JP2004506648A - Non-covalent inhibitors of urokinase and angiogenesis - Google Patents

Non-covalent inhibitors of urokinase and angiogenesis Download PDF

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JP2004506648A
JP2004506648A JP2002519486A JP2002519486A JP2004506648A JP 2004506648 A JP2004506648 A JP 2004506648A JP 2002519486 A JP2002519486 A JP 2002519486A JP 2002519486 A JP2002519486 A JP 2002519486A JP 2004506648 A JP2004506648 A JP 2004506648A
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ウェインハウス, マイケル アイ.
センプレ, ジョセフ エドワード
タミズ, アミール ピー.
マディソン, エドウィン エル.
レヴィー, オディル エスザー
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コーバス インターナショナル, インコーポレイテッド
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Priority to EP20000126874 priority patent/EP1182207B1/en
Priority to US09/733,645 priority patent/US6586405B2/en
Application filed by コーバス インターナショナル, インコーポレイテッド filed Critical コーバス インターナショナル, インコーポレイテッド
Priority to PCT/US2001/025337 priority patent/WO2002014349A2/en
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    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K5/00Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06Dipeptides
    • C07K5/06139Dipeptides with the first amino acid being heterocyclic
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K5/00Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06Dipeptides
    • C07K5/06008Dipeptides with the first amino acid being neutral
    • C07K5/06017Dipeptides with the first amino acid being neutral and aliphatic
    • C07K5/06026Dipeptides with the first amino acid being neutral and aliphatic the side chain containing 0 or 1 carbon atom, i.e. Gly or Ala
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K5/00Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06Dipeptides
    • C07K5/06008Dipeptides with the first amino acid being neutral
    • C07K5/06017Dipeptides with the first amino acid being neutral and aliphatic
    • C07K5/0606Dipeptides with the first amino acid being neutral and aliphatic the side chain containing heteroatoms not provided for by C07K5/06086 - C07K5/06139, e.g. Ser, Met, Cys, Thr
    • C07K5/06069Ser-amino acid
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K38/00Medicinal preparations containing peptides

Abstract

Novel compounds are provided that have activity as non-covalent inhibitors of urokinase and have activity in reducing or inhibiting angiogenesis. These compounds have a group having an amidino or guanidino moiety at P1 or a derivative thereof. These compounds are useful for monitoring plasminogen activator levels in vitro, and for treating conditions that are restored by inhibition or reduced activity of urokinase in vivo, as well as pathological changes in angiogenesis. Useful for treating a pathological condition associated with the condition.

Description

[0001]
(Cross-reference of related applications)
This application is a continuation-in-part of U.S. Patent Application No. 09 / 733,645 (filed on December 7, 2000), which is a continuation of U.S. Application No. 09 / 637,483 (filed on August 11, 2000). (These disclosures are incorporated herein by reference).
[0002]
(Field of the Invention)
Urokinase is an enzyme involved in tumor cell metastasis, neovascularization, and other activities. One object of the present invention is to provide novel compounds that are active as urokinase inhibitors that can be used to inhibit the activity of urokinase and thereby attenuate its deleterious effects. It is another object of the present invention to provide new compounds that inhibit angiogenesis, especially those associated with pathological conditions.
[0003]
(Background and introduction of the invention)
Urinary plasminogen activator (uPA; urokinase) is a serine protease within the trypsin / chymotrypsin family. In its pathological condition, three forms of uPA are seen: single-stranded pro-uPA, double-stranded uPA, and low-molecular-weight uPA (lacking the N-terminal domain). The pro-enzyme, pro-uPA, is converted to u-PA by cleaving the peptide bond at K158-I159. The resulting double-stranded uPA is linked by disulfide bonds and has an M of about 50 kD. r , And a C-terminal serine proteinase domain.
[0004]
uPA activity is concentrated on the cell surface upon binding to its receptor, uPAR. uPAR is a single-chain glycosylphosphatidylinositol (GPI) -immobilized membrane receptor. The N-terminal 92 amino acids of uPAR play a major role in binding to uPA and pro-uPA. Receptors for uPA are found on T cells, NK cells, monocytes, and neutrophils, as well as on vascular endothelial cells, fibroblasts, smooth muscle cells, keratinocytes, placental trophoblasts, hepatocytes, and a wide variety of tumor cells. Are located in
[0005]
After conversion of pro-uPA to uPA, which occurs primarily with uPAR on the cell surface, uPA activates prosminogen to plasmin. Activation occurs upon cleavage at residue PGR-VV for human plasminogen or residue SGR-IV for bovine plasminogen. Since plasminogen is also present on the cell surface, this activation cascade concentrates u-PA and plasmin activity on the plasma membrane. Plasmin has many roles, including the activity of additional uPA and other enzymes, digestion of fibrin, and digestion of components of the extracellular matrix (ECM). Digestion of the ECM surrounding the tumor removes the ECM as a physical barrier to metastatic cells, and the metastatic cells are then free to leave the primary tumor and invade the second site. For a review of the role of the uPA / uPAR system in cancer metastasis, see "The Urokinase-type Plasminogen Activator System in Cancer Metastasis: A Review", Andreasen et al., Int. J. Canc. 72: 1-22 (1997).
[0006]
The correlation between poor levels of uPA and high rates of metastasis and poor prognosis has been described in certain tumors, particularly breast cancer [Quax et al. Cell Biol. 115: 191-199 (1991); Duffy et al., Cancer Res. 50: 6827-6829 (1990)]. For example, the following tumors showed high levels of uPA and / or uPA activity and high rates of metastasis: lung [Oka et al., Cancer Res. 51: 3522-3525 (1991)], bladder [Hasui et al., Int. J. Cancer 50: 871-873 (1992)], stomach [Nekarda et al., Lancet 343: 117 (1994)], uterine cancer [Kobayashi et al., Cancer. Res. 54: 6539-6548 (1994)], ovaries [Kuhn et al., Gynecol. Oncol. 55: 401-409 (1994)], kidney [Hofman et al., Cancer 78: 487-492 (1996)], brain [Bindahl et al. Neuro-Oncol. 22: 101-110 (1994)], and soft tissue sarcomas [Chong et al., Int. J. Cancer (Pred. Oncol.) 69: 268-272 (1996)]. Overproduction of uPA has been reported to result in increased skeletal metastasis by prostate cancer cells in vivo [Achbarou et al., Cancer Res. 54: 2372-377 (1994)].
[0007]
Inhibition or reduction of uPA activity, or disruption / inhibition of the interaction between uPA and its receptor (uPAR), has been shown to have a positive effect on the maintenance of extracellular matrix and an inhibitory effect on metastasis [Ossowski and Reich Ossowski, Cell 52: 321-328 (1988); Ossowski, J. et al., Cell 35: 611-619 (1983); Cell Biol. 107: 2437-2445 (1988); Wilhelm et al., Clin. Exp. Metastasis 13: 296-302 (1995); Achbarou et al., Cancer Res. 54: 2372-377 (1994); Crowley et al., Proc. Natl. Acad. Sci. ScL USA 90: 5021-5025 (1993); Kook et al., EMBO J. Clin. 13: 3983-3991 (1994)]. The results of such experimental studies suggest that uPA-catalyzed plasminogen activity is rate-limiting for tumor progression, local tumor invasion and / or distant metastasis formation. [Andreasen et al., Int. J. Canc. 72: 1-22 (1997)].
[0008]
The effects of the uPA system on cell migration and invasion are thought to be due both to the photolytic effect of plasmin-mediated degradation of the extracellular matrix and to the more direct interaction of the uPA receptor with components of the extracellular matrix. Degradation of the extracellular matrix allows metastatic cells to enter the matrix, while the interaction between the uPA receptor and the matrix itself assists the cells in its migration. Localization of the uPA / plasmin system on the cell surface or on the leading edge of metastatic cells is consistent with the putative role of uPA in metastasis [Plesner et al., Stem Cells 15: 398-408 (1997). )].
[0009]
The interaction of uPA with vitronectin, a component of the extracellular matrix, mediates cell adhesion and can be enhanced if uPAR is bound by uPA. Cell surface adhesion molecules (integrins) also appear to be involved in this adhesion function (particularly β-1 and β-2 integrins) [Paysant et al., Br. J. Haematol. 100: 45-51 (1998); Simon et al., Blood 88: 3185-3194 (1996)]. CD11b / CD18 integrins can associate with the uPA-uPAR complex and promote adhesion of cells bearing these receptors (eg, neutrophils, leukocytes).
[0010]
The uPA / uPAR system is also involved in establishing new vasculature, ie, neovascularization.
[0011]
Establishment of new vasculature is needed to sustain primary and metastatic tumor growth. Pathological neovascularization also includes retinal disease, rubeosis iritis, proliferative vitreoretinopathy inflammatory disease, diabetic retinopathy, chronic uveitis, Fuchs iris heterochromic iridocyclitis, neovascular glaucoma, Characterization of corneal or ocular nerve neovascularization, vascular disease, pterygium, glaucoma surgery bleb failure, hyperkeratosis, keloids and polyps formation (see EP 451,130). . Unwanted angiogenesis can also occur in the following conditions, or can be the result of the following activities: macular degradation, retinopathy of prematurity, corneal transplant rejection, post-lenopathyosis, pandemic keratoconjunctivitis, vitamin A Deficiency, contact lens overwear, atrophic keratoconjunctivitis, upper limbal keratitis, pterygium keratoconjunctivitis, sogren's disease, rosacea acne, fryctenulosis, syphilis, leprosy Non-Mycobacteria infection, lipid degeneration, chemical burn, bacterial or fungal ulcer, herpes simplex or shingles infection, protozoal infection, kaposi's sarcoma, Mohren's ulcer, terien marginal degeneration, marginal keratolysis, trauma, chronic joint Rheumatism, systemic lupus, polyarteritis, Wegener sarcoidosis, Adenitis, Stevens-Johnson disease, radial keratotomy, sickle cell anemia, sarcoid, fibropseudoxanthoma, Paget disease, venous or arterial occlusion, carotid occlusive disease, chronic uveitis, chronic choriotitis , Lyme disease, Eales disease, Bechets disease, Myopia, congenital structural deficiency of the optic nerve head (Optic pit), Stargardt's disease, Flatitis, Chronic retinal detachment, Hyperviscosity syndrome, Toxoplasmosis , Laser complications, abnormal growth of fibrous muscular tissue, hemangiomas, Ausler-Weber-Randue, solid tumors, blood borne tumors, AIDS, ocular neovascular disease ), Deformability Node disease, chronic inflammation, Crohn's disease, ulcerative colitis, rhabdomyosarcoma tumor, retinoblastoma tumor, Ewing sarcoma tumor, neuroblastoma tumor, osteosarcoma tumor, leukemia, psoriasis, atherosclerosis Disease, pemphigoid [described in US Pat. No. 5,712,291].
[0012]
Antagonists of uPA / uPAR binding (the EGF-like domain of uPA fused to IgG Fc) were stated to inhibit neovascularization and proliferation of murine B16 melanoma [Min et al., Cancer Res. 56: 2428-2433 (1996)]. The correlation stated between microvascular density, vascular invasion and uPA levels in breast cancer is consistent with this finding [Hildenbrand et al., Brit. J. Cancer 72: 818-823 (1995)]. The known uPA inhibitor amiloride has also been described to inhibit various neovascular pathologies [Glaser et al., EP 451,130; Avery et al., Arch. Ophthamol. 108: 1474-1476 (1990)].
[0013]
There are two major physiological inhibitors of uPA, PAI-1 and PAI-2, which are members of the serpin family of proteinase inhibitors. The binding of serpins to their cognate proteases involves a number of interactions between the amino acids of each protein, resulting in a separin-reactive loop (Ser-Ala-Arg-Met-Ala (SEQ ID NO: 1 for PAI-1), 2 includes those in Thr-Gly-Arg-Thr-Gly (SEQ ID NO: 2). Introduction of exogenous PAI-2 into laboratory animals has been reported to inhibit the rate of lung metastasis [Evans and Lin, Amer. Surg. 61: 692-697 (1995); Mueller et al., Proc. Natl. Acad. Sci. ScL USA 92: 205-209 (1995)]. The ability of PAI-1 to inhibit metastasis has not yet been consistently demonstrated. The gene for PAI-1 and means for its recombinant expression are disclosed in Loscutoff et al., US Pat. No. 4,952,512. Recombinant and native human PAI-2 are disclosed in Stephens et al., US Pat. No. 5,422,090.
[0014]
The most widely studied uPA inhibitors may be within the class of the inhibitors 4-substituted benzo [b] thiophene-2-carboxamidines, B428 (4-iodo-benzo [b] thiophene-2-carboxamidine) and B623 And its members [Tole et al., Cancer Res. 53: 2555-2559 (1993); Bridges et al., Bioorg. Med. Chem. 1: 403-410 (1993); Bridges et al., U.S. Patent No. 5,340,833]. Infusion of B428 in experimental rats seeded with tumor cells was stated to inhibit uPAR gene expression, reduce primary tumor volume, and reduce metastasis [Xing et al., Cancer Res. 57: 3585-3593 (1997)]. Daily intraperitoneal treatment of tumor-bearing mice with B428 or B623 blocks metastasis to muscle and fat but does not inhibit tumor-induced angiogenesis or reduce the rate of sudden lung metastasis Was. In fact, B623 enhanced the formation of lung metastases (Alonso et al., Breast Cancer Res. Treat. 40: 209-223 (1996)). Infusion of B248 in a syngeneic model of rat prostate cancer also led to a reduction in primary tumor volume and weight, and metastasis [Rabbani et al., Int. J. Cancer 63: 840-845 (1995)].
[0015]
Other known inhibitors of uPA include p-aminobenzamidine, which is a competitive inhibitor of uPA, and 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)]. Recently, epigallo-catechin-3 gallate (EGCG), a polyphenol found in green tea, was reported to bind to uPA and inhibit its activity [Jankun et al., Nature 387: 561 (1997)]. These investigators concluded that EGCG was a weaker inhibitor of uPA than amiloride, but suggested that EGCG could be consumed at much higher doses than amiloride without toxic effects. A competitive inhibitor of uPA, α-N-benzylsulfonyl-p-aminophenylalanine, is disclosed in US Pat. No. 4,165,258 by Pye et al.
[0016]
Other approaches for inhibiting the uPA / uPAR system include the development of bifunctional hybrid molecules consisting of the uPAR-binding domain of uPA and PAI-2, which inhibit uPA and bind uPAR in vitro. [Ballance et al., Eur. J. Biochem. 207: 177-183 (1992)]. Antagonists of uPAR have also been studied [Doyle and Rosenberg, US Pat. No. 5,656,726; Min et al., Cancer Res. 56: 2428-2433 (1996)], antisense oligonucleotides complementary to uPA were similarly studied [Wilhelm et al., Clin. Exp. Metast. 13: 296-302 (1995); Inversen and Scholar, US 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. in US Pat. No. 5,519,120. Small molecules that are said to inhibit urokinase, along with various other serine proteases, include those disclosed by Abe et al. In US Pat. Nos. 5,508,385 and 5,153,176, and Takano et al. J. Pharmacol. Exp. Therapeut. 271: 1027-1033 (1994).
[0017]
Compounds that directly inhibit u-PA binding to uPAR have been developed (Crowley et al., Proc. Natl. Acad. Sci. USA 90: 5021-5025 (1993); Goodson et al., Proc. Natl. Acad. USA 91: 7129-7133 (1994); Kobayashi et al., Brit. J. Cancer 67: 537-544 (1993), and Int. J. Cancer 57: 727-733 (1994), and J. Biol. Chem. 270: 8361-8366 (1995); Lu et al., FEBS Lett. 356: 56-59 (1994) and FEBS Lett. 380: 21-24 (1996)).
[0018]
In addition, prohepatocyte growth factor (HGF), a cell migration stimulating protein, is a substrate for uPA [Naldinie et al. 11: 4825-4833 (1992)]. Direct cleavage of the 66 kDa extracellular matrix protein and fibronectin by uPA has also been reported, suggesting a more direct role for uPA in promoting cell migration [Quigley et al., Proc. Natl. Acad. Sci. 84: 2776-2780 (1987)]. Thus, inhibition of uPA can affect these activities as well.
[0019]
(Summary of the Invention)
The present invention relates to novel peptidic non-covalent urokinase inhibitors. This compound has an arginine mimetic at P1. These compounds have activity as potent inhibitors of urokinase, and are therefore useful in reducing their harmful cavitation. The compounds of the present invention are active in inhibiting angiogenesis, especially in inhibiting angiogenesis associated with pathological processes.
[0020]
Accordingly, in one aspect, the invention relates to the following compounds of formula (I) and pharmaceutically acceptable salts thereof:
[0021]
Embedded image
here,
(A) X is -S (O) 2 -, -N (R ')-S (O) 2 -,-(C = O)-, -OC (= O)-, -NH-C (= O)-, -P- (O) (R ')-, and a direct bond. Wherein R ′ is independently hydrogen, alkyl of 1 to about 4 carbon atoms, aryl of about 6 to about 14 carbon atoms, or aralkyl of about 7 to about 16 carbon atoms, provided that X is -P In the case of (O) (R ')-, R' is not hydrogen;
(B) R 1 Is selected from the group consisting of:
(1) Unsubstituted or Y 1 And Y 2 Alkyl of 1 to about 12 carbon atoms, substituted with one or two substituents selected from the group consisting of:
(2) alkyl of 1 to about 3 carbon atoms substituted with cycloalkyl of about 3 to about 8 carbon atoms, wherein the cycloalkyl is unsubstituted or Y 1 , Y 2 , And Y 3 Alkyl substituted with 1 to 3 substituents selected from the group consisting of
(3) a cycloalkyl of 3 to about 15 carbon atoms, wherein the cycloalkyl is unsubstituted or has 1 , Y 2 , And Y 3 A cycloalkyl, mono-, di- or trisubstituted with 1 to 3 substituents selected from the group consisting of
(4) 4 to about 10 ring atom heterocycloalkyl having a ring atom selected from carbon and heteroatoms, wherein the heteroatom comprises oxygen, nitrogen, and S (O) i Wherein i is 0, 1 or 2 and the heterocycloalkyl is unsubstituted or Y 1 , Y 2 , And Y 3 A heterocycloalkyl, mono-, di- or tri-substituted with 1 to 3 substituents selected from the group consisting of
(5) 4 to about 10 ring atom heterocyclo having a ring atom selected from carbon and heteroatoms, wherein the heteroatom comprises oxygen, nitrogen, and S (O) i Wherein i is 0, 1, or 2, and the heterocyclo is
[0022]
Embedded image
And where
[0023]
Embedded image
Is a 5-7 membered heterocycle having 3-6 ring carbon atoms, wherein V is -CH 2 -, -O-, -S (= O)-, -S (O) 2 -Or -S-, wherein the heterocyclo is unsubstituted or has 1 , Y 2 , And Y 3 Heterocyclo, monosubstituted, disubstituted, or trisubstituted with 1 to 3 substituents selected from the group consisting of
(6) alkenyl of from 2 to about 6 carbon atoms, unsubstituted or substituted with cycloalkyl of from about 3 to about 8 carbon atoms, wherein the cycloalkyl is unsubstituted or has Y on the ring. 1 , Y 2 , And Y 3 Alkenyl, mono-, di- or trisubstituted with 1 to 3 substituents selected from the group consisting of
(7) aryl of about 6 to about 14 carbon atoms, wherein the aryl is unsubstituted or Y 1 , Y 2 , And Y 3 An aryl, mono-, di- or trisubstituted with 1 to 3 substituents selected from the group consisting of
(8) about 5 to about 14 ring atom heteroaryl having a ring atom selected from carbon and a heteroatom, wherein the heteroatom is selected from oxygen, nitrogen, and sulfur; Heteroaryl is unsubstituted or Y 1 , Y 2 , And Y 3 Heteroaryl, mono-, di- or tri-substituted with 1 to 3 substituents selected from the group consisting of
(9) an aralkyl of about 7 to about 15 carbon atoms, wherein the aralkyl is unsubstituted or substituted on the alkyl chain with hydroxy or halogen, and the aralkyl is unsubstituted, Or Y on this aryl ring 1 , Y 2 , And Y 3 Aralkyl, mono-, di- or tri-substituted with 1 to 3 substituents selected from the group consisting of
(10) a heteroaralkyl of about 5 to about 14 ring atoms having a ring atom selected from carbon and a heteroatom, wherein the heteroatom is selected from oxygen, nitrogen, and sulfur; Is unsubstituted or substituted on the alkyl chain with hydroxy or halogen, and the heteroaralkyl is unsubstituted on the ring or Y 1 , Y 2 , And Y 3 Heteroaralkyl, monosubstituted, disubstituted, or trisubstituted with 1 to 3 substituents selected from the group consisting of
(11) an aralkenyl of about 8 to about 16 carbon atoms, wherein the aralkenyl is unsubstituted or has Y on the aryl ring 1 , Y 2 , And Y 3 Aralkenyl, mono-, di- or trisubstituted with 1 to 3 substituents selected from the group consisting of
(12) about 5 to about 14 ring atom heteroaralkenyl having a ring atom selected from carbon and heteroatoms, wherein the heteroatom is selected from oxygen, nitrogen, and sulfur; The heteroaralkenyl can be unsubstituted or substituted on this ring carbon with Y 1 , Y 2 , And Y 3 Heteroaralkenyl, monosubstituted, disubstituted, or trisubstituted with 1 to 3 substituents selected from the group consisting of
[0024]
Embedded image
(17) a fused carbocyclic alkyl of about 9 to about 15 carbon atoms,
(18) difluoromethyl or perfluoroalkyl of 1 to about 12 carbon atoms,
(19) perfluoroaryl of about 6 to about 14 carbon atoms,
(20) a perfluoroaralkyl of about 7 to about 15 carbon atoms, and
(21) hydrogen when X is a direct bond;
Here, each Y 1 , Y 2 , And Y 3 Are independently selected, and
(I) halogen, cyano, nitro, tetrazolyl, guanidino, amidino, methylguanidino, -CF 3 , -CF 2 CF 3 , -CH (CF 3 ) 2 , -C (OH) (CF 3 ) 2 , -OCF 3 , -OCF 2 H, -OCF 2 CF 3 , -OC (O) NH 2 , -OC (O) NHZ 1 , -OC (O) NZ 1 Z 2 , -NHC (O) Z 1 , -NHC (O) NH 2 , -NHC (O) NZ 1 , -NHC (O) NZ 1 Z 2 , -C (O) OH, -C (O) OZ 1 , -C (O) NH 2 , -C (O) NHZ 1 , -C (O) NZ 1 Z 2 , -P (O) 3 H 2 , -P (O) 3 (Z 1 ) 2 , -S (O) 3 H, -S (O) m Z 1 , -Z 1 , -OZ 1 , -OH, -NH 2 , -NHZ 1 , -NZ 1 Z 2 , -C (= NH) NH 2 , -C (= NOH) NH 2 , -N-morpholino, and -S (O) m (CF 2 ) q CF 3 Wherein m is 0, 1 or 2; q is an integer from 0 to 5; 1 And Z 2 Is independently 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 about 5 to about 15 carbon atoms. Selected from the group consisting of heteroaralkyls of about 14 ring atoms, or
