fibrinolysis activators (prostate, uterus). In trauma patients antifibrinolytic agents can reduce all- cause mortality and death due to bleeding. Further, the antifibrinolytic agents of the invention can also be used to control bleeding in trombolytic therapy, e.g. in cases of acute heart attack and ischemic stroke, and major or intracraneal hemorrhages. Moreover, the antifibrinolytic agents of the invention are useful in the treatment of local hemorrhages, e.g. after teeth extraction, in particular in patients with congenital coagulopathies, such as hemophilia, or patients with diabetes; in the treatment of menorrhage in women associated with congenital or acquired coagulopathies, as well as in post-partum haemorrhage, and in the treatment of hemorrhages of
gastrointestinal and urologic origin, including prostatectomy. Throughout the description and claims the word "comprise" and variations of thereof, are not intended to exclude other technical features, additives, components, or steps. Furthermore, the word "comprise" encompasses the case of "consisting of. Additional objects, advantages and features of the invention will become apparent to those skilled in the art upon examination of the description or may be learned by practice of the invention. The following examples are provided by way of illustration, and they are not intended to be limiting of the present invention. Furthermore, the present invention covers all possible combinations of particular and preferred embodiments described herein.
EXAMPLES
General Procedure for Prep-HPLC purification method:
The HPLC measurement was performed using Gilson 281 from 233 pump (binary), an autosampler, and a UV detector. The fractions was detected by
LC-MS. The MS detector was configured with an electrospray ionization source. The source temperature was maintained at 300-350 oC.
Method 1
Reverse phase HPLC was carried out on Luna C18 (100x30 mm; 4um).
Solvent A: water with 0.075% trifluoroacetic acid; Solvent B: acetonitrile with 0.075% trifluoroacetic acid. Gradient: At room temperature, 20% of B to 40% of B within 6 min at 25 mL/min; then 40% B at 25 mL/min over 2 min, UV detector.
The following abbreviations have been used in the examples:
Boc: tert-butoxycarbonyl DCM: dichloromethane
DMF: dimethylformamide
Et3N: triethylamine
TLC: Thin Layer Chromatography
PE: Petroleum ether
AE/EtOAc: ethyl acetate
ACN: acetonitrile
r.t: room temperature
Rt: retention time
THF: tetrahydrofuran
LDA Lithium diisopropylamide
EDC.HCI: 1 -ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride HOBt: Hydroxybenzotriazole
THP-O-NH2: N-(tetrahydro-2H-pyran-2-yloxy)amine
MeOH: methanol
NMM: N-Methylmorpholine
m-CPBA: m-chloroperbenzoic acid
DCM: dichloromethane
DMSO: dimethylsulfoxide
Synthetic route 1 a
Figure imgf000036_0001
1-13 Conditions: a. K2C03 (2 eq), CH3I (1.5 eq) in DMF, r.t.; b. LDA (1 .25 M) in THF, 1 hour at -78 °C; then, R-02 (2 eq), 1 hour at -78 °C; c. LiOH.H20 (10 eq) in THF/MeOH/H20 (3/3/2), r.t.; d. EDC.HCI (2 eq), HOBt (2 eq), THP-0-NH2 (1.5 eq), NMM (3 eq) in DMF, r.t.; e. R-04 (1.5 - 3 eq), Cs2C03 (2 - 3 eq), 12 hours at 90 °C; e'. m-CPBA (4 eq) in DCM, r.t. (Q=S to Q'=S02); f.- HCI / Dioxane, r.t. for 1 hour.
In the scheme above Q is O or S, Q' is Q or SO2 and Cy is phenyl or a 5- to 6- membered heteroaryl and can be optionally subtituted.
Preparation of intermediate 1-01 a: 8-tert-butoxycarbonyl-8- azaspiro[4.51decane-3-carboxylic acid methyl ester
To a solution of 8-tert-butoxycarbonyl-8-azaspiro[4.5]decane-3-carboxylic acid (20 g, 0.071 mol), commercially available from Wuxi Apptec, in DMF (200ml_) was added K2CO3 (18.59 g, 2 eq), then compound CH3I (14.34 g, 1 .5 eq) was added dropwise. The reaction mixture was stirred at room
temperature for 2 h. After TLC (PE/AE 5:1 ) showed the starting material was consumed, the mixture was quenched with water and extracted with EtOAc, the organic layer was washed with brine, dried over anhydrous Na2SO4, concentrated to give the crude compound 8-tert-butoxycarbonyl-8- azaspiro[4.5]decane-3-carboxylic acid methyl ester (20.38 g, 97.09%) as a pale yellow oil which was used for the next step without further purification. ESI-MS (M+1 ): 298 calc. for Ci6H27NO4.
Preparation of reagent R-02a: 4-Fluorobenzenesulfonyl fluoride
To a solution of compound 4-Fluorobenzenesulfonyl chloride (50 g, 0.256 mol) in ACN (500 ml_) was added KF (74.36 g, 5 eq) and 18-crown-6 (2 g) at r.t., then the mixture was stirred at r.t. overnight. The mixture was detected by LC-MS, then saturated aqueous NaHCO3 was added and the mixture was extracted with EtOAc, the organic layer was washed with saturated aqueous NaHCO3, brine, dried over anhydrous Na2SO4, concentrated to give the crude compound 4-Fluorobenzenesulfonyl fluoride (46.70 g) as pale yellow oil which was used for the next step without further purification. ESI-MS (M+1 ): 179 calc. for C6H4F2O2S.
Preparation of intermediate l-02a: 8-tert-butoxycarbonyl-3-(4- fluorophenylsulfonyl)-8-azaspiro[4.51decane-3-carboxylic acid methyl ester To a solution of compound 1-01 a (18.38 g, 0.062 mol) in THF (200ml_) was added LDA (102 ml_, 1 .25 M) at -78°C. After stirring at -78 °C for 1 h, the compound 4-Fluorobenzenesulfonyl fluoride (22.23 g, 2 eq) was added to the solution, the reaction was stirred at -78 °C for 1 hour, and then the mixture was stirred at r.t. overnight. After TLC (PE/AE 5:1 ) showed the starting material was consumed, the mixture was quenched with aqueous NH4CI and extracted with EtOAc, the organic layer was washed with brine, dried over anhydrous Na2SO4, concentrated to give the crude product which was purified by column chromatography (eluting with EA PE= 100:1 to 10: 1 ) to give pure compound l-02a (18.94 g, 67.03%) as a pale yellow oil. ESI-MS (M+1 ): 456 calc. for C22H3oFNO6S.
Preparation of intermediate l-03a: 8-tert-butoxycarbonyl-3-(4- fluorophenylsulfonyl)-8-azaspiro[4.51decane-3-carboxylic acid
To a solution of compound l-02a (10 g, 0.022 mol) in THF/MeOH/H2O (3/3/2, 80ml_) was added LiOH H2O (9.23 g, 10 eq). The resulting mixture was stirred at r.t. for 4 h, after TLC (PE/AE 5:1 ) showed the staring materials were consumed completely, then the mixture was diluted with water and adjusted pH to 2-3 with 1 N HCI and the mixture was extracted with EtOAc, washed with brine, dried over anhydrous Na2SO4, concentrated to give the crude product l-03a (9.60 g, 98.97%) as a pale yellow oil which was used in the next step. ESI-MS (M+1 ): 442 calc. for C2iH28FNO6S. Preparation of intermediate l-04a: 8-tert-butoxycarbonyl-3-(4- fluorophenylsulfonyl)-N-(tetrahvdro-2H-pyran-2-yloxy)-8-azaspiro[4.51decane- 3-carboxamide
To a solution of compound l-03a (9.60 g, 0.022 mol) in DMF (100 mL) was added EDC.HCI (8.40 g, 2 eq), HOBt (5.85 g, 2 eq), THP-O-NH2 (3.86 g, 1 .5 eq), NMM (6.67 g, 3 eq) at r.t., then the mixture was stirred at room
temperature overnight. The mixture was quenched with aqueous water and extracted with EtOAc, the organic layer was washed with brine, dried over anhydrous Na2SO4, concentrated to give the crude product which was purified by column chromatography (eluting with EA/PE = 50:1 to 2:1 ) to give pure compound l-04a (9.0 g, 75.76%) as a pale yellow solid. ESI-MS (M+1 ): 541 calc. for C26H37FN2O7S.
Preparation of intermediate l-05a: 8-tert-butoxycarbonyl-3-[4-(4- methoxyphenoxy)phenylsulfonyl1-N-(tetrahvdro-2H-pyran-2-yloxy)-8- azaspiro[4.51decane-3-carboxamide
To a solution of compound l-04a (200 mg, 0.37 mmol) in DMF (5 mL) was added p-methoxyphenol (69 mg, 1 .5 eq) and Cs2CO3 (241 mg, 2eq), then the reaction mixture was stirred at 90 °C overnight. After TLC (PE/AE 2:1 ) showed the starting material was consumed, the mixture was quenched with water and extracted with EtOAc, the organic layer was washed with brine, dried over anhydrous Na2SO4, concentrated to give the crude compound which was purified by Prep-TLC (PE/AE 2:1 ) to give pure compound l-05a (150 mg, 63.03%) as a pale yellow oil. ESI-MS (M+1 ): 645 calc. for
Figure imgf000039_0001
Preparation of compound 1 -03: N-hvdroxy-3-[4-(4- methoxyphenoxy)phenylsulfonyl1-8-azaspiro[4.51decane-3-carboxamide
A solution of intermediate l-05a (150 mg, 0.233 mmol) in HCI/dioxane (4 M/L, 5ml_) was stirred at r.t. for 1 h, then concentrated to give the crude product which was purified by prep-HPLC (General procedure, Method 1 ) to obtain pure compound 1 -03 (50 mg, 46.73 %) as a red solid. Rt: 2.63, ESI-MS (M+1 ): 461 calc. for C23H28N2O6S.
Preparation of compound 1 -59: 3-(4-fluorophenylsulfonyl)-N-hvdroxy-8- azaspiro[4.51decane-3-carboxamide
Following the same process as described for compound 1 -03 but starting from of intermediate l-04a, compound 1 -59 was obtained. Rt: 2.1 1 , ESI-MS (M+1 ): 356 calc. for Ci6H2iFN2O4S.
