WO2002014349A2 - Inhibiteurs non covalents de l'urokinase et de la formation de vaisseaux sanguins - Google Patents

Inhibiteurs non covalents de l'urokinase et de la formation de vaisseaux sanguins Download PDF

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WO2002014349A2
WO2002014349A2 PCT/US2001/025337 US0125337W WO0214349A2 WO 2002014349 A2 WO2002014349 A2 WO 2002014349A2 US 0125337 W US0125337 W US 0125337W WO 0214349 A2 WO0214349 A2 WO 0214349A2
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carbon atoms
substituted
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PCT/US2001/025337
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WO2002014349A3 (fr
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Odile Esther Levy
Edwin L. Madison
Joseph Edward Semple
Amir P. Tamiz
Michael I. Weinhouse
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Corvas International, Inc.
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Priority claimed from US09/733,645 external-priority patent/US6586405B2/en
Application filed by Corvas International, Inc. filed Critical Corvas International, Inc.
Priority to NZ518195A priority Critical patent/NZ518195A/en
Priority to JP2002519486A priority patent/JP2004506648A/ja
Priority to AU83347/01A priority patent/AU785260B2/en
Priority to IL14904201A priority patent/IL149042A0/xx
Priority to CA002387002A priority patent/CA2387002A1/fr
Publication of WO2002014349A2 publication Critical patent/WO2002014349A2/fr
Priority to IL149042A priority patent/IL149042A/en
Publication of WO2002014349A3 publication Critical patent/WO2002014349A3/fr
Priority to AU2006235835A priority patent/AU2006235835B2/en