(Ii) Y 1 And Y 2 Are together -O [C (Z 3 ) (Z 4 )] r O- or -O [C (Z 3 ) (Z 4 )] r + 1 -Where r is an integer from 1 to 4 and Z 3 And Z 4 Is independently 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 about Selected from the group consisting of heteroaralkyl of 5 to about 14 ring atoms;
(C) R 2 Is -CH 3 , -C 2 H 5 ,-(CH 2 ) 2 OH,-(CH 2 ) 2 OA 1 , -CH (R 5 ) OH, -CH (R 5 ) OA 1 , And -CH 2 NH-X'-R 6 Selected from the group consisting of 1 Is -C (= O) OR 6 , -C (= O) R 6 Or -C (= O) NR 5 R 6 X ′ is —S (O) 2 -, -S (O) 2 -N (R ")-,-(C = O)-, -C (= O) -O-, -C (= O) -NH-, -P (O) (R")-, and a direct bond Wherein R "is hydrogen, alkyl of 1 to about 4 carbon atoms, aryl of about 6 to about 14 carbon atoms, or aralkyl of about 7 to about 16 carbon atoms, with the proviso that When X ′ is —P (O) (R ″) —, R ″ is not hydrogen; 5 Is selected from the group consisting of:
(1) Unsubstituted or Y 1 And Y 2 Alkyl of 1 to about 4 carbon atoms, substituted with 1-2 substituents selected from the group consisting of
(2) alkyl of 1 to about 3 carbon atoms substituted with cycloalkyl of 3 to about 6 carbon atoms, wherein the cycloalkyl is unsubstituted or 1 , Y 2 , And Y 3 An alkyl, mono-, di- or trisubstituted with 1 to 3 substituents selected from the group consisting of
(3) a cycloalkyl of 3 to about 6 carbon atoms, wherein the cycloalkyl is unsubstituted or has a Y 1 , Y 2 , And Y 3 A cycloalkyl, mono-, di- or trisubstituted with 1 to 3 substituents selected from the group consisting of
(4) 4 to about 6 ring atom heterocycloalkyl having a ring atom selected from carbon and heteroatoms, wherein the heteroatom comprises oxygen, nitrogen, and S (O) i Wherein i is 0, 1 or 2 and the heterocycloalkyl is unsubstituted or Y is 1 , Y 2 , And Y 3 A heterocycloalkyl, mono-, di- or tri-substituted with 1 to 3 substituents selected from the group consisting of
(5) 4 to about 6 ring atom heterocyclo having a ring atom selected from carbon and heteroatoms, wherein the heteroatom comprises oxygen, nitrogen, and S (O) i Wherein i is 0, 1, or 2, and the heterocyclo is
[0025]
Embedded image
And where
[0026]
Embedded image
Is a 5-7 membered heterocycle having 3-6 ring carbon atoms, wherein V is -CH 2 -, -O-, -S (= O)-, -S (O) 2 -Or -S-, wherein the heterocyclo is unsubstituted or 1 , Y 2 , And Y 3 Heterocyclo, monosubstituted, disubstituted, or trisubstituted with 1 to 3 substituents selected from the group consisting of
(6) alkenyl of from 2 to about 6 carbon atoms, unsubstituted or substituted with cycloalkyl of from about 3 to about 6 carbon atoms, wherein the cycloalkyl is unsubstituted or has Y on the ring. 1 , Y 2 , And Y 3 Alkenyl, mono-, di- or trisubstituted with 1 to 3 substituents selected from the group consisting of
(7) phenyl which is unsubstituted or Y 1 , Y 2 , And Y 3 Phenyl, mono-, di- or tri-substituted with 1 to 3 substituents selected from the group consisting of
(8) about 5 to about 6 ring atom heteroaryl having a ring atom selected from carbon and heteroatoms, wherein the heteroatom is selected from oxygen, nitrogen, and sulfur; Heteroaryl is unsubstituted or Y 1 , Y 2 , And Y 3 Heteroaryl, mono-, di- or tri-substituted with 1 to 3 substituents selected from the group consisting of
(9) alkyl of 1 to about 4 carbon atoms substituted with phenyl, wherein the phenyl is unsubstituted or has Y on the phenyl ring 1 , Y 2 , And Y 3 An alkyl, mono-, di- or trisubstituted with 1 to 3 substituents selected from the group consisting of
(10) a heteroaralkyl of about 5 to about 6 ring atoms having a ring atom selected from carbon and a heteroatom, wherein the heteroatom is selected from oxygen, nitrogen, and sulfur; The aralkyl is unsubstituted or substituted on the alkyl chain with hydroxy or halogen, and the heteroaralkyl is unsubstituted on the ring or Y on the ring. 1 , Y 2 , And Y 3 Heteroaralkyl, monosubstituted, disubstituted, or trisubstituted with 1 to 3 substituents selected from the group consisting of
(11) an aralkenyl of about 8 to about 12 carbon atoms, wherein the aralkenyl is unsubstituted or has Y on the aryl ring 1 , Y 2 , And Y 3 Aralkenyl, mono-, di- or trisubstituted with 1 to 3 substituents selected from the group consisting of
(12) about 5 to about 6 ring atom heteroaralkenyl having a ring atom selected from carbon and heteroatoms, wherein the heteroatom is selected from oxygen, nitrogen, and sulfur; The heteroaralkenyl can be unsubstituted or substituted on this ring carbon with Y 1 , Y 2 , And Y 3 Heteroaralkenyl, mono-, di-, or tri-substituted with 1 to 3 substituents selected from the group consisting of
(13) hydrogen; and
R 6 Is selected from the group consisting of:
(1) Unsubstituted or Y 1 And Y 2 Alkyl of 1 to about 12 carbon atoms, substituted with 1-2 substituents selected from the group consisting of:
(2) alkyl of 1 to about 3 carbon atoms substituted with cycloalkyl of 3 to about 8 carbon atoms, wherein the cycloalkyl is unsubstituted or 1 , Y 2 , And Y 3 An alkyl, mono-, di- or trisubstituted with 1 to 3 substituents selected from the group consisting of
(3) a cycloalkyl of 3 to about 15 carbon atoms, wherein the cycloalkyl is unsubstituted or has 1 , Y 2 , And Y 3 A cycloalkyl, mono-, di- or trisubstituted with 1 to 3 substituents selected from the group consisting of
(4) 4 to about 10 ring atom heterocycloalkyl having a ring atom selected from carbon and heteroatoms, wherein the heteroatom comprises oxygen, nitrogen, and S (O) i Wherein i is 0, 1 or 2 and the heterocycloalkyl is unsubstituted or Y is 1 , Y 2 , And Y 3 A heterocycloalkyl, mono-, di- or tri-substituted with 1 to 3 substituents selected from the group consisting of
(5) 4 to about 10 ring atom heterocyclo having a ring atom selected from carbon and heteroatoms, wherein the heteroatom comprises oxygen, nitrogen, and S (O) i Wherein i is 0, 1, or 2, and the heterocyclo is
[0027]
Embedded image
And where
[0028]
Embedded image
Is a 5-7 membered heterocycle having 3-6 ring carbon atoms, wherein V is -CH 2 -, -O-, -S (= O)-, -S (O) 2 -Or -S-, wherein the heterocyclo is unsubstituted or 1 , Y 2 , And Y 3 Heterocyclo, monosubstituted, disubstituted, or trisubstituted with 1 to 3 substituents selected from the group consisting of
(6) aryl of about 6 to about 14 carbon atoms, wherein the aryl is unsubstituted or 1 , Y 2 , And Y 3 An aryl, mono-, di- or trisubstituted with 1 to 3 substituents selected from the group consisting of
(7) about 5 to about 14 ring atom heteroaryl having a ring atom selected from carbon and heteroatoms, wherein the heteroatom is selected from oxygen, nitrogen, and sulfur; Heteroaryl is unsubstituted or Y 1 , Y 2 , And Y 3 Heteroaryl, mono-, di- or tri-substituted with 1 to 3 substituents selected from the group consisting of
(8) an aralkyl of about 7 to about 15 carbon atoms, wherein the aralkyl is unsubstituted or substituted on the alkyl chain with hydroxy or halogen, and the aralkyl is unsubstituted, Or Y on this aryl ring 1 , Y 2 , And Y 3 Aralkyl, mono-, di- or tri-substituted with 1 to 3 substituents selected from the group consisting of
(9) about 5 to about 14 ring atom heteroaralkyl having a ring atom selected from carbon and heteroatoms, wherein the heteroatom is selected from oxygen, nitrogen, and sulfur; The aralkyl is unsubstituted or substituted on the alkyl chain with hydroxy or halogen, and the heteroaralkyl is unsubstituted on the ring or Y on the ring. 1 , Y 2 , And Y 3 Hetero-aralkyl, mono-, di- or tri-substituted with 1 to 3 substituents selected from the group consisting of
(10) Hydrogen, provided that A 1 Is -C (= O) OR 6 If R 6 Is not hydrogen;
(D) R 3 Is selected from H or methyl, or R 3 And R 4a And R 4b Are selected together as described in (f);
(E) (i) R 4a Is the configuration of S, and H, -CH 2 -S-CH 3 , -CH 2 OH, -CH 2 CN, lower alkyl of 1 to about 3 carbon atoms, -CH 2 C≡CH, -CH 2 CH = CH 2 And -CH = CH 2 And R is selected from the group consisting of 4b Is hydrogen;
(Ii) R 4a And R 4b Is independently lower alkyl of 1-3 carbon atoms;
(Iii) R 4a And R 4b Are selected together and-(CH 2 ) k Wherein k is 5 or 6 to provide spirocycloalkyl; or
(Iv) R 3 , And R 4a And R 4b Are selected together as described in (f);
(F) Or R 3 And R 4a Are selected together to be in the S configuration to form prolyl, pipecolyl, azetidine-2-carbonyl, 4-hydroxyprolyl, 3-hydroxyprolyl, 4-aminoprolyl, 4-(-CH 2 NH 2 ) -Prolyl, 3,4-methanoprolyl, and 3,4-dehydroprolyl provide a group selected from the group consisting of 4b Is hydrogen;
(G) R 7 Is hydrogen or alkyl of 1 to about 4 carbon atoms;
(H) E is QT,
Here, (i) Q is -C (R Thirteen R 14 ) t -, R 8 And R 9 A phenyl, R 8 Or R 8 And R 9 A 5- or 6-membered heterocyclic ring having 1-2 heteroatoms, substituted with 8 And R 9 Selected from the group consisting of a 9 or 10 membered heterocyclic ring having 1-2 heteroatoms substituted with a heterocyclic atom selected from nitrogen and sulfur;
(Ii) T is -C (= NR 10 ) NHR 11 , -NH-C (= NR 10 ) NHR 11 , And -NHR Fifteen Selected from the group consisting of:
Where R 8 And R 9 Is independently 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 R, and trifluoromethyl; 10 And R 11 Is independently hydrogen, hydroxy, alkoxy of 1 to about 3 carbon atoms, trihydrocarbylsilyl of 3 to about 16 carbon atoms, alkyl of 1 to about 3 carbon atoms, or —C (= O) R 12 Where R 10 And R 11 Are not both hydroxy or both alkoxy; R 12 Is hydrogen, alkyl of 1 to about 6 carbon atoms, alkoxy of 1 to about 6 carbon atoms, or (CF 2 ) j CF 3 Wherein j is 0, 1, 2 or 3; Thirteen And R 14 R is independently selected from the group consisting of hydrogen and lower alkyl of 1 to about 3 carbon atoms; Fifteen Is 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 0-6.
[0029]
The compounds of the present invention have a P as shown in Formula Ia below. 1 , P 2 , P 3 And P 4 It can be divided into parts named:
[0030]
Embedded image
Where X, R 1 , R 2 , R 3 , R 4 , R 7 And E are as defined for formula (I). Thus, the P of the compound of formula (I) 1 Or the part called P1 is the following part:
[0031]
Embedded image
P of the compound of formula (I) 2 Or the part called P2 is the following part:
[0032]
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P of the compound of formula (I) 3 Or the part called P3 is the following part:
[0033]
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The invention is based, among other factors, on the inventors' discovery that the novel compounds of the invention are active as inhibitors of urokinase. The compounds of the present invention show activity in inhibiting angiogenesis.
[0034]
In another aspect, the invention relates to a pharmaceutical composition comprising a therapeutically effective amount of a compound of the invention and a pharmaceutically acceptable carrier.
[0035]
In yet another aspect, the invention relates to methods of using the compounds and pharmaceutical compositions of the invention for inhibiting urokinase.
[0036]
(Definition)
In accordance with the present invention and as used herein, the following terms, unless otherwise indicated, are defined to have the following meanings.
[0037]
The term "alkenyl" refers to an unsaturated aliphatic group having at least one double bond.
[0038]
The term "alkyl" refers to saturated aliphatic groups, including straight-chain, branched-chain and cyclic (including polycyclic) groups.
[0039]
The terms “alkoxy” and “alkoxyl” refer to a group having the formula R—O—, where R is an alkyl group.
[0040]
The term "alkoxycarbonyl" refers to -C (O) OR where R is alkyl.
[0041]
The term "aralkenyl" refers to an alkenyl group substituted with an aryl group. Preferably, the alkenyl group has 2 to about 6 carbon atoms.
[0042]
The term "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 1 to about 6 carbon atoms.
[0043]
The term "aryl" refers to an aromatic group having at least one ring having a conjugated pi-electron system, and includes carbocyclic aryl, heterocyclic aryl, and biaryl groups, all of which are required Can be substituted accordingly.
[0044]
The term "aryloxy" refers to a group having the formula RO-, wherein R is an aryl group.
[0045]
The term "aralkoxy" refers to a group having the formula RO-, wherein R is an aralkyl group.
[0046]
The term "amino acid" refers to both the natural and unnatural amino acids of these D and L stereoisomers, if the stereoisomeric forms of D and L are possible in the structure. Analog. Natural amino acids include alanine (Ala), arginine (Arg), asparagine (Asn), aspartic acid (Asp), cysteine (Cys), glutamine (Gln), glutamic acid (Glu), glycine (Gly), and histidine (His). , Isoleucine (Ile), 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, azetidine carboxylic acid, 2-amino adipic acid, 3-amino adipic acid, β-alanine, amino propionic acid, 2-amino butyric acid, 4-amino Butyric 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, N-ethylglycine, N-ethylasparagine, hydroxylysine, allohydroxylysine, 3-hydroxyproline, 4-hydroxyproline, isodesmosine, alloisoleucine, N-methylglycine, N-methylisoleucine, N-methylba Phosphorus, norvaline, norleucine, ornithine and pipecolic acid. Amino acid analogs include natural and non-natural amino acids, either reversibly or irreversibly chemically blocked or modified at the N-terminal amino group or side group, such as methionine sulfoxide. , Methionine sulfone, S- (carboxymethyl) -cysteine, S- (carboxymethyl) -cysteine sulfoxide and S- (carboxymethyl) -cysteine sulfone.
[0047]
The term "amino acid analog" refers to an amino acid in which either the C-terminal carboxy group, the N-terminal amino group, or a side chain functional group has been chemically modified to another functional group. For example, aspartic acid- (β-methyl ester) is an amino acid analog of aspartic acid; N-ethylglycine is an amino acid analog of glycine; or alanine carboxamide is an amino acid analog of alanine.
[0048]
The term "amino acid residue" refers to a group having the following structure: (1) -C (O) -R-NH-, where R is typically -CH (R ') Wherein R ′ is H or a carbon-containing substituent; or (2)
[0049]
Embedded image
Wherein p is 1, 2 or 3 and represents an azetidine carboxylic acid residue, a proline residue or a pipecolic acid residue, respectively.
[0050]
"Biaryl" refers to phenyl substituted with a carbocyclic or heterocyclic aryl as defined herein, ortho, meta or para to the point of attachment of the phenyl ring.
[0051]
"Brine" refers to a saturated aqueous solution of sodium chloride.
[0052]
"Carbocyclic aryl" refers to an aromatic group wherein the ring atoms of the aromatic ring are carbon atoms. Carbocyclic aryl groups include monocyclic carbocyclic aryl groups and naphthyl groups, all of which can be optionally substituted. Suitable carbocyclic aryl groups include phenyl and naphthyl. Suitable substituted carbocyclic aryl groups include indene and phenyl substituted with one or two substituents. The substituents are advantageously lower alkyl, hydroxy, lower alkoxy, lower alkoxycarbonyl, halogen, trifluoromethyl, difluoromethyl, nitro, cyano, and the like. Substituted (substituted) naphthyl is defined as Y as defined herein above for formula (I). 1 , Y 2 And / or Y 3 By naphthyl, more preferably 1-naphthyl or 2-naphthyl.
[0053]
“Cycloalkenyl” refers to cyclic alkenyl groups. Suitable cycloalkenyl groups include, for example, cyclopentyl and cyclohexyl.
[0054]
“Cycloalkyl” refers to a cyclic alkyl group having at least one ring, and includes a polycyclic group including a condensed cyclic alkyl group. Suitable cycloalkyl groups include, for example, cyclohexyl, cyclopropyl, cyclopentyl, and cycloheptyl.
[0055]
“Cyclohexylmethyl” refers to CH 2 Refers to a cyclohexyl group bonded to.
[0056]
"Fused carbocyclic" refers to a polycyclic fused carbocyclic ring having both aromatic and non-aromatic rings. Suitable fused carbocyclic rings include fluorenyl, tetralin, and the like.
[0057]
“Fused carbocyclic alkyl” refers to an alkyl group substituted with a fused carbocyclic ring moiety (preferably a polycyclic fused carbocyclic ring containing both aromatic and non-aromatic rings) Say. Suitable fused carbocyclic alkyl groups include fluorenylmethyl and the like.
[0058]
The term "halogen" refers to fluorine, chlorine, bromine and iodine.
[0059]
"Heteroaralkenyl" refers to an alkenyl group substituted with a heteroaryl and is described in "Handbook of Chemistry and Physics", 49th edition, 1968, R.A. C. Weast, The Chemical Rubber Co. , Cleveland, OH. In particular, Section C, Rules for Naming Organic Compounds, B.C. See Fundamental Heterocyclic Systems. Preferably, the alkenyl group has 2 to about 6 carbon atoms.
[0060]
"Heteroaralkyl" refers to an alkyl group substituted with a heteroaryl, such as picolyl, and is described in "Handbook of Chemistry and Physics", 49th Edition, 1968, R.A. C. Weast, The Chemical Rubber Co. , Cleveland, OH. In particular, Section C, Rules for Naming Organic Compounds, B.C. See Fundamental Heterocyclic Systems. Preferably, the alkyl group has 1 to about 6 carbon atoms.
[0061]
"Heteroaryl" refers to an aromatic group having from 1 to 14 carbon atoms and the remainder of the ring atoms being heteroatoms, and "Handbook of Chemistry and Physics", 49th edition, 1968, R . C. Weast, The Chemical Rubber Co. , Cleveland, OH. In particular, Section C, Rules for Naming Organic Compounds, B.C. See Fundamental Heterocyclic Systems. Suitable heteroatoms include oxygen, nitrogen, and S (O) i Wherein i is 0, 1 or 2 and suitable heterocyclic aryls include furanyl, thienyl, pyridyl, pyrrolyl, pyrimidyl, pyrazinyl, imidazolyl and the like.
[0062]
“Heterocyclo” refers to a reduced heterocyclic ring system consisting of carbon, nitrogen, oxygen and / or sulfur atoms and is disclosed in “Handbook of Chemistry and Physics”, 49th ed., 1968, R.A. C. Weast, The Chemical Rubber Co. , Cleveland, OH. In particular, Section C, Rules for Naming Organic Compounds, B.C. See Fundamental Heterocyclic Systems.
[0063]
"Heterocycloalkyl" refers to an alkyl group substituted with a heterocyclo group and is described in "Handbook of Chemistry and Physics", 49th edition, 1968, R.A. C. Weast, The Chemical Rubber Co. , Cleveland, OH. Heterocyclic systems described in. In particular, Section C.I. Rules for Naming Organic Compounds, B.C. See Fundamental Heterocyclic Systems. Preferably, the alkyl group has about 1 to about 6 carbon atoms.
[0064]
The term "lower" as referred to herein in connection with an organic radical or organic group refers to one or more carbon atoms, preferably four or less, and advantageously one or two carbon atoms. Have a radical or group defined. Such radicals or groups may be straight-chain or branched.
[0065]
“Perfluoroalkyl” refers to an alkyl group in which all hydrogens have been replaced by fluorine.
[0066]
“Perfluoroaryl” refers to an aryl group in which all hydrogens have been replaced by fluorine.
[0067]
"Perfluoroarylalkyl" refers to an aralkyl group in which all hydrogens in the aryl moiety have been replaced with fluorine.
[0068]
“Pharmaceutically acceptable salts” include salts of the compounds of the present invention, which are derived from combinations of the compounds of the present invention with organic or inorganic acids. In practice, use of the salt form is the same as use of the base form. The compounds of the present invention are useful in both free base and salt form, and both forms are considered to be within the scope of the present invention.
[0069]
"AcN", "CH 3 “CN” or “MeCN” refers to acetonitrile.
[0070]
"AIBN" refers to 2,2'-azobisisobutyronitrile.
[0071]
"Bn" refers to benzyl.
[0072]
"Boc" refers to t-butoxycarbonyl.
[0073]
"Boc 2 "O" refers to Boc anhydride (di-tert-butyl carbonate).
[0074]
"BOC-ON" refers to 2- (tert-butoxycarbonyloxyamino) -2-phenylacetonitrile.
[0075]
"BzlSO 2 "Refers to benzylsulfonyl.
[0076]
“Cbz” or “CBz” refers to benzyloxycarbonyl.
[0077]
"CNNH 2 "Or" H 2 "NCN" refers to cyanamide.
[0078]
"CsCO 3 "Refers to cesium carbonate.
[0079]
"DCA" refers to dichloroacetic acid.
[0080]
"DCC" refers to N, N'-dicyclohexylcarbodiimide.
[0081]
"DCM" or "CH 2 Cl 2 "Refers to dichloromethane.
[0082]
"DIEA" refers to diisopropylethylamine.
[0083]
"DMF" refers to N, N-dimethylformamide.
[0084]
"DMSO" refers to dimethyl sulfoxide.
[0085]
"DMAP" refers to 4-N, N-dimethylaminopyridine.
[0086]
"EDC" refers to 1-ethyl-3- (3-dimethylamino-propyl) carbodiimide hydrochloride.
[0087]
"Et 3 "N" or "TEA" refers to triethylamine.
[0088]
"EtOAc" refers to ethyl acetate.
[0089]
"EtOH" refers to ethanol.
[0090]
“HATU” refers to O- (7-azabenzotriazol-1-yl) -1,1,3,3-tetramethyluronium hexafluorophosphate.
[0091]
"HBTU" means 2- (1H-benzotriazol-1-yl) -1,1,3,3-tetramethyluronium hexafluorophosphate.
[0092]
“HCl” means hydrochloric acid.
[0093]
"HOAc" means acetic acid.
[0094]
"HOAt or HOAT" means 1-hydroxy-7-azabenzotriazole.
[0095]
"HOBt" means 1-hydroxybenzotriazole monohydrate.
[0096]
“I-BuOCOCl” means isobutyl chloroformate.
[0097]
"HPLC" means high performance liquid chromatography.
[0098]
"LiAlH 4 "Means lithium aluminum hydride.
[0099]
"LiAlH 2 (OEt) 2 "Means lithium aluminum hydride diethoxide.
[0100]
"Me" means methyl.
[0101]
“MeOH” means methanol.
[0102]
"NMM" means N-methylmorpholine.
[0103]
"NBS" means N-bromosuccinimide.
[0104]
"PhB (OH) 2 "Means phenylboronic acid.
[0105]
"Ph 3 P "or" PPh 3 "Means triphenylphosphine.
[0106]
"PyBOP" means benzotriazol-yl (ly) -oxy-tris-pyrrolidino-phosphonium hexafluorophosphate.
[0107]
"RP-HPLC" means reverse phase high performance liquid chromatography.
[0108]
"TFA" means trifluoroacetic acid.
[0109]
"THF" means tetrahydrofuran.
[0110]
"TLC" means thin layer chromatography.