Following the same synthetic route 1 a, and using the same reagents as for compound 1 -03 unless otherwise indicated in the table below, the following compounds were obtained:
LC-MS
Example Rt [M+1]+ Starting materials
Method
2.51 and 4-tert-butoxy-carbonyl-4-azaspiro[5.5]-
1-01 475.2
2.71 1 undecane-9-carboxylic acid (1 )
1-06 2.74 489.2 1 4-(methoxycarbonyl) phenol (R-04)
1-07 2.29 475.2 1 4-hydroxybenzoic acid (R-04)
1-08 3.04 488.2 1 4-hydroxy-N-methylbenzamide (R-04)
1-09 3.30 487.3 1 4-tert-butylphenol (R-04)
1-12 3.23 462.2 1 6-methoxypyridin-3-ol (R-04)
1-13 3.06 488.2 1 N-(4-hydroxyphenyl)acetamide (R-04)
1-15 3.77 477.2 1 4-methoxybenzenethiol (R-04)
1-17 3.31 509.2 1 4-methoxybenzenethiol (R-04) via 1-13 LC-MS
Example Rt [M+1]+ Starting materials
Method
1-19 3.53 445.2 1 p-cresol (R-04)
1-22 2.58 488.3 1 4-((Dimethylamino)methyl) phenol (R-04)
1-21 2.50 499.2 1 p-trifluoromethylphenol (R-04)
6-tert-butoxycarbonyl-6-
1-43 3.38 433.2 azaspiro[2.5]octane-2-carboxylic acid (1) and p-methoxyphenol (R-04)
1-54 2.31 461.2 1 p-methoxyphenol (R-04)
1-55 2.18 461.2 1 p-methoxyphenol (R-04)
4-hydroxyphenyl-pyrrolidin-1-yl-methanone
1-62 3.28 528.2
(R-04)
1-63 3.06 502.3 1 4-hydroxy-N,N-dimethyl-benzamide (R-04)
1-64 2.82 474.2 1 4-hydroxybenzamide (R-04)
1-65 3.24 516.2 1 4-hydroxy-N-isopropyl-benzamide (R-04)
1-68 3.44 461.2 1 m-methoxyphenol (R-04)
1-69 3.31 461.2 1 o-methoxyphenol (R-04)
1-70 3.08 467.2 1 4-methoxycyclohexanol (R-04)
1-71 3.35 432.1 1 Phenol (R-04)
1-72 3.81 489.2 1 4-isopropoxyphenol (R-04)
1-73 3.90 515.2 1 4-(trifluoromethoxy)phenol (R-04)
1-74 3.49 489.3 1 Methyl 3-hydroxybenzoate (R-04)
1-75 3.13 475.2 1 3-hydroxybenzoic acid (R-04)
1-76 3.72 466.1 1 4-chlorophenol (R-04)
1-85 2.82 488.1 1 3-hydroxy-N-methylbenzamide (R-04)
1-88 1.89 493.2 1 2-fluoro-4-hydroxy benzoic acid (R-04)
2-fluoro-4-hydroxy-N-methyl benzamide
1-89 1.78 506.2
(R-04)
1-91 3.41 488.1 1 2-hydroxy-N-methylbenzamide (R-04)
3-fluoro-4-hydroxy-N-methylbenzamide
1-93 2.10 506.2
(R-04)
1-94 2.58 507.2 1 Methyl 3-fluoro-4-hydroxybenzoate (R-04)
1-95 3.48 493.2 1 3-fluoro-4-hydroxy-benzoic acid (R-04)
4-hydroxy-2-methoxy-N-methyl benzamide
1-110 2.69 518.3
(R-04)
1-114 2.78 549.2 1 3-chloro-4-(trifluoromethoxy)phenol (R-04)
1-115 2.60 515.2 1 3-(trifluoromethoxy)phenol (R-04)
1-117 2.37 497.1 1 4-(difluoromethoxy)phenol (R-04)
1-118 2.66 533.2 1 3-fluoro-4-(trifluoromethoxy)phenol (R-04)
1-123 3.39 515.1 1 4-(trifluoromethoxy)phenol (R-04)
1-126 1.71 488.1 1 4-hydroxy-N-methyl benzamide (R-04)
1-127 1.85 475.2 1 4-hydroxy benzoic acid (R-04) Synthetic route 2a
Figure imgf000041_0001
Conditions: c. R-04 (1.5 - 3 eq), Cs2C03 (2 - 3 eq), 12 hours at 90 °C; d. HCI in dioxane (4M), r.t.; e. K2C03(2 eq), R-08 (1.5 eq) in ACN, r.t.; f. LiOH.H20 (10 eq) in THF/MeOH/H20 (3/3/2), r.t.; g.
EDC.HCI (2 eq), HOBt (2 eq), THP-0-NH2 (1.5 eq), NMM (3 eq) in DMF, r.t.; h.- HCI / Dioxane, r.t. for 1 hour.
In the scheme above Q is O or S; X is halogen; R' is a hydrocarbon chain optionally substituted; and Cy is phenyl or a 5- to 6-membered heteroaryl and can be optionally subtituted. Preparation of intermediate l-06a: 8-tert-butoxycarbonyl-3-[4-(4- methoxyphenoxy)phenylsulfonyl1-8-azaspiro[4.51decane-3-carboxylic acid methyl ester
To a solution of compound l-02a (900mg, 1 .97 mmol) in DMF (120 ml_) was added Cs2CO3 (1 .92 g, 5.9 mol) and 4-methoxyphenol (490 mg, 3.95 mmol), the reaction mixture was stirred at 60 °C for 2h. The mixture was diluted with EtOAc and washed with brine, dried over anhydrous Na2SO4 and
concentrated to give the crude product which was purified by column chromatography (EA:PE=1 :30-1 :5) to give the compound l-06a (600 mg, 54.0%) as a pale yellow oil. ESI-MS (M+1 ): 560 calc. for C^Hb/NOsS. Preparation of intermediate l-07a: 3-[4-(4-methoxyphenoxy)phenylsulfonyl1-8- azaspiro[4.51decane-3-carboxylic acid methyl ester
A solution of compound l-06a (500 mg, 0.89 mmol) in HCI/dioxane (4M, 6 mL) was stirred at r.t. for 3h. The reaction mixture was concentrated to give the compound l-07a (500mg, crude). ESI-MS (M+1 ): 461 calc. for C24H29NO6S.
Preparation of intermediate l-08a: 3-[4-(4-methoxyphenoxy)phenylsulfonyl1-8- methyl-8-azaspiro[4.51decane-3-carboxylic acid methyl ester
To a solution of intermediate l-07a (181 mg, 0.39 mmol) in CH3CN (10 mL) was added K2CO3 (107 mg, 0.78 mmol), Mel (28 mg, 0.19 mmol). The reaction mixture was stirred at room temperature for 30 min. After TLC showed that most of starting material was consumed, the mixture was diluted with EtOAc and washed with brine, dried over anhydrous Na2SO4 and concentrated to give the crude compound l-08a (1 10 mg, 59.7%) as a pale yellow oil. ESI-MS (M+1 ): 588 calc. for C^sNOeS.
Preparation of intermediate l-09a: 3-[4-(4-methoxyphenoxy)phenylsulfonyl1-8- methyl-8-azaspiro[4.51decane-3-carboxylic acid
To a solution of intermediate 1-08 (1 10 mg, 0.23 mmol) in THF/MeOH/H2O (3/3/2, 8 mL) was added LiOH H2O (195 mg, 4.6 mmol). The resulting mixture was refluxed overnight. After TLC showed that most of the staring materials were consumed completely, the mixture was diluted with water and adjusted pH to 2-3. The mixture was extract with EtOAc and washed with brine, dried over anhydrous Na2SO4 and concentrated to give the crude product l-09a (100 mg, 94.0%) as a pale yellow solid. ESI-MS (M+1 ): 460 calc. for
C24H2gNOeS.
Preparation of intermediate 1-10a: 3-[4-(4-methoxyphenoxy)phenylsulfonyl1-8- methyl-N-(tetrahvdro-2H-pyran-2-yloxy)-8-azaspiro[4.51decane-3- carboxamide
To a solution of intermediate l-09a (100 mg, 0.22 mmol) in DMF (10 mL) was added EDCI (84 mg, 0.44 mmol), HOBt (59 mg, 0.44 mmol), THP-O-NH2 (51 .5 mg, 0.44 mmol), NMM (66 mg, 0.66 mmol). The mixture was stirred at room temperature overnight. The mixture was diluted with EtOAc and washed with brine, dried over anhydrous Na2SO4 and concentrated to give the crude product which was purified by column chromatography (EA:PE=1 :50-1 :4) to give the compound 1-10a (78 mg, 64.0%) as a pale yellow solid. ESI-MS (M+1 ): 559 calc. for C29H38N2O7S.
Preparation of compound 1 -04: N-hvdroxy-3-[4-(4-methoxyphenoxy)- phenylsulfonyl1-8-methyl-8-azaspiro[4.51decane-3-carboxamide
A solution of intermediate 1-10a (78 mg, 0.14 mmol) in HCI/dioxane (4 M/L,10 ml_) was stirred at r.t. for 3 h, the reaction mixture was concentrated to give the crude product which was purified by prep-HPLC (General procedure, Method 1 ) to obtain pure compound 1 -04 (14.7 mg, 22.0%) as a pale yellow solid. Rt: 2.66, ESI-MS (M+1 ): 475 calc. for C24H30N2O6S.
Following the same synthetic route 2a, and using the same reagents as for compound 1 -04 unless otherwise indicated in the table below, the following compounds were obtained:
LC-MS
Example Rt [M+1]+ Starting materials
Method
2.59 and 4-tert-butoxy-carbonyl-4-azaspiro[5.5]-
1 -02 489.3
2.74 1 undecane-9-carboxylic acid (1 )
1 -28 2.56 517.2 1 1 -chloropropan-2-one (R-08)
1 -29 2.52 519.2 1 tert-butyl 2-bromoacetate (R-08)
1 -30 2.96 557.2 1 3,3,3-Trifluoropropyl Bromide (R-08)
1 -31 2.80 503.2 1 1 -Bromopropane (R-08)
1 -32 2.79 501 .3 1 Cyclopropyl bromide (R-08)
1 -33 3.09 518.2 1 Methylcarbamic chloride (R-08)
1 -34 2.39 537.3 1 lodobenzene (R-08)
1 -35 2.26 538.3 1 2-Bromopyridine (R-08)
1 -36 3.12 551 .3 1 Benzyl bromide (R-08)
1 -37 2.52 552.3 1 4-(Bromomethyl) pyridine (R-08)
1 -39 2.47 541 .2 1 4-(Bromomethyl)-1 H-imidazole (R-08)
1 -58 3.21 503.2 1 Acetyl chloride (R-08)
1 -124 3.53 555.2 1 Cyclopropyl bromide (R-08) Synthetic route 1 b
Figure imgf000044_0001
Conditions: e. R-05 (4 eq), NaH (0 - 4 eq), after 30 min at r.t., 12h at 90 °C; f.- HCI / Dioxane, r.t. for 1 hour
In the scheme above R is a hydrocarbon chain which can be optionally subtituted.