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Definitions

  • Urokinase is an enzyme involved in the metastasis of tumor cells, neovascularization, and other activities.
  • One purpose of the present invention is to provide novel compounds which are active as inhibitors of urokinase that can be used to inhibit the activity of urokinase and thereby attenuate its deleterious effects.
  • Another purpose of the present invention is to provide novel compounds which inhibit blood vessel formation, particularly blood vessel formation related to a pathologic condition.
  • Urinary-type plasminogen activator is a serine protease within the trypsin/chymotrypsin family. In its physiological state, uPA is found in three forms: single chain pro-uPA, two chain uPA, and low molecular weight uPA (lacks N-terminal domains). The zymogen, pro- uPA, is converted to u-PA by cleavage of the peptide bond at K158-I159. The resultant two chain uPA is linked by disulfide bridges, has an M r of about 50 kD, and a C-terminal serine proteinase domain.
  • uPA The activity of uPA is focused to cell surfaces upon binding to its receptor, uPAR.
  • uPAR is a single-chain glycosyl phosphatidyl inositol (GPI) -anchored membrane receptor.
  • GPI glycosyl phosphatidyl inositol
  • the N-terminal 92 amino acids of uPAR play a dominant role in binding to uPA and pro-uPA.
  • Receptor for uPA has been located on T-cells, NK cells, monocytes, and neutrophils, as well as vascular endothelial cells, fibroblasts, smooth muscle cells, keratinocytes, placental trophoblasts, hepatocytes, and a wide variety of tumor cells.
  • uPA After conversion of pro-uPA to uPA, which occurs primarily at the uPAR on the cell surface, uPA activates plasminogen to plasmin. Activation occurs upon cleavage at residues PGR-VV for human plasminogen, or at residues SGR-IV for bovine plasminogen. Because plasminogen also is present on the cell surface, this activation cascade focuses the activity of u-PA and plasmin on the plasma membrane. Plasmin has many roles, including activation of additional uPA and other enzymes, digestion of fibrin, and digestion of components of the extracellular matrix (ECM) .
  • ECM extracellular matrix
  • the effects of the uPA system on cell migration and invasion are thought to be due to both a proteolytic effect of plasmin-mediated degradation of the extracellular matrix, as well as more a direct interaction of the uPA receptor with components of the extracellular matrix. Degradation of the extracellular matrix permits a metastasizing cell to invade the matrix, whereas interaction between uPA receptor and the matrix itself assists a cell in its migration. Localization of the uPA/plasmin system on the cell surface, or the leading edge of metastasizing cells, is consistent with postulated role of uPA in metastasis [Plesner et al . , Stem Cells 15:398-408 (1997)].
  • uPAR Interaction of uPAR with vitronectin, a component of the extracellular matrix, mediates cell adhesion and can be enhanced when uPAR is bound by uPA.
  • Cell surface adhesion molecules, integrins also appear to be involved in this adhesion function, particularly beta-1 and beta-2 integrins [Paysant et al . , Br. J. Haematol. 100:45-51 (1998); Simon et al . , Blood 8:3185-3194 (1996)].
  • the CDllb/CD18 integrin can associate with the uPA-uPAR complex and promote adhesion of cells bearing these receptors, e.g., neutrophils, leukocytes.
  • the uPA/uPAR system also is involved in the establishment of new vasculature, or neovascularization.
  • Pathological neovascularization also is a characteristic of retinal disease, rubeosis ulceris, proliferative vitreo retinopathy inflammatory disease, diabetic retinopathy, chronic uveitis, Fuch' s heterochromic iridocyclitis, neovascular glaucoma, corneal or optic nerve neovascularization, vascular disease, pterygium, glaucoma surgery bleb failure, hyperkeratosis, cheloid and polyp formation (see EP 451,130).
  • Undesired angiogenesis also can occur in the following conditions or can be a result of the following activities: macular degeneration, retinopathy of prematurity, corneal graft rejection, retrolental fibroplasia, epidemic keratoconjunctivitis, Vitamin A deficiency, contact lens overwear, atopic keratitis, superior limbic keratitis, pterygium keratitis sicca, sogrens disease, acne rosacea, phylectenulosis, syphilis, Mycobacteria infections other than leprosy, lipid degeneration, chemical burns, bacterial or fungal ulcers, Herpes simplex or zoster infections, protozoan infections, Kaposi's sarcoma, Mooren ulcer, Terrien' s marginal degeneration, marginal keratolysis, trauma, rheumatoid arthritis, systemic lupus, polyarteritis, Wegeners sarcoidosis,
  • An antagonist of uPA/uPAR binding (EGF-like domain of uPA fused to Fc of IgG) was said to inhibit neovascularization and growth of the murine B16 melanoma. [Min et al . , Cancer Res. 5_6: 2428-2433 (1996)]. Consistent with this finding is the correlation noted between microvessel density, vascular invasion and uPA levels in breast carcinomas [Hildenbrand et al . , Brit. J. Cancer 7_2: 818-823 (1995)].
  • the known uPA inhibitor amiloride also was said to inhibit a variety of neovascularization pathologies [Glaser et al . , EP 451,130; Avery et al . , Arch. Ophthalmol. 108:1474-1476 (1990)].
  • PAI-1 and PAI-2 are members of the serpin family of proteinase inhibitors.
  • the binding of serpins to their cognate proteases involves a large number of interactions between amino acids of each protein, including those in the serpin reactive loop (Ser-Ala-Arg-Met-Ala (SEQ. ID. NO. 1) for PAI-1, Thr-Gly-Arg-Thr-Gly (SEQ. ID. NO. 2) for PAI-2) .
  • Serpin reactive loop Ser-Ala-Arg-Met-Ala
  • Thr-Gly-Arg-Thr-Gly SEQ. ID. NO. 2
  • Introduction of exogenous PAI-2 into experimental animals was reported to inhibit the rate of lung metastasis [Evans and Lin, Amer. Surg. 61:692-697 (1995); Mueller et al . , Proc. Natl. Acad.
  • uPA inhibitors may be within the 4-substituted benzo [b] thiophene-2-carboxamidine class of inhibitors, of which B428 (4-iodo-benzo [b] thiophene-2- carboxamidine) and B623 are members [Towle et al . , Cancer Res. 53:2553-2559 (1993); Bridges et al . , Bioorg. Med. Chem. 1:403-410 (1993); Bridges et al . , U.S. Patent No. 5,340,833].
  • uPA uPA
  • p-aminobenzamidine which is a competitive inhibitor of uPA
  • amiloride Both compounds have been shown to reduce tumor size in experimental animals [Jankan et al . , Cancer Res. 57:559-563 (1997); Billstrom et al . , Int. J. Cancer 61:542-547 (1995)].
  • EGCG epigallo-cathecin-3 gallate
  • EGCG is a weaker inhibitor of uPA than amiloride, but suggested EGCG can be consumed in much higher doses than amiloride without toxic effect.
  • Antagonists of uPAR also have been studied- [Doyle and Rosenberg, U.S. Patent No. 5,656,726; Min et al . , Cancer Res. 56:2428-2433 (1996)], as have antisense oligonucleotides complementary to uPA [ ilhelm et al . , Clin. Exp. Metast. 13:296-302 (1995); Iversen and Scholar, U.S. Patent No. 5,552,390].
  • Antibodies directed against uPAR, and said to inhibit the binding of uPA to UPAR are disclosed by Dano et al .
  • pro-hepatocyte growth factor (HGF) , a cell migration stimulating protein, is a substrate of uPA [Naldinie et al . , EMBO J. 11:4825-4833 (1992)]. Direct cleavage of a 66kDa extracellular matrix protein and fibronectin by uPA also has been reported, which suggests a more direct role for uPA in facilitating cell migration [Quigley et al . , Proc. Natl. Acad. Sci. 84:2776-2780 (1987)]. Thus, inhibition of uPA may affect these activities, as well. Summary of the Invention
  • the present invention is directed to novel pepitidic non-covalent urokinase inhibitors.
  • the compounds have an arginine mimic at PI. These compounds have activity as potent inhibitors of urokinase and thereby are useful in decreasing its deleterious effects.
  • Compounds of the present invention are active in inhibiting blood vessel formation, particularly that related to a pathologic process .
  • the present invention is directed to compounds of the formula (I) :
  • Ri is selected from the group consisting of:
  • alkyl of 1 to about 12 carbon atoms which is unsubstituted or substituted with 1 or 2 substituents selected from the group consisting of Yi and Y 2 ,
  • heteroaryl of about 5 to about 14 ring atoms with the ring atoms selected from carbon and heteroatoms, wherein the heteroatoms are selected from oxygen, nitrogen, and sulfur, and which is unsubstituted or mono-, di- or tri- substituted with 1 to 3 substituents selected from the group consisting of Yi, Y 2 , and Y 3 ,
  • aralkyl of about 7 to about 15 carbon atoms which is unsubstituted or substituted on the alkyl chain with hydroxy or halogen and which is unsubstituted or mono-, di-, or tri-substituted on the aryl ring with 1 to 3 substituents selected from the group consisting of Yi, Y 2 , and Y 3 , (10) heteroaralkyl of about 5 to about 14 ring atoms with the ring atoms selected from carbon and heteroatoms, wherein the heteroatoms are selected from oxygen, nitrogen, and sulfur, which is unsubstituted or substituted on the alkyl chain with hydroxy or halogen and which is unsubstituted on the ring or mono-, di- or tri-substituted on the ring with 1 to 3 substituents selected from the group consisting of Yi, Y 2 , and
  • heteroaralkenyl of about 5 to about 14 ring atoms with the ring atoms selected from carbon and heteroatoms, wherein the heteroatoms are selected from oxygen, nitrogen, and sulfur, and which is unsubstituted or mono-, di- or tri-substituted on the ring carbons with 1 to 3 substituents selected from the group consisting of Yi, Y 2 , and Y 3 ,
  • Yi and Y 2 are selected together to be -0[C(Z 3 ) (Z 4 )] r 0- or -0[C(Z 3 ) (Z 4 )] r+ ⁇ -, wherein r is an integer from 1 to 4 and Z 3 and Z 4 are independently selected from the group consisting of hydrogen, alkyl of 1 to about 12 carbon atoms, aryl of about 6 to about 14 carbon atoms, heteroaryl of about 5 to about 14 ring atoms, aralkyl of about 7 to about 15 carbon atoms, and heteroaralkyl of about 5 to about 14 ring atoms; (c) R 2 is selected from the group consisting of -CH 3 , -C 2 H 5 , -(CH 2 ) 2 0H, -(CH 2 ) 2 0A ⁇ , -CH(R 5 )OH, -CH(R 5 )OA ⁇ and -CH 2 NH-X'-R 6 wherein Ai is -C
  • heterocyclo of 4 to about 6 ring atoms with the ring atoms selected from carbon and heteroatoms, wherein the heteroatoms are selected from the group consisting of oxygen, nitrogen, and S(0)i, wherein i is 0, 1, or 2, including —N V, wherein —N V is a 5 to 7 member heterocycle having 3 to 6 ring carbon atoms, where V is -CH 2 -, -0-, -S( 0)-, -S(0) 2 - or -S-, which is unsubstituted or mono-, di-, or tri-substituted on the ring carbons with 1 to 3 substituents selected from the group consisting of Yi, Y 2 , and Y 3 ,
  • heteroaryl of about 5 to about 6 ring atoms with the ring atoms selected from carbon and heteroatoms, wherein the heteroatoms are selected from oxygen, nitrogen, and sulfur, and which is unsubstituted or mono-, di- or tri- substituted with 1 to 3 substituents selected from the group consisting of Yi, Y 2 , and Y 3 , (9) alkyl of 1 to about 4 carbon atoms substituted with phenyl and which is unsubstituted or mono-, di-, or tri-substituted on the phenyl ring with 1 to 3 substituents selected from the group consisting of Yi, Y 2 , and Y 3 , (10) heteroaralkyl of about 5 to about 6 ring atoms with the ring atoms selected from carbon and heteroatoms, wherein the heteroatoms are selected from oxygen, nitrogen, and sulfur, and which is unsubstituted or substituted on the alkyl chain with hydroxy or halogen and which is unsubstit
  • (11) aralkenyl of about 8 to about 12 carbon atoms which is unsubstituted or mono-, di-, or tri-substituted on the aryl ring with 1 to 3 substituents selected from the group consisting of Yi, Y 2 , and Y 3 ,
  • heteroaralkenyl of about 5 to about 6 ring atoms with the ring atoms selected from carbon and heteroatoms, wherein the heteroatoms are selected from oxygen, nitrogen, and sulfur, and which is unsubstituted or mono-, di- or tri-substituted on the ring carbons with 1 to 3 substituents selected from the group consisting of Yi, Y 2 , and Y 3 , and
  • R 6 is selected from the group consisting of: (1) alkyl of 1 to about 12 carbon atoms, which is unsubstituted or substituted with 1 or 2 substituents selected from the group consisting of Yi, and
  • heteroaryl of about 5 to about 14 ring atoms with the ring atoms selected from carbon and heteroatoms, wherein the heteroatoms are selected from oxygen, nitrogen, and sulfur, and which is unsubstituted or mono-, di- or tri-substituted with 1 to 3 substituents selected from the group consisting of Yi, Y 2 , and Y 3 ,
  • aralkyl of about 7 to about 15 carbon atoms which is unsubstituted or substituted on the alkyl chain with hydroxy or halogen and which is unsubstituted or mono-, di-, or tri-substituted on the aryl ring with 1 to 3 substituents selected from the group consisting of Y x , Y 2 , and Y 3 , (9) heteroaralkyl of about 5 to about 14 ring atoms with the ring atoms selected from carbon and heteroatoms, wherein the heteroatoms are selected from oxygen, nitrogen, and sulfur, and which is unsubstituted or substituted on the alkyl chain with hydroxy or halogen and which is unsubstituted on the ring or mono-, di- or trisubstituted on the ring with 1 to 3 substituents selected from the group consisting of Yi, Y 2 , and Y 3 , and
  • R 3 is selected from H or methyl, or R 3 , and R 4a and R 4b are selected together as set forth in (f) ;
  • R a and R 4b are independently lower alkyl of 1 to
  • R 4a and R 4b are selected together and are - (CH 2 ) k - wherein k is 5 or 6 to give a spirocycloalkyl group; or (iv) R 3 , and R 4a and R 4b are selected together as set forth in (f ) ;
  • R 3 and R 4 are selected together to be in the S configuration to give a group at P2 selected from the group consisting of prolyl, pipecolyl, azetidine-2-carbonyl, 4-hydroxyprolyl, 3-hydroxyprolyl, 4- aminoprolyl, 4- (-CH 2 NH 2 ) -prolyl, 3, 4-methanoprolyl and 3, 4-dehydroprolyl and R 4b is hydrogen;