[0111]
(Detailed description of the invention)
The following compounds and pharmaceutically acceptable salts thereof:
[0112]
Embedded image
here,
(A) X is -S (O) 2 -, -N (R ')-S (O) 2 -,-(C = O)-, -OC (= O)-, -NH-C (= O)-, -P- (O) (R ')-, and a direct bond. Wherein R ′ is independently hydrogen, alkyl of 1 to about 4 carbon atoms, aryl of about 6 to about 14 carbon atoms, or aralkyl of about 7 to about 16 carbon atoms, provided that X is -P In the case of (O) (R ')-, R' is not hydrogen;
(B) R 1 Is selected from the group consisting of:
(1) Unsubstituted or Y 1 And Y 2 Alkyl of 1 to about 12 carbon atoms, substituted with one or two substituents selected from the group consisting of:
(2) alkyl of 1 to about 3 carbon atoms substituted with cycloalkyl of about 3 to about 8 carbon atoms, wherein the cycloalkyl is unsubstituted or Y 1 , Y 2 , And Y 3 Alkyl substituted with 1 to 3 substituents selected from the group consisting of
(3) a cycloalkyl of 3 to about 15 carbon atoms, wherein the cycloalkyl is unsubstituted or has 1 , Y 2 , And Y 3 A cycloalkyl, mono-, di- or trisubstituted with 1 to 3 substituents selected from the group consisting of
(4) 4 to about 10 ring atom heterocycloalkyl having a ring atom selected from carbon and heteroatoms, wherein the heteroatom comprises oxygen, nitrogen, and S (O) i Wherein i is 0, 1 or 2 and the heterocycloalkyl is unsubstituted or Y 1 , Y 2 , And Y 3 A heterocycloalkyl, mono-, di- or tri-substituted with 1 to 3 substituents selected from the group consisting of
(5) 4 to about 10 ring atom heterocyclo having a ring atom selected from carbon and heteroatoms, wherein the heteroatom comprises oxygen, nitrogen, and S (O) i Wherein i is 0, 1, or 2, and the heterocyclo is
[0113]
Embedded image
And where
[0114]
Embedded image
Is a 5-7 membered heterocycle having 3-6 ring carbon atoms, wherein V is -CH 2 -, -O-, -S (= O)-, -S (O) 2 -Or -S-, wherein the heterocyclo is unsubstituted or has 1 , Y 2 , And Y 3 Heterocyclo, monosubstituted, disubstituted, or trisubstituted with 1 to 3 substituents selected from the group consisting of
(6) alkenyl of from 2 to about 6 carbon atoms, unsubstituted or substituted with cycloalkyl of from about 3 to about 8 carbon atoms, wherein the cycloalkyl is unsubstituted or has Y on the ring. 1 , Y 2 , And Y 3 Alkenyl, mono-, di- or trisubstituted with 1 to 3 substituents selected from the group consisting of
(7) aryl of about 6 to about 14 carbon atoms, wherein the aryl is unsubstituted or Y 1 , Y 2 , And Y 3 An aryl, mono-, di- or trisubstituted with 1 to 3 substituents selected from the group consisting of
(8) about 5 to about 14 ring atom heteroaryl having a ring atom selected from carbon and a heteroatom, wherein the heteroatom is selected from oxygen, nitrogen, and sulfur; Heteroaryl is unsubstituted or Y 1 , Y 2 , And Y 3 Heteroaryl, mono-, di- or tri-substituted with 1 to 3 substituents selected from the group consisting of
(9) an aralkyl of about 7 to about 15 carbon atoms, wherein the aralkyl is unsubstituted or substituted on the alkyl chain with hydroxy or halogen, and the aralkyl is unsubstituted, Or Y on this aryl ring 1 , Y 2 , And Y 3 Aralkyl, mono-, di- or tri-substituted with 1 to 3 substituents selected from the group consisting of
(10) a heteroaralkyl of about 5 to about 14 ring atoms having a ring atom selected from carbon and a heteroatom, wherein the heteroatom is selected from oxygen, nitrogen, and sulfur; Is unsubstituted or substituted on the alkyl chain with hydroxy or halogen, and the heteroaralkyl is unsubstituted on the ring or Y 1 , Y 2 , And Y 3 Heteroaralkyl, monosubstituted, disubstituted, or trisubstituted with 1 to 3 substituents selected from the group consisting of
(11) an aralkenyl of about 8 to about 16 carbon atoms, wherein the aralkenyl is unsubstituted or has Y on the aryl ring 1 , Y 2 , And Y 3 Aralkenyl, mono-, di- or trisubstituted with 1 to 3 substituents selected from the group consisting of
(12) about 5 to about 14 ring atom heteroaralkenyl having a ring atom selected from carbon and heteroatoms, wherein the heteroatom is selected from oxygen, nitrogen, and sulfur; The heteroaralkenyl can be unsubstituted or substituted on this ring carbon with Y 1 , Y 2 , And Y 3 Heteroaralkenyl, monosubstituted, disubstituted, or trisubstituted with 1 to 3 substituents selected from the group consisting of
[0115]
Embedded image
(17) a fused carbocyclic alkyl of about 9 to about 15 carbon atoms,
(18) difluoromethyl or perfluoroalkyl of 1 to about 12 carbon atoms,
(19) perfluoroaryl of about 6 to about 14 carbon atoms,
(20) a perfluoroaralkyl of about 7 to about 15 carbon atoms, and
(21) hydrogen when X is a direct bond;
Here, each Y 1 , Y 2 , And Y 3 Are independently selected, and
(I) halogen, cyano, nitro, tetrazolyl, guanidino, amidino, methylguanidino, -CF 3 , -CF 2 CF 3 , -CH (CF 3 ) 2 , -C (OH) (CF 3 ) 2 , -OCF 3 , -OCF 2 H, -OCF 2 CF 3 , -OC (O) NH 2 , -OC (O) NHZ 1 , -OC (O) NZ 1 Z 2 , -NHC (O) Z 1 , -NHC (O) NH 2 , -NHC (O) NZ 1 , -NHC (O) NZ 1 Z 2 , -C (O) OH, -C (O) OZ 1 , -C (O) NH 2 , -C (O) NHZ 1 , -C (O) NZ 1 Z 2 , -P (O) 3 H 2 , -P (O) 3 (Z 1 ) 2 , -S (O) 3 H, -S (O) m Z 1 , -Z 1 , -OZ 1 , -OH, -NH 2 , -NHZ 1 , -NZ 1 Z 2 , -C (= NH) NH 2 , -N-morpholino, and -S (O) m (CF 2 ) q CF 3 Wherein m is 0, 1 or 2; q is an integer from 0 to 5; 1 And Z 2 Is independently 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 about 5 to about 15 carbon atoms. Selected from the group consisting of heteroaralkyls of about 14 ring atoms, or
(Ii) Y 1 And Y 2 Are together -O [C (Z 3 ) (Z 4 )] r O- or -O [C (Z 3 ) (Z 4 )] r + 1 -Where r is an integer from 1 to 4 and Z 3 And Z 4 Is independently 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 about Selected from the group consisting of heteroaralkyl of 5 to about 14 ring atoms;
(C) R 2 Is -CH 3 , -C 2 H 5 ,-(CH 2 ) 2 OH,-(CH 2 ) 2 OA 1 , -CH (R 5 ) OH, -CH (R 5 ) OA 1 , And -CH 2 NH-X'-R 6 Selected from the group consisting of 1 Is -C (= O) OR 6 , -C (= O) R 6 Or -C (= O) NR 5 R 6 X ′ is —S (O) 2 -, -S (O) 2 -N (R ")-,-(C = O)-, -C (= O) -O-, -C (= O) -NH-, -P (O) (R")-, and a direct bond Wherein R "is hydrogen, alkyl of 1 to about 4 carbon atoms, aryl of about 6 to about 14 carbon atoms, or aralkyl of about 7 to about 16 carbon atoms, with the proviso that When X ′ is —P (O) (R ″) —, R ″ is not hydrogen; 5 Is selected from the group consisting of:
(1) Unsubstituted or Y 1 And Y 2 Alkyl of 1 to about 4 carbon atoms, substituted with 1-2 substituents selected from the group consisting of
(2) alkyl of 1 to about 3 carbon atoms substituted with cycloalkyl of 3 to about 6 carbon atoms, wherein the cycloalkyl is unsubstituted or 1 , Y 2 , And Y 3 An alkyl, mono-, di- or trisubstituted with 1 to 3 substituents selected from the group consisting of
(3) a cycloalkyl of 3 to about 6 carbon atoms, wherein the cycloalkyl is unsubstituted or has a Y 1 , Y 2 , And Y 3 A cycloalkyl, mono-, di- or trisubstituted with 1 to 3 substituents selected from the group consisting of
(4) 4 to about 6 ring atom heterocycloalkyl having a ring atom selected from carbon and heteroatoms, wherein the heteroatom comprises oxygen, nitrogen, and S (O) i Wherein i is 0, 1 or 2 and the heterocycloalkyl is unsubstituted or Y is 1 , Y 2 , And Y 3 A heterocycloalkyl, mono-, di- or tri-substituted with 1 to 3 substituents selected from the group consisting of
(5) 4 to about 6 ring atom heterocyclo having a ring atom selected from carbon and heteroatoms, wherein the heteroatom comprises oxygen, nitrogen, and S (O) i Wherein i is 0, 1, or 2, and the heterocyclo is
[0116]
Embedded image
And where
[0117]
Embedded image
Is a 5-7 membered heterocycle having 3-6 ring carbon atoms, wherein V is -CH 2 -, -O-, -S (= O)-, -S (O) 2 -Or -S-, wherein the heterocyclo is unsubstituted or 1 , Y 2 , And Y 3 Heterocyclo, monosubstituted, disubstituted, or trisubstituted with 1 to 3 substituents selected from the group consisting of
(6) alkenyl of from 2 to about 6 carbon atoms, unsubstituted or substituted with cycloalkyl of from about 3 to about 6 carbon atoms, wherein the cycloalkyl is unsubstituted or has Y on the ring. 1 , Y 2 , And Y 3 Alkenyl, mono-, di- or trisubstituted with 1 to 3 substituents selected from the group consisting of
(7) phenyl which is unsubstituted or Y 1 , Y 2 , And Y 3 Phenyl, mono-, di- or tri-substituted with 1 to 3 substituents selected from the group consisting of
(8) about 5 to about 6 ring atom heteroaryl having a ring atom selected from carbon and heteroatoms, wherein the heteroatom is selected from oxygen, nitrogen, and sulfur; Heteroaryl is unsubstituted or Y 1 , Y 2 , And Y 3 Heteroaryl, mono-, di- or tri-substituted with 1 to 3 substituents selected from the group consisting of
(9) alkyl of 1 to about 4 carbon atoms substituted with phenyl, wherein the phenyl is unsubstituted or has Y on the phenyl ring 1 , Y 2 , And Y 3 An alkyl, mono-, di- or trisubstituted with 1 to 3 substituents selected from the group consisting of
(10) a heteroaralkyl of about 5 to about 6 ring atoms having a ring atom selected from carbon and a heteroatom, wherein the heteroatom is selected from oxygen, nitrogen, and sulfur; The aralkyl is unsubstituted or substituted on the alkyl chain with hydroxy or halogen, and the heteroaralkyl is unsubstituted on the ring or Y on the ring. 1 , Y 2 , And Y 3 Heteroaralkyl, monosubstituted, disubstituted, or trisubstituted with 1 to 3 substituents selected from the group consisting of
(11) an aralkenyl of about 8 to about 12 carbon atoms, wherein the aralkenyl is unsubstituted or has Y on the aryl ring 1 , Y 2 , And Y 3 Aralkenyl, mono-, di- or trisubstituted with 1 to 3 substituents selected from the group consisting of
(12) about 5 to about 6 ring atom heteroaralkenyl having a ring atom selected from carbon and heteroatoms, wherein the heteroatom is selected from oxygen, nitrogen, and sulfur; The heteroaralkenyl can be unsubstituted or substituted on this ring carbon with Y 1 , Y 2 , And Y 3 Heteroaralkenyl, mono-, di-, or tri-substituted with 1 to 3 substituents selected from the group consisting of
(13) hydrogen; and
R 6 Is selected from the group consisting of:
(1) Unsubstituted or Y 1 And Y 2 Alkyl of 1 to about 12 carbon atoms, substituted with 1-2 substituents selected from the group consisting of:
(2) alkyl of 1 to about 3 carbon atoms substituted with cycloalkyl of 3 to about 8 carbon atoms, wherein the cycloalkyl is unsubstituted or 1 , Y 2 , And Y 3 An alkyl, mono-, di- or trisubstituted with 1 to 3 substituents selected from the group consisting of
(3) a cycloalkyl of 3 to about 15 carbon atoms, wherein the cycloalkyl is unsubstituted or has 1 , Y 2 , And Y 3 A cycloalkyl, mono-, di- or trisubstituted with 1 to 3 substituents selected from the group consisting of
(4) 4 to about 10 ring atom heterocycloalkyl having a ring atom selected from carbon and heteroatoms, wherein the heteroatom comprises oxygen, nitrogen, and S (O) i Wherein i is 0, 1 or 2 and the heterocycloalkyl is unsubstituted or Y is 1 , Y 2 , And Y 3 A heterocycloalkyl, mono-, di- or tri-substituted with 1 to 3 substituents selected from the group consisting of
(5) 4 to about 10 ring atom heterocyclo having a ring atom selected from carbon and heteroatoms, wherein the heteroatom comprises oxygen, nitrogen, and S (O) i Wherein i is 0, 1, or 2, and the heterocyclo is
[0118]
Embedded image
And where
[0119]
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Is a 5-7 membered heterocycle having 3-6 ring carbon atoms, wherein V is -CH 2 -, -O-, -S (= O)-, -S (O) 2 -Or -S-, wherein the heterocyclo is unsubstituted or 1 , Y 2 , And Y 3 Heterocyclo, monosubstituted, disubstituted, or trisubstituted with 1 to 3 substituents selected from the group consisting of
(6) aryl of about 6 to about 14 carbon atoms, wherein the aryl is unsubstituted or 1 , Y 2 , And Y 3 An aryl, mono-, di- or trisubstituted with 1 to 3 substituents selected from the group consisting of
(7) about 5 to about 14 ring atom heteroaryl having a ring atom selected from carbon and heteroatoms, wherein the heteroatom is selected from oxygen, nitrogen, and sulfur; Heteroaryl is unsubstituted or Y 1 , Y 2 , And Y 3 Heteroaryl, mono-, di- or tri-substituted with 1 to 3 substituents selected from the group consisting of
(8) an aralkyl of about 7 to about 15 carbon atoms, wherein the aralkyl is unsubstituted or substituted on the alkyl chain with hydroxy or halogen, and the aralkyl is unsubstituted, Or Y on this aryl ring 1 , Y 2 , And Y 3 Aralkyl, mono-, di- or tri-substituted with 1 to 3 substituents selected from the group consisting of
(9) about 5 to about 14 ring atom heteroaralkyl having a ring atom selected from carbon and heteroatoms, wherein the heteroatom is selected from oxygen, nitrogen, and sulfur; The aralkyl is unsubstituted or substituted on the alkyl chain with hydroxy or halogen, and the heteroaralkyl is unsubstituted on the ring or Y on the ring. 1 , Y 2 , And Y 3 Hetero-aralkyl, mono-, di- or tri-substituted with 1 to 3 substituents selected from the group consisting of
(10) Hydrogen, provided that A 1 Is -C (= O) OR 6 If R 6 Is not hydrogen;
(D) R 3 Is selected from H or methyl, or R 3 And R 4a And R 4b Are selected together as described in (f);
(E) (i) R 4a Is the configuration of S, and H, -CH 2 -S-CH 3 , -CH 2 OH, -CH 2 CN, lower alkyl of 1 to about 3 carbon atoms, -CH 2 C≡CH, -CH 2 CH = CH 2 And -CH = CH 2 And R is selected from the group consisting of 4b Is hydrogen;
(Ii) R 4a And R 4b Is independently lower alkyl of 1-3 carbon atoms;
(Iii) R 4a And R 4b Are selected together and-(CH 2 ) k Wherein k is 5 or 6 to provide spirocycloalkyl; or
(Iv) R 3 , And R 4a And R 4b Are selected together as described in (f);
(F) Or R 3 And R 4a Are selected together to be in the S configuration to form prolyl, pipecolyl, azetidine-2-carbonyl, 4-hydroxyprolyl, 3-hydroxyprolyl, 4-aminoprolyl, 4-(-CH 2 NH 2 ) -Prolyl, 3,4-methanoprolyl, and 3,4-dehydroprolyl provide a group selected from the group consisting of 4b Is hydrogen;
(G) R 7 Is hydrogen or alkyl of 1 to about 4 carbon atoms;
(H) E is QT,
Here, (i) Q is -C (R Thirteen R 14 ) t -, R 8 And R 9 A phenyl, R 8 Or R 8 And R 9 A 5- or 6-membered heterocyclic ring having 1-2 heteroatoms, substituted with 8 And R 9 Selected from the group consisting of a 9 or 10 membered heterocyclic ring having 1-2 heteroatoms substituted with a heterocyclic atom selected from nitrogen and sulfur;
(Ii) T is -C (= NR 10 ) NHR 11 , -NH-C (= NR 10 ) NHR 11 , And -NHR Fifteen Selected from the group consisting of:
Where R 8 And R 9 Is independently 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 R, and trifluoromethyl; 10 And R 11 Is independently hydrogen, hydroxy, alkoxy of 1 to about 3 carbon atoms, trihydrocarbylsilyl of 3 to about 16 carbon atoms, alkyl of 1 to about 3 carbon atoms, or —C (= O) R 12 Where R 10 And R 11 Are not both hydroxy or both alkoxy; R 12 Is hydrogen, alkyl of 1 to about 6 carbon atoms, alkoxy of 1 to about 6 carbon atoms, or (CF 2 ) j CF 3 Wherein j is 0, 1, 2 or 3; Thirteen And R 14 R is independently selected from the group consisting of hydrogen and lower alkyl of 1 to about 3 carbon atoms; Fifteen Is 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 0-6.
[0120]
According to another aspect of the present invention, preferred compounds are those wherein E is -C (R Thirteen R 14 ) t And T is -NHR Fifteen Wherein the compound of formula (I) is not included. According to a more preferred aspect, preferred compounds of formula I are those wherein Q is R 8 And R 9 A phenyl, R 8 Or R 8 And R 9 Selected from the group consisting of a 5- or 6-membered heterocyclic ring having 1-2 heteroatoms and a 9- or 10-membered heterocyclic ring having 1-2 heteroatoms substituted with Wherein the heteroatoms include compounds selected from the group consisting of nitrogen and sulfur. More preferably, Q is R 8 And R 9 Is phenyl substituted with More preferably, T is -C (= NR 10 ) NHR 11 Or -NHC (= NR 10 ) NHR 11 It is.
[0121]
According to one aspect of the present invention, preference is given to compounds of the formula I, wherein E is selected from the group consisting of:
[0122]
Embedded image
[0123]
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[0124]
Embedded image
[0125]
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Preferred X groups include -S (O) 2 -, -OC (= O)-, -NH-C (= O)-, and direct bonds. Particularly preferred is -S (O) 2 -And -OC (= O)-.
[0126]
Preferred R 1 The groups include alkyl, especially isobutyl, 2-ethylhexyl, methyl, n-butyl, isopropyl, cyclohexylmethyl and cyclohexylpropyl; cycloalkyl, especially (-) methyl, (+) methyl, and cyclohexyl; aryl, In particular, naphthyl and phenyl; aralkyl, especially benzyl, 3-phenylpropyl, and 2-phenylethyl; and fused carbocyclic alkyl, especially fluorenylmethyl. Particularly preferred R 1 Groups include phenyl, benzyl, 2-phenylethyl, isobutyl, n-butyl and 3-phenylpropyl.
[0127]
R 1 A preferred combination of -X- is phenyl-S (O) 2 -, Benzyl-S (O) 2 -, 2-phenylethyl-S (O) 2 -, 3-phenylpropyl-S (O) 2 -, N-butyl-S (O) 2 -, Benzyl-OC (= O)-, and isobutyl-OC (= O)-.
[0128]
Preferred R 2 As a group, -CH 3 , -C 2 H 5 , -CH 2 NH-X'-R 5 And -CH (R 5 ) OH, where R 5 Is hydrogen, alkyl, especially methyl, or alkyl. The preferred chirality at the α-carbon is R. When chiral, the preferred chirality at the β carbon is R. Preferred R 2 The group is d-seryl (-CH (R 5 ) OH, where R 5 Is H), (R, R) d-allothrenyl (-CH (R 5 ) OH, where R 5 Is methyl), d-2-aminobutyryl, N-β-methyloxycarbonyl-d-2,3-diaminopropionyl (—CH 2 NH-X'-R 5 , Where R 5 Is CH 3 And X ′ is (—C = O) O—), N-β- (2-phenylethylcarbonyl) -d-2,3-diaminopropynyl (—CH 2 NH-R 5 , Where R 5 Is 2-phenylethyl and X ′ is — (C = O) —), N-β-benzyloxycarbonyl-d-2,3-diaminopropionyl (—CH 2 NH-X'-R 5 , Where R 5 Is benzyl, and X ′ is — (C = O) O—) and d-alanyl (—CH 2 R defining the P3 position 2 Group. Particularly preferred R 2 The group is P 3 With d-seryl (R 5 Is H) or (R, R) d-allothrenyl (R 5 Is methyl) 2 Group.
[0129]
Alternative preferred R 2 As a group,-(CH 2 ) 2 OA 1 And -CH (R 5 ) OA 1 , More preferably -CH (R 5 ) OA 1 And preferably R 5 Is H. More preferred R 2 Is selected such that P3 is defined as an acyl or carbonate of d-seryl. R 2 Is-(CH 2 ) 2 OA 1 Or -CH (R 5 ) OA 1 Can act as prodrugs.
[0130]
R 3 And R 4 Are preferred when both are not selected 3 The group is hydrogen. R 3 And R 4 Are preferred when both are not selected 4 The group is methyl, vinyl, allyl or propargyl. R 3 And R 4 Are selected together, prolyl, 3-hydroxy-prolyl, 4-hydroxyprolyl, 3,4-dehydroprolyl, 3,4-methanoprolyl, and azetidine-2-carbonyl- define the group at P2 Is the preferred choice for
[0131]
Preferred R 7 The groups include hydrogen.
[0132]
Preferred E groups are those wherein Q is R 8 And R 9 A phenyl, R 8 And R 9 Substituted with pyridyl, or R 8 And T is -C (= NR 10 ) NHR 11 Or -NH-C (= NR 10 ) NHR 11 And an E group.
[0133]
Preferred E groups include 4-amidinophenyl, 4-guanidinophenyl, 3-amidinopropyl, and 5- (2-amidino-thienyl). Particularly preferred E groups include 4-amidinophenyl and 4-guanidinophenyl.
[0134]
Among the compounds of the present invention, a preferred compound is R which defines d-serine or d-allothreonine or acyl or a carbonate thereof at the P3 position of the compound. 2 An element and R at P1 defining an amidinophenyl, guanidinophenyl or amidinothienyl group 2 Includes compounds with elements. Particular preference is given to i) R 3 And hydrogen and R 4 (P2 is alanine), or ii) R such that P2 is prolyl, azetidine-2-carbonyl, 3,4-methanoprolyl or 3,4-dehydroprolyl. 3 And R 4 Is any of the compounds selected together.
[0135]
Preferred compounds of the present invention include those shown in FIGS. 10A-10F and FIGS. 13A-13C. Particularly preferred are compounds D, F, I, J, K, L, O, R, T, U, V, AE, AH, AJ, AN and AV of FIGS. 10A-10F and the BG of FIGS. 13A-13C. , BJ, BK and BQ.
[0136]
Also particularly preferred are the compounds of the following formula (I): compound AX (X = S (O) 2 , R 1 = 4-chlorobenzyl, R 2 = -CH 2 OH, R 3 = H, R 4 = CH 3 , R 7 = H and E = 4-amidinophenyl), AY (X = SO 2 , R 1 = 3-chlorobenzyl, R 2 = -CH 2 OH, R 3 = H, R 4 = CH 3 , R 7 = H and E = 4-amidinophenyl) and AZ (X = SO 2 , R 1 = 2-fluorobenzyl, R 2 = -CH 2 OH, R 3 = H, R 4 = CH 3 , R 7 = H and E = 4-amidinophenyl).
[0137]
(2. Preparation of preferred compounds)
FIGS. 1-5 show synthetic schemes for the synthesis of intermediates that can be used in the preparation of certain compounds of the present invention.
[0138]
FIG. 1 shows a solution phase synthesis for the synthesis of intermediates useful in preparing compounds of the present invention. See Examples 60-62. See also Examples 95-97.
[0139]
Examples 63-66, 67-70 and 71-73 show solution phase syntheses for the synthesis of intermediates useful in the synthesis of certain compounds of the present invention.
[0140]
FIG. 2 illustrates an alternative synthetic route for preparing intermediates useful in preparing compounds of the present invention using solution phase synthesis.
[0141]
FIG. 6 shows a reaction scheme for the preparation of compounds of the invention having a hydroxyl esterified to P3.
[0142]
FIG. 7 shows a reaction scheme for the preparation of compounds of the invention having 4-guanidinophenyl at P1.
[0143]
FIG. 8 shows a reaction scheme for the preparation of compounds of the invention having 3-guanidinophenyl at P1.
[0144]
FIG. 9 shows a reaction scheme for the preparation of compounds of the invention having 2-guanidinophenyl at P1.
[0145]
FIG. 11 shows a reaction scheme for the preparation of compounds of the invention having 3-amidinopyridyl at P1.
[0146]
FIG. 12 shows a reaction scheme for the synthesis of compounds of the invention having 4-amidinopyridyl at P1.
[0147]
FIG. 14 shows a reaction scheme for the synthesis of compounds of the invention having a bicyclic heterocyclic group at P1.
[0148]
FIG. 15 shows a reaction scheme for the synthesis of compounds of the invention having an amino-imidazolyl group at P1.
[0149]
FIG. 16 shows a reaction scheme for compounds of the present invention having a 2-chloro-4-guanidinophenyl group at P1.
[0150]
FIG. 17 shows a reaction scheme for the synthesis of compounds of the invention having a guanidino-pyridyl group at P1.
[0151]
A preferred method of chemically coupling (eg, for an amide linkage functional group) involves the formation of a peptide bond by using conventional coupling reagents known in the art. See, for example, Bodanszky, N.M. See Peptide Chemistry, pp. 55-73, Springer-Verlag, New York (1988) and the references cited herein. Chemical coupling is by either a one-step or two-step method. In one-step coupling, two coupling partners are directly coupled. Preferred coupling reagents for the coupling partner of the one-step coupling include DCC with HOBt, EDC with HOBt, HOAt, HBTU or EDC with TBTU. In a two-step coupling, an activated ester or anhydride of the C-terminal carboxyl group of one coupling partner is formed before it is coupled to the other coupling partner.
[0152]
For the preparation of certain compounds having hydrogenation sensitive substituents, it is preferred to avoid using hydrogen gas with palladium on carbon. Hydrogen, cyano, nitro, or -SZ 1 Another preferred method of preparing compounds of the present invention that contain a hydrogenation sensitive group such as an alkenyl or allyl moiety substituted with boron tris (trifluoroacetic acid), B (OOCOCF 3 ) 3 , Using the N of the arginine group g -Cleavage of the nitro. This reagent is used at 0 ° C. for BBr in dichloroamine. 3 And CF 3 Prepared by reaction of COOH. This reagent is also commercially available. In general, N g -The nitro compound is treated at 0 ° C with boron tris (trifluoroacetic acid) in trifluoroacetic acid. For example, Fieser, M .; And Fieser, L .; F. , Reagents for Organic Synthesis, p. 46, John Wiley & Sons, New York (1974); And Bauer, W.C. Angew. Chem. , Internet. Ed., 12, 147 (1973).