Preparation of intermediate 1-1 1 a: 8-tert-butoxycarbonyl-3-(4-[2-(piperidin-1 - yl)ethoxylphenylsulfonyl)-N-(tetrahvdro-2H-pyran-2-yloxy)-8- azaspiro[4.51decane-3-carboxamide
To a solution of 2-(piperidin-1 -yl)ethanol (191 mg, 4 eq) in DMF (5ml_) was added NaH (36 mg, 4 eq), then the mixture was stirred at r.t. for 0.5h, intermediate l-04a (200 mg, 0.37 mmol) was added, then the reaction mixture was stirred at 90 °C overnight. After TLC (PE/AE 2:1 ) showed the starting material was consumed, the mixture was quenched with water and extracted with EtOAc, the organic layer was washed with brine, dried over anhydrous Na2SO4, concentrated to give the crude compound which was purified by Prep-TLC (PE/AE = 2:1 ) to give pure intermediate 1-1 1 a (184 mg, 76.67%) as a pale yellow oil. ESI-MS (M+1 ): 650 calc. for C33H51 N3O8S.
Preparation of compound 1 -1 1 : N-hvdroxy-3-(4-[2-(piperidin-1 - yl)ethoxylphenylsulfonyl)-8-azaspiro[4.51decane-3-carboxamide
A solution of intermediate 1-1 1 a (180 mg, 0.277 mmol) in HCI/dioxane (5ml_) was stirred at r.t. for 1 h, then concentrated to give the crude product by prep-
HPLC (General procedure, Method 1 ) to obtain pure compound 1 -1 1 (58.40 mg, 45.27%) as a pale yellow solid. Rt: 2.14. ESI-MS (M+1 ): 466 calc. for
Figure imgf000044_0002
Following the same synthetic route 1 b, and using the same reagents as for compound 1 -1 1 unless otherwise indicated in the table below, the following compounds were obtained: LC-MS
Example Rt [M+1]+ Starting materials
Method
1-05 2.80 425.2 1 3-methylbutan-1-ol (R-05)
1-10 2.50 466.2 1 1 -(2-hyd roxyethyl)pyrrol id i n-2-one (R-05)
1-18 3.67 437.2 1 Cyclohexanol (R-05)
1-20 3.50 445.2 1 Benzyl alcohol (R-05)
1-92 2.65 451 .2 1 Cyclohexylmethanol (R-05)
4-(trifluoromethoxy)phenyl methanol (R-
1-99 3.30 529.0
05)
1-100 2.96 446.1 1 Pyridin-4-yl methanol (R-05)
4-(hydroxymethyl)-N-methyl-benzamide
1-108 1.98 502.3
(R-05)
1-1 12 2.57 435.1 1 1 H-Pyrazol-4-ylmethanol (R-05)
1-1 13 2.56 369.1 1 Sodium methoxyde (R-05)
4-(trifluoromethoxy)phenyl methanol (R-
1-130 2.97 529.2
05)
Figure imgf000045_0001
Conditions: e. R-07 (10 eq), 12h at 100 °C; f.- HCI / Dioxane, r.t. for 1 hour In the scheme above R' is R or H, and R is a hydrocarbon chain which can be optionally subtituted.
Preparation of intermediate 1-12a: 8-tert-butoxycarbonyl-3-[4-(4- methoxypiperidin-1 -yl)phenylsulfonyl1-N-(tetrahvdro-2H-pyran-2-yloxy)-8- azaspiro[4.51decane-3-carboxamide
A solution of intermediate l-04a (100 mg, 0.185 mmol) and 4- methoxypiperidine (213 mg, 10 eq) was stirred at 100 °C overnight. After TLC (PE/AE 2:1 ) showed the starting material was consumed, the mixture was quenched with water and extracted with EtOAc, the organic layer was washed with brine, dried over anhydrous Na2SO4, concentrated to give the crude compound which was purified by prep-TLC (PE/AE 1 : 1 ) to give pure intermediate 1-12a (50 mg, 42.74%) as a pale yellow oil. ESI-MS (M+1 ): 636 calc. for C32H49N3O8S. Preparation of compound 1 -16: N-hvdroxy-3-[4-(4-methoxypiperidin-1 - yl)phenylsulfonyl1-8-azaspiro[4.51decane-3-carboxamide
A solution of intermediate 1-12a (50 mg, 0.079 mmol) in HCI/dioxane (3 mL) was stirred at r.t. for 1 h, then concentrated to give the crude product which was purified by prep-HPLC (General procedure, Method 1 ) to obtain pure compound 1 -16 (25.0 mg, 69.44%) as a pale yellow solid. ESI-MS (M+1 ): 452 calc. for C22H33N3O5S.
Following the same synthetic route 1 c, and using the same reagents as for compound 1 -16 unless otherwise indicated in the table below, the following compounds were obtained:
LC-MS
Example Rt [M+1]+ Starting materials
Method
1 -14 3.57 436.3 1 Cyclohexylamine (R-07)
1 -61 1 .99 354.2 1 Benzylamine (R-07)
1 -67 2.51 444.3 1 Benzylamine (R-07)
1 -77 3.00 466.3 1 4-methoxycyclohexanamine (R-07)
1 -78 1 .83 436.3 1 Cyclohexylnamine (R-07)
1 -79 2.15 436.3 1 Cyclohexylnamine (R-07)
1 -82 2.02 450.1 1 Cyclohexylmethanamine (R-07)
4-amino-N-methyl-
1 -83 2.94 493.2 1 cyclohexanecarboxamide (R-07)
1 -84 2.40 480.2 1 4-aminocyclohexanecarboxylic acid (R-07)
1 -96 1 .47 438.1 1 Tetrahydropyran-4-amine (R-07)
1 -97 2.08 422.1 1 Cyclopentanamine (R-07)
4-(trifluoromethoxy)phenyl-methanamine
1 -103 2.58 528.2 1 (R-07)
4-(aminomethyl)-N-methyl benzamide (R-
1 -104 1 .56 501 .2 1 07)
1 -105 3.30 502.2 1 Methyl 4-(aminomethyl) benzoate (R-07)
1 -106 1 .75 488.2 1 4-(aminomethyl) benzoic acid (R-07)
1 -109 3.65 472.2 1 4-4-difluorocyclohexylamine (R-07)
(4-4-difluorocyclohexyl)methanamine (R-
1 -125 2.20 486.2
07) Synthetic route 1 d
Figure imgf000047_0001
Conditions: e. Benzyl alcohol (4 eq), NaH (4 eq), after 30 min at r.t., 12 h at 90 °C; f.- Pd/C (0.5 g) at H2 atmosphere, r.t. for 1 hour; g.- R-09 (1.2 eq), Et3N (3 eq) in DCM, r.t. overnight; h.- HCI / Dioxane, r.t. for 1 hour
In the scheme above R is a hydrocarbon chain optionally substituted, a carbocyclic or heteroaliphatic ring optionally substituted, a phenyl or 5- to 6- membered heteroaryl optionally substituted. Preparation of intermediate 1-13a: 3-[4-(benzyloxy)phenylsulfonyl1-8-tert- butoxycarbonyl-N-(tetrahvdro-2H-pyran-2-yloxy)-8-azaspiro[4.51decane-3- carboxamide
To a solution of phenylmethanol (500 mg, 5eq) in THF (5ml_) was added NaH (1 10 mg, 5eq), then the mixture was stirred at r.t for 0.5 h, intermediate l-04a (500 mg, 0.926 mmol) was added, then the reaction mixture was stirred at 80 °C overnight. After TLC (PE/AE 2:1 ) showed the starting material was consumed, the mixture was quenched with water and extracted with EtOAc, the organic layer was washed with brine, dried over anhydrous Na2SO4, concentrated to give the crude compound which was purified by prep-TLC (PE/AE 2:1 ) to give pure intermediate 1-13a (380 mg, 65.29%) as a pale yellow oil. ESI-MS (M+1 ): 629 calc. for C33H44N2O8S.
Preparation of intermediate 1-14a: 8-tert-butoxycarbonyl-3-[4- hvdroxyphenylsulfonyl1-N-(tetrahydro-2H-pyran-2-yloxy)-8- azaspiro[4.51decane-3-carboxamide To a solution of intermediate 1-13a (380 mg, 0.605 mmol) in MeOH (20 ml_) was added Pd/C (0.5 g) at room temperature at H2 atmosphere, then the mixture was stirred at room temperature for 1 h until TLC (PE/AE 2:1 ) showed the starting material was consumed, then filtered and the filtrate was concentrated to give the crude product 1-14a (180 mg, 55.38%) which was used for the next step without further purification. ESI-MS (M+1 ): 539 calc. for
Figure imgf000048_0001
Preparation of intermediate 1-15a: 8-tert-butoxycarbonyl-3-[4-(4- methoxybenzoyloxy)phenylsulfonyl1-N-(tetrahvdro-2H-pyran-2-yloxy)-8- azaspiro[4.51decane-3-carboxamide
To a solution of intermediate 1-14a (60 mg, 0.151 mmol) and Et3N (46 mg, 3eq) in DCM (5ml_) was added 4-methoxybenzoyl chloride (31 mg, 1 .2eq) at room temperature, then the mixture was stirred at r.t overnight. After TLC (PE/AE 2:1 ) showed the starting material was consumed, the mixture was extracted with DCM, the organic layer was washed with brine, dried over anhydrous Na2SO4, concentrated to give the crude compound which was purified by prep-TLC (PE/AE 2:1 ) to give pure compound 1-15a (38 mg, 37.62%) as a pale yellow oil. ESI-MS (M+1 ): 673 calc. for C34H44N2Oi0S.