  • R 7 is hydrogen or alkyl of 1 to about 4 carbon atoms; .and (h) E is Q-T wherein (i) Q is selected from the group consisting of -C (R ⁇ 3 R ⁇ ) t ⁇ r phenyl subtituted with Rs and R 9 , a 5- or 6- membered heterocyclic ring having 1 to 2 heteroatoms substituted with R 8 or R 8 and R 9 , and a 9- or 10-membered heterocyclic ring having 1 to 2 heteroatoms substituted with Rs and R 9 , wherein heteroatoms are selected from nitrogen and sulfur; and
  • R ⁇ 2 is hydrogen, alkyl of 1 to about 6 carbon atoms, alkoxy of 1 to about 6 carbon atoms or (CF 2 )jCF 3 wherein j is 0, 1, 2 or 3; each of R ⁇ 3 and R i4 is independently selected from the group consisting of hydrogen and lower alkyl of 1 to about 3 carbon atoms;
  • R ⁇ 5 is selected from the group consisting of hydrogen, alkyl of 1 to about 6 carbon atoms and -(CF 2 ) h CF
  • the present invention is based on our finding that the novel compounds of our invention are active as inhibitors of urokinase.
  • Compounds of the present invention exhibit activity in inhibiting angiogenesis.
  • the present invention is directed to pharmaceutical compositions comprising a therapeutically effective amount of a compound of the present invention and a pharmaceutically acceptable carrier.
  • the present invention is directed to methods of using the compounds and pharmaceutical compositions of the present invention for inhibition of urokinase.
  • alkenyl refers to unsaturated aliphatic groups having at least one double bond.
  • alkyl refers to saturated aliphatic groups including straight-chain, branched-chain and cyclic
  • alkoxy and alkoxyl refer to a group having the formula, R-0-, wherein R is an alkyl group.
  • alkoxycarbonyl refers to -C(0)0R wherein R is alkyl.
  • alkenyl refers to an alkenyl group substituted with an aryl group. Preferably the alkenyl group has from 2 to about 6 carbon atoms.
  • aralkyl refers to an alkyl group substituted with an aryl group. Suitable aralkyl groups include benzyl, phenethyl, and the like, all of which may be optionally substituted. Preferably the alkyl group has from 1 to about 6 carbon atoms.
  • aryl refers to an aromatic group which has at least one ring having a conjugated pi electron system and includes a carbocyclic aryl, heterocyclic aryl and biaryl groups, all of which may be optionally substituted.
  • aryloxy refers to a group having the formula, R-0-, wherein R is an aryl group.
  • aralkoxy refers to a group having the formula, R-0-, wherein R is an aralkyl group.
  • amino acid refers to both natural, unnatural amino acids in their D and L stereo isomers if their structures allow such stereoisomeric forms, and their analogs.
  • Natural amino acids include alanine (Ala), arginine (Arg) , asparagine (Asn) , aspartic acid (Asp) , cysteine (Cys) , glutamine (Gin) , glutamic acid (Glu) , glycine (Gly), histidine (His), isoleucine (lie), leucine (Leu) , lysine (Lys) , methionine (Met) , phenylalanine (Phe) , proline (Pro), serine (Ser), threonine (Thr), tryptophan (Trp) , tyrosine (Tyr) and valine (Val) .
  • Unnatural amino acids include, but are not limited to azetidinecarboxylic acid, 2-aminoadipic acid, 3-aminoadipic acid, beta-alanine, aminopropionic acid, 2-aminobutyric acid, 4-aminobutyric acid, 6-aminocaproic acid, 2-aminoheptanoic acid, 2-aminoisobutyric acid, 3-aminoisobutyric acid, 2-aminopimelic acid, 2,4 diaminoisobutyric acid, demosine, 2, 2 ' -diaminopimelic acid, 2, 3-diaminopropionic acid,
  • Amino acid analogs include the natural and unnatural amino acids which are chemically blocked, reversibly or irreversibly, or modified on their N-terminal amino group or their side-chain groups, as for example, methionine sulfoxide, methionine sulfone, S- (carboxymethyl) -cysteine, S- (carboxymethyl) -cysteine sulfoxide and S- (carboxymethyl) -cysteine sulfone.
  • amino acid analog refers to an amino acid wherein either the C-terminal carboxy group, the N-terminal amino group or side-chain functional group has been chemically modified to another functional group.
  • aspartic acid- (beta-methyl ester) is an amino acid analog of aspartic acid
  • N-ethylglycine is an amino acid analog of glycine
  • alanine carboxamide is an amino acid analog of alanine.
  • amino acid residue refers to radicals having the structure: (1) -C (0) -R-NH-, wherein R typically is -CH(R')-, wherein R' is H or a carbon containing
  • aryl refers to phenyl substituted by carbocyclic or heterocyclic aryl as defined herein, ortho, meta or para to the point of attachment of the phenyl ring.
  • Carbocyclic aryl refers to aromatic groups wherein the ring atoms on the aromatic ring are carbon atoms.
  • Carbocyclic aryl groups include monocyclic carbocyclic aryl groups and naphthyl groups, all of which may be optionally substituted.
  • Suitable carbocyclic aryl groups include phenyl and naphthyl.
  • Suitable substituted carbocyclic aryl groups include indene and phenyl substituted by one to two substituents such being advantageously lower alkyl, hydroxy, lower alkoxy, lower alkoxycarbonyl, halogen, trifluoromethyl, difluoromethyl, nitro, and cyano.
  • Substituted naphthyl refers to naphthyl, more preferably 1- or 2-naphthyl, substituted by Yi, Y 2 and/or Y 3 as defined in connection with formula (I) hereinabove.
  • Cycloalkenyl refers to a cyclic alkenyl group. Suitable cycloalkenyl groups include, for example, cyclopentenyl and cyclohexenyl .
  • Cycloalkyl refers to a cyclic alkyl group having at least one ring and includes polycyclic groups, including fused ring cyclic alkyl groups. Suitable cycloalkyl groups include, for example, cyclohexyl, cyclopropyl, cyclopentyl, and cycloheptyl.
  • Cyclohexylmethyl refers to a cyclohexyl group attached to CH 2 .
  • fused carbocyclic refers to a multicyclic fused carbocyclic ring having both aromatic and non-aromatic rings. Suitable fused carbocyclic rings include fluorenyl, tetralin and the like.
  • fused carbocyclic alkyl refers to an alkyl group substituted with a fused carbocyclic ring moiety, preferably a multicyclic fused carbocyclic ring including both aromatic and non-aromatic rings. Suitable fused carbocyclic alkyl groups include fluorenylmethyl, and the like.
  • halogen refers to fluorine, chlorine, bromine and iodine.
  • Heteroaralkenyl refers to an alkenyl group substituted with a heteroaryl, and includes those heterocyclic systems described in "Handbook of Chemistry and Physics", 49th edition, 1968, R.C. east, editor; The Chemical Rubber Co., Cleveland, OH. See particularly Section C, Rules for Naming Organic Compounds, B. Fundamental Heterocyclic Systems.
  • the alkenyl group has from 2 to about 6 carbon atoms.
  • Heteroaralkyl refers to an alkyl group substituted with a heteroaryl, such as picolyl, and includes those heterocyclic systems described in "Handbook of Chemistry and Physics", 49th edition, 1968, R.C. Weast, editor; The Chemical Rubber Co., Cleveland, OH. See particularly Section C, Rules for Naming Organic Compounds, B. Fundamental Heterocyclic Systems.
  • the alkyl group has from 1 to about 6 carbon atoms .
  • Heteroaryl refers to aromatic groups having from 1 to 14 carbon atoms and the remainder of the ring atoms are heteroatoms, and includes those heterocyclic systems described in "Handbook of Chemistry and Physics", 49th edition, 1968, R.C. Weast, editor; The Chemical Rubber Co., Cleveland, OH. See particularly Section C, Rules for Naming Organic Compounds, B. Fundamental Heterocyclic Systems.
  • Suitable heteroatoms include oxygen, nitrogen, and S(0) ⁇ , wherein i is 0, 1 or 2, and suitable heterocyclic aryls include furanyl, thienyl, pyridyl, pyrrolyl, pyrimidyl, pyrazinyl, imidazolyl, and the like.
  • Heterocyclo refers to a reduced heterocyclic ring system comprised of carbon, nitrogen, oxygen and/or sulfur atoms, and includes those heterocyclic systems described in "Handbook of Chemistry and Physics", 49th edition, 1968, R.C. Weast, editor; The Chemical Rubber Co., Cleveland, OH. See particularly Section C, Rules for Naming Organic Compounds, B. Fundamental Heterocyclic Systems.
  • Heterocycloalkyl refers to an alkyl group substituted with a heterocyclo group, and includes those heterocyclic systems described in "Handbook of Chemistry and Physics",- 49th edition, 1968, R.C. Weast, editor; The Chemical Rubber Co., Cleveland, OH. See particularly Section C, Rules for Naming Organic Compounds, B. Fundamental Heterocyclic
  • the alkyl group has from about 1 to about 6 carbon atoms .
  • radicals or groups with one and up to and including 5 carbon atoms, preferably up to and including 4 carbon atoms, and advantageously one or two carbon atoms.
  • Such radicals or groups may be straight chain or branched chain.
  • Perfluoroalkyl refers to an alkyl group which has every hydrogen replaced with fluorine.
  • Perfluoroaryl refers to an aryl group which has every hydrogen replaced with fluorine.
  • Perfluoroarylalkyl refers to an aralkyl group in which every hydrogen on the aryl moiety is replaced with fluorine.
  • “Pharmaceutically acceptable salt” includes salts of the compounds of the present invention derived from the combination of such compounds and an organic or inorganic acid. In practice the use of the salt form amounts to use of the base form. The compounds of the present invention are useful in both free base and salt form, with both forms being considered as being within the scope of the present invention.
  • AIBN refers to 2, 2 ' -azobisisobutyronitrile.
  • Bn refers to benzyl.
  • Boc refers to t-butoxycarbonyl .
  • Boc 2 0 refers to Boc anhydride (di-tert-butyl carbonate) .
  • BOC-ON refers to 2- (tert-butoxycarbonyloxyamino) -2- phenylacetonitrile .
  • BzlS0 2 refers to benzylsulfonyl .
  • Cbz or “CBz” refers to benzyloxycarbonyl .
  • CNNH 2 or "H 2 NCN” refers to cyanamide.
  • CsCo 3 refers to cesium carbonate.
  • DCA dichloroacetic acid
  • DCC N, N' -dicyclohexylcarbodiimide
  • DCM dichloromethane
  • DIEA diisopropylethylamine
  • DMF N,N-dimethylformamide
  • DMSO dimethyl sulfoxide
  • DMAP 4-N,N-dimethylaminopyridine
  • EDC refers to l-ethyl-3- (3-dimethylamino-propyl) carbodiimide hydrochloride salt.
  • Et 3 N or “TEA” refers to triethylamine .
  • EtOAc refers to ethyl acetate.
  • EtOH refers to ethanol.
  • HATU refers to 0- (7-azabenzotriazol-l-yl) -1, 1, 3, 3- tetramethyluromium hexafluorophosphate .
  • HBTU refers to 2- (lH-benzotriazol-1-yl) -1, 1, 3, 3- tetramethyluronium hexafluorophosphate .
  • HCl refers to hydrochloric acid
  • HOAc refers to acetic acid.
  • HOAt or “HOAT” refers to l-hydroxy-7- azabenzotriazole .
  • HOBt refers to 1-hydroxybenzotriazole monohydrate.
  • i-BuOCOCl refers to isobutylchloroformate .
  • HPLC refers to high pressure liquid chromatography.
  • LiAlH 4 refers to lithium aluminum hydride.
  • LiAlH 2 (OEt) 2 refers to lithium aluminum hydride diethoxide.
  • Me refers to methyl
  • MeOH refers to methanol .
  • NMM refers to N-methylmorpholine .
  • NBS N-bromosuccinimide
  • PhB(OH) 2 refers to phenylboronic acid.
  • PyBOP refers to benzotriazole-ly-oxy-tris- pyrrolidino-phosphonium hexafluorophosphate .
  • RP-HPLC refers to reverse phase high pressure liquid chromatography.
  • TFA trifluoroacetic acid
  • THF tetrahydrofuran
  • TLC thin layer chromatography
  • Figure 1 depicts a reaction scheme for solution phase synthesis of an intermediate useful in synthesizing a compound of the present invention.
  • Compound 1-1 is N- -Cbz- D-serine (O-t-butyl)
  • compound 1-2 is alanine methyl ester, hydrochloride salt.
  • i through “iv” are defined as i) EDC, 1-hydroxybenzotriazole and acetonitrile, diisopropylethylamine to give Cbz-D-Ser (O-t-butyl) -Ala-OMe; (ii) ethanol/acetic acid/water (4:1:1), 10% Pd on carbon, 45 psi H 2 for 2 hours, 95% yield after work-up; iii) acetonitrile, benzenesulfonyl chloride, diisopropylethylamine, 43% yield after work-up; and iv) methanol, 1.0 M lithium hydroxide, acidification on DOWEX ion exchange resin, eluting with methanol/water, 95% yield after work-up.
  • Figure 2 depicts a reaction scheme for a solution phase synthetic route which may be used to prepare an intermediate useful in the preparation of a compound of the present invention.
  • "i” through “iii” are defined as: i) isobutyl chloroformate, sodium carbonate, water, 99.5% yield after workup; ii) alanine t-butyl ester, hydrochloride salt, EDC, and hydroxybenzotriazole in acetonitrile; diisopropylethylamine, quantitative yield after workup; and iii) TFA, DCM, quantitative yield after workup. See also Examples 74 to 76.
  • Figure 3 depicts a reaction scheme for the synthesis of intermediates which may be used in the preparation of compounds of the present invention.
  • "i” through “xi” are defined as follows: i) CuCN, DMF, reflux (4 hours) ; ii) EtOAc, 10% aqueous NaCN; iii) N-bromosuccinimide, 2,2' -azo-bisisobutyronitrile, CC1 4 , reflux (5 hours); iv)
  • Figure 4 depicts a reaction scheme for the synthesis of intermediates which may be used in the preparation of compounds of the present invention.
  • “i” through “iv” are defined as follows: i) NaN 3 , DMF; ii) 10% Pd/C, EtOAc, 45 psi H 2 (11 hours) ; iii) hydroxylamine HC1, NMM, MeOH; and iv) 10% Pd/C, MeOH, 45 psi H 2 (48 hours) .
  • Figure 5 depicts a reaction scheme for the synthesis of intermediates which may be used in the preparation of compounds of the present invention.
  • i Cu(I)CN, DMF; ii) NBS, benzoylperoxide, CC1 4 , 80°C (14 hours); iii) NaN 3 , DMF, stirring (20 hours) ; iv) hydroxylamine HCl, NMM, MeOH, stirring (3 days); v) CsC0 3 , iodopropane, DMF, 50°C (20 hours); vi) triphenylphosphine, THF, stirring (20 hours); and vii) 3M NaOH to pH 14.
  • "i" is defined as: i) pyridine, R 6 C0C1.
  • Figure 7 depicts a reaction scheme for the synthesis of certain compounds of the present invention.
  • "i” through “vi” are defined as follows: i) trifluoracetic anhydride, 0°C, stir overnight; ice, CH 2 C1 2 , Na 2 S0 4 ; ii) Pd/C (10%) in MeOH (overnight); iii) N-N ' -di-Boc-N"- trifluoro ethanesulfonyl-guanidine, TEA, CH 2 C1 2 , 6 hours; HCl, brine, Na 2 S0 2 ; column chromatography (CH 2 Cl 2 /MeOH 99:1); iv) potassium carbonate, H 2 0/MeOH (2:15), overnight;
  • Figure 8 depicts the reaction scheme for the synthesis of certain compounds of the present invention.