[0153]
Further, another preferred reagent for selective nitro group cleavage is titanium trichloride. This reagent is commercially available. N g The nitro compound was treated with titanium trichloride in aqueous methanol containing ammonium acetate buffer, followed by exposing the reaction to air or dimethyl sulfoxide. For example, Freidinger, R.A. M. Hirschmann, R .; And Veber, D .; F. , J. et al. org. Chem. 43, 4800 (1978).
[0154]
FIG. 6 shows a reaction scheme for the synthesis of compounds of the present invention, where R 2 Is -CH 2 OA 1 And A 1 Is -C (= O) R 6 Is:
[0155]
Embedded image
An intermediate such as 6-1 (the compound of Example 9) can be prepared by acid chloride R in the presence of a base such as pyridine. 6 Reacts with COCl. R 2 Is-(CH 2 ) 2 OA 1 Or -CH (R 5 ) OA 1 And A 1 Is -C (= O) R 6 Are preferably in the presence of a base such as pyridine with a suitable acid chloride derivative (R 6 COCl) and an appropriate intermediate corresponding to 6-1.
[0156]
R 2 Is-(CH 2 ) 2 OA 1 Or -CH (R 5 ) OA and A 1 Is -C (= O) OR 6 Is a compound represented by R 2 Is-(CH 2 ) 2 OH or -CH (R 5 )) Can be conveniently prepared by treating the corresponding compound, which is OH, with the appropriate chloroform derivative. P 1 In preparing compounds having an amidino or guanidino group, it may be preferable to cap the P3 hydroxyl with a carbonate group before deprotecting the amidino or guanidino group. Therefore, it is preferred to treat the corresponding intermediate with a chloroform derivative (see, eg, Example 8). The product was then hydrogenated and optionally treated under hydrolysis conditions to yield the product (see, for example, Examples 16, 21, 28, 35 and 40).
[0157]
(3. Selection of preferred compounds)
According to one aspect of the present invention, preferred compounds of the present invention are selected for their potency and selectivity for inhibiting serine proteases, especially urokinase. Such an evaluation is routinely performed in vitro according to the procedure as set forth in Example A. As described herein and generally known, the target serine protease and its substrate are combined under assay conditions that allow the reaction of the protease with the substrate. The assay is performed in the absence of the test compound and in the presence of increasing concentrations of the test compound. The concentration of the test compound at which 50% serine protease activity is inhibited by the test compound is determined by the IC 50 Value (inhibitory concentration) or EC 50 (Effective density) value. Relatively low IC within a series of test compounds or groups of test compounds 50 Value or EC 50 These compounds with high values have relatively high IC 50 Value or EC 50 It is believed that they are more potent inhibitors of serine proteases than those compounds that have values. IC 50 Measurements are often used for simpler assays, except that EC 50 Is often used for more complex assays (eg, cell-based assays). K i Is an IC 50 Is calculated from
[0158]
Preferred compounds according to this aspect of the invention have a K less than or equal to 100 nM as measured in an in vitro assay for inhibition of urokinase activity. i Has a value. Particularly preferred compounds have a K less than 30 nM. i Has a value.
[0159]
Test compounds are also evaluated for selectivity for serine proteases. As described in the Examples and generally known, test compounds are assayed for their potency against a panel of serine proteases or other enzymes, and the IC 50 Value or EC 50 A value is determined for each test compound in each assay system. Low IC for target enzymes (eg, urokinase) 50 Value or EC 50 Value or corresponding low K i Relatively high ICs for compounds that show values and other enzymes in the test panel (eg, tissue plasminogen activator, thrombin, factor Xa) 50 Value or EC 50 Compounds that show a value are considered to be selective for the target enzyme. Generally, a compound will have its IC in a targeted enzyme assay. 50 Value or EC 50 Value (or K i Value) is the next smallest IC measured in the enzyme selectivity panel. 50 Value or EC 50 If it is at least one order of magnitude smaller than the value, it is considered selective.
[0160]
Preferred compounds of the invention have a K of less than 100 nM as measured in an in vitro assay for inhibition of urokinase activity. i Has a value. Particularly preferred compounds have an IC measured in an in vitro tPA inhibition assay. 50 K in in vitro urokinase inhibition assay that is at least one order of magnitude smaller than i Has a value. IC greater than 100 50 tPA assay: K i Compounds having a selectivity ratio in the urokinase assay are particularly preferred.
[0161]
The compounds of the present invention are also evaluated for their activity in vivo. The type of assay chosen for the evaluation of a test compound will depend on the pathological condition being treated or prevented by the use of the compound, as well as the route of administration being evaluated for the test compound.
[0162]
For example, to evaluate the activity of compounds of the present invention to reduce tumor growth via inhibition of urokinase, Jankun et al. [Canc. Res. 57: 559-563, 1997] can be used. Briefly, SCID mice are injected with DU145, an ATCC cell line that expresses high levels of uPA, and LnCaP that does not express uPA. After the tumor is established, the mice are provided with the test compound according to a dosing regime determined from the in vitro characteristics of the compound. The Jankun et al. Compound was administered in water. Tumor volume measurements are taken twice a week for about 5 weeks. A compound is considered active if the animal receiving the compound exhibits a reduced tumor volume as compared to an animal receiving the appropriate control compound. Furthermore, a comparison of the effect of the compound in animals injected with DU145 cells versus LnCaP cells may indicate whether the effect of the compound was due to inhibition of urokinase or not.
[0163]
Another in vivo experimental model designed to evaluate the effect of p-aminobenzamidine (a compound referred to as a urokinase inhibitor compound) on decreasing tumor volume is described in Billstrom et al. [Int. J. Cancer 61: 542-547, 1995].
[0164]
To assess the ability of the compounds of the present invention to reduce the occurrence of metastasis or inhibit metastasis, Kobayashi et al. [Int. J. Canc. 57: 727-733d, 1994] can be used. Briefly, mouse xenografts selected for high lung colony forming ability are injected intravenously into C57B1 / 6 mice (experimental metastasis) or subcutaneously into the abdominal wall (spontaneous injection). Metastasis). Various concentrations of the compound to be tested can be mixed with the tumor cells in Matrigel prior to injection. Daily intraperitoneal injections of test compound are given either 1-6 days or 7-13 days after tumor inoculation. Animals are sacrificed approximately 3-4 weeks after tumor inoculation and lung tumor colonies are counted. Evaluation of the resulting data will allow decisions regarding the effects of the test compound, optimal dosing and routes of administration.
[0165]
The activity of the compounds of the invention on decreasing tumor volume and metastasis may be assessed by Rabbani et al. [Int. J. Cancer 63: 840-845, 1995]. There were Mat LyLu tumor cells overexpressing uPA injected into the flank of Copenhagen rats. The animals were implanted using an osmotic minipump to continuously administer various doses of the test compound for up to three weeks. Tumor mass and tumor volume of experimental and control animals were evaluated during the experiment, as were metastatic growth. Evaluation of the resulting data allowed decisions regarding test compound efficacy, optimal dosing, and routes of administration. Some of these authors are described by Xing et al. [Canc. Res. 57: 3585-3593, 1997] describes a related protocol.
[0166]
To evaluate the inhibitory activity of the compounds of the present invention on neovascularization, a rabbit corneal neovascularization model could be used. Avery et al. [Arch. Ophthalmol. 108: 1474-1475, 1990] describe anesthetizing a New Zealand white rabbit, then making a central corneal incision, and forming a radical corneal pocket. A slow release prostaglandin pellet was placed in this pocket to induce neovascularization. Test compounds were administered intraperitoneally for 5 days, and on day 5, animals were sacrificed. The effect of this test compound is to examine periodic pictures taken of the limbus, which can be used to calculate the area of neovascular response and thus the limbal neovascularization Is evaluated by Areas of reduced neovascularization as compared to appropriate controls indicate that the test compound was effective in reducing or inhibiting neovascularization.
[0167]
An angiogenesis model used to evaluate the effects of test compounds in preventing angiogenesis is described by Min et al. [Canc. Res. 56: 2428-2433, 1996]. C57BL6 mice receive a subcutaneous injection of Matrigel mixture containing bFGF (as an angiogenesis inducer) with or without test compound. Five days later, the animals are sacrificed and Matrigel plugs are photographed where neovascularization can be visualized. Experimental animals receiving Matrigel and an effective amount of the test compound show less angiogenesis than control animals or experimental animals receiving little or no effective dose of the compound.
[0168]
In vivo systems in which a test compound is designed for its ability to limit the spread of a primary tumor are described in Crowley et al. [Proc. Natl. Acad. Sci. 90: 5021-5025, 1993]. Nude mice are injected with tumor cells (PC3) engineered to express CAT (chloramphenicol acetyltransferase). The cells secrete large amounts of uPA and exhibit a saturating amount of uPA activity that binds to uPAR on the cell surface. The compound to be tested for its ability to reduce tumor size and / or metastasis is administered to the animal and a subsequent measurement of tumor size and / or metastatic growth is made. In addition, CAT levels detected in various organs provide an indication of the ability of the test compound to inhibit metastasis; the detection of less CAT in tissues of treated animals relative to control animals indicates that CAT expression transferred to that tissue Indicates fewer cells.
[0169]
An in vivo experimental model designed to evaluate the ability of test compounds to inhibit urokinase using the tumor cell line F3II (referred to as highly invasive) is described in Alonso et al. [Breast Canc. Res. Treat. 40: 209-223, 1996]. This group describes in vivo studies on toxicity determination, tumor growth, invasiveness, spontaneous metastasis, experimental lung metastasis and angiogenesis assays.
[0170]
The CAM model (chick embryo chorioallantoic membrane model) (first described by L. Ossowski in 1998 [J. Cell Biol. 107: 2437-2445, 1988]) was used to evaluate the urokinase inhibitory activity of test compounds. Provide another way. In this CAM model, tumor cell invasion through the chorioallantoic membrane depends on the presence of catalytically active uPA. Contacting the CAM with the tumor cells in the presence of an urokinase inhibitor causes little or no invasion of the tumor cells through the membrane. Thus, CAM assays are performed with CAM and tumor cells in the presence and absence of various concentrations of the test compound. Tumor cell invasion is measured under conditions that provide an indication of the urokinase inhibitory activity of the compound. Compounds with urokinase inhibitory activity correlate with less tumor invasion.
[0171]
This CAM model is also used in standard assays of angiogenesis (ie, effects on new blood vessel formation) (Brooks, PC; Montgomery, AMP; and Cheresh, D. A., Methods in Molecular Biology 129: 257-269 (1999)). According to this model, a filter disk containing an angiogenesis-inducing factor (eg, basic fibroblast growth factor (bFGF)) is placed on the CAM. Diffusion of cytokines into the CAM induces local angiogenesis, which is accomplished in several ways, for example by counting the number of vessel branch points in the CAM just below the filter disk. Can be measured. The ability of the compounds of the invention to inhibit cytokine-induced angiogenesis can be tested using this model. The test compound can be added to a filter disk containing an angiogenesis-inducing factor, placed directly on a membrane, or administered entirely. The extent of neovascularization in the presence and / or absence of a test compound can be compared using this model. The formation of fewer new blood vessels in the presence of the test compound is an indicator of anti-angiogenic activity. Since some of the compounds of the present invention are active as inhibitors of urokinase, anti-angiogenic activity against such compounds may suggest that urokinase plays a significant role in angiogenesis.
[0172]
(4. Pharmaceutical composition)
In another aspect, the invention includes a pharmaceutical composition prepared for storage or administration, comprising a therapeutically effective amount of a compound of the invention in a pharmaceutically acceptable carrier.
[0173]
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 particular mammal under consideration. These factors and their relationship to determine this amount are well known to those skilled in the medical arts. The amount and method of administration may 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.
[0174]
Therapeutically effective amounts of the compounds of the present invention can vary widely depending on the desired effect and therapeutic index. Typically, dosages will be between about 0.01 mg / kg body weight and 100 mg / kg body weight, preferably between about 0.01 mg / kg body weight and 10 mg / kg body weight.
[0175]
Pharmaceutically acceptable carriers for therapeutic use are well known in the pharmaceutical arts, and are described, for example, in Remington's Pharmaceutical Sciences, Mack Publishing Co. (A. R., edited by Gennaro, 1985). For example, sterile saline and phosphate buffered saline at physiological pH can be used. Preservatives, stabilizers, dyes and even flavoring agents may be provided in the pharmaceutical composition. For example, sodium benzoate, sorbic acid, esters of p-hydroxybenzoic acid may be added as preservatives. Ibid, 1449. In addition, antioxidants and suspending agents may be used. Ibid.
[0176]
The pharmaceutical compositions of the present invention may be formulated and used as tablets, capsules or elixirs for oral administration; suppositories for rectal administration; sterile compositions and suspensions for injection administration. Dosage and manner of administration may be varied to achieve optimal efficacy, but will depend on such factors as weight, diet, concomitant medications and other factors which those skilled in the medical arts will recognize.
[0177]
Where administration is parenteral (eg, daily intravenous), the injectable pharmaceutical composition may be in a conventional form (either as a liquid solution or suspension), a solution in liquid prior to infusion or It may be prepared in a solid form suitable for suspension, or as an emulsion. Suitable excipients are, for example, water, saline, dextrose, mannitol, lactose, lecithin, albumin, sodium glutamate and the like. In addition, if desired, injectable pharmaceutical compositions can contain small amounts of non-toxic auxiliary substances such as wetting agents, pH buffering agents, and the like. If desired, absorption enhancing preparations (eg, liposomes) can be utilized.
[0178]
(5. Usefulness)
Compounds of the invention having urokinase inhibitory activity and / or an activity of reducing or inhibiting angiogenesis (including angiogenesis and neovascularization) can be used both in vitro and in vivo for many applications, Some of these are described herein below.
[0179]
The compounds of the present invention are active as inhibitors of urokinase and specifically bind to urokinase. Thus, these compounds containing sites suitable for ligation to a solid / gel support can be used to purify urokinase from a sample or to remove urokinase from a sample using conventional affinity chromatography procedures. For chromatography, it can be used in vitro. These compounds can be attached or linked to affinity chromatography, either directly or through a suitable linker support, using conventional methods. See, for example, Current Protocols in Protein Science, John Wiley & Sons (edited by JE Colligan et al., 1997) and Protein Purification Protocols, Humana Press (ed. S. Doonan, 1966) and references therein.
[0180]
Compounds of the present invention having urokinase inhibitory activity are useful in in vitro activity to measure tPA activity in a sample. In assays measuring total plasminogen activating activity in blood samples, compounds of the invention having urokinase inhibitory activity inactivate a portion of plasminogen activation due to uPA (knock out), which indicates that tPA activity Allows for the calculation of the portion of total plasminogen activation due to UPA activity and total plasminogen activity due to uPA activity. Use of such assays to monitor tPA activity allows for better dose control in patients receiving tPA. These assays also monitor uPA / tPA activity in tissue samples (eg, from biopsies) or for any clinical condition where the measurement of plasminogen activator activity is assisted. Can be used to These assays can also be used to monitor plasminogen activator activity, where the patient has been treated with a non-endogenous compound having plasminogen activator activity, such as streptokinase or staphylokinase.
[0181]
The compounds of the present invention are used in vivo for the treatment of pathological conditions ameliorated by decreased urokinase activity. For example, these compounds inhibit metalloprotease activation by the uPA-plasmin cascade in synovial fluid and can therefore be used in the treatment of arthritis.
[0182]
These compounds are believed to be useful in reducing or inhibiting metabolism, neovascularization and extracellular matrix degradation in tumors and other neoplasms. These compounds are characterized by pathological neovascularization, such as retinal diseases, retinopathies, and other conditions, including those described in the Background and Introduction of the Invention herein. It is useful as a therapeutic agent in treating a given condition.
[0183]
Another use for compounds of the invention having urokinase inhibitory activity is as an antidote if large amounts of exogenous urokinase are given to a patient for other purposes (eg, to dissolve thrombus). is there.
[0184]
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 blocking mediators of cell adhesion or migration. Such anti-inflammatory applications include treatment of stroke and complications of organ transplantation.
[0185]
The invention includes a method for preventing or treating a condition in a mammal suspected of having a condition that is attenuated by inhibition of urokinase, the method comprising providing a therapeutically effective amount of a compound or pharmaceutical composition of the invention. And administering to the mammal.
[0186]
The compounds or pharmaceutical compositions of the invention are usually administered in vivo in a mammal, preferably a human. In using them in vivo, the compounds or pharmaceutical compositions can be administered in a variety of modes (orally, parenterally, intravenously, intravenously, subcutaneously, intramuscularly) using different modes of administration. (Colonally, rectally, nasally, or intraperitoneally). Administration is preferably oral, for example by tablets, capsules or elixir taken daily.
[0187]
In practicing the methods of the present invention, the compounds or pharmaceutical compositions of the present invention are administered alone or in combination with each other, or in combination with other therapeutic or in vivo diagnostic agents.
[0188]
As will be apparent to those skilled in the medical arts, a "therapeutically effective amount" of a compound or pharmaceutical composition of the invention will vary with the age, weight, and species of mammal used, the particular compound used, the particular It will vary depending on the mode and effect desired and the therapeutic indication. As these factors and their relationship to determining this amount are well known in the medical arts, determining a therapeutically effective dosage level (the amount necessary to achieve the desired result of inhibiting uPA activity) Is within the skill of the art. Typically, administration of a compound or pharmaceutical composition of the invention will be initiated at a lower dosage level, which will be increased until the desired effect of inhibiting uPA activity is achieved to the desired extent. Defines a therapeutically effective amount. For the compounds of the present invention, alone or as part of a pharmaceutical composition, such doses will be between about 0.01 mg / kg body weight and 100 mg / kg body weight, preferably about 0.01 mg / kg body weight. It is between body weight and 10 mg / kg body weight.
[0189]
To assist in understanding, the invention will now be further illustrated by the following examples. These examples should, of course, not be construed as specifically limiting the invention, as they pertain to the present invention, and are hereby deemed known or later developed and within the purview of those skilled in the art. Certain such variations of the invention are considered to be within the scope of the invention as described herein and claimed above.
[0190]
(Example)
(A. Synthesis of specific compound of the present invention)
(Example 1)
(Preparation of n-butylsulfonyl-D-serine (tert-butyl ether) -methyl ester (1))
[0191]
Embedded image
A solution of HCl.H-dSer (tBu) -OMe (2 g, 9.44 mmol) and n-butylsulfonyl chloride (1.1 ml, 8.50 mmol) in tetrahydrofuran (38) was stirred at room temperature for 10 minutes. . Then diisopropylethylamine (5.75 ml, 33.07 mmol) was added and the cloudy yellow solution was stirred overnight at ambient temperature. The reaction mixture was then diluted with ethyl acetate (200ml) and washed with 1N HCl then brine (20ml each). After drying over anhydrous sodium sulfate, the solvent was removed under reduced pressure. The scaly yellow solid (1.56 g, 62%) was determined to be pure by tlc (5% ethyl acetate in hexane).
[0192]
(Example 2)
(Preparation of n-butylsulfonyl-D-serine (tert-butyl ether) (2))
[0193]
Embedded image
To a solution of compound 1 (1.46 g, 4.94 mmol) in dioxane (32.95 ml) was added 2.0 N LiOH (5.44 ml, 10.87 mmol) dropwise. The cloudy yellow solution was stirred overnight at ambient temperature. When starting material was no longer observed by tlc (5% ethyl acetate / hexane), excess dioxane was removed under reduced pressure. The reaction mixture was diluted with a 1: 1 mixture of water and ethanol and passed through a previously washed DOWEX (50 × 8-400) ion exchange resin (30 ml). The resin was thoroughly rinsed with methanol and water. The combined filtrate was concentrated under reduced pressure to give 1.44 g of the title compound as a creamy solid in quantitative yield.
[0194]
(Example 3)
(Preparation of n-butylsulfonyl-D-serine (tert-butyl ether) -alanine tert-butyl ester (3))
[0195]
Embedded image
Compound of Example 2 (0.50 g, 1.79 mmol), alanine tert-butyl ester hydrochloride (0.65 g, 3.58 mmol), EDC (0.68 g, 3.57 mmol), N-hydroxybenzotriazole (0 A solution of .27 g, 1.79 mmol) and diisopropylethylamine (1.56 ml, 8.94 mmol) was stirred in acetonitrile (18 ml) at ambient temperature. After 18 hours, the solvent was removed under reduced pressure and the resulting residue was resuspended in ethyl acetate (50 ml) and 1N HCl (10 ml). The ethyl acetate layer was washed with 1N HCl (10 ml), saturated sodium bicarbonate (2 × 15 ml) and brine (15 ml), then dried over sodium sulfate. The solvent was removed under reduced pressure and the crude was purified by flash column chromatography (eluting with 50% ethyl acetate / hexane) to give 429 mg (59%) of the product. This product was purified by reverse layer (C18) HPLC (0.1% trifluoroacetic acid in 5-90% aqueous acetonitrile over 20 minutes). R = 9 min).
[0196]
(Example 4)
(Preparation of n-butylsulfonyl-D-serine-alanine (4))
[0197]
Embedded image
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 under reduced pressure. The residue was resuspended in 10 ml acetonitrile and 50 ml n-heptane and concentrated under reduced pressure to give 410 mg of the product.
[0198]
(Example 5)
(Preparation of α-azido-4-cyanotoluene (5))
[0199]
Embedded image
Sodium azide (Aldrich, 3.5 g, 54 mmol) is added to a solution of α-bromotoluene nitrile (Aldrich, 10 g, 51 mmol) in DMF (100 ml) and the resulting mixture is allowed to stand at ambient temperature for 5 hours Stirred. The reaction mixture was then diluted with water (350ml) and extracted with ether (2x100ml). The combined organic layers were washed with brine and dried (MgSO 4 ). Removal of the solvent gave the title compound (8 g, 96%). 1 H NMR (CDCl 3 ): Δ 4.42 (s, 2H), 7.41 (d, 2H, J = 8.1 Hz), 7.65 (d, 2H, J = 8.1 Hz).
[0200]
(Example 6)
(Preparation of tolyl (6) in 4- (aminomethyl) benzyl)
[0201]
Embedded image
A catalyst of 10% Pd on carbon (Aldrich, 800 mg) was added to a solution of α-azido-4-cyanotoluene (Compound 5, 8 g, 51 mmol) in EtOAc (150 ml). The reaction mixture was hydrogenated on a Parr apparatus for 11 hours (H 2 , 45 psi). The catalyst was filtered off and the solvent was removed under reduced pressure to give the title compound (6.3 g, 93%). 1 H NMR (CDCl 3 ): Δ 3.85 (s, 2H), 7.45 (d, 2H, J = 8.1), 7.60 (d, 2H, J = 8.1 Hz), 7.78 (s, 2H, NH 2 ).
[0202]
(Example 7)
(Preparation of n-butylsulfonyl-D-serine-alanine-4-cyanobenzylamide (7))
[0203]
Embedded image
Compound 4 (150 mg, 0.51 mmol), 4- (aminomethyl) benzylnitrile (Compound 6, 171 mg, 1.02 mmol), EDC (195 mg, 1.02 mmol), and N-hydroxy in acetonitrile (5.1 ml) A solution of benzotriazole (78 mg, 0.51 mmol) was stirred at ambient temperature for 10 minutes. Then 2,4,6-collidine (0.34 ml, 2.54 mmol) was added and the reaction mixture was stirred overnight at ambient temperature. The solvent was removed under reduced pressure and the resulting residue was resuspended in ethyl acetate (100ml) and 0.5M HCl (10ml). The ethyl acetate layer was washed with water, followed by 0.5M HCl (10 ml), saturated sodium bicarbonate (2 × 10 ml) and brine (15 ml), then dried over sodium sulfate. The solvent was removed under reduced pressure to give 237 mg of product. The product eluted at 9.5 minutes by reverse phase (C18) HPLC (0.1% trifluoroacetic acid in 5-75% aqueous acetonitrile over 20 minutes).
[0204]
(Example 8)
(Preparation of n-butylsulfonyl-D-serine-alanine-4-hydroxyamidinobenzylamide (8))
[0205]
Embedded image
To a solution of the product of Example 7 (117 mg, 0.285 mmol) in methanol (1.14 ml) was added hydroxyamine hydrochloride (33.7 mg, 0.485 mmol) followed by N-methylmorpholine ( 53 μl, 0.485 mmol) were added. The reaction mixture was stirred overnight at ambient temperature, then at 50 ° for 6 hours. The reaction mixture was concentrated under reduced pressure. The crude product was used for the next step (Example 9) without further purification. The product eluted at 6.5 minutes by reverse phase (C18) HPLC (0.1% trifluoroacetic acid in 5-50% aqueous acetonitrile over 20 minutes).
[0206]
(Example 9)
(Preparation of n-butylsulfonyl-D-serine-alanine-4-amidinobenzylamide (9))
[0207]
Embedded image
To the product of Example 8 (126 mg, 0.285 mmol) in acetic acid (2.85 ml) and water (0.28 ml) was added active zinc powder (185 mg). The reaction mixture was stirred overnight at room temperature. The zinc powder was filtered using a glass funnel and the filtrate was purified by preparative HPLC. Product containing fractions were eluted with 5-20% aqueous acetonitrile containing 0.1% TFA and pooled and lyophilized to give 35 mg of the title compound as a white powder. The product eluted at 6.0 minutes by reverse phase (C18) HPLC (0.1% trifluoroacetic acid in 5-50% aqueous acetonitrile over 20 minutes).