Preparation of compound 1 -26: N-hvdroxy-3-[4-(4- methoxybenzoyloxy)phenylsulfonyl1-8-azaspiro[4.51decane-3-carboxamide A solution of intermediate 1-15a (38 mg, 0.057 mmol) in HCI/dioxane (5 mL) was stirred at r.t for 1 h, then concentrated to give the crude product by prep- HPLC (General procedure, Method 1 ) to obtained pure compound I-26 (22 mg, 78.57 %) as a pale yellow solid. Rt: 2.71 , ESI-MS (M+1 ): 489 calc. for
Figure imgf000048_0002
Preparation of compound 1 -60: N-hvdroxy-3-(4-hvdroxyphenylsulfonyl)-8- azaspiro[4.51decane-3-carboxamide
Following the same process as described for compound 1 -26 but starting from of intermediate 1-14a, compound 1 -60 was obtained. Rt: 2.1 1 , ESI-MS (M+1 ): 354 calc. for Ci6H22N2O5S. Following the same synthetic route 1 d, and using the same reagents as for compound 1 -26 unless otherwise indicated in the table below, the following compounds were obtained: LC-MS
Example Rt [M+1]+ Starting materials
Method
1-86 3.12 503.1 1 4-chlorocarbonyl benzoic acid (R-09)
4-(methylcarbamoyl)benzoyl chloride (R-
1-87 2.79 516.2
09)
1-98 2.47 465.3 1 Cyclohexanecarbonyl chloride (R-09)
3-fluoro-4-(methylcarbamoyl)benzoyl
1-101 1.84 534.2
chloride (R-09)
4-(trifluoromethoxy)benzoyl chloride (R-
1-102 2.75 543.1
09)
1-1 19 2.20 489.0 1 4-methoxybenzoyl chloride (R-09)
4-(trifluoromethoxy)benzoyl chloride (R-
1-120 2.61 543.1
09)
Synthetic route 1 e
Figure imgf000049_0001
IT 1-19 1-18
Conditions: a. LDA (2.0 M) in THF, 1 hour at -78 °C; then, R-10 (1.2 eq), 1 hour at -78 °C; b.
Pd(dppf)CI2 (0.1 eq) and Na2C03 (2 eq) in dioxane/ H20, r.t. overnight; c. LiOH.H20 (10 eq) in THF/MeOH/H20 (3/3/2), r.t.; d. EDC.HCI (2 eq), HOBt (2 eq), THP-0-NH2 (1.5 eq), NMM (3 eq) in DMF, r.t.; e.- HCI / Dioxane, r.t. for 1 hour.
In the scheme above B is boronic acids, boronate esters or trifluoroborate salts and R is phenyl or a 5- to 6- membered heteroaryl or a 3- to 7- heterocyclic or carbocyclic aliphatic ring or a hydrocarbon chain which can be optionally substituted. Preparation of reagent R-10a: 4-bromobenzenesulfonyl fluoride
To a solution of 4-bromobenzenesulfonyl chloride (2.29 g, 9 mmol) in CH3CN (45 mL) was added KF (2.1 g, 36 mmol) and 18-crown-6 (0.5 g), then the mixture was stirred at room temperature overnight. The mixture was
quenched with aqueous water and extracted with EtOAc, the organic layer was washed with brine, dried over anhydrous Na2SO4, concentrated to give the crude product which was purified by column to give the reagent R-10a (1 .6 g, 74.4%) as a pale yellow solid. ESI-MS (M+1 ): 239.2 calc. for
C6H4BrFO2S.
Preparation of intermediate 1-16a: 8-tert-butoxycarbonyl-3-(4- bromophenylsulfonyl)-8-azaspiro[4.51decane-3-carboxylic acid methyl ester To a solution of compound 1-01 a (230 mg, 0.77 mmol) in THF (20 mL) was added LDA (0.6 mL, 2.0 M, 1 .2 mmol) at -78 °C. After stirring at -78 °C for 1 hour, the reagent R-10a (214 mg, 0.9 mmol) was added to the solution, the reaction was stirred at -78 °C for 1 hour, and then the mixture was stirred at r.t overnight. After TLC (PE/AE 5:1 ) showed the starting material was consumed, the mixture was quenched with aqueous NH4CI and extracted with EtOAc, the organic layer was washed with brine, dried over anhydrous Na2SO4, concentrated to give the crude product which was purified by column chromatography (eluting with EA PE= 100:1 to 10: 1 ) to give pure compound 1-16a (150 mg, 38%) as a pale yellow oil. ESI-MS (M-55): 459.9 calc. for C22H30BrNO6S. Preparation of intermediate 1-17a: 8-tert-butoxycarbonyl-3-(4-(4- methoxyphenyl)phenylsulfonyl)-8-azaspiro[4.51decane-3-carboxylic acid methyl ester
To a suspension of compound 1-16a (50 mg, 0.1 mmol) and the commercially available 4-methoxyphenyl boronic acid (20 mg, 0.13 mmol, R-1 1 ), in dioxane (10 mL)/ H2O (2 mL) was added Pd(dppf)CI2 (10 mg) and Na2CO3 (22 mg, 0.2 mmol). The mixture was stirred at reflux under N2 overnight. The resulting mixture was cooled to room temperature and water was added. The organic layer was separated and the aqueous layer was extracted 3 times with EtOAc. The combined organic layer was washed with brine, dried over sodium sulfate and concentrated under reduced pressure and purified by prep-TLC to afford the desired product 1-17a (52 mg, 95%). ESI-MS (M-55): 488.0 calc. for C29H37NO7S. Preparation of intermediate 1-18a: 8-tert-butoxycarbonyl-3-(4-(4- methoxyphenyl)phenylsulfonyl)-8-azaspiro[4.51decane-3-carboxylic acid
To a solution of compound 1-17a (52 mg, 0.096 mmol) in THF/MeOH/H2O (3/3/2, 8 mL) was added LiOH.H2O (42 mg, 10 eq). The resulting mixture was stirred at r.t. for 4 h, after TLC (PE/AE 5:1 ) showed the staring materials were consumed completely, then the mixture was diluted with water and adjusted pH to 3-4 with 1 N HCI and the mixture was extracted with EtOAc, washed with brine, dried over anhydrous Na2SO4, concentrated to give the crude product 1-18a (50 mg, ~100%) as a pale yellow oil which was used directly in the next step. ESI-MS (M-55): 473.9; calc. for C28H35NO7S
Preparation of intermediate 1-19a: 8-tert-butoxycarbonyl-3-(4-(4- methoxyphenyl)phenylsulfonyl)-N-(tetrahvdro-2H-pyran-2-yloxy)-8- azaspiro[4.51decane-3-carboxamide
To a solution of compound 1-18a (50 mg, 0.095 mmol) in DMF (15 mL) was added EDC.HCI (40 mg, 0.2 mmol), HOBt (27 mg, 0.2 mmol), THPONH2 (24 mg, 0.2 mmol), NMM (40 mg, 0.4 mmol), then the mixture was stirred at room temperature overnight. The mixture was quenched with aqueous water and extracted with EtOAc, the organic layer was washed with brine, dried over anhydrous Na2SO4, concentrated to give the crude product which was purified by prep-TLC to give the compound 1-19a (45 mg, 72%) as a pale yellow solid. ESI MS (M-139): 488.9; calc. for C33H44N2O8S. Preparation of compound 1 -41 : N-hvdroxy-3-(4-(4- methoxyphenyl)phenylsulfonyl)-8-azaspiro[4.51decane-3-carboxamide
A solution of intermediate 1-19a (45 mg, 0.072 mmol) in HCI/dioxane (10 mL, 2 N) was stirred at r.t for 2 h, then concentrated to give the crude product which was purified by prep HPLC twice (General procedure, Method 1 ) to obtained pure compound 1 -41 (6.6 mg, 16.8 %) as a yellow solid. ESI-MS
(M+1 ): 445.2 calc. for C23H28N2O5S. Rt is 2.01 .
Following the same synthetic route 1 e, and using the same reagents as for compound 1 -41 unless otherwise indicated in the table below, the following compounds were obtained: LC-MS
Example Rt [M+1]+ Starting materials
Method
1 ,3,2-Dioxaborolane, 4,4,5,5-tetramethyl-
1 -66 2.26 429.2 1
2-(phenylmethyl)- (R-1 1 )
Synthetic route 1f
Figure imgf000052_0001
1-12 I -20
Conditions: a. R-09 (1.2 eq), Et3N (2 eq) in DCM, r.t. overnight; b. HCI / Dioxane, r.t. for 1 hour.
In the scheme above R and R' are hydrogen, X is a halogen and R" is phenyl or a 5- to 6- membered heteroaryl or a 3- to 7- heterocyclic or carbocyclic aliphatic ring or a hydrocarbon chain which can be optionally substituted.
Preparation of intermediate l-20a: 8-tert-butoxycarbonyl-3-(4-
(cvclohexanecarbonylamino)phenylsulfonyl)-N-(tetrahvdro-2H-pyran-2-yloxy)-
8-azaspiro[4.51decane-3-carboxamide
To a solution of compound 1-12c (80 mg, 0.149 mmol), intermediate obtained to synthesize 1 -61 (following synthetic route 1 c), in DCM (3 ml_) was added cyclohexanecarbonyl chloride (26.2mg, 1 .2eq) and Et3N (30.1 mg, 0.298 mmol), then the reaction mixture was stirred at r.t overnight. After TLC (PE/AE 5:1 ) showed the starting material was consumed, the mixture was quenched with water and extracted with EtOAc, the organic layer was washed with brine, dried over anhydrous Na2SO4, concentrated to give the crude compound which was purified by Prep-TLC (PE/AE 5:1 ) to give pure intermediate l-20a (65 mg, 67.50%) as a white solid. ESI-MS (M+1 ): 648.3 calc. for C33H49N3O8S. Preparation of compound 1 -81 : N-hvdroxy-3-(4-
(cvclohexanecarbonylamino)phenylsulfonyl)-8-azaspiro[4.51decane-3- carboxamide
A solution of compound l-20a (65 mg, 0.1 mmol) in HCI/dioxane (3 ml_) was stirred at r.t for 1 h, then concentrated to give the crude product which was purified by prep-HPLC (General procedure, Method 1 ) to obtained pure compound 1 -81 (22.0 mg, 45.74 %) as a yellow solid. ESI-MS (M+1 ): 464.2 calc. for C23H33N3O5S. Rt is 1 .89. Synthetic route 3a
Figure imgf000053_0001
Ih
Conditions: a. LDA (1.20 M) in THF, 1 hour at -78 °C; then, R-12 (1.2 eq), 1 hour at -78 °C; b. R-13 (3 - 10 eq), NaH (0 - 3 eq) in THF, r.t. - reflux overnight; c. Cul (2 eq), MeOH/MeONa (25%, 20 eq) in DMF, 20 minutes at 1 10 °C; d. LiOH.H20 (10 eq) in THF/MeOH/H20 (3/3/2), r.t.; e. EDC.HCI (2 eq), HOBt (2 eq), THP-0-NH2 (1.5 eq), NMM (3 eq) in DMF, r.t.; f. HCI / Dioxane, r.t. for 1 hour.