  • "i” throught “v” are defined as: i) trifluoroacetic anyhydride, stir overnight; ice, CH 2 C1 2 , Na 2 S0 4 ; ii) Pd/C (10%) in MeOH (overnight); iii) N-N'-Boc-N"- trifluoromethanesulfonyl-guanidine, TEA, CH 2 C1 2 , 24 hours; HCl, brine, Na 2 S0 4 ; column chromatography (CH 2 Cl 2 /MeOH 98:2); iv) potassium carbonate, H 2 0/MeOH (1:1), overnight; H 2 0, CH 2 Cl 2 /MeOH (9:1), Na 2 S0 4 ; and v) benzylsulfonyl-D-serine-L- alanine carboxylate, HATU, HOAT, DIEA, in
  • Figure 9 -depicts a reaction scheme for the synthesis of a compound of the present invention.
  • "i” through “vi” are defined as follows: i) trifluoroacetic anhydride, 0°C, one hour; ii) KN0 3 , -20°C, stir overnight;
  • FIGS 10A to 10F depict certain preferred compounds of the present invention.
  • Figure 11 depits a reaction scheme for the synthesis of a compound of the present invention.
  • "i” through “xii” are defined as follows: i) CC1 4 , N- bromosuccinimide; N 2 , AIBN, stirring, flash chromatography; ⁇ ) DMF, NaN 3 , stir overnight, diethylether, water, brine, MgS0 4 , filter; (iii) MeOH, TEA, 65°C, 4 hours; EtOAc, H 2 0, brine, MgS0 ; iv) THF/water,- Ph 3 P, stir overnight; IN HCl, H 2 0, DCM, ph ⁇ 9; v) dioxane/water, K 2 C0 3 , Boc 2 0, stir overnight; EtOAc, aqueous NAHC0 3 , brine, Na 2 S0 4 , flash column chromatography; vi) DMF, 2-iodopropane, CsC0 3 ; EtOAc, a
  • Figure 12 depicts a reaction scheme for the synthesis of a compound of the present invention.
  • "i” through “vi” are defined as follows: i) EDC, HOBt, DIEA and CH 3 CN; ii) TFA, CH 2 C1 2 ; iii) BNS0 2 -dSer (tBu) -OH, EDC, HOBt, 2, 4, 6-collidine, CH 3 CN; iv) hydroxylamine hydrochloride; v) Zn/acetic acid; and vi) TFA, CH 2 C1 2
  • Figures 13A to 13C depict certain preferred compounds of the present invention.
  • Figure 14 depicts a reaction scheme for the synthesis of a compound of the present invention.
  • "i” through “v” are defined as follows: i) BH 3 , THF, 78% yield; ii) BOC-ON, 95% yield; iii) Pd/C, H 2 , 81% yield; iv) CNBr, NaHC0 3 , CH 3 CN, H 2 0; v) TFA, CH 2 C1 2 ; and vi) benzylsulfonyl-D- Serine-L-alanine carboxylate, HATU, HOAT, DIEA, CH 3 CN.
  • Figure 15 depicts a reaction scheme for the synthesis of a compound of the present invention.
  • "i” through “iii” are defined as follows: i) thionylchloride, MeOH; ii) Na/Hg (5%), CNNH 2 , reflux, 62% yield; and iii) benzylsulfonyl-D-serine-L-alanine carboxylate, HATU, HOAT, DIEA, CH 3 CN.
  • Figure 16 depicts a reaction scheme for the synthesis of a compound of the present invention.
  • i through “vi” are defined as follows: i) BH 3 , THF; ii) cl/Dioxane, quantitative yield; iii) BOC-ON, THF; iv) TEA, CH 2 C1 2 ; v) TFA/ CH 2 C1 2 ; and vi) benzylsulfonyl-D-serine-L- alanine carboxylate, HATU, HOAT, DIEA, CH 3 CN.
  • Figure 17 depicts a reaction scheme for the synthesis of a compound of the present invention.
  • "i through “vi” are defined as follows: i) BH 3 , THF; ii) HCl, quantitative yield; iii) BOC-ON, THF; iv) TEA, CH 2 C1 2 ; v) TFA, CH 2 C1 2 ; and vi) benzylsulfonyl-D-serine-L-alanine carboxylate, HATU, HOAT, DIEA, CH 3 CN.
  • Ri is selected from the group consisting of: (1) alkyl of 1 to about 12 carbon atoms which is unsubstituted or substituted with 1 or 2 substituents selected from the group consisting of Y x and Y 2 ,
  • aryl of about 6 to about 14 carbon atoms which is unsubstituted or mono-, di- or tri-substituted with 1 to 3 substituents selected from the group consisting of Y x , Y 2 , and Y 3 ,
  • heteroaryl of about 5 to about 14 ring atoms with the ring atoms selected from carbon and heteroatoms, wherein the heteroatoms are selected from oxygen, nitrogen, and sulfur, and which is unsubstituted or mono-, di- or trisubstituted with 1 to 3 substituents selected from the group consisting of Yi, Y 2 , and Y 3 ,
  • aralkyl of about 7 to about 15 carbon atoms which is unsubstituted or substituted on the alkyl chain with hydroxy or halogen and which is unsubstituted or mono-, di-, or tri-substituted on the aryl ring with 1 to 3 substituents selected from the group consisting of Yi, Y 2 , and Y 3 , (10) heteroaralkyl of about 5 to about 14 ring atoms with the ring atoms selected from carbon and heteroatoms, wherein the heteroatoms are selected from oxygen, nitrogen, and sulfur, which is unsubstituted or substituted on the alkyl chain with hydroxy or halogen and which is unsubstituted on the ring or mono-, di- or tri-substituted on the ring with 1 to 3 substituents selected from the group consisting of Yi, Y 2 , and
  • -N-morpholino and -S (0) m (CF 2 ) q CF 3 , wherein m is 0, 1 or 2, q is an integer from 0 to 5, and Zi and Z 2 are independently selected from the group consisting of alkyl of 1 to about 12 carbon atoms, aryl of about 6 to about 14 carbon atoms, heteroaryl of about 5 to about 14 ring atoms, aralkyl of about 7 to about 15 carbon atoms, and heteroaralkyl of about 5 to about 14 ring atoms, or
  • Yi and Y 2 are selected together to be -0[C(Z 3 ) (Z 4 )] r 0- or -0[C(Z 3 ) (Z 4 )] r+ ⁇ -, wherein r is an integer from 1 to 4 and Z 3 and Z 4 are independently selected from the group consisting of hydrogen, alkyl of 1 to about 12 carbon atoms, aryl of about 6 to about 14 carbon atoms, heteroaryl of about 5 to about 14 ring atoms, aralkyl of about 7 to about 15 carbon atoms, and heteroaralkyl of about 5 to about 14 ring atoms;
  • alkyl of 1 to about 4 carbon atoms which is unsubstituted or substituted with 1 to 2 substituents selected from the group consisting of Yi and Y 2 ,
  • heterocycloalkyl of 4 to about 6 ring atoms with the ring atoms selected from carbon and heteroatoms, wherein the heteroatoms are selected from the group consisting of oxygen, nitrogen, and S(0)i, wherein i is 0, 1 or 2, which is unsubstituted or mono-, di-, or trisubstituted on the ring with 1 to 3 substituents selected from the group consisting of Yi, Y 2 , and Y 3 ,
  • heterocyclo of 4 to about 6 ring atoms with the ring atoms selected from carbon and heteroatoms, wherein the heteroatoms are selected from the group consisting of oxygen, nitrogen, and S(0) ⁇ , wherein i is 0, 1, or 2, including —N V, wherein —N v is a 5 to 7 member heterocycle having 3 to 6 ring carbon atoms, where V is -CH 2 -, -0-, -S( 0)-, -S(0) 2 - or -S-, which is unsubstituted or mono-, di-, or tri-substituted on the ring carbons with 1 to 3 substituents selected from the group consisting of Yi, Y 2 , and Y 3 ,
  • alkenyl of 2 to about 6 carbon atoms which is unsubstituted or substituted with cycloalkyl of 3 to about 6 carbon atoms, which is unsubstituted or mono-, di-, or trisubstituted on the ring with 1 to 3 substituents selected from the group consisting of Yi, Y 2 , and Y 3 ,
  • phenyl which is unsubstituted or mono-, di- or tri-substituted with 1 to 3 substituents selected from the group consisting of Yi, Y 2 , and Y 3 ,
  • heteroaryl of about 5 to about 6 ring atoms with the ring atoms selected from carbon and heteroatoms, wherein the heteroatoms are selected from oxygen, nitrogen, and sulfur, and which is unsubstituted or mono-, di- or tri- substituted with 1 to 3 substituents selected from the group consisting of Yi, Y 2 , and Y 3 ,
  • heteroaralkyl of about 5 to about 6 ring atoms with the ring atoms selected from carbon and heteroatoms, wherein the heteroatoms are selected from oxygen, nitrogen, and sulfur, and which is unsubstituted or substituted on the alkyl chain with hydroxy or halogen and which is unsubstituted on the ring or mono-, di- or trisubstituted on the ring with 1 to 3 substituents selected from the group consisting of Yi, Y 2 , and Y 3 , (11) aralkenyl of about 8 to about 12 carbon atoms which is unsubstituted or mono-, di-, or tri-substituted on the aryl ring with 1 to 3 substituents selected from the group consisting of Yi, Y 2 , and Y 3 ,
  • heteroaralkenyl of about 5 to about 6 ring atoms with the ring atoms selected from carbon and heteroatoms, wherein the heteroatoms are selected from oxygen, nitrogen, and sulfur, and which is unsubstituted or mono-, di- or tri-substituted on the ring carbons with 1 to 3 substituents selected from the group consisting of Yi, Y 2 , and Y 3 , and (13) hydrogen; and
  • R 6 is selected from the group consisting of:
  • alkyl of 1 to about 12 carbon atoms which is unsubstituted or substituted with 1 or 2 substituents selected from the group consisting of Yi, and Y 2 ,
  • aralkyl of about 7 to about 15 carbon atoms which is unsubstituted or substituted on the alkyl chain with hydroxy or halogen and which is unsubstituted or mono-, di-, or tri-substituted on the aryl ring with 1 to 3 substituents selected from the group consisting of Y X f Y 2 , and Y 3 ,
  • heteroaralkyl of about 5 to about 14 ring atoms with the ring atoms selected from carbon and heteroatoms, wherein the heteroatoms are selected from oxygen, nitrogen, and sulfur, and which is unsubstituted or substituted on the alkyl chain with hydroxy or halogen and which is unsubstituted on the ring or mono-, di- or trisubstituted on the ring with 1 to 3 substituents selected from the group consisting of Yi, Y 2 , and Y 3 , and
  • R 3 is selected from H or methyl, or R 3/ and R 4a and R 4b are selected together as set forth in (f) ;
  • R a and R 4b are independently lower alkyl of 1 to 3 carbon atoms;
  • R 4a and R 4b are selected together and are - (CH 2 ) k - wherein k is 5 or 6 to give a spirocycloalkyl group; or
  • R 3 , and R a and R 4b are selected together as set forth in (f) ;
  • R 3 and R 4 are selected together to be in the S configuration to give a group at P2 selected from the group consisting of prolyl, pipecolyl, azetidine-2-carbonyl, 4-hydroxyprolyl, 3-hydroxyprolyl, 4- aminoprolyl, 4- (-CH 2 NH 2 ) -prolyl 3, 4-methanoprolyl and 3, 4-dehydroprolyl and R 4b is hydrogen;
  • R 7 is hydrogen or alkyl of 1 to about 4 carbon atoms;
  • (h) E is Q-T wherein (i) Q is selected from the group consisting of - C (R 1 3R 1 4) -v phenyl substituted with R 8 and R 9 , a 5- or 6- membered heterocyclic ring having 1 to 2 heteroatoms substituted with R 8 or R 8 and R 9 and a 9- or 10-membered heterocyclic ring having 1 to 2 heteroatoms substituted with Rs and R 9 , wherein heteroatoms are selected from nitrogen and sulfur; and
  • R 8 and R 9 are independently selected from the group consisting of hydrogen, hydroxy, halogen, alkyl of 1 to about 4 carbon atoms, alkyl of 1 to about 4 carbon atoms substituted with alkoxy of 1 to about 4 carbon atoms, alkoxy of 1 to about 6 carbon atoms, and trifluoromethyl;
  • R ⁇ 2 is hydrogen, alkyl of 1 to about 6 carbon atoms, alkoxy of 1 to about 6 carbon atoms or (CF 2
  • each of R ⁇ 3 and R X4 is independently selected from the group consisting of hydrogen and lower alkyl of 1 to about 3 carbon atoms;
  • R 15 is selected from the group consisting of hydrogen, alkyl of 1 to about 6 carbon atoms and -(CF 2 ) h CF 3 wherein h is 0, 1, 2 or 3 and t is an integer from 0 to 6; and pharmaceutically acceptable salts thereof.
  • preferred compounds include those of formula (I) with the proviso that, when E is -C (R 13 R 14 ) t ⁇ , then T is not -NHR1 5 .
  • preferred compounds of formula I include those compounds where Q is selected from the group consisting of phenyl substituted with R 8 and R 9 , a 5- or 6-membered heterocyclic ring having 1 to 2 heteroatoms substituted with R 8 or R 8 and R 9 and a 9- or 10-membered heterocyclic ring having 1 to 2 heteroatoms, wherein the heteroatoms are selected from the group consisting of nitrogen and sulfur.
  • Q is phenyl substituted with R 8 and R 9 .
  • R x groups include alkyls, especially isobutyl, 2-ethylhexyl, methyl, n-butyl, isopropyl, cyclohexylmethyl, and cyclohexylpropyl; cycloalkyl, especially (-)menthyl, (+)menthyl, and cyclohexyl; aryls, especially naphthyl and phenyl; aralkyls, especially benzyl, 3-phenylpropyl, and 2- phenylethyl; and fused carbocyclic alkyls, especially fluorenylmethyl.
  • Ri groups include phenyl, benzyl, 2-phenylethyl, isobutyl, n-butyl and 3-phenylpropyl .
  • R 2 groups include -CH 3 , -C 2 H 5 , -CH 2 NH-X'-R 5 and -CH(R 5 )0H, wherein R 5 is hydrogen, alkyl, especially methyl, or aralkyl.
  • R 5 is hydrogen, alkyl, especially methyl, or aralkyl.
  • Preferred chirality at the alpha carbon is R.
  • preferred chirality at the beta carbon is R.
  • R 2 groups are those which define P 3 as d-seryl (R 5 is H) or (R, R) d-allothreonyl (R 5 is methyl).
  • Alternate preferred R 2 groups include -(CH 2 ) 2 0A ⁇ and -CH(R 5 )0A ⁇ , more preferably -CH(R 5 )0A ⁇ ; preferably R 5 is H. More preferably R 2 is selected so that P 3 is defined as an acyl or carbonate ester of d-seryl.
  • Compounds wherein R 2 is -(CH 2 ) 2 OA ⁇ or -CH(R 5 )OA ⁇ may act as prodrugs .
  • a preferred R 3 group, when R 3 and R 4 are not selected together, is hydrogen.
  • a preferred R 4 group, when R 3 and R 4 are not selected together, is methyl, vinyl, allyl or propargyl .
  • prolyl, 3-hydroxy-prolyl, 4-hydroxyprolyl, 3, 4-dehydroprolyl, 3, 4-methanoprolyl, and azetidine-2-carbonyl- are preferred selections to define a group at P2.
  • Preferred R 7 groups include hydrogen.
  • Preferred E groups include 4-amidinophenyl, 4- guanidinophenyl, 3-amidinopropyl, and 5- (2-amidino-thienyl) .
  • Especially preferred E groups include 4-amidinophenyl and 4- guanidinophenyl .
  • preferred compounds include those having an R 2 element that defines d-serine or d-allothreonine or an acyl or carbonate ester thereof at the P3 position of the compound and an amidinophenyl, guanidinophenyl or amidinothienyl group at PI .
  • such compounds also having either i) a hydrogen at R 3 and methyl at R 4 (P2 is alanine) , or ii) having R 3 and R 4 selected together so that P2 is prolyl, azetidine-2-carbonyl, 3, 4-methanoprolyl or 3,4- dehydroprolyl .
  • Preferred compounds of the present invention include those depicted in Figures 10A to 10F and Figures 13A to 13C. Especially preferred are Compounds D, F, I, J, K, L. 0, R, T, U, V, AE, AH, AJ, AN and AV of Figures 10A to 10F and BG, BJ, BK and BQ of Figures 13A to 13C.
  • Figures 1 to 5 depict synthetic schemes for synthesis of intermediates which may be used in preparation of certain compounds of the present invention.
  • Figure 1 depicts solution phase synthesis of intermediates useful in the . preparation of compounds of the present invention. See Examples 60 to 62. See also
  • Examples 63 to 66, 67 to 70 and 71 to 73 describe solution phase syntheses of intermediates useful in the synthesis of compounds of the present invention.