[0208]
(Example 10)
(Preparation of benzylsulfonyl-D-serine (tert-butyl ether) -methyl ester (10))
[0209]
Embedded image
A solution of HCl.H-dSer (tBu) -OMe (1 g, 4.72 mmol) and phenethylsulfonyl chloride (1.45 g, 7.08 mmol) in acetonitrile (19 ml) was stirred at room temperature for 10 minutes. Then diisopropylethylamine (1.53 ml, 11.81 mmol) was added and the clear yellow solution was stirred at ambient temperature for 18 hours. The reaction mixture was then diluted with ethyl acetate (100ml) and washed with 1N HCl (10ml) followed by brine (10ml). After drying over sodium sulfate, the solvent was removed under reduced pressure. The crude product was purified by flash column chromatography (eluting with dichloromethane followed by a gradient consisting of 1-5% ethyl acetate in dichloromethane) to give 840 ml (52%) of the product. . The final product Tlc in 5% ethyl acetate in dichloromethane showed one spot with an Rf of 0.52.
[0210]
(Example 11)
(Preparation of benzylsulfonyl-D-serine (tert-butyl ether) (11))
[0211]
Embedded image
To a solution of the compound of Example 10 (1.0 g, 3.03 mmol) in dioxane (20 ml) was added 2.0 N LiOH (3.33 ml, 6.67 mmol) dropwise. The solution was stirred overnight at ambient temperature. Excess dioxane was removed under reduced pressure. The reaction mixture was diluted with a 1: 1 mixture of water and methanol and passed through a pre-washed DOWEX (50 × 8-400) ion exchange resin (30 ml). The resin was thoroughly rinsed with methanol and water. The combined filtrate was concentrated under reduced pressure to give 1.10 g of the title compound as a yellow paste.
[0212]
(Example 12)
(Preparation of tert-butyloxycarbonyl-3,4-dehydroproline p-cyanobenzylamide (12))
[0213]
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Tert-Butyloxycarbonyl-3,4-dehydroproline (0.4 g, 1.88 mmol), 4- (aminomethyl) benzylnitrile (compound 6, 0.47 g, 2.82 mmol) in acetonitrile (7.5 ml). ), EDC (0.54 g, 2.82 mmol), N-hydroxybenzotriazole (0.29 g, 1.88 mmol), and diisopropylethylamine (1.64 ml, 9.39 mmol) were stirred at ambient temperature overnight. did. The solvent was removed under reduced pressure and the resulting residue was resuspended in ethyl acetate (25ml) and 0.5M HCl (5ml). The ethyl acetate layer was washed with water followed by 0.5 M HCl (5 ml), saturated sodium bicarbonate (2 × 5 ml), and brine (10 ml), and then dried over sodium sulfate. The solvent was removed under reduced pressure and the crude product was purified by flash column chromatography eluting with 4/1 ethyl acetate / hexane to give 561 mg of pure product (91.3%). The product eluted at 10.5 minutes by reverse phase (C18) HPLC (0.1% trifluoroacetic acid in 5-75% aqueous acetonitrile over 20 minutes).
[0214]
(Example 13)
(Preparation of 3,4-dehydroproline-4-cyanobenzylamide hydrochloride (13))
[0215]
Embedded image
To a solution of the compound of Example 12 (0.47 g, 1.44 mmol) in ethyl acetate (5.7 ml) was added 5M anhydrous HCl in ethyl acetate (1.44 ml) and the reaction was allowed to proceed at ambient temperature. Stirred overnight. The reaction mixture was concentrated under reduced pressure to give 363 mg (95%) of a white solid.
[0216]
(Example 14)
(Preparation of benzylsulfonyl-D-serine (tert-butyl ether) -proline (dehydro) -4-cyanobenzylamide (14))
[0219]
Embedded image
Compound of Example 11 (0.10 g, 0.32 mmol), Compound of Example 13 (0.092 g, 0.34 mmol), EDC (0.091 g, 0.48 mmol) in acetonitrile (1.2 ml), And a solution of N-hydroxybenzotriazole (0.053 g, 0.35 mmol) was stirred for 10 minutes. Then 2,4,6-collidine (0.209 ml, 1.58 mmol) was added and the reaction mixture was stirred at ambient temperature overnight. The solvent was removed under reduced pressure. The residue obtained was resuspended in ethyl acetate (50 ml) and 1N HCl (10 ml). The ethyl acetate layer was washed with 1N HCl (10 ml), saturated sodium bicarbonate (2 × 15 ml), and brine (15 ml), then dried over sodium sulfate to give a yellow syrup (160 mg, 94%). The product was a single peak on reversed-phase (C18) HPLC (t over 20 minutes in 5-90% aqueous 0.1% trifluoroacetic acid in acetonitrile at t R = 11 minutes).
[0218]
(Example 15)
(Preparation of benzylsulfonyl-D-serine-3,4-dehydroproline-4-cyanobenzylamide (15))
[0219]
Embedded image
To a solution of the compound of Example 14 (0.16 g, 0.30 mmol) in dichloromethane (0.6 ml) was added trifluoroacetic acid (0.6 ml). The reaction mixture was stirred at ambient temperature for 1 hour. The reaction mixture was diluted with 10 ml of n-heptane and concentrated under reduced pressure. The residue was resuspended in acetonitrile (5 ml) and n-heptane (10 ml) and concentrated under reduced pressure to give 183 mg of the product.
[0220]
(Example 16)
(Preparation of benzylsulfonyl-D-serine-3,4-dihydroproline-4-hydroxyamidinobenzylamide (16))
[0221]
Embedded image
To a solution of the product of Example 15 (143 mg, 0.305 mmol) in methanol (1.22 ml) was added hydroxyamine hydrochloride (0.036 mg, 0.519 mmol) followed by N-methylmorpholine ( 57 μl, 0.519 mmol) was added. The reaction mixture was stirred overnight at ambient temperature. Analytical HPLC indicated that the reaction was not complete. Additional hydroxyamine hydrochloride (0.036 mg, 0.519 mmol) and N-methylmorpholine (57 μl, 0.519 mmol) were added and stirring continued at ambient temperature overnight. The reaction mixture was concentrated under reduced pressure. The crude product was purified by preparative HPLC. The fractions containing the product were eluted with 5-20% aqueous acetonitrile containing 0.1% TFA and pooled and lyophilized to give 16 mg of the title compound as a white powder.
[0222]
(Example 17)
(Preparation of benzylsulfonyl-D-serine-3,4-dihydroproline-p-amidinobenzylamide (17))
[0223]
Embedded image
To the product of Example 16 (15 mg, 0.030 mmol) in acetic acid (0.30 ml) and water (0.03 ml) was added activated zinc powder (19 mg). The reaction mixture was stirred overnight at room temperature. The zinc powder was filtered using a glass funnel and the filtrate was purified by preparative HPLC. The fractions containing the product were eluted with 5-20% aqueous acetonitrile containing 0.1% TFA and pooled and lyophilized to give 7 mg of the title compound as a white powder. The product eluted in minutes by reverse phase (C18) HPLC (0.1% trifluoroacetic acid in 5-50% aqueous acetonitrile over 20 minutes).
[0224]
(Example 18)
(Preparation of bis (benzyloxycarbonyl) guanidine (18))
[0225]
Embedded image
The synthesis of the title compound was carried out as described in the literature (tetrahedron Letters, Vol. 35, No. 7, pages 977-980) and as described below.
[0226]
N, N′-bis (benzyloxycarbonyl) -S-methylisothiourea (1 g, 2.79 mmol) in 7N anhydrous ammonia in methanol (5.5 ml) was stirred overnight at ambient temperature. The reaction mixture was concentrated and the residue was diluted with ethyl acetate (10ml). The organic phase was washed twice with saturated sodium bicarbonate and once with brine (10 ml each). After drying over sodium sulfate, the crude product was subjected to flash column chromatography, eluting with ethyl acetate / hexane (3/2) to give 0.87 g of a white solid. The product was then recrystallized in an ethyl acetate: hexane (1: 1) solvent system to give 337 mg (37%) of pure product (mp = 151 ° C.).
[0227]
(Example 19)
(Preparation of tert-butyloxycarbonyl-4-amino-1-butanol (19))
[0228]
Embedded image
To a solution of 4-amino-1-butanol (1 g, 11.22 mmol) in tetrahydrofuran (45 ml) was added anhydrous Boc (2.20 g, 10.10 mmol) and triethylamine (2.84 g, 28.04 mmol). The reaction mixture was stirred overnight at room temperature. The reaction mixture was concentrated and the residue was diluted with ethyl acetate (250ml) and HCl (50ml). The phases were separated and the organic phase was washed with 1N HCl, water and brine (50 ml each). After drying over sodium sulfate, the product (1.9 g (99%)) gave a clear oil.
[0229]
(Example 20)
(Synthesis of N, N'-bis (benzyloxycarbonyl) agmatine trifluoroacetate (20))
[0230]
Embedded image
A solution of compound 18 (the product of Example 18 (0.2 g, 0.61 mmol)) and triphenylphosphine (0.12 g, 0.46 mmol) in dry toluene (6.6 ml) was added with a syringe under nitrogen. Compound 19 (product of Example 19) was added via. The mixture was cooled to 0 ° C. and diethyl azodicarboxylate (0.080 g, 0.46 mmol) was added dropwise over 15 minutes. The reaction mixture was stirred overnight at room temperature and tlc with ethyl acetate / hexane (3/2) confirmed the completion of the reaction. Five drops of water were added and the solvent was evaporated under reduced pressure. The crude preparation was purified by flash column chromatography, eluting with hexane / ethyl acetate (9/1) then hexane / ethyl acetate (3/2) to give the pure product (83 mg (74%)). The product was then treated with dichloromethane and trifluoroacetic acid (1 ml each) at ambient temperature for 1 hour. Removal of the solvent under reduced pressure provided the product 20 (80 mg).
[0231]
(Example 21)
(Preparation of n-butylsulfonyl-D-serine-agmatine-gN, N′-bis (benzyloxycarbonyl (21))
[0232]
Embedded image
The compound of Example 4 (0.05 g, 0.17 mmol) and the compound of Example 20 (0.065 g, 0.17 mmol), EDC (0.065 g, 0.34 mmol), and hydroxybenzotriazole (0. (026 g, 0.17 mmol) was stirred in acetonitrile (0.67 ml) for 10 minutes. Then, 2,4,6-collidine (0.11 ml, 0.84 mmol) was added and the reaction mixture was stirred overnight at ambient temperature. The solvent was removed under reduced pressure and the resulting residue was resuspended in ethyl acetate and 1N HCl (5 ml each). The ethyl acetate phase was washed with 1N HCl (5 ml), saturated sodium bicarbonate (2 × 5 ml) and brine (5 ml), then dried over sodium sulfate to a solid (114 mg, 94%).
[0233]
(Example 22)
(Preparation of n-butylsulfonyl-D-serine-alanine-agmatine (22))
[0234]
Embedded image
The compound of Example 21 (114 mg, 0.17 mmol) was dissolved in methanol (15 ml) and hydrogenated on a Parr shaker (40 psi) overnight in the presence of palladium on carbon (15 mg). The catalyst was removed by filtration. The reaction mixture was diluted to 35 ml with water and purified by preparative HPLC. The fractions containing the product were eluted with 5-20% aqueous acetonitrile containing 0.1% TFA and pooled and lyophilized to give 16 mg of the title compound as a white powder.
[0235]
(Example 23)
(Preparation of 2-cyano-5-methylthiophene (23))
[0236]
Embedded image
A solution of 2-bromo-5-methylthiophene (TCI chemicals) (5 g, 28 mmol) and copper (I) (Aldrich, 2.53 g, 28 mmol) in DMF (10 ml) was heated at its reflux temperature for 4 hours. . After cooling to ambient temperature, ethyl acetate (500 ml) and 10% aqueous NaCN (500 ml) were added. After separation, the aqueous phase was extracted with ethyl acetate (300ml) and the combined organic phases were concentrated to an oil. The oil was further purified by flash column chromatography (ethyl acetate) to give the title compound (3.03 g, 87%). TLC: Rf 0.30 (1: 1 hexane / ethyl acetate); 1 H NMR (CDCl 3 ): Δ 2.55 (m, 3H), 6.76 (d, 1H, J = 3.6 Hz), 7.42 (d, 1H, J = 3.6 Hz).
[0237]
(Example 24)
(Preparation of 2-cyano-5- (bromomethyl) thiophene (24))
[0238]
Embedded image
CCl 4 (Aldrich, 60 ml), 2-cyano-5-thiophene (compound 23, 3.0 g, 24 mmol), N-bromosuccinimide (Aldrich, 4.8 g, 27 mmol), and 2,2′-azobisisomid Butyronitrile (Aldrich, 0.4 g, 2.4 mmol) was heated at its reflux temperature for 5 hours. After cooling to ambient temperature, the solvent was removed to give a yellow oil. The oil was purified by flash column chromatography (hexane / ethyl acetate (1: 1)) to give the title compound (4.5 g, 91%). TLC: Rf 0.91 (1: 1 hexane / ethyl acetate); 1 H NMR (CDCl 3 ): Δ 4.66 (s, 2H), 7.01 (d, 1H, J = 3.7 Hz), 7.55 (d, 1H, J = 3.7 Hz).
[0239]
(Example 25)
(Preparation of 2-cyano-5- (azidomethyl) thiophene (25))
[0240]
Embedded image
2-Cyano-5- (bromomethyl) thiophene (compound 24, 3.5 g, 17.3 mmol) and sodium azide (Aldrich, 1.7 g, 26 mmol) in DMF (Aldrich, 60 ml) at ambient temperature for 10 hours Stirred. Flash column chromatography (20% ethyl acetate in hexane) provided the title compound (2.35 g, 83%). 1 H NMR (CDCl 3 ): Δ 4.56 (s, 2H), 7.01 (d, 1H, J = 3.7 Hz), 7.55 (d, 1H, J = 3.7 Hz).
[0241]
(Example 26)
(Preparation of 2-cyano-5- (aminomethyl) thiophene (26))
[0242]
Embedded image
In THF (Aldrich, 40 ml) and in water (10 ml), triphenylphosphine (Aldrich, 5.7 g) at 0 ° C. was added 2-cyano-5- (azidomethyl) thiophene (compound 25, 2.5 g, 10 mmol). Added to the solution. The resulting solution was warmed to room temperature and stirred at ambient temperature for 10 hours. RP HPLC purification provided the title compound (2.3 g, 94%). MS (electrospray): 139 (M + 1); 1 H NMR (CDCl 3 ): Δ 4.01 (s, 2H), 4.75 (br s, 2H, NH 2 ), 6.82 (d, 1H, J = 3.5 Hz), 7.08 (d, 1H, J = 3.5 Hz).
[0243]
(Example 27)
(Preparation of n-butylsulfonyl-D-serine-alanine-2-cyano-5- (methylamido) thiophene (27))
[0244]
Embedded image
The compound of Example 4 (4, 860 mg, 2.9 mmol), the compound of Example 26 (26, 400 mg, 2.9 mmol), EDC (556 mg, 2.94 mmol), N-hydroxybenzotriazole (488 mg, 3 .19 mmol) and a solution of diisopropylethylamine (1.5 mmol, 8.7 mmol) was stirred overnight at ambient temperature. The solvent was removed under reduced pressure and the resulting residue was resuspended in ethyl acetate (50ml) and 1N sodium bicarbonate (10ml). The ethyl acetate phase was washed with 1N sodium bicarbonate (10 ml), saturated sodium bicarbonate (2 × 15 ml) and brine (15 ml), then dried over sodium sulfate to a solid (114 mg, 94%) to a yellow oil Got. The crude product was subjected to flash column chromatography and eluted with 4/5/1 ratio succinethyl / hexane / methanol. The product was purified by reverse phase (C18) HPLC (0.1% trifluoroacetic acid in 5-75% aqueous acetonitrile over 30 minutes, t R = 8.5), resulting in a 3: 1 mixture of diastereomers. The desired mass was confirmed by low resolution mass spectrometry (MH + 417).
[0245]
(Example 28)
(Synthesis of n-butylsulfonyl-D-serine-alanine-2-hydroxyamidino-5- (methylamido) thiophene (28))
[0246]
Embedded image
To a solution of the product of Example 27 (220 mg, 0.53 mol) in methanol (5 ml) was added hydroxyamine hydrochloride (184 mg, 2.64 mmol) followed by N-methylmorpholine (290 μl, 2.64 mmol). Was added. The reaction mixture was stirred overnight at room temperature. The methanol mixture was concentrated under reduced pressure, and the remaining residue was diluted with ethylene acetate and washed with saturated sodium bicarbonate and brine (10 ml each). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a yellow oil (120mg, 50%). This product is obtained on reversed phase (C18) HPLC (t R = 5 min (5-75% aqueous 0.1% trifluoroacetic acid in acetonitrile for 20 min) gave a 3: 1 mixture of diastereomers. The low-resolution mass spectrum confirmed the desired mass (MH + 450.5).
[0247]
(Example 29)
(Synthesis of n-butylsulfonyl-D-serine-alanine-2-amidino-5- (methylamido) thiophene (29))
[0248]
Embedded image
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 of 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 this product eluted in 0-20% aqueous acetonitrile containing 0.1% TFA. These were kept and lyophilized to give 7 g of the title compound as a white powder. The 3: 2 mixture of diastereomers of the product eluted at 13 minutes by reverse phase (C18) HPLC (5% to 50% aqueous 0.1% trifluoroacetic acid in acetonitrile for 20 minutes). The low-resolution mass spectrum confirmed the desired mass (MH + 434).
[0249]
(Example 30)
(Synthesis of tert-butyloxycarbonyl-3,4-methanoproline (30) (the following steps A to E))
(Step A. Synthesis of N-benzyl-3,4-methanoprolinol (b))
[0250]
Embedded image
A mixture of benzylidene initiator (a) (J. Org. Chem. 1999, 64 (2), 547) (4.6 g, 21.4 mmol) and lithium aluminum hydride (1.0 M in THF), 64 ml, 64 mmol ) Was heated at reflux for 5 hours. After cooling to 0 ° C., the remaining aluminum hydride was quenched by careful dropwise addition of saturated aqueous sodium sulfate (5 ml) over 15 minutes. The mixture was diluted with ethyl acetate (200ml) and then filtered through celite. The filtrate was dried over sodium sulfate, filtered, and concentrated to give crude N-benzylamino alcohol (3.45 g), which was carried on to the next step without further purification.
[0251]
(Step B. Synthesis of N-benzyloxycarbonyl-3,4-methanoprolinol)
[0252]
Embedded image
A solution of crude N-benzyl-3,4-methanoprolinol (Step A, (b)) (3 g, 14.76 mmol) in methanol (120 ml) containing 10% Pd / C (300 mg) and concentrated HCl (1 0.5 ml) was hydrogenated at 50 psi for 16 hours. The reaction mixture was filtered to remove the carbon-supported catalyst and the filtrate was concentrated. This 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. The reaction mixture was concentrated, dissolved in 1M HCl (100 ml) and eluted with ethyl acetate (3 × 200 ml). The combined organic phases were dried over magnesium sulfate, filtered and concentrated. The residue was purified by flash chromatography using 1: 3 ethyl acetate / hexane to give N-Cbz-3,4-methanoprolinol (2.4 g).
[0253]
(Step C. Synthesis of N-benzyloxycarbonyl-3,4-methanoprolinol)
[0254]
Embedded image
To a solution of N-benzyloxycarbonyl-3,4-methanoprolinol (2.2 g, 8.90 mmol) in acetone (68 ml) stirred at 0 ° C., Jones reagent (6.6 ml) was added over 5 minutes. [Jones reagent: prepared by diluting chromium trioxide (13.4 g) and concentrated sulfuric acid (11.5 ml) with water to a total volume of 50 ml]. After stirring at 0 ° C. for 3 hours, isopropanol (11 ml) was added and stirring was continued for another 10 minutes. The reaction mixture was diluted with water (400ml) and extracted with ethyl acetate (3x500ml). The combined organic phases were dried over magnesium sulfate, filtered and concentrated to give N-Cbz-3,4-methanoproline (2.25 g, 96 g).
[0255]
(Step D. Synthesis of 3,4-methanoproline hydrochloride salt)
[0256]
Embedded image
A solution of N-benzyloxycarbonyl-3,4-methanoproline (obtained from step C above) (1.23 g, 4.71 mmol) in ethanol (47 ml) and 0.5 M HCL (9.42 ml). To this was added 10% palladium on carbon catalyst. The reaction mixture was hydrogenated at normal pressure overnight. The catalyst was filtered off and the filtrate was concentrated under reduced pressure to give 767 mg of pure product. The product eluted at 14 minutes by reverse phase (C18) HPLC (5-75% 0.1% trifluoroacetic acid in aqueous acetonitrile for 20 minutes).
[0257]
(Step E. Preparation of tert-butyloxycarbonyl-3,4-methanoproline)
[0258]
Embedded image
To a solution of 3,4-methanoproline hydrochloride salt (from step D above) (1.04 g, 6.38 mmol) in dioxane and water (25 ml each), potassium carbonate (1.76 g, 12.7 mmol) and Boc-anhydride (2.78 g, 12.76 mmol) was added. The reaction mixture was stirred overnight at room temperature. The dioxane was removed under reduced pressure and the remaining residue was diluted with diethyl ether. The phases were separated and the aqueous phase was extracted with diethyl ether (1 × 25 ml). The aqueous phase was acidified with 1N hydrochloric acid to pH <3 and extracted with ethyl acetate (3 × 25 ml). The organic phase was dried over sodium sulfate, decanted and concentrated under reduced pressure to give 745 mg (50%) of the product. The product eluted at 12.5 minutes by reverse phase (C18) HPLC (5-75% 0.1% trifluoroacetic acid in aqueous acetonitrile for 20 minutes).
[0259]
(Example 31)
(Preparation of tert-butyloxycarbonyl-3,4-methanoproline-4-cyanobenzylamide (31))
[0260]
Embedded image
Compound of Example 30 (Step E) (0.70 g, 3.082 mmol), p-cyanobenaylamine hydrochloride salt (0.784 g, 4.64 mmol), EDC (0.88 g, 4.62 mmol), N A solution of -hydroxybenzotriazole (0.47 g, 3.082 mmol) and diisopropylethylamine (2.68 ml, 15.41 mmol) was stirred in acetonitrile (12 ml) at room temperature. After 18 hours, the solvent was removed under reduced pressure, and the remaining residue was resuspended in ethyl acetate (150 ml) and 0.5 M HCl (2 × 15 ml), brine (1 × 15 ml), saturated Washed sequentially with sodium carbonate (2 × 15 ml) and brine (1 × 15 ml). The organic phase was dried over sodium sulfate, decanted and concentrated under reduced pressure. The crude product was purified by flash column chromatography eluting with 4/1 ethyl acetate / hexane to give 822 mg (70%) of the product.
[0261]
(Example 32)
(Preparation of 3,4-methanoproline-4-cyanobenzylamide hydrochloride salt (32))
[0262]
Embedded image
To a solution of the compound of Example 31 (1 g, 2.93 mmol) in ethyl acetate (11.7 ml) was added 5M anhydrous HCl in ethyl acetate (2.93 ml) and the mixture was allowed to stand at room temperature overnight. Stirred. The reaction mixture was concentrated under reduced pressure to give 645 mg (79%) of a white solid.
[0263]
(Example 33)
(Preparation of benzylsulfonyl-D-serine (tert-butyl ether) -3,4-methanoproline-4-cyanobenzylamine (33))
[0264]
Embedded image
Compound of Example 11 (0.10 g, 0.32 mmol), Compound 32 (0.105 g, 0.38 mmol), EDC (0.073 g, 0.38 mmol), N-hydroxybenzotriazole (0.049 g, 0.32 mmol) and a solution of diisopropylethylamine (0.28 ml, 1.59 mmol) was stirred at room temperature overnight. The solvent was removed under reduced pressure and the remaining residue was resuspended in ethyl acetate (50 ml) and 1N HCl (10 ml). The ethyl acetate phase was washed with 1N HCl (10 ml), saturated sodium bicarbonate (2 × 15 ml) and brine (15 ml), then dried over sodium sulfate. The organic phase was decanted and concentrated under reduced pressure to give 171 mg of the product.
[0265]
(Example 34)
(Preparation of benzylsulfonyl-D-serine-3,4-methanoproline-4-cyanobenzylamine (34))
[0266]
Embedded image
To a solution of the compound of Example 33 (0.17 g, 0.32 mmol) in dichloromethane (4 ml) was added trifluoroacetic acid (4 ml). The mixture was stirred at room temperature for 1 hour. The reaction mixture was diluted with 10 ml of n-heptane and concentrated under reduced pressure. The residue was resuspended in 5 ml of acetonitrile and 10 ml of n-heptane and concentrated under reduced pressure to give 154 mg of the product.