In the scheme above Q is O or NH, R is a 3- to 7- heterocyclic or carbocyclic aliphatic ring or a hydrocarbon chain which can be optionally substituted and R' is halogen or alcoxy.
Preparation of reagent R-12a: 3-chloro-4-fluorobenzenesulfonyl fluoride To a solution of the commercially available 3-chloro-4-fluorobenzenesulfonyl chloride (7 g, 30 mmol) in CH3CN (40 mL) was added KF (7 g, 120 mmol) and 18-crown-6 (0.5 g), then the mixture was stirred at room temperature overnight. The mixture was quenched with aqueous water and extracted with EtOAc, the organic layer was washed with brine, dried over anhydrous
Na2SO4, concentrated to give the crude product which was purified by column to give the reagent R-12a (4.2 g, 95.5%) as a pale yellow solid. ESI-MS (M+1 ): 213.2 calc. for C6H3CIF2O2S Preparation of intermediate 1-21 a: 8-tert-butoxycarbonyl-3-(3-chloro-4- fluorophenylsulfonyl)-8-azaspiro[4.51decane-3-carboxylic acid methyl ester To a solution of compound 1-01 a (743 mg, 2.5 mmol) in THF (20 mL) was added LDA (4.2 mL, 1 .2 M, 5 mmol) at -78 °C. After stirring at-78 °C for 1 hour, the reagent R-12a (640 g, 3.0 mmol) was added to the solution, the reaction was stirred at -78 °C for 1 hour, and then the mixture was stirred at r.t overnight. After TLC (PE/AE 5:1 ) showed the starting material was consumed, the mixture was quenched with aqueous NH4CI and extracted with EtOAc, the organic layer was washed with brine, dried over anhydrous Na2SO4, concentrated to give the crude product which was purified by column chromatography (eluting with EA PE= 100:1 to 10: 1 ) to give pure
intermediate 1-21 a (349 mg, 29%) as a pale yellow oil. ESI-MS (M-55): 434.1 calc. for C22H29CIFNO6S.
Preparation of intermediate l-22a: 8-tert-butoxycarbonyl-3-(3-chloro-4- (cvclohexylamino)phenylsulfonyl)-8-azaspiro[4.51decane-3-carboxylic acid methyl ester
The intermediate 1-21 a (240 mg, 0.51 mmol) was dissolved in
cyclohexylamine (505 mg, 5.1 mmol), R13a. The solution was stirred at 85 °C overnight. The mixture was quenched with aqueous water and extracted with EtOAc, the organic layer was washed with brine, dried over anhydrous
Na2SO4, concentrated to give the crude product which was purified by prep- TLC to give the compound l-22a (1 10 mg, 38%) as a pale yellow solid. ESI- MS (M-55): 513; calc. for C28H4iCIN2O6S. Preparation of intermediate l-24a: 8-tert-butoxycarbonyl-3-(3-methoxy-4- (cvclohexylamino)phenylsulfonyl)-8-azaspiro[4.51decane-3-carboxylic acid The compound l-22a (179 mg, 0.326 mmol), Cul (124.5 mg, 0.652 mmol) and MeOH/MeONa (25%, 1 .41 g, 6.52 mmol) was dissolved in DMF (10 ml_). The solution was stirred at 1 10 °C for 20 min. The mixture was quenched with aqueous water and extracted with EtOAc, the organic layer was washed with brine, dried over anhydrous Na2SO4, concentrated to give the crude product which was purified by prep-TLC to give the compound l-24a (160 mg, 88%) as a pale yellow solid. The hydrolysis of the methyl ester to the corresponding carboxylic acid occurred simultaneously together with the methoxylation; thus, compound l-23a was not isolated and the intermediate l-24a was directly obtained from l-22a. ESI-MS (M-55): 495.2; calc. for C28H42N2O7S.
Preparation of intermediate l-25a: 8-tert-butoxycarbonyl-3-(3-methoxy-4-
(cvclohexylamino)phenylsulfonyl)-N-(tetrahvdro-2H-pyran-2-yloxy)-8- azaspiro[4.51decane-3-carboxamide
To a solution of compound l-24a (160 mg, 0.29 mmol) in DMF (15 ml_) was added EDC.HCI (1 12 mg, 0.58 mmol), HOBt (79 mg, 0.58 mmol), THPONH2 (68 mg, 0.58 mmol) and NMM (88 mg, 0.87 mmol) at r.t, then the mixture was stirred at room temperature overnight. The mixture was quenched with aqueous water and extracted with EtOAc, the organic layer was washed with brine, dried over anhydrous Na2SO4, concentrated to give the crude product which was purified by prep-TLC to give the intermediate l-25a (100 mg, 53%) as a pale yellow solid. ESI-MS (M-139): 510.1 ; calc. for C33H51 N3O8S.
Preparation of compound 1 -90: N-hvdroxy-3-(3-methoxy-4- (cvclohexylamino)phenylsulfonyl)-8-azaspiro[4.51decane-3-carboxamide
A solution of compound l-25a (100 mg, 0.15 mmol) in HCI/dioxane (10 ml_) was stirred at r.t for 2 h, then concentrated to give the crude product which was purified by prep HPLC (General procedure, Method 1 ) to obtained pure compound 1 -90 (39.7 mg, 58 %) as a white solid. ESI-MS (M+1 ): 466.1 calc. for CZ3H35N3O5S. Rt is 2.48.
Preparation of compound 1 -1 16: N-hvdroxy-3-(3-chloro-4- fluorophenylsulfonyl)-8-azaspiro[4.51decane-3-carboxamide
Following the same process as described for compound 1 -90 but skipping the second and third step (b and c) from synthetic route described above (3a), compound 1 -1 16 was obtained. Rt is 2.96, ESI-MS (M+1 ): 391 .2 calc. for Ci6H2oCIFN2O4S. Following the same synthetic route 3a, and using the same reagents as for compound 1 -90 unless otherwise indicated in the table below, the following compounds were obtained:
Figure imgf000056_0002
Synthetic route 3b
Figure imgf000056_0001
li I-28
Conditions: a. R-04 (1.5 - 3 eq), Cs2C03 (2 - 3 eq), 12 hours at 90 °C; b. LiOH.H20 (10 eq) in THF/MeOH/H20 (3/3/2), r.t.; c. EDC.HCI (2 eq), HOBt (2 eq), THP-0-NH2 (1.5 eq), NMM (3 eq) in DMF, r.t.; d. HCI / Dioxane, r.t. for 1 hour.
In the scheme above Q is O or S and Cy is phenyl or a 5- to 6- membered heteroaryl and can be optionally substituted.
Preparation of intermediate l-26a: 8-tert-butoxycarbonyl-3-(3-chloro-4-(4- methoxyphenoxy)phenylsulfonyl)-8-azaspiro[4.51decane-3-carboxylic acid methyl ester
To a suspension of compound 1-21 a (420 mg, 0.86 mmol) and p- methoxyphenol (214 mg, 1 .72 mmol), in DMF (5 mL) was added Cs2CO3 (700 mg, 2.2 mmol). The mixture was stirred at reflux under N2 overnight. The resulting mixture was cooled to room temperature and water was added. The organic layer was separated and the aqueous layer was extracted 3 times with EtOAc. The combined organic layers were washed with brine, dried over sodium sulfate and concentrated under reduced pressure and purified by prep TLC to afford the desired product l-26a (380 mg, 75%). ESI-MS (M-55): 538.2 calc. for CzjHaeCINOeS.
Preparation of intermediate l-27a: 8-tert-butoxycarbonyl-3-(3-chloro-4-(4- methoxyphenoxy)phenylsulfonyl)-8-azaspiro[4.51decane-3-carboxylic acid To a solution of compound l-26a (310 mg, 0.52 mmol) in THF/MeOH/H2O (3/3/2, 16 mL) was added LiOH.H2O (225 mg, 10 eq). The resulting mixture was stirred at r.t. for 4 h, after TLC (PE/AE 5:1 ) showed the staring materials were consumed completely, then the mixture was diluted with water and adjusted pH to 3-4 with 1 N HCI and the mixture was extracted with EtOAc, washed with brine, dried over anhydrous Na2SO4, concentrated and purified by prep-TLC to afford the desired product l-27a (120 mg, 40%). ESI-MS (M- 55): 524.2; calc. for C28H34CINO8S. Preparation of intermediate l-28a: 8-tert-butoxycarbonyl-3-(3-chloro-4-(4- methoxyphenoxy)phenylsulfonyl)-N-(tetrahvdro-2H-pyran-2-yloxy)-8- azaspiro[4.51decane-3-carboxamide
To a solution of intermediate l-27a (120 mg, 0.21 mmol) in DMF (10 mL) was added EDC.HCI (81 mg, 0.42 mmol), HOBt (57 mg, 0.42 mmol), THPONH2 (50 mg, 0.42 mmol), NMM (64 mg, 0.63 mmol), then the mixture was stirred at room temperature overnight. The mixture was quenched with aqueous water and extracted with EtOAc, the organic layer was washed with brine, dried over anhydrous Na2SO4, concentrated to give the crude product l-28a (160 mg, -100%) as a pale yellow oil which was used directly in the next step. ESI- MS (M-139): 539.2; calc. for
Figure imgf000057_0001
Preparation of compound 1 -56: N-hvdroxy-3-(3-chloro-4-(4- methoxyphenoxy)phenylsulfonyl)-8-azaspiro[4.51decane-3-carboxamide
A solution of intermediate l-28a (160 mg, -0.21 mmol) in HCI/dioxane (10 mL, 1 N) was stirred at r.t for 2 h, then concentrated to give the crude product which was purified by prep-HPLC (General procedure, Method 1 ) to obtained pure compound 1 -56 (30.8 mg, 29.7 %) as a yellow solid. ESI-MS (M+1 ): 495.2 calc. for C23H27CIN2O6S. Rt is 2.66.
Following the same synthetic route 3b, and using the same reagents as for compound 1 -56 unless otherwise indicated in the table below, the following compounds were obtained:
Figure imgf000058_0002
Synthetic route 3c
Figure imgf000058_0001
1-25 1-29 Ij
Conditions: a. R-09 (1.2 eq), Et3N (4 eq) in DCM, r.t. overnight; b. HCI / Dioxane, r.t. for 1 hour.