  • Figure 2 depicts an alternate synthetic route to prepare an intermediate useful in the preparation of compounds of the present invention using solution phase synthesis. See also Examples 74 to 76.
  • Figure 6 depicts a reaction scheme for the preparation of a compound of the present invention having an esterified hydroxyl at P3.
  • Figure 7 depicts a reaction scheme for the preparation of a compound of the present invention having a 4- guanidinophenyl at PI.
  • Figure 8 depicts a reaction scheme for the preparation of a compound of the present invention having a 3- guanidinophenyl at PI.
  • Figure 9 depicts a reaction scheme for the preparation of a compound of the present invention having 2- guanidinothiophenyl at PI.
  • Figure 11 depicts a reaction scheme for the preparation of a compound of the present invention having a 3- amidinopyridyl at Pi.
  • Figure 12 depicts a reaction scheme for the synthesis of a compound of the present invention having 4- amidinophenyl at Pi .
  • Figure 14 depicts a reaction scheme for the synthesis of a compound of the present invention having a bicyclic heterocyclic group at PI.
  • Figure 15 depicts a reaction scheme for the synthesis of a compound of the present invention having an amino- imidazolyl group at PI.
  • Figure 16 depicts a reaction scheme for a compound of the present invention having a 2-chloro-4-guanidinophenyl group at Pi .
  • Figure 17 depicts a reaction scheme for the synthesis for a compound of the present invention having a guanidino- pyridyl group at PI .
  • Preferred means of chemically coupling include formation of a peptide bond by using conventional coupling reagents known in the art. See Bodanszky, N. Peptide Chemistry, pp. 55-73, Springer-Verlag, New York (1988) and references cited therein.
  • the chemical coupling may be either by means of one-step or two-step coupling. In one-step coupling, the two coupling partners are coupled directly.
  • Preferred coupling reagents for one- step coupling of the coupling partners include DCC with HOBt, EDC with HOBt, EDC with HOAt, HBTU or TBTU.
  • an activated ester or anhydride of the C- ter inal carboxy group of one coupling partner is formed prior to its coupling to the other coupling partner.
  • Another preferred method for preparing compounds of the present invention containing hydrogenation sensitive groups such as alkenyl or aryl moieties substituted with halogen, cyano, nitro, or -S-Zi is to use boron tris (trifluoroacetate) , B(OCOCF 3 ) 3 , to cleave the N 9 -nitro of the arginine group.
  • the reagent is prepared by the reaction of BBr 3 and CF 3 COOH in dichloromethane at 0°C.
  • the reagent is also commercially available.
  • the N g -nitro compound is treated with boron tris (trifluoroacetate) in trifluoroacetic acid at 0°C. See, e . g.
  • An intermediate such as 6-1 (the compound of Example 9) is reacted with an acid chloride R 6 C0C1 in the presence of a base such as pyridine.
  • the appropriate chloroformate derivative In the preparation of compounds having an amidino or guanidino group at Pi, it may be preferred to cap the P3 hydroxyl with the carbonate group prior to deprotecting the amidino or guanidino group. Accordingly, it is preferred to treat the corresponding intermediate with the chloroformate derivative. (See, e . g. , Example 8). The product is then hydrogenated and optionally treated under hydrolysis conditions to yield the
  • preferred compounds of the present invention are selected for their potency and selectivity toward inhibition of serine proteases, especially urokinase.
  • Such evaluations are routinely performed in vi tro, following procedures such as those set forth in Example A.
  • a target serine protease and its substrate are combined under assay conditions permitting reaction of the protease with its substrate.
  • the assay is performed in the absence of test compound, and in the presence of increasing concentrations of the test compound.
  • the concentration of test compound at which 50% of the serine protease activity is inhibited by the test compound is the IC 50 value (Inhibitory Concentration) or EC50 (Effective Concentration) value for that compound.
  • IC 50 or EC 50 values are considered more potent inhibitors of the serine protease than those compounds having higher IC 50 or EC 50 values.
  • the IC 50 measurement is often used for more simplistic assays, whereas the EC 50 is often used for more complicated assays, such as those employing cells.
  • Kj . is calculated from the IC 50 .
  • Preferred compounds according to this aspect of the present invention have a K ⁇ value of lOOnM or less as measured in an in vi tro assay for inhibition of urokinase activity. Especially preferred compounds have a Ki value of less than 30nM.
  • test compounds also are evaluated for selectivity toward a serine protease.
  • a test compound is assayed for its potency toward a panel of serine proteases and other enzymes and an IC 50 value or EC 50 value is determined for each test compound in each assay system.
  • a compound is deemed selective if its IC 50 value or EC 50 value (or i value) in the target enzyme assay is at least one order of magnitude less than the next smallest IC 50 value or EC 50 value measured in the selectivity panel of enzymes .
  • Preferred compounds of the present invention have a Ki value of lOOnM or less as measured in an in vitro assay for inhibition of urokinase activity.
  • Especially preferred compounds have a Ki value in the in vitro urokinase inhibition assay that is at least one order of magnitude smaller than the IC 50 value measured in the in vi tro tPA inhibition assay.
  • Compounds having a selectivity ratio of IC 50 tPA assay: K urokinase assay of greater than 100 are especially preferred.
  • Compounds of the present invention also are evaluated for their activity in vivo .
  • the type of assay chosen for evaluation of test compounds will depend on the pathological condition to be treated or prevented by use of the compound, as well as the route of administration to be evaluated for the test compound.
  • the procedures described by Jankun et al . [Cane. Res. 57:559-563, 1997] to evaluate PAI-1 can be employed. Briefly, the ATCC cell lines DU145, which expresses a high level of uPA, and LnCaP, which does not express uPA, are injected into SCID mice. After tumors are established, the mice are given test compound according to a dosing regime determined from the compound's in vitro characteristics. The Jankun et al . compound was administered in water. Tumor volume measurements are taken twice a week for about five weeks.
  • a compound is deemed active if an animal to which the compound was administered exhibited decreased tumor volume, as compared to animals receiving appropriate control compounds. Furthermore, a comparison of a compound's effect in animals injected with DU145 cells versus LnCaP cells can indicate whether the compound' s effect was due to inhibition of urokinase or otherwise.
  • a murine xenograft selected for high lung colonization potential is injected into C57B1/6 mice i.v. (experimental metastasis) or s.c. into the abdominal wall (spontaneous metastasis).
  • Various concentrations of the compound to be tested can be admixed with the tumor cells in Matrigel prior to injection.
  • Daily i.p. injections of the test compound are made either on days 1-6 or days 7-13 after tumor inoculation.
  • the animals are killed about three or four weeks after tumor inoculation, and the lung tumor colonies are counted. Evaluation of the resulting data permits a determination as to efficacy of the test compound, optimal dosing and route of administration.
  • the activity of the compounds of the present invention toward decreasing tumor volume and metastasis can be evaluated in the model described by Rabbani et al . [Int. J. Cancer 63:840-845, 1995] to evaluate their inhibitor.
  • Mat LyLu tumor cells over-expressing uPA were injected into the flank of Copenhagen rats.
  • the animals were implanted with osmotic minipumps to continuously administer various doses of test compound for up to three weeks.
  • the tumor mass and volume of experimental and control animals were evaluated during the experiment, as were metastatic growths.- Evaluation of the resulting data permits a determination as to efficacy of the test compound, optimal dosing, and route of administration.
  • a rabbit cornea neovascularization model can be employed.
  • Avery et al . describe anesthetizing New Zealand albino rabbits and then making a central corneal incision and forming a radial corneal pocket.
  • a slow release prostaglandin pellet was placed in the pocket to induce neovascularization.
  • Test compound was administered i.p. for five days, at which time the animals were killed. The effect of the test compound is evaluated by review of periodic photographs taken of the limbus, which can be used to calculate the area of neovascular response and, therefore, limbal neovascularization.
  • a decreased area of neovascularization as compared with appropriate controls indicates the test compound was effective at decreasing or inhibiting neovascularatization.
  • An angiogenesis model used to evaluate the effect of a test compound in preventing angiogenesis is described by Min et al . [Cane. Res. 56:2428-2433, 1996].
  • C57BL6 mice receive subcutaneous injections of a Matrigel mixture containing bFGF, as the angiogenesis-inducing agent, with and without test compound. After five days, the animals are killed and the Matrigel plugs, in which neovascularization can be visualized, are photographed.
  • An experimental animal receiving Matrigel and an effective dose of test compound will exhibit less vascularization than a control animal or an experimental animal receiving a less- or non-effective dose of compound.
  • CAM CAM-induced CAM-induced CAM-induced CAM-induced CAM-induced CAM-induced CAM-induced CAM-induced CAM-induced CAM-induced CAM-induced CAM-induced CAM-induced CAM-induced CAM-induced CAM-induced CAM-induced CAM-induced CAM-induced CAM-induced CAM-induced CAM-induced CAM-induced CAM-induced cytoplasmic cytoplasmic cytoplasmic cytoplasmic cytoplasmic cytoplasmic cytoplasmic cytoplasmic cytoplasmic cytoplasmic cytoplasmic cytoplasmic cytoplasmic cytoplasmic cytoplasmic cytoplasmic cytoplasmic cytoplasmic cytoplasmic cytoplasmic cytoplasmic cytoplasmic cytoplasmic cytoplasmic cytoplasmic cytoplasmic cytoplasmic cytoplasmic cytoplasmic cytoplasmic cytoplasmic cytoplasmic cytoplasmic cytoplasmic cytoplasmic cytoplasmic cytoplasmic cytoplasmic cytoplasmic
  • the CAM model is also used in a standard assay of angiogenesis (i.e., effect on formation of new blood vessels (Brooks, P.C.; Montgomery, A.M. P.; and Cheresh, D.A., Methods in Molecular Biology 129: 257-269 (1999)).
  • a filter disc containing an angiogenesis inducer such as basic fibroblast growth factor (bFGF) is placed onto the CAM. Diffusion of the cytokine into the CAM induces local angiogenesis, which may be measured in several ways such as by counting the number of blood vessel branch points within the CAM directly below the filter disc.
  • the ability of compounds of the present invention to inhibit cytokine-induced angiogenesis can be tested using this model.
  • a test compound can either be added to the filter disc that contains the angiogenesis inducer, be placed directly on the membrane or be administered systemically. The extent of new blood vessel formation in the presence and/or absence of test compound can be compared using this model.
  • compositions prepared for storage or administration which comprise a therapeutically effective amount of a compound of the present invention in a pharmaceutically acceptable carrier.
  • the therapeutically effective amount of a compound of the present invention will depend on the route of administration, the type of mammal being treated, and the physical characteristics of the specific mammal under consideration. These factors and their relationship to determining this amount are well known to skilled practitioners in the medical arts. This amount and the method of administration can be tailored to achieve optimal efficacy but will depend on such factors as weight, diet, concurrent medication and other factors which those skilled in the medical arts will recognize.
  • the therapeutically effective amount of the compound of the present invention can range broadly depending upon the desired affects and the therapeutic indication. Typically, dosages will be between about 0.01 mg/kg and 100 mg/kg body weight, preferably between about 0.01 and 10 mg/kg, body weight .
  • Pharmaceutically acceptable 'carriers for therapeutic use are well known in the pharmaceutical art, and are described, for example, in Remington's Pharmaceutical Sciences, Mack Publishing Co. (A.R. Gennaro edit. 1985).
  • sterile saline and phosphate-buffered saline at physiological pH may be used.
  • Preservatives, stabilizers, dyes and even flavoring agents may be provided in the pharmaceutical composition.
  • sodium benzoate, sorbic acid and esters of p-hydroxybenzoic acid may be added as preservatives. Id. at 1449.
  • antioxidants and suspending agents may be used. Id.
  • compositions of the present invention may be formulated and used as tablets, capsules or elixirs for oral administration; suppositories for rectal administration; sterile solutions and suspensions for injectable administration; and the like.
  • the dose and method of administration can be tailored to achieve optimal efficacy but will depend on such factors as weight, diet, concurrent medication and other factors which those skilled in the medical arts will recognize.
  • injectable pharmaceutical compositions can be prepared in conventional forms, either as liquid solutions or suspensions, solid forms suitable for solution or suspension in liquid prior to injection, or as emulsions.
  • Suitable excipients are, for example, water, saline, dextrose, mannitol, lactose, lecithin, albumin, sodium glutamate, or the like.
  • the injectable pharmaceutical compositions may contain minor amounts of nontoxic auxiliary substances, such as wetting agents, pH buffering agents, and the like.
  • absorption enhancing preparations e.g., liposomes may be utilized.