[0267]
(Example 35)
(Preparation of benzylsulfonyl-D-serine-3,4-methaneproline-4-hydroxyamidinobenzylamide (35))
[0268]
Embedded image
To a solution of the product of 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 μl, 1 .902 mmol) was added. The reaction mixture was stirred overnight at ambient temperature. The reaction mixture was concentrated under reduced pressure. The crude product was resuspended in 25% acetonitrile in water and purified by preparative HPLC. Fractions containing product eluting with 0-20% aqueous acetonitrile containing 0.1% TFA solution were pooled and lyophilized to give 19.5 mg of the title compound as a white powder. By low-resolution mass spectroscopy, the desired mass (MH + 516) was confirmed.
[0269]
(Example 36)
(Preparation of benzylsulfonyl-D-serine-3,4-methanoproline-p-amidinobenzylamide (36))
[0270]
Embedded image
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 of activated zinc dust. The reaction mixture was stirred overnight at room temperature. Zinc powder. Filtration was performed using a glass funnel and the filtrate was purified by preparative HPLC. Fractions containing the product eluting with 0-20% aqueous acetonitrile containing 0.1% TFA were pooled and lyophilized to give 15 mg of the title compound as a white powder. The product eluted at 10.5 minutes by reverse phase (C18) HPLC with 5% 0.1% trifluoroacetic acid in aqueous 50% acetonitrile for 20 minutes. Low resolution mass spectroscopy confirmed the desired mass (MH + 500).
[0271]
(Example 37)
(Preparation of 4-trifluoroacetamidomethylaniline (7-2))
[0272]
Embedded image
4-Nitrobenzylamine (7-1) (4.0 g, 21 mmol) was added to trifluoroacetic anhydride (15 ml) while cooling this mixture on ice. The mixture was warmed to room temperature and stirred overnight. The suspension is poured on ice (about 200 g) and the cloudy suspension is 2 Cl 2 (2 × 100 ml) and extracted with Na 2 SO 4 , And the solvent was removed to give a clear oil. This oil was shaken in a Parr flask overnight with Pd / C (10% / 300 mg) in MeOH (50 ml). The solid was removed by filtration and the solvent was removed under reduced pressure to give a white solid corresponding to the title compound (7-2) (4.5 g, quantitative yield).
[0273]
(Example 38)
(Preparation of N-[(4-trifluoroacetamidomethyl) phenyl] -N′-N ″ -bis (tert-butoxycarbonyl) guanidine (7-3))
[0274]
Embedded image
4-trifluoroacetamidomethylaniline (7-2) (279 mg, 1.28 mmol) was added to CH 2 Cl 2 N-N'-di-Boc-N "-trifluoromethanesulfonyl-guanidine (prepared according to the procedure described in J. Org. Chem. 1998, 63, 3804-3805) (500 mg, 1.28 mmol), and a stirred mixture of TFA (108 μl, 1.28 mmol). The reaction mixture was stirred for 6 hours. This mixture is converted to CH 2 Cl 2 (30 ml) and washed with HCl (1M, 20 ml), brine (20 ml), 2 SO 4 And the solvent was removed under reduced pressure to give a solid. Column chromatography (CH 2 Cl 2 / MeOH, 99: 1) to give a white solid corresponding to the title compound (350 mg, 52%).
[0275]
(Example 39)
(Preparation of N-[(4-aminomethyl) phenyl] -N′-tert-butoxycarbonylguanidine (7-4))
[0276]
Embedded image
Potassium carbonate (500 mg) 2 N-[(4-trifluoroacetamidomethyl) phenyl] -N′-N ″ -bis (tert-butoxycarbonyl) guanidine (7-3) in O / MeOH (2:15, 17 ml) (300 mg, 0 .833 mmol) and the mixture was stirred overnight. The solvent is removed under reduced pressure and the resulting residue is 2 O (10 ml) and CH 2 Cl 2 / MeOH (9: 1, 3 × 10 ml). Organic layer 2 SO 4 And removed under reduced pressure to give a white solid corresponding to the title compound (150 mg, 68%).
[0277]
(Example 40)
(Preparation of benzylsulfonyl-D-serine-L-alanine- (4- (N-tert-butoxycarbonylguanidino) benzylamide (7-5))
[0278]
Embedded image
N-[(4-aminomethyl) phenyl] -N'-tert-butoxycarbonylguanidine (7-4) (36 mg, 0.14 mmol) in acetonitrile (2.0 ml), benzylsulfonyl-D-serine-L- A solution of alanine carboxylate (50 mg, 0.13 mmol), HATU (74 mg, 0.20 mmol), HOAT (27 mg, 0.020 mmol), and DIEA (68 μl, 0.39 mmol) was stirred at room temperature overnight. The solution was diluted with EtOAc (20 ml), HCl (1M, 10 ml), NaHCO 3 (Saturated, 10 ml), washed with brine (10 ml) and removed under reduced pressure to give an oil. HPLC purification (CH 3 CN, H 2 O, 0.1% TFA) gave a fluffy white solid as the title compound (35 mg, 45%), MS (electron spray) 577 (M + 1).
[0279]
(Example 41)
(Preparation of benzylsulfonyl-D-serine-L-alanine-4-guanidinobenzylamide (7-6))
[0280]
Embedded image
CH 2 Cl 2 / TFA (1: 1, 600 μl) in a mixture of benzylsulfonyl-D-serine-L-alanine- (4- (N-tert-butoxycarbonylguanidino) benzylamide (7-5) (9.0 mg, 0.1 mg). 016 mmol) was stirred for 90 minutes at room temperature.The solvent was removed under reduced pressure to give a clear oil.HPLC purification (CH 3 CN, H 2 O, 0.1% TFA) gave a fluffy white solid as the title compound (5.0 mg, 66%), MS (electron spray) 477 (M + 1).
[0281]
(Example 42)
(Preparation of 3-trifluoroacetamidomethylaniline (8-2))
[0282]
Embedded image
3-Nitrobenzylamine (8-1) (3.0 g, 16 mmol) was added to trifluoroacetic anhydride (30 ml) while cooling the mixture on ice and the mixture was stirred overnight. The suspension is poured on ice (about 200 g) and the cloudy suspension is washed with CH 2 Cl 2 (2 × 100 ml) and extracted with Na 2 SO 4 And dried to give a clear oil. This oil was shaken in a Parr flask overnight with Pd / C (10%, 300 mg) in MeOH (30 ml). The solid was removed by filtration and the solvent was removed under reduced pressure to give a solid corresponding to the title compound (3.3 g, 95%).
[0283]
(Example 43)
(Preparation of N-[(3-trifluoroacetamidomethyl) phenyl] -N′-N ″ -bis (tert-butoxycarbonyl) guanidine (8-3))
[0284]
Embedded image
3-trifluoroacetamidomethylaniline (8-2) (product of Example 42) (500 mg, 2.29 mmol) was added to CH 2 Cl 2 NN′-di-Boc-N ″ -trifluoromethanesulfonyl-guanidine (prepared according to the procedure described in J. Org. Chem. 1998, 63, 3804-3805) (1086 ml) (986 mg, 2.52 mmol) and TEA (642 μl, 4.58 mmol) were added and the mixture was stirred for 24 hours. The mixture was washed with HCl (1 M, 10 ml), brine (10 ml) and Na 2 SO 4 And the solvent was removed under reduced pressure to give a solid. Column chromatography (CH 2 Cl 2 / MeOH, 98: 2) to give a white solid corresponding to the title compound (479 mg, 55%). MS (electron spray) 461 (M + 1).
[0285]
(Example 44)
(Preparation of N-[(3-aminomethyl) phenyl] -N′-tert-butoxycarbonylguanidine (8-4))
[0286]
Embedded image
Potassium carbonate (1.0 g) 2 N-[(3-trifluoroacetamidomethyl) phenyl] -N′-N ″ -bis (tert-butoxycarbonyl) guanidine (8-3) in O / MeOH (1: 1, 4 ml) (Example 43) (450 mg, 0.978 mmol) was added and the mixture was stirred overnight. The solvent is removed under reduced pressure and the resulting residue is 2 O (10 ml) and CH 2 Cl 2 / MeOH (9: 1, 3 × 10 ml). Organic layer 2 SO 4 And removed under reduced pressure to give a white solid corresponding to the title compound (232 mg, 90%).
[0287]
(Example 45)
(Preparation of benzylsulfonyl-D-serine-L-alanine-3-guanidinobenzylamide (8-5))
[0288]
Embedded image
N-[(3-Aminomethyl) phenyl] -N'-tert-butoxycarbonylguanidine (8-4) (product of Example 44) in acetonitrile (2.0 ml) (130 mg, 0.492 mmol), benzyl A solution of sulfonyl-D-serine-L-alanine carboxylate (190 mg, 0.492 mmol), HATU (380 mg, 0.739 mmol), HOAT (136 mg, 0.739 mmol), and DIEA (258 μl, 1.48 mmol) Stirred overnight at room temperature. The solution was diluted with EtOAc (20 ml), HCl (1M, 10 ml), NaHCO 3 (Saturated, 10 ml), washed with brine (10 ml) and removed under reduced pressure to give an oil. This oil is CH 2 Cl 2 / TFA (1: 1, 3 ml) and stirred at room temperature for 2 hours. The solvent was removed under reduced pressure to give a clear oil. HPLC purification (CH 3 CN, H 2 O, 0.1% TFA) gave a fluffy white solid as the title compound (80 mg, 34%), MS (electron spray) 477 (M + 1).
[0289]
(Example 46)
(Preparation of 2-nitro-5- (4-trifluoroacetamidomethyl) -thiophene (9-2))
[0290]
Embedded image
2-Aminomethylthiophene (9-1) (5.0 g, 44 mmol) was added in portions to stirring trifluoroacetic anhydride (20 ml) while cooling the mixture on ice, and the mixture was stirred for 1 hour. Add KNO to this solution 3 (8.9 g, 88 mmol) was added in portions at -20 <0> C. The homogeneous solution was warmed to ambient temperature and stirred overnight. The resulting suspension is poured onto ice (about 200 g) and CH 2 Cl 2 (2 × 100 ml) and extracted with Na 2 SO 4 And dried to remove the solvent to give a solid. Column chromatography (CH 2 Cl 2 ) Provided a white solid corresponding to the title compound (3.3 g, 29%). MS (electron spray) 255 (M + 1).
[0291]
(Example 47)
(Preparation of N- [2- (5-trifluoroacetamidomethyl) thiophenyl] -N′-N ″-(bis-tert-butoxycarbonyl) guanidine (9-3))
[0292]
Embedded image
2-Nitro-5- (4-trifluoroacetamidomethyl) -thiophene (9-2) (1.0 g, 3.9 mmol) was added to a saturated solution of HCl in MeOH at 0 <0> C. SnCl 2 (4.4 g) was added dropwise over 15 minutes and the mixture was stirred for 30 minutes. The solution is concentrated, then NaHCO 3 (10 ml). This solution is CH 2 Cl 2 (2 × 20 ml), Na 2 SO 4 And the solvent was removed under reduced pressure to give 2-amino-5- (4-trifluoroacetamidomethyl) -thiophene as a yellow oil (purity by NMR> 95%, 0.80 g, 95%). This compound was used immediately in the next step without further purification.
[0293]
2-Amino-5- (4-trifluoroacetamidomethyl) -thiophene (0.80 g, 3.6 mmol) was added to CH 2 Cl 2 (20 ml) was added to a stirred solution of NN′-di-Boc-N ″ -trifluoromethanesulfonyl-guanidine (1.5 g, 3.8 mmol) and TEA (1.1 ml, 7.8 mmol). The mixture was stirred for 24 hours. This mixture is converted to CH 2 Cl 2 (20 ml) and washed with HCl (1M, 20 ml), brine (20 ml), 2 SO 4 And dried. The solvent was removed under reduced pressure to give an oil. Column chromatography (CH 2 Cl 2 / MeOH, 98: 2) to give an oil (150 mg, 9%) corresponding to the title compound (9-3). MS (electrospray): 467 (M + l).
[0294]
(Example 48)
(Preparation of (N- [5-4-aminomethyl) thiophenyl] -N '-(tert-butoxycarbonyl) guanine (9-4))
[0295]
Embedded image
Potassium carbonate (500 mg) 2 N- [2- (5-trifluoroacetamidomethyl) thiophenyl] -N′-N ″-(bis-tert-butoxycarbonyl) guanidine (9-3) in O / MeOH (1: 1, 4 ml) 150 mg, 0.322 mmol) was added to the stirred solution and the mixture was stirred overnight. The solvent was removed under reduced pressure. The remaining residue is H 2 Dissolved in O (10 mL) and CH 2 Cl 2 / MeOH (95: 5, 3 x 10 ml). The organic layer is Na 2 SO 4 And dried. The solvent was removed under reduced pressure to give a yellow solid corresponding to the title compound (150mg, 68%).
[0296]
(Example 49)
(Preparation of benzylsulfonyl-D-serine-L-alanine- [5- (2-guanidino) thiophene] (9-5))
[0297]
Embedded image
N- [5- (4-Aminomethyl) thiophenyl] -N ′-(tert-butoxycarbonyl) guanine (9-4) (77 mg, 0.29 mmol), benzylsulfonyl-D in acetonitrile (3.0 mL). A solution of -serine-L-alanine carboxylate (110 mg, 0.285 mmol), HATU (162 mg, 0.427 mmol), HOAT (58 mg, 0.42 mmol) and DIEA (199 μL, 1.13 mmol) was added at ambient temperature. Stirred overnight. The solution was diluted with EtOAc (20 ml), HCl (1M, 10 ml), NaHCO 3 (Saturated, 10 mL), and brine (10 ml). The organic layer is Na 2 SO 4 And the solvent was removed under reduced pressure to give an oil. TFA / CH 2 Cl 2 (1: 1, 2 ml) was added; the mixture was stirred for 3 hours. HPLC purification (CH 3 CN, H 2 O, 0.1% THF) gave a fluffy white solid (3 mg) corresponding to the title compound. MS (electrospray): 483 (M + 1).
[0298]
(Example 50)
(Preparation of 6-bromomethylnicotinonitrile (11-2))
[0299]
Embedded image
To a solution of 6-methylnicotinonitrile (11-1) (Lancaster) (15 g, 127 mmol) in carbon tetrachloride (300 ml) was added N-bromosuccinimide (27.12 g, 152.4 mmol). The resulting solution was degassed and purged with nitrogen, and AIBN (2,2'-azobisisobutyronitrile) (2.08 g, 12.6 mmol) was added. After 7 hours at 85 ° C., another batch of AIBN (1.04 g) was added and stirring continued for an additional hour. After removal of the solvent, the crude product was subjected to flash column chromatography in ethyl acetate / hexane to give 10 g of pure material (40%).
[0300]
(Example 51)
(Preparation of 6-azidomethylnicotinonitrile (11-3))
[0301]
Embedded image
To a solution of 6-bromomethylnicotinonitrile (11-2) (8.0 g, 40.6 mmol) in DMF (100 ml) was added sodium azide (3.17 g, 48.8 mmol). The reaction mixture was stirred overnight at room temperature. The reaction mixture was diluted with 400 ml of diethyl ether and 100 ml of water, and the layers were separated. The aqueous layer was re-extracted with diethyl ether (2 × 100 ml). The combined ether extracts were washed with brine (2 × 100 ml) and then MgSO 4 And filtered to give 6.53 g of the product as a white solid.
[0302]
(Example 52)
(Preparation of 3-hydroxyamidino-6-azidomethylpyridine (11-4))
[0303]
Embedded image
To a solution of the azide (11-3) of Example 51 (6.46 g, 40.6 mmol) in methanol (100 ml) was added hydroxyamine hydrochloride (3.95 g, 56.84 mmol) followed by triethylamine (9.9. 6 ml) was added. The solution was heated at 65 ° C. for 4 hours. The solvent was removed under reduced pressure, and the remaining residue was extracted with ethyl acetate (300ml) and water (100ml). The aqueous layer was re-extracted with ethyl acetate (2 × 200 ml). The combined organic extracts were washed with brine (2 × 100 ml) and MgSO 4 And evaporated to give the title product (11-4) (7.8 g).
[0304]
(Example 53)
(Preparation of 3-hydroxyamidino-6-aminomethylpyridine (11-5))
[0305]
Embedded image
To a solution of the azide (11-4) of Example 52 (7.8 g, 40.6 mmol) in THF / water (80 ml / 5 ml) was added triphenylphosphine (12.8 g, 48.72 mmol). The reaction mixture was stirred overnight at room temperature. The solvent was removed to dryness and the remaining residue was diluted with 100 ml 1N HCl and 100 ml water. The aqueous layer was extracted several times with dichloromethane. The aqueous layer was made basic (pH about 9) with an alkali resin (Bio Rad AG 1-X8). The resin was filtered and washed thoroughly with water. The combined washes were dried under reduced pressure to give a white solid (6.60 g).
[0306]
(Example 54)
(Preparation of 3-hydroxyamidino-6-aminomethyl (Boc) pyridine (11-6))
[0307]
Embedded image
To a solution of the product of Example 53 (11-5) (0.5 g, 3.01 mmol) in dioxane (6 ml) and water (6 ml) was added potassium carbonate (832 mg, 6.02 mmol) followed by , Boc-anhydride (0.66 g, 3.01 mmol) was added. The reaction mixture was stirred overnight at room temperature. The reaction mixture was concentrated and the remaining residue was dissolved in ethyl acetate and washed with sodium bicarbonate (sat.) And brine. The organic layer was dried over sodium sulfate, filtered, and concentrated. Flash column chromatography (8/2 ethyl acetate / hexane followed by ethyl acetate and 9/1 dichloromethane / methanol) gave 0.43 g of the product as a white solid. NMR δ (ppm) CDCl 3 : 8.8 (d, 1H), 7.9 (dd, 1H), 7.3 (d, 1H), 6.6 (bs, 1H), 5.5 (bs, 1H), 4.8 ( bs, 1H), 4.4 (d, 2H), 1.4 (s, 1H).
[0308]
(Example 55)
(Preparation of 3-isopropyloxyamidino-6-aminomethyl (Boc) pyridine (11-7))
[0309]
Embedded image
To a solution of the product of Example 54 (11-6) (0.43 g, 1.39 mmol) in dimethylformamide (5 ml) was added 2-iodopropane (210 μl, 2.1 mmol) followed by cesium carbonate (0.68 g, 2.1 mmol) was added. The reaction mixture was stirred overnight at room temperature. The reaction mixture was diluted with ethyl acetate and washed with sodium hydrogen carbonate (sat.). The organic layer was dried over sodium sulfate, filtered, and concentrated to give an orange oil. Flash column chromatography (1/1 ethyl acetate / hexane followed by ethyl acetate) provided 229 mg (53%) of the product as a crystalline solid. NMR δ (ppm) CDCl 3 : 8.8 (d, 1H), 7.9 (dd, 1H), 7.3 (d, 1H), 5.5 (bs, 1H), 4.7 (bs, 2H), 4.4 ( d, 1H), 1.4 (s, 9H), 1.3 (d, 6H).
[0310]
(Example 56)
(Preparation of Boc-Ala-3-isopropyloxyamidino-6-aminomethylpyridine (11-8))
[0311]
Embedded image
Compound (11-7) of Example 55 (229 mg, 0.74 mmol) was dissolved in dioxane (1 ml) and treated with a solution of 4N HCl in dioxane (1 ml) at room temperature for 1 hour. The solvent was removed under reduced pressure to give 290 mg of a white solid. This product was then dissolved in acetonitrile (4.12 ml), neutralized with diisopropylethylamine (538 μl, 3.09 mmol), and as coupling agents EDC (197 mg, 1.03 mmol) and HOBt (157.6 mg). , 1.03 mmol) to Boc-alanine (195 mg, 1.03 mmol). After stirring at room temperature overnight, the solvent was removed under reduced pressure and the remaining residue was diluted with ethyl acetate. The organic layer was washed several times with sodium bicarbonate (saturated) and brine, dried over sodium sulfate, filtered, and concentrated to give a yellowish oil. Flash column chromatography (9/1 ethyl acetate / hexane followed by ethyl acetate) provided 181 mg of a white oil (46%). NMR δ (ppm) CDCl 3 : 8.7 (d, 1 H), 7.9 (dd, 1 H), 7.3 (d, 1 H), 7.1 (bs, 1 H), 5.0 (bs, 1 H), 4.7 ( bs, 2H), 4.6 (d, 2H), 4.3 (m, 1H), 4.1 (m, 1H), 1.4 (s, 9H), 1.39 (d, 3H), 1.3 (d, 6H).
[0312]
(Example 57)
(Preparation of benzylsulfonyl-dSer (tBu) -Ala-3-isopropyloxyamidino-6-aminomethylpyridine (11-9))
[0313]
Embedded image
Compound (11-8) of Example 56 (181 mg, 0.48 mmol) was dissolved in dioxane (1 ml) and treated with 4N HCl in dioxane (1 ml) at room temperature for 1.5 hours. The solvent was removed under reduced pressure to give 200 mg of a white solid. This product was then dissolved in acetonitrile (5 ml), neutralized with diisopropylethylamine (298 μl, 1.71 mmol), and EDC (109 mg, 0.57 mmol) and HOBt (96 gm, 0.63 mmol) as coupling agents Using BnSO 2 -Coupled to dSer (tBu) OH (180 mg, 0.57 mmol). After stirring overnight at room temperature, the solvent was removed under reduced pressure and the resulting residue was diluted with ethyl acetate. The organic layer was washed several times with sodium bicarbonate (saturated) and brine, dried over sodium sulfate, filtered, and concentrated to give a solid (250 mg, 76%). The product eluted at 11.5 minutes by reverse phase (C18) HPLC (0.1% trifluoroacetic acid in 5-75% aqueous acetonitrile over 20 minutes). MS (electrospray): 577 (M + 1).
[0314]
(Example 58)
(Preparation of benzylsulfonyl-dSer-Ala-3-isopropyloxyamidino-6-aminomethylpyridine (11-10))
[0315]
Embedded image
Compound (11-9) of Example 57 (137 mg, 0.24 mmol) was treated with 2 ml each of dichloromethane and trifluoroacetic acid at room temperature for 1 hour. Removal of the solvent under reduced pressure gave 169 mg of an orange oil. The product eluted at 9 minutes by reverse phase (C18) HPLC (0.1% trifluoroacetic acid in 5-90% aqueous acetonitrile over 20 minutes).
[0316]
(Example 59)
(Preparation of benzylsulfonyl-dSer-Ala-3-amidino-6-aminomethylpyridine (11-11))
[0317]
Embedded image
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 active zinc (91 mg) for 45 minutes. The solution was filtered and the filtrate was purified by reverse phase HPLC (C18). The product eluted at 11 minutes by reverse phase HPLC (0.1% trifluoroacetic acid in 5-25% aqueous acetonitrile over 20 minutes). MS (electrospray): 463 (M + l).
[0318]
(B. Synthesis route of specific intermediate compound)
(I) Examples 60-97 describe the synthesis of certain intermediates used in preparing compounds of the present invention. See also FIGS.
(Example 60)
(Preparation for synthesis of D-Ser (Ot-Bu) -Ala-OMe acetate (1-3))
[0319]
Embedded image
N-α-Cbz-D-serine (1-1) (Bachem, 4.97 g, 16.8 mmol), alanine methyl ester hydrochloride (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. After stirring as a slurry for 10 minutes, diisopropylethylamine (14.4 ml, 84 mmol) was added and the resulting clear mixture was stirred for another 18 hours. The solvent was removed under reduced pressure and the residue was suspended in ethyl acetate (500ml). The solution was washed with 0.5M HCl (2 × 100 ml), followed by saturated sodium bicarbonate (2 × 100 ml) and brine (100 ml). The organic layer was then dried over sodium sulfate and the solvent was removed under reduced pressure to give Cbz-D-Ser (Ot-Bu) -Ala-OMe by reversed phase (C18) HPLC (over 20 min. In 0.1% trifluoroacetic acid in 5-75% aqueous acetonitrile, t R = 16.9) as a single peak in quantitative yield.
[0320]
Cbz-D-Ser (Ot-Bu) -Ala-OMe was then dissolved in ethanol / acetic acid / water (150 ml of a 4: 1: 1 mixture). The flask was filled with nitrogen and 10% palladium on carbon (1.5 g) was added. The mixture was hydrogenated at 45 psi for 2 hours. The palladium catalyst was filtered and the solvent was removed under reduced pressure to give 4.58 g (95% yield) of the title compound by reverse phase (C18) HPLC (5-75% aqueous acetonitrile over 20 min. In 1% trifluoroacetic acid, t R = 8.0 min) as a single peak. MS (M + H = 247.2).
[0321]
(Example 61)
(Preparation of benzenesulfonyl-D-Ser (Ot-Bu) -Ala-OMe (1-4))
[0322]
Embedded image
To a stirred slurry of compound (1-3) of Example 60 (1.0 g, 3.3 mmol) in acetonitrile (13 ml) was added benzenesulfonyl chloride (0.87 g, 4.9 mmol). To this mixture was added diisopropylethylamine (1.67 ml, 9.8 mmol) in 5 portions over 1 hour. The mixture was stirred for another hour. The solvent was removed under reduced pressure and the residue was suspended in ethyl acetate (100ml). The solution was washed with 0.5M HCl (2 × 10 ml), followed by saturated sodium bicarbonate (2 × 10 ml) and brine (1 × 10 ml). The organic layer was then dried over sodium sulfate and the solvent was removed under reduced pressure. The residue was purified by flash chromatography (eluting with 50% hexane / ethyl acetate) to give 0.54 g, 1.4 mmol of product in 43% yield. The product was purified by reverse phase (C18) HPLC (0.1% trifluoroacetic acid in 5-75% aqueous acetonitrile over 20 min. R = 20.2 min). 1 H NMR (CD 3 OD): 7.5-7.9 ppm (m, 5H), 4.3 ppm (q, 1H), 3.9 ppm (t, 1H), 3.7 (s, 3H), 3.4 ppm (m, 1H). ), 3.5 (m, 1H), 1.3 ppm (d, 3H), 1.05 ppm (s, 9H).