In the scheme above Q is O or NH, R is hydrogen, X is a halogen and R' is phenyl or a 5- to 6- membered heteroaryl or a 3- to 7- heterocyclic or carbocyclic aliphatic ring or a hydrocarbon chain which can be optionally substituted.
Preparation of intermediate l-29a: 8-tert-butoxycarbonyl-3-(3-chloro-4-(4- (trifluoromethoxy)benzoyloxy)phenylsulfonyl)-N-(tetrahvdro-2H-pyran-2- yloxy)-8-azaspiro[4.51decane-3-carboxamide
To a solution of compound l-25e (1 15 mg, 0.2 mmol), intermediate obtained to synthesize 1 -121 (following synthetic route 3a), and Et3N (61 mg, 0.6 mmol) in DCM (10 ml_) was added dropwise the commercially available 4- (Trifluoromethoxy)benzoyl chloride, R-09, (0.3 mmol) in DCM (5 mL) with stirring at 0 °C under N2. The mixture solution was stirred at r.t overnight. The mixture solution was added water and extracted with EtOAc, the organic layer was washed with brine, dried over anhydrous Na2SO4, concentrated to give the crude product which was purified by prep-TLC to give the intermediate l-29a (40 mg, 26%) as a yellow solid. ESI-MS (M-139): 621 .1 ; calc. for
Figure imgf000059_0001
Preparation of compound 1 -128: N-hvdroxy-3-(3-chloro-4-(4- (trifluoromethoxy)benzoyloxy)phenylsulfonyl)-8-azaspiro[4.51decane-3- carboxamide
A solution of intermediate l-29a (40 mg, 0.053 mmol) in HCI/dioxane (15 ml_, 1 N) was stirred at r.t for 4 h, then concentrated to give the crude product which was purified by prep-HPLC (General procedure, Method 1 ) to obtained pure compound 1 -128 (1 1 .2 mg, 37 %) as a yellow solid. ESI-MS (M+1 ):
577.2 calc. for C24H24CI F3N2O7S. Rt is 2.96.
Following the same synthetic route 3c, and using the same reagents as for compound 1 -128 unless otherwise indicated in the table below, the following compounds were obtained:
Figure imgf000059_0002
Synthesized compounds are obtained as racemic mixtures. Corresponding isomers are purified by supercritical fluid chromatography (SFC) to obtain two enantiomers from each racemic compound.
Procedure for Supercritical Fluid Chromatography (SFC):
Details about the preparative SFC separation methods utilized are as follows: Method 1 :
- Instrument: Thar SFC Pre-80
- Column: ChiralPak AD-H, 250x30 mml.D.
- Mobile phase: A for CO2 and B for Ethanol(0.1 %ΝΗ3·Η2Ο)
- Gradient: B 50%
- Flow rate: 60 mL /min
- Back pressure: 100 bar
- Column temperature: 40 °C
- Wavelength: 220 nm
- Cycletime: -20 min - Sample preparation: Compound was dissolved in ethanol to -12 mg/mL
- Injection: 2.0 mL per injection.
Method 2:
- Instrument: Mg II preparative SFC
- Column: ChiralPak AD-H, 250x30mml.D.
- Mobile phase: A for CO2 and B for Methanol(0.1 %ΝΗ3·Η2Ο)
- Gradient: B 50%
- Flow rate: 40ml_ /min
- Back pressure: 100bar
- Column temperature: 38 °C
- Wavelength: 220 nm
- Cycletime: ~20min
- Sample preparation: Compound was dissolved in methanol to
~13mg/ml
- Injection: 3.5 mL per injection.
Method 3:
- Instrument: Mg II preparative SFC
- Column: ChiralPak AD-H, 250x30mml.D.
- Mobile phase: A for CO2 and B for IPA(0.1 %ΝΗ3·Η2Ο)
- Gradient: B 35%
- Flow rate: 55ml /min
- Back pressure: 100bar
- Column temperature: 38 °C
- Wavelength: 254 nm
- Cycletime: ~8 min
- Sample preparation: Compound was dissolved in methanol to ~ 5 mg/mL
- Injection: 1 .0 mL per injection.
After separation, the fractions were dried off via rotary evaporator at bath temperature 40 °C to get the desired isomers. Then, after concentration, enantiomeric excess (e.e.) for both enantiomers were tested under the analytical separation methods described below: Method 1 :
- Instrument: SHIMADZU-20A UFLC
- Column: ChiralPak AD-3, 1 00 4.6 mm
- Mobile phase: A for Hexane (0.1 %IPAm) and B for
Ethanol(0.05%IPAm)
- Gradient: B 40%
- Flow rate: 1 .0 mL/min
- Column temperature: 30 °C
- Wavelength: 220 nm
Method 2:
- Instrument: Thar analytical SFC
- Column: ChiralPak AD-H, 250x4.6mm
- Mobile phase: A for CO2 and B for Methanol (0.05%DEA)
- Gradient: B 50%
- Flow rate: 2.0mL/min
- Back pressure: 100bar
- Column temperature: 35 °C
- Wavelength: 220nm
Method 3:
- Instrument: Thar analytical SFC
- Column: ChiralPak AD-H, 250x4.6mm
- Mobile phase: A for CO2 and B for IPA (0.1 %Ethanolamine)
- Gradient: B 30%
- Flow rate: 2.4 mL/min
- Back pressure: 1 00 bar
- Column temperature: 35 °C
- Wavelength: 220 nm
After concentration, as previously reported, LC-MS was used to test the purity for both enantiomers. Then, optical rotation was measured at 20 °C using the Autopol V polarimeter. The following isomers were obtained after chiral separation: Chiral Optical
Starting SFC e.e. by chiral
Example Rt (SFC) HPLC Rotation material Method HPLC (%)
Method [«]D
1-107 1-131 2.42 1 100 1 + 19.59°
1-107 1-132 3.15 1 95.3 1 - 19.71°
1-08 1-133 2.98 2 100 2 + 16.66°
1-08 1-134 5.88 2 100 2 - 17.10°
1-55 1-135 4.44 1 100 1 —
1-55 1-136 6.08 1 97.9 1 —
1-79 1-137 5.85 3 96.1 3 —
1-79 1-138 7.63 3 97.7 3 —
Antifibrinolytic effect on whole blood clot formation and lysis
Thromboelastometry is a viscoelastometric method for haemostasis testing in whole blood. TEM® measures the interactions of coagulation factors, inhibitors and cellular components during the phases of clotting and
subsequent lysis over time. The rheological conditions of this method mimic the sluggish flow of blood in veins. Detection method:
Blood samples were obtained between 8-9 a.m. from healthy volunteers and mice in tubes containing citrate solution (0.129 M sodium citrate, Vacutainer BD) and ROTEM tests were performed following the technical details of the ROTEM® analyser (Pentapharm GmbH, Munich, Germany). A modification of in-tem test as described below was used for the examination of antifibrinolytic effects of tested compounds and its interaction with platelets in citrated blood. Kits: START-TEM assay as a recalcification reagent (ref#503-01 ) and IN-TEM assay for activation of intrinsic coagulation pathway (ref #503-02).
Procedure:
In a pre-warmed cuvette and holder 1 μΙ_ of Tissue plasminogen activator (tPA) (150,000 U/mL, Actylise), 20 μΙ_ of start-tern reagent (CaCI2), 20 μΙ_ of in-tem reagent (activators of coagulation system), 3 μΙ_ of DMSO (control) or tested compounds (CMs) in DMSO and 300 μΙ_ of citrated blood pre-warmed were pipetted. The cup holder containing the sample mixture was placed immediately on the appropriate channel. The measurement was recorded for 60 min to allow clot formation and lysis. Table 3 shows the results in human blood as effective concentration to delay lysis time by 50% (EC50LT); where, EC50LT≥ 25 μΜ (+), 1 0 μΜ < EC50LT < 25 μΜ (++), 1 μΜ < ECSOLT < 1 0 μΜ (+++), EC50LT < 1 μΜ (++++) at all the assayed concentrations (1 000-0.2 μΜ).
Example ECSOLT Example ECSOLT
TXA +++ 1-58 ++++
1-01 ++++ 1-59 +
1-02 ++ 1-60 +
1-03 ++++ 1-21 ++++
1-04 ++++ 1-41 +
1-05 ++ 1-43 ++++
1-06 + 1-54 +++
1-07 ++++ 1-55 ++++
1-08 ++++ 1-56 ++++
1-09 ++++ 1-61 +
1-10 ++++ 1-62 ++++
1-1 1 ++++ 1-63 +++
1-12 ++ 1-64 +
1-13 ++++ 1-65 +++
1-14 ++++ 1-66 +
1-15 + 1-67 ++++
1-16 +++ 1-68 +++
1-17 +++ 1-69 +++
1-18 +++ 1-70 ++++
1-19 ++++ 1-71 +++
1-20 ++++ 1-72 +++
1-22 +++ 1-73 ++++
1-26 ++++ 1-74 +++
1-28 +++ 1-75 +++
1-29 +++ 1-76 +
1-30 ++++ 1-77 +++
1-31 ++++ 1-78 +
1-32 +++ 1-79 ++++
1-33 + 1-80 ++++
1-34 + 1-81 ++++
1-35 +++ 1-82 ++++
1-36 +++ 1-83 +
1-37 + 1-84 ++++
1-39 + 1-85 +++ Example EC50LT Example EC50LT
1-86 ++++ 1-1 13 ++++
1-87 +++ 1-1 14 ++
1-88 ++++ 1-1 15 ++++
1-89 + 1-1 16 ++++
1-90 + 1-1 17 ++++
1-91 + 1-1 18 ++++
1-92 +++ 1-1 19 ++++
1-93 + 1-120 +
1-94 + 1-121 +++
1-95 + 1-122 +
1-96 ++++ 1-123 ++++
1-97 +++ 1-124 +++
1-98 + 1-125 ++++
1-99 ++++ 1-126 +
1-100 ++++ 1-127 ++++
1-101 +++ 1-128 +
1-102 + 1-129 +++
1-103 ++++ 1-130 +++
1-104 ++++ 1-131 ++++
1-105 + 1-132 ++++
1-106 ++++ 1-133 ++++
1-107 ++++ 1-134 ++++
1-108 ++++ 1-135 ++++
1-109 + 1-136 ++++
1-1 10 ++++ 1-137 ++++
1-1 1 1 + 1-138 ++++
1-1 12 ++++
Table 3
Table 4 shows the results in mice blood as effective concentration to delay lysis time by 50% (EC50LT); where, EC50LT≥10μΜ (+),1 μΜ < EC50LT < 0μΜ (++),1 nM < EC50LT < 1 μΜ (+++) and EC50LT < 1 nM (++++) for all the assayed concentrations (1000-0.2 μΜ).