  • the compounds of the present invention having urokinase inhibitory activity and/or activity in reducing or inhibiting blood vessel formation, including angiogenesis and neovascularization, may be used both in vitro and in vivo for a number of applications, some of which are described herein below.
  • the compounds of the present invention are active as inhibitors of urokinase and specifically bind urokinase. Accordingly those compounds that contain sites suitable for linking to a solid/gel support may be used in vitro for affinity chromatography to purify urokinase from a sample or to remove urokinase from a sample using conventional affinity chromatography procedures. These compounds are attached or coupled to an affinity chromatography either directly or through a suitable linker support using conventional methods. See, e.g. Current Protocols in
  • the compounds of the present invention having urokinase inhibitory activity are useful in in vitro assays to measure tPA activity in a sample.
  • assays which measure the total plasminogen activation activity in a blood sample a compound of the present invention having urokinase inhibiting activity will knock out that portion of plasminogen activation attributable to uPA, which will allow for calculation of the portion of the total plasminogen activation due to tPA activity as well as that due to uPA activity.
  • Use of such assays to monitor tPA activity would allow better dosage control in patients receiving tPA.
  • These assays could also be used to monitor uPA activity levels in tissue samples, such as from biopsy or to monitor uPA/tPA activities for any clinical situation where measurement of plasminogen activation activity is of assistance. These assays may also be used to monitor plasminogen activator activity where a patient has been treated with a non-endogenous compound having plasminogen activator activity, such as streptokinase and staphlyokinase .
  • the compounds of the present invention are useful in vivo for treatment of pathologic conditions which would be ameliorated by decreased urokinase activity.
  • these compounds will inhibit the activation of metalloproteases by the uPA-plasmin cascade in synovial fluid and thus, may be used in treatment of arthritis. It is believed these compounds will be useful in decreasing or inhibiting metastasis, neovascularization, and degradation of the extracellular matrix in tumors and other neoplasms.
  • These compounds will be useful as therapeutic agents in treating conditions characterized by pathological neovascularation such as retinal ' disease, retinopathies and other conditions, including those described hereinabove in the Background and Introduction to the Invention.
  • Another use for the compounds of the present invention having urokinase inhibitory activity is as an antidote if too much exogenous urokinase has been given to a patient for therewith purposes, such as for dissolving a blood clot.
  • the compounds of the present invention may be used in treating conditions characterized by inflammation due to their anti-inflammatory effects from inhibition of urokinase, thereby interfering with mediators of cell adhesion or migration.
  • Such anti-inflammatory applications include treatment of stroke and complications of organ transplants.
  • the present invention includes methods for preventing or treating a condition in a mammal suspected of having a condition which will be attenuated by inhibition of urokinase activity comprising administering to said mammal a therapeutically effective amount- of a compound or a pharmaceutical composition of the present invention.
  • the compounds or pharmaceutical compositions of the present invention are administered in vivo, ordinarily in a mammal, preferably in a human.
  • the compounds or pharmaceutical compositions can be administered to a mammal in a variety of ways, including orally, parenterally, intravenously, subcutaneously, intramuscularly, colonically, rectally, nasally or intraperitoneally, employing a variety of dosage forms.
  • Administration is preferably oral, such as by tablets capsules or elixirs taken on a daily basis.
  • the compounds or pharmaceutical compositions of the present invention are administered alone or in combination with one another, or in combination with other therapeutic or in vivo diagnostic agents.
  • a "therapeutically effective amount" of the compounds or pharmaceutical compositions of the present invention will vary depending upon the age, weight and mammalian species treated, the particular compounds employed, the particular mode of administration and the desired affects and the therapeutic indication. Because these factors and their relationship to determining this amount are well known in the medical arts, the determination of therapeutically effective dosage levels, the amount necessary to achieve the desired result of inhibiting uPA activity, will be within the ambit of one skilled in these arts. Typically, administration of the compounds or pharmaceutical composition of the present invention is commenced at lower dosage levels, with dosage levels being increased until the desired effect of inhibiting uPA activity to the desired extent is achieved, which would define a therapeutically effective amount. For the compounds of the present invention, alone or as part of a pharmaceutical composition, such doses are between about 0.01 mg/kg and 100 mg/kg body weight, preferably between about 0.01 mg/kg and 10 mg/kg, body weight .
  • Example 5 To a solution of the compound of Example 3 (0.42 g, 1.02 mmol) in dichloromethane (4.2 ml) was added trifluoroacetic acid (4.2 ml). The reaction mixture was stirred at ambient temperature for 1 hour. The reaction mixture was diluted with 50 ml n-heptane and concentrated in vacuo . The residue was resuspended in 10 ml acetonitrile and 50 ml n-heptane and concentrated in vacuo to yield 410 mg product.
  • Example 5 To a solution of the compound of Example 3 (0.42 g, 1.02 mmol) in dichloromethane (4.2 ml) was added trifluoroacetic acid (4.2 ml). The reaction mixture was stirred at ambient temperature for 1 hour. The reaction mixture was diluted with 50 ml n-heptane and concentrated in vacuo . The residue was resuspended in 10 ml acetonitrile and 50 ml n
  • Example 7 To a solution of the product in Example 7 (117 mg, 0.285 mmol) in 1.14 ml methanol was added hydroxylamine hydrochloride (33.7 mg, 0.485 mmol), followed by N- methylmorpholine (53 ⁇ l, 0.485 mmol). The reaction mixture was stirred overnight at ambient temperature and then at 50°C for six hours. The reaction mixture was concentrated in vacuo . The crude product was taken to the next step (Example 9) without further purification. The product eluted at 6.5 minutes by reverse phase (C18) HPLC at 0.1% trifluoroacetic acid in 5-50% aqueous acetonitrile over 20 minutes.
  • hydroxylamine hydrochloride 33.7 mg, 0.485 mmol
  • N- methylmorpholine 53 ⁇ l, 0.485 mmol
  • Example 8 To the product of Example 8 (126 mg, 0.285 mmol) in acetic acid (2.85 ml) and water (0.28 ml) was added 185 mg activated zinc dust. The reaction mixture was stirred overnight at room temperature. The zinc dust was filtered using a glass funnel and the filtrate was purified by preparative HPLC. The fractions containing the product eluted in a 5-20% aqueous acetonitrile containing 0.1% TFA and were pooled and lyophilized yielding 35 mg of the title compound as a white powder. The product eluted at 6.0 minutes by reverse phase (C18) HPLC at 0.1% trifluoroacetic acid in 5-50% aqueous acetonitrile over 20 minutes.
  • C18 reverse phase
  • Example 14 To a solution of the compound of Example 12 (0.47 g, 1.44 mmol) in ethylacetate (5.7 ml) was added 5M anhydrous HCl in ethylacetate (1.44 ml) and the reaction was stirred at ambient temperature overnight. The reaction mixture was concentrated in vacuo to yield 363 mg (95%) of a white solid.
  • Example 14 To a solution of the compound of Example 12 (0.47 g, 1.44 mmol) in ethylacetate (5.7 ml) was added 5M anhydrous HCl in ethylacetate (1.44 ml) and the reaction was stirred at ambient temperature overnight. The reaction mixture was concentrated in vacuo to yield 363 mg (95%) of a white solid.
  • Example 14 To a solution of the compound of Example 12 (0.47 g, 1.44 mmol) in ethylacetate (5.7 ml) was added 5M anhydrous HCl in ethylacetate (1.44 ml
  • the ethylacetate layer was washed with IN HCl (10 ml) , saturated sodium bicarbonate (2 x 15 ml) and brine (15 ml) , then dried with sodium sulfate to a yellow syrup (160 mg, 94%) .
  • Example 15 To a solution of the product in Example 15 (143 mg, 0.305 mmol) in 1.22 ml methanol was added hydroxylamine hydrochloride (0.036 mg, 0.519 mmol) followed by N- methylmorpholine (57 ⁇ l, 0.519 mmol). The reaction mixture was stirred over night at ambient temperature. Analytical HPLC suggested that the reaction was not complete. Additional hydroxylamine hydrochloride (0.036 mg, 0.519 mmol) and N-methylmorpholine (57 Dl, 0.519 mmol) were added and stirring continued at ambient temperature over night. The reaction mixture was concentrated in vacuo and the crude was purified by preparative HPLC. The fractions containing the product eluting in 5-20% aqueous acetonitrile containing 0.1% TFA solution were pooled and lyophilized yielding 16 mg of the title compound as a white powder. / 6
  • Example 16 To the product of Example 16 (15 mg, 0.030 mmol) in acetic acid (0.30 ml) and water (0.03 ml) was added 19 mg activated zinc dust. The reaction mixture was stirred over night at room temperature. The zinc dust was filtered using a glass funnel and the filtrate was purified by preparative HPLC. The fractions containing the product eluted in a 5- 20% aqueous acetonitrile containing 0.1% TFA, and were pooled and lyophilized yielding 7 mg of the title compound as a white powder. The product eluted at minutes by reverse phase (C18) HPLC at 0.1% trifluoroacetic acid in 5-50% aqueous acetonitrile over 20 minutes.
  • C18 reverse phase
  • Example 21 The compound of Example 21 (114 mg, 0.17 mmol) was dissolved in methanol (15 ml) and was hydrogenated on a Parr shaker overnight at 40 psi in the presence of 15 mg palladium on charcoal. The catalyst was filtered off. The reaction mixture was diluted to 35 ml with water and purified by preparative HPLC. The fractions containing the product eluted in 0-25% aqueous acetonitrile containing 0.1% TFA and were pooled and lyophilized yielding 16 mg of the title compound as a white powder.
  • Triphenylphosphine (Aldrich, 5.7 g) was added to a solution of 2-cyano-5- (azidomethyl) thiophene (compound 25, 2.5 g, 10 mmol) in THF (Aldrich, 40 ml) and water (10 ml) at 0°C. The resulting solution was allowed to warm to room temperature and stirred at ambient temperature for 10 hours. RP HPLC purification led to the title compound (2.3 g, 94%) .
  • Low resolution mass spectrum confirmed the desired mass (MH + 450.5) .
  • Example 28 To the product of Example 28 (60 mg, 0.13 mmol) in acetic acid (1.3 ml) and water (0.13 ml) was added 87 mg activated zinc dust. The reaction mixture was stirred overnight at ambient temperature. The zinc dust was filtered using a glass funnel and the filtrate was purified by preparative HPLC. The fractions containing the product eluted in 0-20% aqueous acetonitrile containing 0.1% TFA. They were pooled and lyophilized yielding 7 mg of the title compound as a white powder.
  • step A (b) A solution of crude N-benzyl-3, 4-methanoprolinol (step A, (b) ) (3 grams, 14.76 mmol) in methanol (120 ml) and concentrated HCl (1.5 ml) with 10% Pd/C (300 mg) was hydrogenated at 50 psi for 16 hours. The reaction mixture was filtered to remove the carbon-based catalyst and the filtrate was concentrated. The residue was dissolved in water/dioxane (100 ml) and diisopropylethylamine (3.2 ml) was added. Benzyl chloroformate (2.76 ml, 16.2 mmol) was added and the reaction mixture was stirred overnight.
  • step E A solution of the compound of Example 30 (step E) (0.70 g, 3.082 mmol), p-cyanobenzylamine hydrochloride salt (0.784 g, 4.62 mmol), EDC (0.88 g, 4.62 mmol), N- hydroxybenzotriazole (0.47 g, 3.082 mmol) and diisopropylethylamine (2.68 ml, 15.41 mmol) was stirred in acetonitrile (12 ml) at ambient temperature.
  • Example 33 To a solution of the compound of Example 31 (1 g, 2.93 mmol) in ethylacetate (11.7 ml) was added 5M anhydrous HCl in ethylacetate (2.93 ml) and the reaction was stirred at ambient temperature overnight. The reaction mixture was concentrated in vacuo to yield 645 mg (79%) of a white solid.
  • Example 33 To a solution of the compound of Example 31 (1 g, 2.93 mmol) in ethylacetate (11.7 ml) was added 5M anhydrous HCl in ethylacetate (2.93 ml) and the reaction was stirred at ambient temperature overnight. The reaction mixture was concentrated in vacuo to yield 645 mg (79%) of a white solid.
  • Example 33 To a solution of the compound of Example 31 (1 g, 2.93 mmol) in ethylacetate (11.7 ml) was added 5M anhydrous HCl in ethylacetate (2.93 ml) and the reaction was stir
  • Example 34 To a solution of the product in Example 34 (154 mg, 0.32 mmol) in 1.3 ml methanol was added hydroxylamine hydrochloride (0.11 g, 1.58 mmol) followed by N- methylmorpholine (209 ⁇ l, 1.902 mmol). The reaction mixture was stirred overnight at ambient temperature. The reaction mixture was concentrated in vacuo. The crude product was resuspended in 25% acetonitrile in water and purified by preparative HPLC. The fractions containing the product eluting in 0-20% aqueous acetonitrile containing 0.1% TFA solution were pooled and lyophilized yielding 19.5 mg of the title compound as a white powder. Low resolution mass spectroscopy confirmed the desired mass (MH + 516) .
  • Example 35 To the product of Example 35 (19.5 mg, 0.041 mmol) in acetic acid (0.41 ml) and water (0.041 ml) was added 27 mg activated zinc dust. The reaction mixture was stirred over night at room temperature. The zinc dust was filtered using a glass funnel and the filtrate was purified by preparative HPLC. The fractions containing the product eluting in 0-20% aqueous acetonitrile containing 0.1% TFA were pooled and lyophilized yielding 15 mg of the title compound as a white powder. The product eluted at 10.5 minutes by reverse phase (C18) HPLC at 0.1% trifluoroacetic acid in 5-50% aqueous acetonitrile over 20 minutes. Low resolution mass spectroscopy confirmed the desired mass (MH + 500).