[0323]
(Example 62)
(Preparation of benzenesulfonyl-D-Ser (Ot-Bu) -Ala-OH (1-5))
[0324]
Embedded image
To compound (1-4) of Example 61 (0.53 g, 1.4 mmol) in methanol (9 ml) was added 1.0 M lithium hydroxide (3.0 ml, 3 mmol). After stirring for 18 hours, the reaction mixture was poured onto a column of DOWEX (5 × 8-400) ion exchange resin and eluted with methanol / water (60 ml of a 1: 1 mixture). The methanol was removed under reduced pressure and the remaining water was lyophilized to give 0.49 g, 1.3 mmol (95%) of the title compound by reverse phase (C18) HPLC (5-75% aqueous over 20 min. In 0.1% trifluoroacetic acid in acetonitrile, t R = 13.5 min). 1 H NMR (CD 3 OD): 7.9 ppm (d, 2H), 7.6 ppm (t, 1H), 7.5 ppm (t, 2H), 4.25 ppm (q, 1H), 3.9 ppm (t, 1H), 3. 5 ppm (m, 1H), 3.4 ppm (m, 1H), 1.3 ppm (d, 3H), 1.075 ppm (s, 9H).
[0325]
(Example 63)
(Preparation of Benzylsulfonyl-D-Ser (Ot-Bu) -OMe)
[0326]
Embedded image
To a stirred solution of D-serine (Ot-Bu) methyl ester hydrochloride (2.07 g, 9.8 mmol) in acetonitrile (39 ml) was added α-toluenesulfonyl chloride (1.86 g, 9.8 mmol). did. To this mixture was added diisopropylethylamine (3.7 ml, 21.5 mmol) in 5 portions over 1 hour. The mixture was stirred for another hour. The solvent was removed under reduced pressure and the residue was suspended in ethyl acetate (100ml). The solution was washed with 0.5 M HCl (2 × 100 ml), followed by saturated sodium bicarbonate (2 × 10 ml) and brine (1 × 10 ml). The organic layer was dried over sodium sulfate and the solvent was removed under reduced pressure to give 2.84 g of the title compound in 88% yield. R f = 0.4 (4: 1 ethyl acetate: hexane).
[0327]
(Example 64)
(Preparation of benzylsulfonyl-D-Ser (Ot-Bu) -OH)
[0328]
Embedded image
To a stirred solution of the compound of Example 63 (2.66 g, 8.1 mmol) in methanol (54 ml) was added 1.0 M lithium hydroxide (17.8 ml, 17.8 mmol). The reaction mixture was stirred for 18 hours, then poured onto a column of 10 ml DOWEX (50 × 8-400) ion exchange resin and eluted with methanol: water (60 ml of a 1: 1 mixture). The methanol was removed under reduced pressure and the remaining aqueous solution was lyophilized to give 2.47 g of the title compound in 97% yield. t R = 14.8 minutes (0.1% trifluoroacetic acid in 5-75% aqueous acetonitrile over 20 minutes).
[0329]
(Example 65)
(Preparation of Benzylsulfonyl-D-Ser (Ot-Bu) -Ala-OMe)
[0330]
Embedded image
The compound of Example 64 (1.0 g, 3.2 mmol), alanine methyl ester hydrochloride (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 another 18 hours. The solvent was removed under reduced pressure and the residue was suspended in ethyl acetate (100ml). The solution was washed with 0.5M HCl (2 × 10 ml), followed by saturated sodium bicarbonate (2 × 10 ml) and brine (1 × 10 ml). The organic layer was then dried over sodium sulfate and the solvent was removed under reduced pressure to give 1.22 g of the title compound in 97% yield. t R = 16.2 minutes (0.1% trifluoroacetic acid in 5-75% aqueous acetonitrile over 20 minutes).
[0331]
(Example 66)
(Preparation of benzylsulfonyl-D-Ser (Ot-Bu) -Ala-OH)
[0332]
Embedded image
To the compound of Example 65 (1.22 g, 3.1 mmol) in methanol (22 ml) was added 1 M lithium hydroxide (7.2 ml, 7.2 mmol). After stirring for 18 hours, the reaction mixture was poured onto a column of 10 ml DOWEX (50 × 8-400) ion exchange resin and eluted with methanol: water (60 ml of a 1: 1 mixture). The methanol was removed under reduced pressure and the aqueous solution was lyophilized to give 1.16 g of the title compound in 91% yield. t R = 13.2 minutes (0.1% trifluoroacetic acid in 5-75% aqueous acetonitrile over 20 minutes).
[0333]
(Example 67)
(Preparation of i-butoxycarbonyl-D-Ser (Ot-Bu) -OMe)
[0334]
Embedded image
To a stirred, homogeneous solution of HCl-D-Ser (Ot-Bu) -OMe (15 g, 71 mmol, Bachem) in tetrahydrofuran (200 ml) was added saturated sodium bicarbonate (80 ml), followed by isobutyl chloroformate. (19.45 g, 142 mmol) was added. The layers were separated and the aqueous layer was washed with ethyl acetate (50ml). The organic phases were combined and the solvent was removed under reduced pressure. The residue was suspended in ethyl acetate (100ml) and washed with 1M HCl (100ml), saturated sodium bicarbonate (100ml), and brine (100ml). The organic layer was dried over magnesium sulfate, treated with decolorizing carbon (eg, sold under the trade name Darco), filtered, and the solvent was removed under reduced pressure to give a quantitative yield. The title compound was obtained. R f = 0.3 (20% ethyl acetate / hexane).
[0335]
(Example 68)
(Preparation of i-butoxycarbonyl-D-Ser (Ot-Bu) -OH)
[0336]
Embedded image
To a stirred solution of the compound of Example 67 (19.51 g, 70 mmol) in tetrahydrofuran (78 ml) was added lithium hydroxide (78 mmol, 3.28 g). The reaction mixture was stirred vigorously for 3 hours until no starting material was observed by TLC (20% ethyl acetate / hexane). The solution was acidified to about pH 2 with concentrated HCl and the solvent was removed under reduced pressure. The crude product was suspended in ethyl acetate and extracted with saturated sodium hydrogen carbonate (2x, 75ml). The combined sodium bicarbonate washes were acidified with 6M HCl, and the separated oil was extracted with ethyl acetate (2 × 100 ml). The combined organic layers were dried over magnesium sulfate, treated with Darco, filtered, and the solvent was removed under reduced pressure to give a quantitative yield of the title compound. R f = 0.01 (20% ethyl acetate in hexane).
[0337]
(Example 69)
(Preparation of i-butoxycarbonyl-D-Ser (Ot-Bu) -Ala-OMe)
[0338]
Embedded image
Compound of Example 68 (16.5 g, 63 mmol), HCl-Ala-OMe (10.6 g, 76 mmol), 1-hydroxybenzotriazole (10.2 g, 76 mmol) in acetonitrile (280 ml), and EDC (16. To a solution (0 ° C.) of (33 g, 85 mmol) was added 4-methylmorpholine (35 ml, 315 mmol). The mixture was stirred at 0 ° C. for 1 hour and then at ambient temperature for 72 hours. The solvent was removed under reduced pressure and the resulting residue was suspended in ethyl acetate (300ml) and 1M HCl (350ml). The aqueous layer was separated and washed with ethyl acetate (300ml). The combined ethyl acetate layers were washed with 1M HCl (300 ml), saturated sodium bicarbonate (400 ml), and brine (200 ml). The organic layer was dried over magnesium sulfate, treated with Darco decolorizing carbon and filtered. The solvent was removed under reduced pressure to give 21.48 g of the title compound (98% yield).
[0339]
(Example 70)
(Preparation of i-butoxycarbonyl-D-Ser-Ala-OH)
[0340]
Embedded image
The compound of Example 69 (21 g, 58 mmol) was dissolved in trifluoroacetic acid (110 ml); the resulting mixture was stirred for 35 minutes. The solution was cooled in an ice bath and saturated sodium bicarbonate (630 ml) was added, followed by solid sodium bicarbonate (70 g) over a period of 45 minutes until pH = 7. This aqueous solution was extracted with ethyl acetate (3 × 250 ml). The combined organic extracts were combined, dried over magnesium sulfate, treated with Darco decolorizing carbon, and filtered. The solvent was removed under vacuum to give a quantitative yield of i-butoxycarbonyl-D-Ser-Ala-OMe.
[0341]
To a stirred solution of the crude residue in tetrahydrofuran (68 ml) was added lithium hydroxide (2.7 g, 64 mmol, 1.1 eq.) In water (17 ml). The reaction mixture was stirred vigorously for 0.5 h until no more starting material was observed as observed by TLC (9: 1 dichloromethane / isopropanol). The solution was acidified with 6 M HCl (13 ml) to about pH 2 and the solvent was removed under reduced pressure. The crude product was suspended in ethyl acetate (400ml) and water (50ml). The aqueous layer was extracted with ethyl acetate (200ml). The combined organic layers were dried over magnesium sulfate, filtered, and the solvent was removed under vacuum to give 13.94 g of the title compound (86% yield). R f = 0.3 (90: 30: 5 chloroform / methanol / acetic acid).
[0342]
(Example 71)
(Preparation of N-α- (3-phenylpropyl) -D-serine-t-butyl ether methyl ester)
[0343]
Embedded image
Serine Ot-butyl ether methyl ester (1.50 g, 7.1 mmol), hydrocinnamaldehyde (1.40 ml, 10.6 mmol), and triethylamine (1.18 ml, 8.5 mmol) in tetrahydrofuran (70 ml). Refluxed for 4 hours. After allowing the solution to cool to room temperature, sodium borohydride (0.46 g, 12 mmol) was added to the stirring solution in two portions. The reaction mixture was stirred at ambient temperature for 30 minutes; the solution was concentrated under reduced pressure. This residue was partitioned between ethyl acetate 1.0 M HCl. The organic layer was washed with 1.0N HCl. The aqueous layer was basified with 40% NaOH to pH 10, then extracted with ethyl acetate (2x). Layers were dried over sodium sulfate; the solvent was removed under vacuum. The residue was purified by flash chromatography on silica gel (1 × 6 inch column) eluting with 10-30% ethyl acetate / hexane to give 150 mg (7% yield) of the title compound. R f = 0.60 (50% ethyl acetate / hexane).
[0344]
(Example 72)
(Preparation of N-α-t-butoxycarbonyl-N-α- (3-phenylpropyl) -D-serine-t-butyl ether methyl ester)
[0345]
Embedded image
Example 71 (150 mg, 0.51 mmol), di-t-butyl dicarbonate (167 mg, 0.77 mmol) and diisopropylethylamine (0.13 ml, 0.77 mmol) in tetrahydrofuran (2 ml) at ambient temperature And stirred overnight. The reaction mixture was concentrated under reduced pressure. The residue was diluted with ethyl acetate (20 ml) and washed sequentially with 1.0 N HCl (2 ×), saturated sodium bicarbonate (2 ×), and brine (1 ×). The solvent was removed under vacuum to give 206 mg of the title compound in quantitative yield. R f = 0.74 (50% ethyl acetate / hexane).
[0346]
(Example 73)
(Preparation of N-α-t-butoxycarbonyl-N-α- (3-phenylpropyl) -D-serine-t-butyl ether)
[0347]
Embedded image
To a solution of the compound of Example 72 (206 mg, 0.52 mmol) in methanol (3.5 ml) was added 1.0 N LiOH (0.63 ml, 0.63 mmol) dropwise. The solution turned cloudy and then became homogenous in 5 minutes. The reaction mixture was allowed to stir at ambient temperature overnight. Additional 1.0 N LiOH (1.47 ml) was added. After 2 hours, an additional 1.0 N LiOH (1.0 ml) was added. After no more starting material was observed by TLC (50% ethyl acetate / hexane), the reaction mixture was acidified to pH 4 using DOWEX (50 × 8-400) ion exchange resin. The solution was filtered, rinsed with methanol and then with water. The solution was concentrated under reduced pressure and then lyophilized to give 189 mg of the title compound (95% yield) as a yellow oil. R f = 0.04 (50% ethyl acetate / hexane).
[0348]
(Example 74)
(Preparation of i-butoxycarbonyl-D-Ser (Ot-butyl) -OH (2-2))
[0349]
Embedded image
To a solution of D-serine-Ot-butyl ether (2-1) (10.5 g, 65.3 mmol) in water (51 ml) was added sodium carbonate (20.1 g, 196.2 mmol), Isobutyl chloroformate (9.8 ml, 75.2 mmol) was added. After 3 hours, the mixture became cloudy and was stirred for a further 3 hours. After 6 hours, the solution was acidified with 6M HCl (about 50 ml). Then the reaction mixture was extracted with ethyl acetate (3 × 200 ml). After the first extraction, the aqueous layer was saturated with sodium chloride and extracted with ethyl acetate (2 × 200 ml). The ethyl acetate layers were combined and dried over sodium sulfate, then the solvent was removed under vacuum to give 17.0 g (99.5% yield) of the title compound as a white solid. HPLC: t = 18.5 min (5% to 75% acetonitrile gradient in 0.1% aqueous TFA buffer), 4.6 × 250 mm, 5 micron particles, 100 Å pore, C18 column, 1 ml / min flow rate ). NMR (CDC1 3 ): 5.5 ppm (m, 1H), 4.45 ppm (bs, 1H), 3.82 to 3.95 ppm (m, 3H), 3.52 to 3.6 ppm (m, 1H), 1.85 1.97 ppm (m, 1H), 1.2 ppm (s, 9H), 0.9 ppm (d, 6H).
[0350]
(Example 75)
(Preparation of i-butoxycarbonyl-D-Ser (t-Bu) -L-Ala-Ot-Bu (2-3))
[0351]
Embedded image
Compound (2-2) of Example 74 (10 g, 38.3 mmol), L-alanine t-butyl ester, HCl salt (10.43 g, 57.4 mmol) in acetonitrile (153 ml), EDC (11.05 g, A solution of 57 mmol) and hydroxybenzotriazole (5.85 g, 38.3 mmol) was stirred at room temperature for 15 minutes. Diisopropylethylamine (32.7 ml, 191 mmol) was added and the reaction mixture was stirred for 18 hours. The solvent was removed under reduced pressure: the resulting residue was resuspended in ethyl acetate (1000 ml) and 1M HCl (100 ml). The ethyl acetate layer was washed with 0.5 M HCl (100 ml), saturated sodium bicarbonate (2 × 100 ml), and brine (100 ml). The ethyl acetate layer was dried over sodium sulfate and the solvent was removed under vacuum to give a quantitative yield of the title compound. HPLC: t r = 18.7 minutes (5% to 90% acetonitrile gradient (in 0.1% aqueous TFA buffer), 4.6 x 250 mm, 5 micron particles, 100 Angstrom pore, C18 column, 1 ml / min flow rate). NMR (CDCl 3 ): 7.15 ppm (bs, 1H), 5.6 ppm (bs, 1H), 4.4 to 4.5 ppm (m, 1H), 4.2 ppm (bs, 1H), 3.87 to 3.95 ppm ( m, 3H), 3.3-3.4 ppm (m, 1H), 1.85-1.95 ppm (m, 1H), 1.45 ppm (s, 9H), 1.39 ppm (d, 3H), 1 .2 ppm (s, 9H), 0.9 ppm (d, 6H).
[0352]
(Example 76)
(Preparation of i-butoxycarbonyl-D-Ser-L-Ala-OH (2-4))
[0353]
Embedded image
To a solution of the compound of Example 75 (2-3) (7.15 g, 18.4 mmol) in dichloromethane (35 ml) was added trifluoroacetic acid (35 ml). The mixture was stirred for 2 hours, then the solvent was removed under reduced pressure. Toluene was added and the solvent was transferred to a vacuum to remove trifluoroacetic acid. The yield of the title compound as a sticky yellow oil was then transferred to the next step. A gradient of 5% to 90% acetonitrile in 0.1% aqueous TFA buffer at 4.6 × 250 mm, 5 micron particles, 100 Å pore, C18 column at a flow rate of 1 ml / min, t r = 10.5 minutes.
[0354]
(Example 77)
(Preparation of 2-cyano-5-methylthiophene (3-2))
[0355]
Embedded image
A solution of 2-bromo-5-methylthiophene (3-1) (TCI chemicals, 5 g, 28 mmol) and copper (I) cyanide (Aldrich, 2.53 mg, 28 mmol) in DMF (10 ml) at reflux for 4 hours Heated. After cooling to ambient temperature, ethyl acetate (500 ml) and 10% aqueous NaCN (500 ml) were added. After separation of the aqueous and organic phases, the aqueous phase was extracted with ethyl acetate (300 ml). The combined organic phases were concentrated to an oil, which was further purified by flash column chromatography (ethyl acetate) to give the title compound (3.03 g, 87%). TLC: Rf 0.30 (1: 1 hexane / ethyl acetate); 1 H NMR (CDCl 3 ): Δ 2.55 (m, 3H), 6.76 (d, 1H, J = 3.6 Hz), 7.42 (d, 1H, J = 3.6 Hz).
[0356]
(Example 78)
(Preparation of 2-cyano-5- (bromomethyl) thiophene (3-3))
[0357]
Embedded image
CCl 4 (Aldrich, 60 ml), 2-cyano-5-methylthiophene (compound 3-2, 3.0 mg, 24 mmol), N-bromosuccinimide (Aldrich, 4.8 g, 27 mmol) and 2,2′-azobisiso A solution of butyronitrile (Aldrich, 0.4 g, 2.4 mmol) was heated at reflux for 5 hours. After cooling to ambient temperature, the solvent was removed under vacuum to give a yellow oil. The oil was purified by flash column chromatography (1: 1 hexane / ethyl acetate) to give the title compound (4.5 g, 91%). TLC: Rf 0.91 (1: 1 hexane / ethyl acetate); 1 H NMR (CDCl 3 ): Δ 4.66 (s, 2H), 7.10 (d, 1H, J = 3.8 Hz), 7.48 (d, 1H, J = 3.8 Hz).
[0358]
(Example 79)
(Preparation of 2-cyano-5- (azidomethyl) thiophene (3-4))
[0359]
Embedded image
A solution of 2-cyano-5- (bromomethyl) thiophene (compound 3-3, 3.5 g, 17.3 mmol) and sodium azide (Aldrich, 1.7 g, 26 mmol) in DMF (Aldrich, 60 ml) was added to the surroundings. Stirred at temperature for 10 hours. Flash column chromatography (20% ethyl acetate in hexane) provided the title compound (2.35 g, 83%). TLC: Rf 0.48 (20% ethyl acetate in hexane); 1 H NMR (CDCl 3 ): Δ 4.56 (s, 2H), 7.01 (d, 1H, J = 3.7 Hz), 7.55 (d, 1H, J = 3.7 Hz).
[0360]
(Example 80)
(Preparation of 2-cyano-5- (aminomethyl) thiophene (3-5))
[0361]
Embedded image
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 aqueous solution was warmed to room temperature and stirred at ambient temperature for 10 hours. RP-HPLC purification provided the title compound (2.3 g, 94%). MS (Electrospray): 139 (M + 1); 1 H NMR (CDCl 3 ): Δ 4.01 (s, 2H), 4.75 (br s, 2H, NH 2 6.82 (d, 1H, J = 3.5 Hz); 7.08 (d, 1H, J = 3.5 Hz).
[0362]
(Example 81)
(Preparation of 2-cyano-5- (t-butoxycarbylaminomethyl) thiophene (3-6))
[0363]
Embedded image
Potassium carbonate (Aldrich, 2 g) was added to 2-cyano-5- (aminomethyl) thiophene (3-5, 0.6 g, 4 mmol) in water (4 ml) and 1,4-dioxane (Aldrich), Boc 2 O (Fluka, 0.95 g, 4 mmol) was added to the solution. The resulting mixture was stirred at ambient temperature for 12 hours. Flash column chromatography (1: 1 hexane / ethyl acetate) provided the title compound (0.58 g, 56%). MS (Electrospray): 239 (M + 1); 1 H NMR (CDCl 3 ): Δ 1.44 (s, 9H), 4.55 (s, 2H), 4.90 (br s, 1H, NH); 6.88 (d, 1H, J = 3.6 Hz); 7.07 (D, 1H, J = 3.6 Hz).
[0364]
(Example 82)
(Preparation of 2- (N-hydroxyamidinyl) -5- (t-butoxycarbonylaminomethyl) thiophene (3-7))
[0365]
Embedded image
2-cyano-5- (t-butoxycarbylaminomethyl) thiophene (compounds 3-6, 560 mg, 2.44 mmol), hydroxylamine hydrochloride (Aldrich, 330 mg, 4.8 mmol) and 4 in methanol (5 ml) A solution of -methylmorpholine (Aldrich, 1 ml, 9.1 mmol) was stirred at ambient temperature for 12 hours. The resulting mixture was stirred at ambient temperature for 12 hours. Flash column chromatography (5: 95: 1 isopropyl alcohol / methylene chloride / triethylamine) provided the title compound (550 mg, 86%). MS (electrospray): 272 (M + 1).
[0366]
(Example 83)
(Preparation of 2-amidinyl-5- (aminomethyl) thiophene (3-8))
[0367]
Embedded image
10% of Pd-on-C (Aldrich, 100 mg) was added to 2- (N-hydroxyamidinyl) -5- (t-butoxycarbonylaminomethyl)) thiophene (Compound 3-7, 900 mg, 3.3 mmol). The resulting mixture was hydrogenated in a Parr apparatus at room temperature for 10 hours (H 2 45 psi). The catalyst was removed by filtration and the solvent was evaporated under vacuum to give the Boc-protected intermediate [900 mg, 94%, MS (electrospray) 256 (M + 1)], which was converted to 1,4-dioxane ( Treatment with 4M HCl in Aldrich (5 ml) at ambient temperature for 3 hours afforded the title compound (460 mg, 84%). MS (electrospray): 56 (M + 1); 1 H NMR (CD 3 OD): δ 4.43 (s, 2H), 7.42 (d, 1H, J = 3.5 Hz) 7.78 (d, 1H, J = 3.5 Hz).
[0368]
(Example 84)
(Purification of 2- [N- (propoxy) amidinyl] -5- (aminomethyl) thiophene (3-9))
[0369]
Embedded image
CsCO 3 (Aldrich, 0.5 g) was treated with 2- (N-hydroxyamidinyl) -5- (t-butoxycarbonylaminomethyl) thiophene (compound 3-7, 271 mg, 1.0 ml) and iodopropane (DMF) in DMF. Aldrich (200 mg, 1.2 mmol). The reaction mixture was stirred at ambient temperature for 10 hours. Flash column chromatography (5: 95: 1 isopropyl alcohol / methylene chloride / triethylamine) gave the Boc protected intermediate [MS (Electrospray) 314 (M + 1)], which was combined with 1,4-dioxane Treatment with 4M HCl in (Aldrich) at ambient temperature for 3 hours afforded the title compound (201 mg, 81%). The title compound (550 mg, 86%) was obtained. MS (electrospray): 214 (M + 1); 1 H NMR (CDCl 3 ): Δ 0.95 (t, 3H, J = 7.5 Hz), 1.55 (brs, 2H, NH 2 ), 1.70 (m, 2H), 4.00 (t, 2H, J = 7.5 Hz), 4.01 (d, 2H, J = 7.5 Hz), 4.70 (brs, 2H, NH 2 ), 6.80 (d, 1H, J = 3.6 Hz), 7.08 (d, 1H, J = 3.6 Hz).
[0370]
(Example 85)
(Preparation of α-azido-4-cyanotoluene (4-2))
[0371]
Embedded image
Sodium azide (Aldrich, 3.5 g, 54 mmol) is added to a solution of p-cyanobenzyl bromide (Aldrich, 10 g, 51 mmol) in DMF (100 ml) and the resulting mixture is stirred at ambient temperature for 5 hours did. The reaction mixture was then diluted with water (350ml) and extracted with ether (2x100ml). The combined organic phases were washed with brine and dried (MgSO 4 ). Removal of the solvent provided the title compound (8 g, 96%). 1 H NMR (CDCl 3 ): Δ 4.42 (s, 2H), 7.41 (d, 2H, J = 8.1 Hz), 7.65 (d, 2H, J = 8.1 Hz).