Example EC50LT
TXA +++
1-03 ++++
1-07 ++++
1-08 +++
1-14 +++ Example EC50LT
1-62 ++++
1-73 ++
Table 4
As observed in the tables above (Tables 3 and 4), compounds of the invention show significant delay in the lysis time, in most cases higher than TXA.
Antifibrinolvtic effect in vivo (tail bleeding assay) Bleeding time was evaluated in 2 months old wild-type C57BI6 (n=10) mice by removing the tail tip. Mice (20-25 g) were anaesthetized with 2.5% isoflurane and maintained at 37 °C on heating pads. The hemostatic efficacy was evaluated in 2 models, traditional bleeding model and hyperfibrinolytic bleeding model.
Traditional bleeding model consisted in intraperitoneal injection of different compounds (4 mg/mouse, 160 mg/Kg) dissolved in 2% carboxymethyl cellulose or vehicle. Thirty minutes after injection, 5 mm of tail tip were removed using a scalpel blade and the tail tip bathed in 1 ml_ of sterile saline at 37 °C. The time to cessation of bleeding was measured up to 30 min. The time of bleeding was defined as the interval between initial transection and the visual cessation of bleeding. The results are shown in Table 5.
Figure imgf000065_0001
*p<0.05 vs vehicle
Table 5
Hyperfibrinolytic bleeding model, consisted in injection of 0.5 mg/kg tPA into the ocular plexus to prolong bleeding time due to excessive fibrinolysis. First, the femoral vein was exposed and cannulated with a saline-filled
polyurethane catheter (Microcannula 72-9030, Harvard Apparatus) for agents administration. The catheter was connected to a syringe pump (AL-1000, WPI) for the infusion of 200 μΙ_ (10% bolus, 90% perfusion during 40 minutes) of tested agents. Then, tPA (0.5 mg/kg) was injected into the ocular plexus and five minutes after tPA administration, saline or the different compounds was infused through the femoral catheter to ensure systemic distribution of all the agents. Reference compounds, TXA and Aprotinin, were administered at 300 and 10 mg/Kg respectively; however, all compounds of the invention were administered at 1 mg/Kg. Five minutes later, 5 mm of tail tip were removed using a scalpel blade and the tail tip bathed in 1 mL of sterile saline at 37 °C.
The time of bleeding was defined as the interval between initial transections and the visual cessation of bleeding, that was measured up to 30 minutes. A value of 30 min was assigned to those animals bleeding longer than the observation period. Table 6 shows the results reporting bleeding time (BT); where BT > 20 minutes (+),10 minutes < BT < 20 minutes (++), 5 minutes <
BT < 10 minutes (+++) and BT < 5 minutes (++++). Bleeding time was determined in wild type mice (C57/BI6), where n > 10 per assayed compound; therefore, BT is reported as a mean value - in the case of saline, BT is reported as mean±ESM.
Example BT
Saline 28.9±0.7
TXA ++**
Aprotinin ++**
1-03*** +++**†
1-07 ++*
1-08 ++++**††
1-14 ++++**††
1-20 ++*
1-26 ++*
1-55 ++++**††
1-62 +++**†
1-73 ++++**††
1-79 ++**
1-80 +++**†
1-82 +++**
1-96 +++**
1-99 ++++**††
1-102 ++*
1-107 +++**
1-108 ++++**†† Example BT
1-1 12 +++**
1-1 13 +++**
*p<0.05;**p<0.01 vs saline; compound administered at 10 mg/Kg;
†p<0.05;††p<0.01 vs TXA
Table 6
As shown table 6, tested compounds of the invention show a very significant reduction of the bleeding time when compared to the control or TXA. In all the cases the dose of tested compounds were lower than TXA or Aprotinin doses.
REFERENCES CITED IN THE APPLICATION
Green and P. G. M. Wuts, Protective Groups in Organic Chemistry, Wiley, 3rd ed. 1999, Chapter 2, pp. 17-200 and Chapter 5, pp. 369-451 .
D. Bouyssi et al., "Rearrangement of oxaspiroheptanes to cyclohexanones mediated by lithium iodide", Svnlett 2000, vol. 5, pp. 749-751 .
Osamu Kitagawa et al., "Stereoselective Iodine Atom Transfer [3 + 2]
Cycloaddition Reaction with Alkenes Using Unsymmetrical Allylated Active Methine Radicals", The Journal of Organic Chemistry 2004, vol. 69, pp. 2607- 2610.

Claims

1 . A compound of formula (I), or a pharmaceutically or veterinary acceptable salt thereof, or any stereoisomer either of the compound of formula (I) or of its pharmaceutically or veterinary acceptable salt
Figure imgf000068_0001
(I)
wherein
A and B form a spirocyclic ring system wherein the spiro atom connecting A and B is a carbon atom and wherein A is a known 3- to 8-membered carbocyclic or heterocyclic monocyclic ring, saturated or partially unsaturated; or alternatively
A is a known 6- to 18-membered carbocyclic or heterocyclic polycyclic ring system, saturated, partially unsaturated, or partially aromatic; and
B is a known 4- to 7-membered carbocyclic or heterocyclic monocyclic ring, saturated or partially unsaturated;
C is phenyl or a known 5- to 6-membered heteroaromatic ring;
R1-R3 are independently selected from H, halogen, -NO2, -CN, Ra, -ORa, -OC(Y)Ra', -OC(Y)ORa', -OC(Y)NR Ra, -OSO2ORa', -NR Ra, -NR C(Y)Ra, -NR C(Y)ORa, -NR C(Y)NR Ra, -NR S(O)2Ra', -NR SO2NR Ra, -SRa', -S(O)Ra', -S(O)ORa', -SO2Ra',-SO2(ORa), -SO2NR Ra, -SC(Y)NR Ra, -C(Y)Ra',
-C(Y)ORa', -C(Y)NR Ra, -C(Y)NR ORa, and -C(O)NR SO2Ra'; R4-R7 are independently selected from halogen, -NO2, -CN, Rc, -ORc, -NRdRc, -NRdC(Y)Rc, -NRdC(Y)ORc, -NRdC(Y)NRdRc, -NRdS(O)2Rc, -NRdSO2NRdRc, -SRC, -S(O)Rc, -S(O)ORc, -SO2Rc, -SO2R(ORc), -SO2NRdRc, -SC(Y)NRdRc, -C(Y)RC, -C(Y)ORc, -C(Y)NRdRc, -C(Y)NRdORc,and -C(O)NRdSO2Rc;
Ra is a saturated or unsaturated (Ci-Ci2)alkyl optionally substituted with one or more substituents Re and/or one Cy1; or alternatively Ra is Cy2;
wherein Cy1 and Cy2 are independently optionally substituted with: one Cy3 and/or one or more substituents Re, and/or one or more saturated or unsaturated (CrC6)alkyl groups optionally substituted with one or more substituents Re and/or one Cy3; and
wherein any Cy3 is optionally substituted with one or more substituents independently selected from Re and saturated or unsaturated (CrC6)alkyl optionally substituted with one or more substituents Re; each Ra and R are independently H or Ra;
Rc and each Rd are independently selected from H, Cy4, and saturated or unsaturated (CrC6)alkyl optionally substituted with one or more substituents Rh and/or one Cy5;
wherein Cy4 is optionally substituted with one or more substituents
independently selected from Rh and saturated or unsaturated (CrC6)alkyl optionally substituted with one or more substituents Rh; and
wherein Cy5 is optionally substituted with one or more substituents
independently selected from Rh and saturated or unsaturated (CrC6)alkyl optionally substituted with one or more substituents Rh; each Re is independently selected from halogen, -NO2, -CN, -ORf, -OC(Y)Rf, -OC(Y)ORf, -OC(Y)NR9Rf, -NR9Rf, -NR9C(Y)Rf, -NR9C(Y)ORf, -NR9C(Y)NR9Rf, -NR9S(O)2Rf, -NR9SO2NR9Rf, -SRf, -S(O)Rf, -S(O)ORf, -SO2Rf, -SO2(ORf),
-SO2NR9Rf, -SC(Y)NR9Rf, -C(Y)Rf, -C(Y)ORf, -C(Y)NR9Rf, -C(Y)NR9ORf and -C(O)NR9SO2Rf;
Rf and each R9 are independently selected from H, Cy6, and saturated or unsaturated (CrC6)alkyl optionally substituted with one or more substituents Rh and/or one Cy7; wherein Cy6 is optionally substituted with: one Cy7, and/or one or more substituents Rh, and/or one or more saturated or unsaturated (CrC6)alkyl groups optionally substituted with one or more substituents Rh and/or one Cy7; and
wherein any Cy7 is optionally substituted with one or more substituents independently selected from Rh and (Ci-C4)alkyl optionally substituted with one or more substituents Rh; each Rh is independently selected from halogen, -NO2, -CN, -OR', -ΟΟ(Ο)^, -00(0)0^, -OCiOJNR'R1, -NR'R', -NR'CiOJR1, -NR^OJOR1, -NR'C(O)NR'R', -NR^O^R1, -NR'SCfeNR'R1, -SR1, -S{0)R -SOzR1, -SOziOR1), -SCfeNR'R1, -C{0)R -0(0)0^, -CiOJNR'R1, and -C(O)NR'OR'; each R1 is independently H or -(Ci-C4)alkyl optionally substituted with one or more halogen atoms;
Y is O, S, or NRg;
Cy1, Cy2, Cy4 and Cy6 are independently a C or N-attached known ring system selected from 3- to 8-membered carbocyclic or heterocyclic monocyclic ring, saturated or partially unsaturated; phenyl; 5- or 6-membered heteroaromatic ring; and 6- to 18-membered carbocyclic or heterocyclic polycyclic ring system, saturated, partially unsaturated, aromatic or partially aromatic; Cy3, Cy5 and Cy7 are independently a C or N-attached known ring system selected from 3- to 8-membered carbocyclic or heterocyclic monocyclic ring, saturated or partially unsaturated; phenyl; and 5- or 6-membered
heteroaromatic ring; wherein in the carbocyclic rings all ring members are carbon atoms; and in the heterocyclic and heteroaromatic rings one or more ring members are selected from N, O, and S; and wherein in all saturated or partially
unsaturated rings one or two members of the rings are optionally C(O) and/or C(NH) and/or C[N(C C4)alkyl].