  • 3-Nitrobenzylamine (8-1) (3.0 g, 16 mmol) was added in portions to stirring trifluoroacetic anhydride (30 ml) while the mixture was being cooled on ice, and the mixture stirred overnight. The suspension was poured onto ice
  • 3-Trifluoracetamidometylaniline (8-2) (the product of Example 42) (500 mg, 2.29 mmol) was added to a stirring mixture of N-N' -Di-Boc-N' ' -trifluoromethanesulfonyl- guanidine (prepared according to the procedure described in J. Org. chem . 1998, 63, 3804-3805) (986 mg, 2.52 mmol), TEA (642 ⁇ l, 4.58 mmol) in CH 2 C1 2 (10 ml), and the mixture was stirred for 24 hours.
  • Example 51 To a solution of the azide of Example 51 (11-3) (6.46 g, 40.6 mmol) in methanol (100 ml) was added hydroxylamine hydrochloride (3.95 g, 56.84 mmol), followed by triethylamine (9.6 ml). The solution was heated to 65°C for four hours. The solvent was removed under reduced pressure and the remaining residue was extracted with ethylacetate (300 ml) and water (100 ml) . The water layer was re- extracted with ethylacetate (2 x 200 ml) . The combined organic extracts were washed with brine (2 x 100 ml) , dried over MgS0 4 -and evaporated to yield the title product (11-4) (7.8 g) .
  • Example 55 The compound of Example 55 (11-7) (229 mg, 0.74 mmol) was dissolved in dioxane (1 ml) and treated with a solution of 4N HCl in dioxane (1 ml) for one hour at room temperature. Removal of the solvent under reduced pressure afforded 290 mg of a white solid. That product was then dissolved in acetonitrile (4.12 ml), neutralized with diisopropylethylamine (538 ⁇ l, 3.09 mmol) and coupled to Boc-alanine (195 mg, 1.03 mmol) using EDC (197 mg, 1.03 mmol) and HOBt (157.6 mg, 1.03 mmol) as coupling reagents.
  • Example 56 The compound of Example 56 (11-8) (181 mg, 0.48 mmol) was dissolved in dioxane (1 ml) and treated with 4N HCl in dioxane (1 ml) for 1.5 hour at room temperature. Removal of the solvent under reduced pressure afforded 200 mg of a white solid. That product was then dissolved in acetonitrile (5 ml) , neutralized with diisopropylethylamine (298 ⁇ l, 1.71 mmol) and coupled to BnS0 2 -dSer (tBu) -OH (180 mg, 0.57 mmol), using EDC (109 mg, 0.57 mmol) and HOBt (96 mg, 0.63 mmol) as coupling reagents.
  • Example 57 The compound of Example 57 (11-9) (137 mg, 0.24 mmol) was treated with 2 ml each dichloromethane and trifluoroacetic acid for one hour at room temperature. Removal of the solvent under reduced pressure afforded 169 mg of an orangish oil. The product eluted at 9 minutes by reverse phase (C18) HPLC at 0.1% trifluoroacetic acid in 5- 90% aqueous acetonitrile over 20 minutes.
  • C18 reverse phase
  • Example 58 The product of Example 58 (11-10) (74 mg, 0.14 mmol) in water (8 ml) and acetic acid (0.8 ml) was treated with activated zinc dust (91 mg) for 45 minutes. The solution was filtered and the filtrate was subjected to purification by reverse phase HPLC (C18) . The product eluted at 11 minutes by reverse phase HPLC at 0.1% trifluoroacetic acid in 5-25% aqueous acetonitrile over 20 minutes. MS (electrospray) : 463 (M + 1) .
  • N- ⁇ -Cbz-D-serine ( 1-1 ) (Bachem, 4 . 97 g, 16. 8 mmol ) , alanine methyl ester, hydrochloride salt ( 1-2 ) (Novabiochem, 4 . 7 g, 33 . 7 mmol ) , EDC ( 6. 5 g, 33 . 7 mmol ) , and 1- hydroxybenzotriazole (2.6 g, 16.8 mmol) were combined, and acetonitrile (67 ml) was added.
  • Example 61 (1-4) To the compound of Example 61 (1-4) (0.53 g, 1.4 mmol) in methanol (9 ml) was added 1.0M lithium hydroxide (3.0 ml, 3 mmol) . After stirring for 18 hours, the reaction mixture was- poured over a column of 10 ml of DOWEX (50 X 8-400) ion exchange resin, and eluted with methanol/water (60 ml of a 1:1 mixture).
  • DOWEX 50 X 8-400
  • Example 64 The compound of Example 64 (1.0 g, 3.2 mmol), alanine methyl ester, hydrochloride salt (Novabiochem, 0.89 g, 6.3 mmol), EDC (1.22 g, 6.3 mmol), and 1-hydroxybenzotriazole (0.49 g, 3.2 mmol) were combined and acetonitrile (13 ml) was added. After stirring the resulting slurry for 10 minutes, diisopropylethylamine (2.71 ml, 15.8 mmol) was added and the resulting clear mixture was stirred for an additional 18 hours. The solvent was removed. under reduced pressure, and the residue was suspended in ethyl acetate (100 ml).
  • the aqueous layer was separated and washed with ethyl acetate (300 ml) .
  • the combined ethyl acetate layers were combined and washed with IM HCl (300 ml) , saturated sodium bicarbonate (400 ml) , and brine (200 ml) .
  • the organic layer was dried with magnesium sulfate, treated with Darco decolorizing charcoal and filtered. The solvent was removed under reduced pressure, yielding 21.48 g of the title compound (98% yield) .
  • the aqueous layer was basified with 40% NaOH to pH 10, then extracted with ethyl acetate (2X) .
  • the combined organic layers were dried over sodium sulfate; the solvent was removed in vacuo.
  • the residue was purified by flash chromatography over silica gel (1 x 6 inch column) eluting with 10-30% ethyl acetate/hexanes to afford 150 mg (7% yield) of the title compound.
  • R f 0.60 (50% ethyl acetate/hexanes).
  • t r 10.5 minutes in a 5% to 90% acetonitrile gradient in 0.1% aqueous TFA buffer on a 4.6 x 250 mm, 5 micron particle, 100 angstrom pore, C18 column at a 1 ml/minute flow rate.
  • Triphenylphosphine (Aldrich, 5.7 g) was added to a solution of 2-cyano-5- (azidomethyl) thiophene (compound 3-4, 2.5 g, 10 mmol) in THF (Aldrich, 40 ml) and water (10 ml) at 0°C. The solution was allowed to warm to room temperature and stirred at ambient temperature for 10 hours. RP-HPLC purification gave the title compound (2.3 g, 94%).
  • Triphenylphosphine (Aldrich, 1.6 g, 6.2 mmol) was added to a solution of 3-fluoro-4- (azidomethyl) phenyl (-N- propyloxy) amidine (compound 5-7, 1.03 g, 4.1 mmol) in THF (15 ml) .
  • the reaction mixture was stirred at ambient temperature for 20 hours.
  • the resulting solution was extracted with EtOAc (2 x 100ml) .
  • the combined organic layers were washed brine and dried (MgS0 4 ) . Removal of solvent under vacuum gave the title compound (825 mg, 77%) .
  • Example 99 Following the protocol of Example 98, with the noted substitution for isopropyl chloroformate, the following compounds are prepared: Compound name Substitution in Example 99 n-butylsulfonyl-D- [ (+) - (+) -menthyl chloroformate menthyloxycarbonyl] -serine- alanine-4-amidinobenzylamide n-butylsulfonyl-D- phenyl chloroformate (phenoxycarbonyl) -serine- alanine-4-amidinobenzylamide
  • rt-PA tissue plasminogen activator
  • the buffer used for all assays was HBSA (lOmM HEPES, pH 7.5, 150mM sodium chloride, 0.1% bovine serum albumin).
  • the assay for IC 50 determinations was conducted by combining in appropriate wells of a Corning microtiter plate, 50 microliters of HBSA, 50 microliters of the test compound at a specified concentration (covering a broad concentration range) diluted in HBSA (or HBSA alone for V 0 (uninhibited velocity) measurement) , and 50 microliters of the enzyme diluted in HBSA.
  • Urokinase catalytic activity was determined using the chromogenic substrate 150mM S-2444 (L-Pyroglutamyl-glycyl-L- arginine-p-nitroaniline hydrochloride) , obtained from DiaPharma Group, Inc.
  • Urokinase (Abbokinase) , manufactured by Abbott Laboratories, was obtained from Priority Pharmaceuticals and diluted to 750pM in the HBSA assay buffer prior to use.
  • the assay buffer was HBS (lO M HEPES, 150mM sodium chloride pH 7.4) with 0.1% BSA. Ki was calculated using the IC 5 o value.
  • Enzyme activity was determined using the chromogenic substrate, Pefachrome t-PA (CH 3 S0 2 -D-hexahydrotyrosine- glycyl-L-Arginine-p-nitroaniline, obtained from Pentapharm Ltd.). The substrate was reconstituted in deionized water prior to use. Purified human ⁇ -thrombin was obtained from Enzyme Research Laboratories, Inc. The buffer used for all assays was HBSA (10 mM HEPES, pH 7.5, 150mM sodium chloride, 0.1% bovine serum albumin).
  • IC 50 determinations were conducted where HBSA (50 ⁇ L) , ⁇ -thrombin (50 ⁇ l) (the final enzyme concentration is 0.5nM) and inhibitor (50 ⁇ l) (covering a broad concentration range) , were combined in appropriate wells and incubated for 30 minutes at room temperature prior to the addition of substrate Pefachrome-t-PA (50 ⁇ l) (the final substrate concentration is 250 ⁇ M, about 5 times Km) .
  • the initial velocity of Pefachrome t-PA hydrolysis was measured by the change in absorbance at 405 nm using a Thermo Max® Kinetic Microplate Reader over a 5 minute period in which less than 5% of the added substrate was utilized.
  • the concentration of added inhibitor which caused a 50% decrease in the initial rate of hydrolysis was defined as the IC50 value.
  • Factor Xa catalytic activity was determined using the chromogenic substrate S-2765 (N-benzyloxycarbonyl-D- arginine-L-glycine-L-arginine-p-nitroaniline ) , obtained from DiaPharma Group (Franklin, OH) . All substrates were reconstituted in deionized water prior to use. The final concentration of S-2765 was 250 ⁇ M (about 5-times Km) . Purified human Factor X was obtained from Enzyme Research
  • rt-PA tissue plasminogen activator
  • the substrate was made up in deionized water followed by dilution in HBSA prior to the assay in which the final concentration was 500 micromolar (about 3-times Km) .
  • Human rt-PA Human rt-PA (Activase®) was obtained from Genentech Inc. The enzyme was reconstituted in deionized water and diluted into HBSA prior to the assay in which the final concentration was l.OnM. Plasmin Assay
  • Plasmin catalytic activity was determined using the chromogenic substrate, S-2366 [L-pyroglutamyl-L-prolyl-L- arginine-p-nitroaniline hydrochloride] , which was obtained from DiaPharma Group.
  • the substrate was made up in deionized water followed by dilution in HBSA prior to the assay in which the final concentration was 300 micromolar (about 2.5-times Km).
  • Purified human plasmin was obtained from Enzyme Research Laboratories, Inc. The enzyme was diluted into HBSA prior to assay in which the final concentration was l.OnM.
  • Activated Protein C (aPC) Assay aPC catalytic activity was. determined using the chromogenic substrate, Pefachrome PC (delta-carbobenzyloxy- D-lysine-L-prolyl-L-arginine-p-nitroaniline dihydrochloride) , obtained from Pentapharm Ltd.). The substrate was made up in deionized water followed by dilution in HBSA prior to the assay in which the final concentration was 400 micromolar (about 3 times Km) . Purified human aPC was obtained from Hematologic
  • the enzyme was diluted into HBSA prior to assay in which the final concentration was l.OnM.
  • Chymotrypsin catalytic activity was determined using the chromogenic substrate, S-2586 (methoxy-succinyl-L- arginine-L-prolyl-L-tyrosyl-p-nitroanilide) , which was obtained from DiaPharma Group.
  • the substrate was made up in deionized water followed by dilution in HBSA prior to the assay in which the final concentration was 100 micromolar (about 9-times Km) .
  • Purified (3X-crystallized; CDI) bovine pancreatic ⁇ -chymotrypsin was obtained from Worthington Biochemical Corp. The enzyme was reconstituted in deionized water and diluted into HBSA prior to assay in which the final concentration was 0.5nM.
  • Trypsin catalytic activity was determined using the chromogenic substrate, S-2222 (benzoyl-L-isoleucine-L- glutamic acid- [gamma-methyl ester] -L-arginine-p- nitroanilide) , which was obtained from DiaPharma Group.
  • the substrate was made up in deionized water followed by dilution in HBSA prior to the assay in which the final concentration was 250 micromolar (about 4-times Km) .
  • Purified (3X-crystallized; TRL3) bovine pancreatic trypsin was obtained from Worthington Biochemical Corp. The enzyme was reconstituted in deionized water and diluted into HBSA prior to assay in which the final concentration was 0.5nM.
  • Tables IA and IB list the determined Ki or IC 50 values for certain of the enzymes listed above for compounds of the present invention that demonstrate a high degree of specificity for the inhibition of urokinase compared to other serine proteases.
  • the chicken CAM (chick embryo chorioallantoic membrane) model a standard angiogenesis assay, is used to evaluate the ability of a test compound to inhibit angiogenesis.
  • This model is an established model for evaluation of activity of a test compound to affect formulation of new blood vessels.
  • a filter disc saturated with a 0.5 ⁇ g/ml solution of basic fibroblast growth factor (bFGF) is placed on the CAM of 10 day old chick embryos to induce angiogenesis. Twenty four hours later, 0 to 1 ⁇ g of Test Compound, in a total volume of 100 ⁇ l of sterile PBS, is injected intravenously into the embryo. Approximately 48 hours later, the embryos are sacrificed and the filter discs and surrounding CAM tissue are excised for analysis.
  • bFGF basic fibroblast growth factor
  • Angiogenesis is quantitated by counting the number of blood vessel branch points within the confined region of the filter [Brooks, P.C., et al, Methods in Molecular Biology 120:257-269 (1999)].
  • the angiogenic index is defined as the difference in the number of blood vessel branch points between an experimental group and the untreated, control embryos. Each experimental group will contain 8 to 10 chicken embryos.
  • a chicken embryo model is used to evaluate the activity of Test Compound to inhibit the growth of human tumor cells in vivo.
  • a single cell suspension of human fibrosarcoma cells (HT 1080) containing 4 X 10 5 cells in a total volume of 40 ⁇ l, is applied to 10 day old chick embryos as described by Brooks, et al ("Brooks, P.C., et al, "Integrin ⁇ v ⁇ 3 Antagonists Promote Tumor Regression by Inducing
  • Test Compound 0 to 10 ⁇ g are injected intravenously into the embryos. Following this single administration of compound, control and treated embryos are incubated for a total of 7 days and then sacrificed. Tumors are excised, trimmed free of surrounding CAM tissue, and weighed. The wet weights for tumors excised in this experiment are tabulated. Each experimental group will contain 10 to 12 chicken embryos.