2. The compound of formula (I) according to claim 1 , wherein A is a known 3- to 8-membered carbocyclic or heterocyclic monocyclic ring or a known 6- to 10-membered carbocyclic or heterocyclic bicyclic ring system.
3. The compound of formula (I) according to claim 2, wherein A is a monocyclic ring selected from a 3- to 6-membered carbocyclic ring and a 5- to 6-membered heterocyclic ring.
4. The compound of formula (I) according to any of the claims 1 -3, wherein B is a 6- to 7-membered carbocyclic or heterocyclic monocyclic ring.
5. The compound of formula (I) according to any of the claims 1 -4, wherein A and B form a spirocyclic ring system selected from the group consisting of:
Figure imgf000071_0001
Figure imgf000072_0001
6. The compound of formula (I) according to any of the claims 1 -5, wherein C is phenyl.
7. The compound of formula (I) according to any of the claims 1 -6, wherein in R1-R3, Rf and each R9 are independently selected from H and saturated or unsaturated (CrC6)alkyl optionally substituted with one or more fluorine atoms.
8. The compound of formula (I) according to any of the claims 1 -7, wherein in R1-R3, Cy1 and Cy2 are independently optionally substituted with one or more substituents selected from Re and saturated or unsaturated (CrC6)alkyl optionally substituted; and Cy6 is optionally substituted with one or more substituents independently selected from Rh and saturated or unsaturated (CrC6)alkyl optionally substituted.
9. The compound of formula (I) according to any of the claims 1 -8, wherein R1-R3 are independently selected from H, halogen, -NO2, -CN, Ra, -ORa, -OC(O)Ra', -OC(O)ORa', -OC(O)NR Ra', -NR Ra, -NR C(O)Ra, -NR C(O)ORa', -NR C(O)NR Ra, -NR S(O)2Ra', -SRa', -S(O)Ra', -SO2Ra', -SO2NR Ra, -C(O)Ra', -C(O)ORa', -C(O)NR Ra, and -C(O)NR ORa'.
10. The compound of formula (I) according to any of the claims 1 -9, wherein R4-R7 are independently selected from halogen, -NO2, -CN, Rc, -ORc, -NRdRc, -NRdC(O)Rc, -NRdC(O)ORc, -NRdC(O)NRdRc, -NRdS(O)2Rc, -SRC, -S(O)Rc, -SO2Rc, -SO2NRdRc, -C(O)Rc, -C(O)ORc, and -C(O)NRdRc.
1 1 . The compound of formula (I) according to any of the claims 1 -10, wherein R2 and R3 are independently selected from H, halogen, Ra, -ORa, and -NR Ra; and R5-R7 are independently selected from H, halogen, Rc, -ORc, and -NRdRc, wherein Ra, Ra, R , Rc and Rd are independently selected from H and
-(Ci-C4)alkyl optionally substituted with one or more fluorine atoms.
12. A pharmaceutical or veterinary composition which comprises an effective amount of a compound of formula (I) as defined in any of the claims 1 -1 1 , or a pharmaceutically or veterinary acceptable salt thereof, or any stereoisomer either of the compound of formula (I) or of its pharmaceutically or veterinary acceptable salt, together with one or more pharmaceutically or veterinary acceptable excipients or carriers.
13. A compound of formula (I) according to any of the claims 1 -1 1 , or a pharmaceutically or veterinary acceptable salt thereof, or any stereoisomer either of the compound of formula (I) or of its pharmaceutically or veterinary acceptable salt, for use as a medicament.
14. A compound of formula (I) according to any of the claims 1 -1 1 , or a pharmaceutically or veterinary acceptable salt thereof, or any stereoisomer either of the compound of formula (I) or of its pharmaceutically or veterinary acceptable salt, for use as antifibrinolytic and antihemorrhagic agent.
15. A compound of formula (III)
Figure imgf000073_0001
wherein A, B, C, R1-R7 are as defined in any of the claims 1 -1 1 , and R' is H or a carboxy protective group selected from the group consisting of (CrC6)alkyl, benzyl, p-methoxyphenyl, trimethylsilyl and [2-(Trimethylsilyl)ethoxy]methyl (SEM), with the proviso that compound (III) is other than 7-methoxycarbonyl- 7-phenylsulphonyl-2-oxaspiro[2.4]heptane and (2S*,4R*)-2-Phenylsulfonyl-4- iodomethyl-6,1 1 -dioxaspiro[4.6]undecane-2-carboxylic acid methyl ester; or a compound of formula (II)
Figure imgf000074_0001
(II)
wherein A, B, C, R1-R7 are as defined in any of the claims 1 -1 1 , and R is an hydroxamic acid protective group selected from the group consisting of tetrahydro-2H-pyran-2-yloxy (THP), benzyl, 1 -naphthylmethyl and
dimethyloxybenzyl (DMB).
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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2016071216A1 (en) * 2014-11-03 2016-05-12 Bayer Pharma Aktiengesellschaft Piperidinylpyrazolopyrimidinones and their use
US10202331B2 (en) 2014-11-03 2019-02-12 Thrombolytics, Llc Antifibrinolytic compounds

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB201216127D0 (en) * 2012-09-11 2012-10-24 Jaguar Cars A method for determining the change in a vehicle battery

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3868411A (en) * 1969-07-22 1975-02-25 En Nom Collectif Science Union Isobutyl cyclohexenyl compounds alpha (p-isobutyl-cyclohexenyl) alkanoic acids
US4483867A (en) * 1981-11-17 1984-11-20 Kabivitrum Ab Antifibrinolytically active derivatives of tranexamic acid
US20020099035A1 (en) * 2001-01-24 2002-07-25 Sandanayaka Vincent P. Method for preparing alpha-sulfonyl hydroxamic acid derivatives

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20000075809A (en) * 1997-02-27 2000-12-26 윌리암 에이취 캘넌, 에곤 이 버그 N-Hydroxy-2-(alkyl, aryl or heteroaryl sulfanyl, sulfinyl or sulfonyl)-3-substituted alkyl, aryl or heteroarylamides as matrix metalloproteinase inhibitors
US6197791B1 (en) * 1997-02-27 2001-03-06 American Cyanamid Company N-hdroxy-2-(alkyl, aryl, or heteroaryl, sulfanyl, sulfinyl or sulfonyl)-3-substituted alkyl, aryl or heteroarylamides as matrix metalloproteinase inhibitors
US6172057B1 (en) * 1997-02-27 2001-01-09 American Cyanamid Company N-Hydroxy-2-(alkyl, aryl, or heteroaryl sulfanyl, sulfinyl or sulfonyl)-3-substituted alkyl, aryl or heteroarylamides as matrix metalloproteinase inhibitors
IL137566A0 (en) * 1998-02-19 2001-07-24 American Cyanamid Co N-hydroxy-2-(alkyl, aryl or heteroaryl sulfanyl, sulfinyl or sulfonyl)-3-substituted-alkyl, aryl or heteroarylamides as matrix metalloproteinase inhibitors
US6225311B1 (en) * 1999-01-27 2001-05-01 American Cyanamid Company Acetylenic α-amino acid-based sulfonamide hydroxamic acid tace inhibitors
US20040024024A1 (en) * 2001-05-11 2004-02-05 Freskos John N. Aromatic sulfone hydroxamates and their use as protease inhibitors

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3868411A (en) * 1969-07-22 1975-02-25 En Nom Collectif Science Union Isobutyl cyclohexenyl compounds alpha (p-isobutyl-cyclohexenyl) alkanoic acids
US4483867A (en) * 1981-11-17 1984-11-20 Kabivitrum Ab Antifibrinolytically active derivatives of tranexamic acid
US20020099035A1 (en) * 2001-01-24 2002-07-25 Sandanayaka Vincent P. Method for preparing alpha-sulfonyl hydroxamic acid derivatives

Non-Patent Citations (8)

* Cited by examiner, † Cited by third party
Title
"PROTECTIVE GROUPS IN ORGANIC CHEMISTRY", pages: 369 - 451
BOUYSSI D ET AL: "Rearrangement of oxaspiroheptanes to cyclohexanones mediated by lithium iodide", SYNLETT 2000 DE, no. 5, 2000, pages 749 - 751, XP008157852, ISSN: 0936-5214 *
D. BOUYSSI ET AL.: "Rearrangement of oxaspiroheptanes to cyclohexanones mediated by lithium iodide", SYNLETT, vol. 5, 2000, pages 749 - 751
GREEN; P. G. M. WUTS: "Protective Groups in Organic Chemistry", 1999, WILEY, pages: 17 - 200
OSAMU KITAGAWA ET AL.: "Stereoselective Iodine Atom Transfer [3 + 2] Cycloaddition Reaction with Alkenes Using Unsymmetrical Allylated Active Methine Radicals", THE JOURNAL OF ORGANIC CHEMISTRY, vol. 69, 2004, pages 2607 - 2610
OSAMU KITAGAWA ET AL: "Stereoselective Iodine Atom Transfer [3 + 2] Cycloaddition Reaction with Alkenes Using Unsymmetrical Allylated Active Methine Radicals", THE JOURNAL OF ORGANIC CHEMISTRY, vol. 69, no. 7, 1 April 2004 (2004-04-01), pages 2607 - 2610, XP055043559, ISSN: 0022-3263, DOI: 10.1021/jo035771a *
T. W. GREEN; P. G. M. WUTS: "Protective Groups in Organic Chemistry", 1999, WILEY, pages: 17 - 200
T. W. GREEN; P. G. M. WUTS: "Protective Groups in Organic Chemistry", 1999, WILEY, pages: 369 - 451

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2016071216A1 (en) * 2014-11-03 2016-05-12 Bayer Pharma Aktiengesellschaft Piperidinylpyrazolopyrimidinones and their use
CN107108638A (en) * 2014-11-03 2017-08-29 拜耳制药股份公司 Piperidyl PyrazolopyrimidinonecGMP and application thereof
JP2017533235A (en) * 2014-11-03 2017-11-09 バイエル ファーマ アクチエンゲゼルシャフト Piperidinylpyrazolopyrimidinones and their use
US10118930B2 (en) 2014-11-03 2018-11-06 Bayer Pharma Aktiengesellschaft Piperidinylpyrazolopyrimidinones and their use
US10202331B2 (en) 2014-11-03 2019-02-12 Thrombolytics, Llc Antifibrinolytic compounds
RU2713937C2 (en) * 2014-11-03 2020-02-11 Байер Фарма Акциенгезельшафт Piperidinylpyrazolopyrimidinones and use thereof

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