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Abstract

L'invention concerne de nouveaux composés présentant une activité d'inhibiteurs non covalents de l'urokinase ainsi qu'une activité de réduction ou d'inhibition de la formation de vaisseaux sanguins. Ces composés comportent en P1 un groupe portant un fragment amidino ou guanidino ou un dérivé de ce fragment. Ces composés servent au contrôle in vivo de niveaux d'activateurs de plasminogène, au traitement in vivo d'affections pouvant être traitées par inhibition ou réduction de l'activité de l'urokinase, ainsi qu'au traitement d'affections pathologiques dans lesquelles la formation de vaisseaux sanguins est liée à une affection pathologique.
PCT/US2001/025337 2000-08-11 2001-08-10 Inhibiteurs non covalents de l'urokinase et de la formation de vaisseaux sanguins WO2002014349A2 (fr)

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NZ518195A NZ518195A (en) 2000-08-11 2001-08-10 Non-covalent inhibitors of urokinase and blood vessel formation
JP2002519486A JP2004506648A (ja) 2000-08-11 2001-08-10 ウロキナーゼおよび血管形成の非共有結合性インヒビター
AU83347/01A AU785260B2 (en) 2000-08-11 2001-08-10 Non-covalent inhibitors of urokinase and blood vessel formation
IL14904201A IL149042A0 (en) 2000-08-11 2001-08-10 Non-covalent inhibitors of urokinase and blood vessel formation
CA002387002A CA2387002A1 (fr) 2000-08-11 2001-08-10 Inhibiteurs non covalents de l'urokinase et de la formation de vaisseaux sanguins
IL149042A IL149042A (en) 2000-08-11 2002-04-09 Non-covalent inhibitors of urokinase and vascular formation
AU2006235835A AU2006235835B2 (en) 2000-08-11 2006-11-03 Non-covalent inhibitors of urokinase and blood vessel formation

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US09/733,645 US6586405B2 (en) 2000-08-11 2000-12-07 Non-covalent inhibitors of urokinase and blood vessel formation
EP00126874A EP1182207B1 (fr) 2000-08-11 2000-12-07 Inhibiteurs non-covalents de l'urokinase et de l'angiogenèse

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Cited By (14)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003053999A2 (fr) * 2001-12-12 2003-07-03 Wilex Ag Inhibiteurs selectifs d'urokinase
WO2003076391A2 (fr) * 2002-03-11 2003-09-18 Curacyte Ag Inhibiteurs de l'urokinase, leur production et leur utilisation
EP1364960A1 (fr) * 2001-02-02 2003-11-26 Chugai Seiyaku Kabushiki Kaisha Derives de peptide
WO2004062657A1 (fr) * 2003-01-15 2004-07-29 Curacyte Chemistry Gmbh 4-amidino- et 4-guanidinobenzylamines acylees utilisees pour inhiber la kallikreine plasmatique
WO2004103984A1 (fr) 2003-05-26 2004-12-02 Wilex Ag Composes comportant des groupes hydroxyamidine et hydroxyguanidine utilises en tant qu'inhibiteurs de l'urokinase
WO2007025718A1 (fr) * 2005-08-29 2007-03-08 Wilex Ag Composes d'oxadiazole utilises comme inhibiteurs d'urokinase
US7262211B2 (en) * 2001-12-04 2007-08-28 Dendreon Corporation Aromatic heterocyclic non-covalent inhibitors of urokinase and blood vessel formation
EP2087885A1 (fr) 2005-06-24 2009-08-12 Wilex AG Utilisation des inhibiteurs d'urokinase pour le traitement et/ou la prévention des maladies neuropathologiques
WO2009133348A1 (fr) * 2008-04-29 2009-11-05 Vantia Limited Dérivés d’aminopyridine
WO2012004678A2 (fr) 2010-07-07 2012-01-12 The Medicines Company (Leipzig) Gmbh Inhibiteurs de la sérine protéase
US8124587B2 (en) 2005-09-16 2012-02-28 The Medicines Company (Leipzig) Gmbh 2-(aminomethyl)-5-chlorobenzylamide derivatives and their use as inhibitors of the clotting factor Xa
US8207378B2 (en) 2006-10-24 2012-06-26 The Medicines Company (Leipzig) Gmbh Trypsin-like serine protease inhibitors, and their preparation and use
US9090658B2 (en) 2003-09-11 2015-07-28 The Medicines Company (Leipzig) Gmbh Base-substituted benzylamine analogs for use as coagulation factor Xa inhibitors, the production and use thereof
CN114957087A (zh) * 2022-04-13 2022-08-30 湖南复瑞生物医药技术有限责任公司 一种帕罗韦德中间体制备方法

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114269431A (zh) * 2019-08-21 2022-04-01 卡尔维斯塔制药有限公司 酶抑制剂

Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0601459A2 (fr) * 1992-12-02 1994-06-15 Bristol-Myers Squibb Company Inhibiteurs hétérocycliques de thrombine contenant un groupe sulfonamid
EP0669317A1 (fr) * 1994-01-27 1995-08-30 Mitsubishi Chemical Corporation Dérivés de la prolinamide
WO1996017860A1 (fr) * 1994-12-06 1996-06-13 Basf Aktiengesellschaft Nouveaux p-amidinobenzylamides dipeptidiques a restes n-terminaux sulfonyle ou aminosulfonyle
US5932567A (en) * 1995-02-10 1999-08-03 Basf Aktiengesellschaft Thrombin inhibitors
WO2000005245A2 (fr) * 1998-07-24 2000-02-03 Corvas International, Inc. Inhibiteurs d'urokinase et de la formation de vaisseaux sanguins
WO2000061608A2 (fr) * 1999-04-09 2000-10-19 Basf Aktiengesellschaft Inhibiteurs de faible poids moleculaire des proteases du complement
WO2001096366A2 (fr) * 2000-06-15 2001-12-20 Curacyte Ag Inhibiteurs du facteur de coagulation xa
WO2001096286A2 (fr) * 2000-06-15 2001-12-20 Curacyte Ag Inhibiteurs de l'urokinase

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0601459A2 (fr) * 1992-12-02 1994-06-15 Bristol-Myers Squibb Company Inhibiteurs hétérocycliques de thrombine contenant un groupe sulfonamid
EP0669317A1 (fr) * 1994-01-27 1995-08-30 Mitsubishi Chemical Corporation Dérivés de la prolinamide
WO1996017860A1 (fr) * 1994-12-06 1996-06-13 Basf Aktiengesellschaft Nouveaux p-amidinobenzylamides dipeptidiques a restes n-terminaux sulfonyle ou aminosulfonyle
US5932567A (en) * 1995-02-10 1999-08-03 Basf Aktiengesellschaft Thrombin inhibitors
WO2000005245A2 (fr) * 1998-07-24 2000-02-03 Corvas International, Inc. Inhibiteurs d'urokinase et de la formation de vaisseaux sanguins
WO2000061608A2 (fr) * 1999-04-09 2000-10-19 Basf Aktiengesellschaft Inhibiteurs de faible poids moleculaire des proteases du complement
WO2001096366A2 (fr) * 2000-06-15 2001-12-20 Curacyte Ag Inhibiteurs du facteur de coagulation xa
WO2001096286A2 (fr) * 2000-06-15 2001-12-20 Curacyte Ag Inhibiteurs de l'urokinase

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
TAMURA^Ð? S Y ET AL: "Synthesis and biological activity of peptidyl aldehyde urokinase inhibitors" BIOORGANIC & MEDICINAL CHEMISTRY LETTERS, OXFORD, GB, vol. 10, no. 9, May 2000 (2000-05), pages 983-987, XP004199057 ISSN: 0960-894X *

Cited By (24)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1364960A1 (fr) * 2001-02-02 2003-11-26 Chugai Seiyaku Kabushiki Kaisha Derives de peptide
EP1364960A4 (fr) * 2001-02-02 2005-05-18 Chugai Pharmaceutical Co Ltd Derives de peptide
US7262211B2 (en) * 2001-12-04 2007-08-28 Dendreon Corporation Aromatic heterocyclic non-covalent inhibitors of urokinase and blood vessel formation
WO2003053999A3 (fr) * 2001-12-12 2003-11-06 Wilex Ag Inhibiteurs selectifs d'urokinase
WO2003053999A2 (fr) * 2001-12-12 2003-07-03 Wilex Ag Inhibiteurs selectifs d'urokinase
WO2003076391A3 (fr) * 2002-03-11 2004-01-22 Curacyte Ag Inhibiteurs de l'urokinase, leur production et leur utilisation
WO2003076391A2 (fr) * 2002-03-11 2003-09-18 Curacyte Ag Inhibiteurs de l'urokinase, leur production et leur utilisation
US8476306B2 (en) 2002-03-11 2013-07-02 The Medicines Company (Leipzig) Gmbh Urokinase inhibitors, production and use thereof
US7838560B2 (en) 2002-03-11 2010-11-23 The Medicines Company (Leipzig) Gmbh Urokinase inhibitors, production and use thereof
JP4898091B2 (ja) * 2002-03-11 2012-03-14 ザ メディシンズ カンパニー （ライプツィヒ） ゲーエムベーハー ウロキナーゼの阻害剤、それらの製造および使用
WO2004062657A1 (fr) * 2003-01-15 2004-07-29 Curacyte Chemistry Gmbh 4-amidino- et 4-guanidinobenzylamines acylees utilisees pour inhiber la kallikreine plasmatique
US9365613B2 (en) * 2003-01-15 2016-06-14 The Medicines Company (Leipzig) Gmbh Acylated 4-amidino- and -4-guanidinobenzylamines for inhibition of plasma kallikrein
EP2340822A1 (fr) * 2003-01-15 2011-07-06 The Medicines Company (Leipzig) GmbH 4-amidino- et 4-guanidinobenzylamine acylée pour inhiber la kallicréine plasmatique
EP2243774A1 (fr) 2003-05-26 2010-10-27 Wilex AG Composés comportant des groupes hydroxyamidine et hydroxyguanidine utilisés en tant qu'inhibiteurs de l'urokinase
WO2004103984A1 (fr) 2003-05-26 2004-12-02 Wilex Ag Composes comportant des groupes hydroxyamidine et hydroxyguanidine utilises en tant qu'inhibiteurs de l'urokinase
US9090658B2 (en) 2003-09-11 2015-07-28 The Medicines Company (Leipzig) Gmbh Base-substituted benzylamine analogs for use as coagulation factor Xa inhibitors, the production and use thereof
EP2087885A1 (fr) 2005-06-24 2009-08-12 Wilex AG Utilisation des inhibiteurs d'urokinase pour le traitement et/ou la prévention des maladies neuropathologiques
WO2007025718A1 (fr) * 2005-08-29 2007-03-08 Wilex Ag Composes d'oxadiazole utilises comme inhibiteurs d'urokinase
US8124587B2 (en) 2005-09-16 2012-02-28 The Medicines Company (Leipzig) Gmbh 2-(aminomethyl)-5-chlorobenzylamide derivatives and their use as inhibitors of the clotting factor Xa
US8207378B2 (en) 2006-10-24 2012-06-26 The Medicines Company (Leipzig) Gmbh Trypsin-like serine protease inhibitors, and their preparation and use
US8410310B2 (en) 2006-10-24 2013-04-02 The Medicines Company (Leipzig) Gmbh Trypsin-like serine protease inhibitors, and their preparation and use
WO2009133348A1 (fr) * 2008-04-29 2009-11-05 Vantia Limited Dérivés d’aminopyridine
WO2012004678A2 (fr) 2010-07-07 2012-01-12 The Medicines Company (Leipzig) Gmbh Inhibiteurs de la sérine protéase
CN114957087A (zh) * 2022-04-13 2022-08-30 湖南复瑞生物医药技术有限责任公司 一种帕罗韦德中间体制备方法

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