[0372]
(Example 86)
(Preparation of p-cyanobenzylamine (4-3))
[0373]
Embedded image
10% Pd-on-c (Aldrich, 800 mg) catalyst was added to a solution of α-azido-4-cyanotoluene (Compound 4-2, 8 g, 51 mmol) in EtOAc (150 ml). The reaction mixture was hydrogenated on a Parr apparatus for 11 hours (H 2 , 45 psi). The catalyst was removed by filtration and the solvent was removed under vacuum to give the title compound (6.3 g, 93%). 1 H NMR (CDCl 3 ): Δ 3.85 (s, 2H), 7.45 (d, 2H, J = 8.1), 7.60 (d, 2H, J = 8.1 Hz), 7.78 (s, 2H, NH) 2 ).
[0374]
(Example 87)
(Preparation of 4- (aminomethyl) phenyl-N-hydroxyamidine (4-4))
[0375]
Embedded image
Hydroxyamine hydrochloride (7 g) was added to a solution of compound 4-3 (7 g) and NMM (4 ml) in methanol (100 ml). The mixture was stirred at ambient temperature for 3 days. This compound was purified by RP HPLC to give the title compound (7 g, 89%). MS (Electrospray): 166 (M + 1).
[0376]
(Example 88)
(Preparation of 4- (aminomethyl) phenylamidine (4-5))
[0377]
Embedded image
10% Pd-on-c (Aldrich, 800 mg) was added to a solution of 4- (aminomethyl) phenyl-N-hydroxyamidine (Compound 4-4, 7 g) in methanol (150 ml). The reaction mixture was hydrogenated in a Parr apparatus for 48 hours (H 2 , 45 psi). The catalyst was removed by filtration and the solvent was removed under vacuum to give the title compound (6.3 g, 99%). MS (electrospray): 150 (M + 1).
[0378]
(Example 89)
(Preparation of 2-fluoro-4-cyanotoluene (5-2))
[0379]
Embedded image
Copper (I) cyanide (Aldrich, 3.6 g, 40 mmol) was added to a solution of 4-bromo-2-fluorotoluene (Aldrich, 5 g, 27 mmol) in DMF (60 ml). The reaction mixture was heated at 150 ° C. for 11 hours. After cooling to room temperature, the mixture was partitioned between water and EtOAc (500 ml each). Dry the organic phase (MgSO 4 4 ) And the solvent was removed under vacuum to give the title compound (2.08, 58%). . 1 H NMR (CDCl 3 ): Δ 2.36 (s, 3H), 7.30 (m, 3H), 7.35 (d, 1H, J = 8.1 Hz).
[0380]
(Example 90)
(Preparation of 3-fluoro-4- (bromomethyl) benzonitrile (5-3))
[0381]
Embedded image
NBS (Aldrich, 3.02 g, 17 mmol) and benzoyl peroxide (Aldrich, 0.37 g, 1.5 mmol) were added to CCl 4 Was added to a solution of 2-fluoro-4-cyanotoluene (compound 5-2, 2.08 g, 15 mmol) therein. The reaction mixture was heated at 80 C for 14 hours. After cooling to ambient temperature, the mixture was diluted with ether (100 ml) and aqueous Na 2 S 3 O 3 And dried (MgSO 4) 4 )did. Removal of the solvent under vacuum led to a yellow oil, which was purified by flash chromatography. The title compound 5-3 (1.4 g, 42%) was obtained with a by-product (3-fluoro-4- (bromomethyl) benzonitrile (5-4) (1.0 g, 30%)). For the title compound (5-3): 1 1 H NMR (CDC1 3 ): Δ 4.46 (s, 2H), 7.35 (d, 1H, J = 8.0 Hz), 7.42 (d, 1H, J = 8.0 Hz), 7.52 (t, 1H, J = 8.0 Hz). By-product (5-4): 1 1 H NMR (CDC1 3 ): Δ 6.90 (s, 1H), 7.35 (d, IH, J = 8.0 Hz), 7.55 (d, 1H, J = 8.0 Hz), 7.96 (t, 1H, J) = 8.0 Hz).
[0382]
(Example 91)
(Preparation of 3-fluoro-4- (azidomethyl) benzonitrile (5-5))
[0383]
Embedded image
Sodium azide (Aldrich, 0.63 g, 9.8 mmol) was added to a solution of 3-fluoro-4- (bromomethyl) benzonitrile (Compound 5-3, 1.4 g, 6.5 mmol) in DMF (15 ml). Was added. After stirring at ambient temperature for 20 hours, the reaction mixture was partitioned between EtOAc and water (100 mL each). The organic layer is then dried (MgSO 4 4 ) And the solvent was removed in vacuo to give the title compound (0.995 g, 86%). 1 1 H NMR (CDC1 3 ): Δ 4.50 (S, 2H), 7.38 (d, 2H, J = 8.1 Hz), 7.52 (m, 2H).
[0384]
(Example 92)
(Preparation of 3-fluoro-4- (azidomethyl) phenyl-N-hydroxyamidine (5-6))
[0385]
Embedded image
Hydroxylamine hydrochloride (Aldrich, 800 mg, 11.6 mmol) was treated with 3-fluoro-4- (azidomethyl) benzonitrile (compound 5-5, 1.2 g, 6.8 mmol) and NMM (2 ml) in methanol (25 ml). ). After stirring at room temperature for 3 days, the reaction mixture was diluted with EtOAc and washed with brine. The solvent was removed under vacuum to yield the title compound (1.38, 82%). 1 H NMR (CD 3 OD): [delta] 4.41 (s, 2H), 7.45 (m, 3H).
[0386]
(Example 93)
(Preparation of 3-fluoro-4- (azidomethyl) phenyl (-N-propyloxy) amidine (5-7))
[0387]
Embedded image
Cesium carbonate (Aldrich, 3.2 g, 9.9 mmol) was added to iodopropane (1 ml, 10 mmol) and 3-fluoro-4- (azidomethyl) phenyl-N-hydroxyamidine (compound 5- 6, 1.38 g, 6.6 mmol). The reaction mixture was heated at 50 C for 20 hours. After cooling to ambient temperature, water was added and the resulting mixture was extracted with ether. The organic layer is washed with brine and dried (MgSO 4 )did. Flash chromatography provided the title compound (1.03 g, 62%). 1 1 H NMR (CDC1 3 ): Δ 0.99 (t, 3H, J = 7.5 Hz), 1.75 (m, 2H), 4.08 (t, 2H, J = 7.5 Hz), 4.40 (s, 2H), 4.78 (br s, 2H), 7.40 (m, 3H).
[0388]
(Example 94)
(Preparation of 3-fluoro-4- (aminomethyl) phenyl (-N-propyloxy) amidine (5-8))
[0389]
Embedded image
Triphenylphosphine (Aldrich, 1.6 g, 6.2 mmol) was added to 3-fluoro-4- (azidomethyl) phenyl (-N-propyloxy) amidine (1.03 g of compound 5-7, THF, 15 ml). 4.1 mmol). The reaction mixture was stirred at ambient temperature for 20 hours. NaOH (3M) was added to the reaction mixture until pH = 14. The resulting solution was extracted with EtOAc (2 × 100 ml). The combined organic layers are washed with brine and dried (MgSO 4 )did. The solvent was removed under vacuum to give the title compound (825mg, 77%). 1 H NMR (CD 3 OD): δ 0.98 (t, 3H, J = 7.5 Hz), 1.72 (m, 2H), 3.82 (s, 2H), 3.95 (t, 2H, J = 7.5 Hz) , 7.35 (d, 1H, J = 8.0 Hz), 7.40 (m, 2H)
(Example 95)
(Preparation of N-α-benzyloxycarbonyl-D-Ser (Ot-butyl) -L-Alat-butyl ester)
[0390]
Embedded image
To a solution of N-α-Cbz-D-serine t-butyl ether (5.02 g, 17 mmol) in acetonitrile (100 ml) was added EDC (4.90 g, 25.5 mmol, 1.5 equivalents) and 1-hydroxybenzo. Triazole (2.60 g, 17 mmol) was added. After stirring for 45 minutes, alanine t-butyl ester, HCl salt (3.55 g, 19.6 mmol, 1.15 eq) and 4-methylmorpholine (7.5 ml, 68 mmol, 4 eq) were added. The reaction mixture was stirred for 1.5 hours. TLC showed the reaction was complete. The reaction mixture was concentrated under reduced pressure. This residue was dissolved in ethyl acetate (100 ml) and then washed successively with 1 N HCl, saturated sodium bicarbonate, water and brine (25 ml each). The solvent was removed in vacuo to give an oil which crystallized on standing. The title compound (6.95 g) was obtained in 97% yield as a pale yellow solid. R f = 0.63 (5% isopropanol in dichloromethane).
[0391]
(Example 96)
(Preparation of D-Ser (Ot-butyl) -L-Alat-butyl ester)
[0392]
Embedded image
A solution of the compound of Example 95 (1.14 g, 2.69 mmol) in methanol was cured over palladium hydroxide (110 mg) at balloon pressure for 1.5 hours. The reaction mixture was filtered through celite and the solvent was removed in vacuo to yield the title compound as a yellow oil in quantitative yield. R f = 0.44 (5% methanol in dichloromethane).
[0393]
(Example 97)
(Preparation of phenethylsulfonyl-D-Ser (Ot-butyl) -L-Alat-butyl ester)
[0394]
Embedded image
To a stirred solution of the compound of Example 96 (4.2 g, 14.6 mmol) and phenethylsulfonyl chloride (3.58 g, 17.5 mmol, 1.2 equiv) in acetonitrile (100 ml) cooled in an ice bath was added , 4,6-collidine (4.8 ml, 36.5 mmol, 2.5 equiv) was added. The reaction mixture was allowed to warm to room temperature and then stirred overnight. The reaction mixture was concentrated under reduced pressure. The residue was dissolved in ethyl acetate (80 ml); the resulting solution was washed successively with 1.0 N HCl, saturated sodium bicarbonate, water and brine, then dried over sodium sulfate. The solvent was removed with Invacuo. The residue was chromatographed over silica gel, eluting with 0-60% ethyl acetate / hexane. The title compound was isolated as a yellow oil in 89% yield. Rf = 0.84 (5% methanol in dichloromethane).
[0395]
(C. Synthesis route of compound having acyl ester or carbonate at P3)
(Example 98)
(Preparation of n-butylsulfonyl-D- (isopropyloxycarbonyl) serine-alanine-4-amidinobenzylamide)
[0396]
Embedded image
To a solution of the compound of Example 9 (2.32 g, 4.3 mmol) in pyridine (100 ml) cooled in an ice bath was added isopropyl chloroformate (21 ml of a 1 M solution in toluene, 21 mml, 5 eq). The reaction was monitored by analytical HPLC until determined to be complete. The reaction mixture was diluted with toluene (300 ml); the solvent was removed under reduced pressure. The residue was dissolved in acetonitrile (400ml) and the solvent was removed under reduced pressure. The residue was dissolved in acetonitrile (30ml) and ethyl acetate (30ml). Ether was added and the precipitate was isolated. The solid was washed with ether and then dried in Invacuo. The solid was washed with ether and ethyl acetate (2 × 200 ml of a 1: 1 mixture) and then dried in vacuo to give the title compound.
[0397]
(Example 99)
Following the protocol of Example 98, using the famous substitution of isopropyl chloroformate, the following compounds were prepared:
[0398]
[Table 1]
(Example 100)
(Preparation of (4-amino-3-nitro-benzyl) -carbamic acid tert-butyl ester)
[0399]
Embedded image
BH in THF (1 M, 40 mL) 3 Was added dropwise over 10 minutes to a stirred solution of 4-amino-3-nitro-5-benzonitrile (1.5 g, 9.2 mmol) in THF (20 mL) while cooling in an ice bath. . The yellow mixture obtained upon warming to room temperature was stirred for 2 hours and quenched at 0 ° C. with HCl (6M, 20 mL). The solvent is removed under vacuum; 2 Dilute with O and adjust the pH to about 10 using NaOH (1M). CH layer 2 Cl 2 (3 × 50 mL). The combined organic layers are 2 SO 4 And dried. The solvent was removed, yielding 1.2 g (78%) of a yellow solid, which was used for the next synthetic step without further purification.
[0400]
The solid (500 mg, 3.0 mmol) was added to a homogeneous mixture of BOC-ON (754 mg, 3.0 mmol) in THF (10 mL). The resulting mixture was stirred overnight at room temperature. The mixture is treated with NaHCO 3 Diluted with aqueous solution, CH 2 Cl 2 (3 × 50 mL). The combined organic layers were washed with NaSO 4 And dried. The solvent was removed under vacuum to give an oil. Column chromatography (CH 2 Cl 2 / MeOH, 99: 1) to give the title compound as a yellow solid which solidified on standing (760 mg, 95%), MS (electrospray) 268 (M + 1).
[0401]
(Example 101)
(Preparation of (3,4-diamino-benzyl) -carbamic acid tert-butyl ester)
[0402]
Embedded image
(4-Amino-3-nitro-benzyl) -carbamic acid tert-butyl ester (500 mg, 1.87 mmol) and Pd (10% on carbon, 100 mg) in MeOH (10 mL) were treated with H 2 (40 psi) for 3 hours. The catalyst was removed by filtration and the solvent was removed under vacuum to give an oil which was used immediately in the next step (Example 102).
[0403]
(Example 102)
(Preparation of (2-amino-1H-benzimidazol-5-ylmethyl) -carbamic acid tert-butyl ester)
[0404]
Embedded image
(3,4-Diamino-benzyl) -carbamic acid tert-butyl ester (500 mg, 2.10 mmol) was added to CH 3 CN / H 2 CNBr (267 mg, 2.52 mmol) and NaHCO 3 in O (1: 1, 10 mL) 3 (354 mg, 4.2 mmol) was added and the mixture was stirred for 48 hours. The solvent was removed under vacuum to give a white solid, which was used for the next synthetic step without further isolation. MS (Electrospray) 263 (M + l).
[0405]
(Example 103)
(Preparation of benzylsulfonyl-D-serine-L-alanine-5- (2-aminobenzimidazolyl) methylamide)
[0406]
Embedded image
(2-Amino-1H-benzimidazol-5-ylmethyl) -carbamic acid tert-butyl ester (50 mg, 0.3703 mmol) was treated with benzylsulfonyl-D-serine-L-alanine carboxylate in acetonitrile (1.0 mL). (111 mg, 0.337 mmol), HATU (192 mg, 0.506 mmol), HOAT (69 mg, 0.506 mmol), and DIEA (174 μL, 1.01 mmol) were added to a stirred solution. The mixture was stirred overnight at room temperature. HPLC purification (CH 3 CN, H 2 O, 0.1% TFA) gave a flocculent white solid as the title compound (25 mg, 24%), MS (electrospray) 475 (M + 1).
[0407]
(Example 104)
(Preparation of benzylsulfonyl-D-serine-L-alanine-4- (2-aminoimidazolyl) -propylamide)
[0408]
Embedded image
4- (2-Aminoimidazolyl) -propylamine HCl (50 mg, 0.236 mmol) (prepared according to the procedure of Olofson et al., J. Org. Chem. 1998, 63, 1248) was prepared in acetonitrile (1.0 mL). A stirred solution of benzylsulfonyl-D-serine-L-alanine carboxylate (78 mg, 0.236 mmol), HATU (179 mg, 0.472 mmol), HOAT (64 mg, 0.472 mmol), and DIEA (164 μL, 0.943 mmol) Added. The mixture was stirred overnight at room temperature. HPLC purification (CH 3 CN, H 2 O, 0.1% TFA) to give a flocculent white solid as the title compound. MS (electrospray) 453 (M + l).
[0409]
(Example 105)
(Preparation of (4-amino-2-chloro-benzyl) -carbamic acid tert-butyl ester)
[0410]
Embedded image
BH in THF (1 M, 36 mL) 3 Was added dropwise over 10 minutes to a stirred solution of 4-amino-2-chloro-5-benzonitrile (2.0 g, 13 mmol) in THF (10 mL) while cooling in an ice bath. The yellow mixture obtained upon warming to room temperature was stirred for 2 hours and quenched at 0 ° C. with HCl (6M, 20 mL). The solvent was removed under vacuum. Residue is H 2 Dilute with O and adjust the pH to about 10 using NaOH (1M). CH layer 2 Cl 2 (3 × 50 mL). The combined organic layers are 2 SO 4 And dried. The solvent was removed to give a yellow oil (2.5 g, 100%) which was used without further purification in the next synthetic step.
[0411]
The oil (500 mg, 2.6 mmol) (from the previous step) was added to a homogeneous mixture of BOC-ON (664 mg, 2.7 mmol) in THF (10 mL) and the resulting mixture was stirred at room temperature overnight. . The mixture is treated with NaHCO 3 Diluted with aqueous solution, CH 2 Cl 2 (3 × 50 mL). The combined organic layers were washed with NaSO 4 And dried. The solvent was removed under vacuum to give a colorless oil. Column chromatography (CH 2 Cl 2 / MeOH, 99: 1) gave the title compound as a yellow solid which solidified on standing. MS (electrospray) 257 (M + l).
[0412]
(Example 106)
(Preparation of [(4-BOC-aminomateomethyl) -3-chlorophenyl] -N′-N ″ -bis (tert-butoxycarbonyl) guanidine)
[0413]
Embedded image
(4-Amino-2-chloro-benzyl) -carbamic acid tert-butyl ester (250 mg, 0.978 mmol) was added to CH 2 Cl 2 (5 mL) was added to a stirred mixture of NN′-di-Boc-N ″ -trifluoromethanesulfonyl-guanidine (343 mg, 0.879 mmol), TFA (108 μL, 1.28 mmol). The mixture was stirred for 24 hours. This mixture is taken to CH 2 Cl 2 (20 mL) and washed with HCl (1 M, 20 mL), brine (10 mL). Organic layer 2 SO 4 And dried. The solvent was removed under vacuum to give a solid. Column chromatography (CH 2 Cl 2 / MeOH, 99: 1) to give an oil (150 mg, 34%) corresponding to the title compound.
[0414]
(Example 107)
(Preparation of benzylsulfonyl-D-serine-L-alanine-4-guanidino-2-chlorobenzylamide)
[0415]
Embedded image
CH 2 Cl 2 [(4-BOC-aminomateomethyl) -3-chlorophenyl] -N′-N ″ -bis (tert-butoxycarbonyl) guanidine (100 mg, 0.201 mmol) in a mixture of / TFA (1: 1, 2 mL) ) Was stirred at room temperature for 90 minutes. The solvent was removed under vacuum to give a clear oil, which was used for the next synthetic step without further purification.
[0416]
The oil (from the previous step) was combined with benzylsulfonyl-D-serine-L-alanine carboxylate (69 mg, 0.209 mmol), HATU (157 mg, 0.418 mmol), HOAT (57 mg) in acetonitrile (5.0 mL). , 0.418 mmol) and DIEA (145 μL, 0.836 mmol). The mixture was stirred overnight at room temperature. HPLC purification (CH 3 CN, H 2 O, 0.1% TFA) gave the title compound as a flocculent white solid (25 mg, 24%), MS (electrospray) 511 (M + 1).
[0417]
(Example 108)
(Preparation of (6-amino-pyridin-3-ylmethyl) -carbamic acid tert-butyl ester)
[0418]
Embedded image
BH in THF (1 M, 40 mL) 3 Was added dropwise over 10 minutes to a stirred solution of 4-amino-2-chloro-5-benzonitrile (2.0 g, 14 mmol) in THF (5 mL) while cooling in an ice bath. The yellow mixture obtained upon warming to room temperature was stirred for 2 hours and quenched at 0 ° C. with HCl (6M, 20 mL). The solvent was removed under vacuum. Residue is H 2 Dilute with O and adjust the pH to about 10 using NaOH (1M). CH layer 2 Cl 2 (3 × 50 mL). The combined organic layers are 2 SO 4 The solvent was removed to give a solid (2.8 g, 100%).
[0419]
This solid (2.0 g, 13 mmol) was added to CH 2 3 To a homogeneous mixture of BOC-ON (3.4 mg, 14 mmol) in CN (10 mL) was added and the mixture was stirred at room temperature overnight. The mixture is treated with NaHCO 3 Diluted with aqueous solution, CH 2 Cl 2 (3 × 50 mL). The combined organic layers were washed with NaSO 4 And dried. The solvent was removed under vacuum to give a colorless oil. Column chromatography (CH 2 Cl 2 / MeOH, 99: 1) gave the title compound as an oil which solidified on standing. MS (Electrospray) 226 (M + l).
[0420]
(Example 109)
(Preparation of [(3-BOC-aminomateomethyl) -pyridinyl] -N′-N ″ -bis (tert-butoxycarbonyl) guanidine)
[0421]
Embedded image
A portion of (6-amino-pyridin-3-ylmethyl) -carbamic acid tert-butyl ester (400 mg, 1.78 mmol) was converted to CH 2 Cl 2 (5 mL) was added to a stirred mixture of NN′-di-Boc-N ″ -trifluoromethanesulfonyl-guanidine (765 mg, 1.96 mmol) and TFA (385 mL, 2.67 mmol). The resulting mixture was stirred for 24 hours. This mixture is taken to CH 2 Cl 2 (20 mL) and washed with HCl (1 M, 20 mL), brine (10 mL). Organic layer 2 SO 4 And dried. The solvent was removed under vacuum to give a solid. Column chromatography (CH 2 Cl 2 / MeOH, 99: 1) to give an oil corresponding to the title compound. MS (electrospray) 466 (M + l).
[0422]
(Example 110)
(Benzylsulfonyl-D-serine-L-alanine-4-guanidino- (2-pyridinyl) methylamide)
[0423]
Embedded image
CH 2 Cl 2 Of [(3-BOC-aminomateomethyl) -pyridinyl] -N'-N "-bis (tert-butoxycarbonyl) guanidine (100 mg, 0.215 mmol) in a mixture of / TFA (1: 1, 2 mL). The solution was stirred at room temperature for 9 minutes. The solvent was removed under vacuum to give a clear oil. This oil was treated with benzylsulfonyl-D-serine-L-alanine carboxylate (63 mg, 0.194 mmol), HATU (147 mg, 0.387 mmol), HOAT (52 mg, 0.387 mmol) in acetonitrile (1.0 mL). , And DIEA (135 μL, 0.774 mmol). The mixture was stirred overnight at room temperature. HPLC purification (CH 3 CN, H 2 O, 0.1% TFA) gave a white foamy solid (15 mg, 16%) as the title compound. MS (electrospray) 478 (M + l).
[0424]
(Example A)
(In vitro enzyme assay for specificity determination)
The ability of the compounds of the present invention to act as selective inhibitors of urokinase catalytic activity was determined by the concentration of test compound that inhibited the activity of this enzyme by 50% (IC 50 ) And by comparing this value to the values determined for all or some of the following related serine proteases: recombinant tissue plasminogen activator (rt-PA), plasmin Activated protein C, chymotrypsin, factor Xa, thrombin and trypsin.
[0425]
The buffer used for all assays was HBSA (10 mM HEPES, pH 7.5, 150 mM sodium chloride, 0.1% bovine serum albumin).
[0426]
IC 50 The assay for the determination was performed by combining the following in appropriate wells of a Corning microtiter plate: 50 μl of HBSA, 50 μl of a specific concentration (covering a wide concentration range) diluted in HBSA Test compound (or V 0 (Non-inhibition rate) HBSA alone for measurement), and 50 μl of enzyme diluted in HBSA. After a 30 minute incubation at ambient temperature, 50 μl of substrate at the concentration specified below was added to the wells to give a final volume of 200 μl (約 4 Km). The initial rate of chromogenic substrate hydrolysis was measured over a 5-minute interval by the change in absorbance at 405 nm using a Terumo Max® kinetic microplate reader, where less than 5% of the added substrate was used. The concentration of added inhibitor that caused a 50% reduction in the initial rate of hydrolysis was determined by the IC 50 Value. Ki is IC 50 It can be calculated from the value.
[0427]
(Urokinase assay)
DiaPharma Group Inc. Urokinase catalytic activity was determined using the chromogenic substrate 150 mM S-2444 (L-pyroglutamyl-glycyl-L-arginine-p-nitroaniline hydrochloride) obtained from Co., Ltd. Urokinase (Abokinae), manufactured by Abbott Laboratories, was obtained from Priority Pharmaceuticals and diluted to 750 pM in HBSA assay buffer before use. The assay buffer was HBS containing 0.1% BSA (10 mM HEPES, 150 mM sodium chloride, pH 7.4). IC 50 Ki was calculated using the values.
[0428]
(Thrombin (fIIa) assay)
The chromogenic substrate Pefachrome t-PA (CHP obtained from Pentapharm Ltd. 3 SO 2 -D-hexahydrotyrosine-glycyl-L-arginine-p-nitroaniline) was used to determine the enzyme activity. This substrate was reconstituted in deionized water before use. Purified human α-thrombin is available from Enzyme Research Laboratories, Inc. Obtained from. The buffer used for all assays was HBSA (10 mM HEPES, pH 7.5, 150 mM sodium chloride, 0.1% bovine serum albumin).
[0429]
IC 50 The determination combines HBSA (50 μl), α-thrombin (50 μl) (final enzyme concentration is 0.5 nM) and inhibitor (50 μl) (covers a wide concentration range) in the appropriate wells and is the substrate. This was performed by incubating at room temperature for 30 minutes before the addition of Pefachrome-t-PA (50 μl) (final substrate concentration is 250 μM, about 5 × Km). The initial rate of Pefachrome t-PA hydrolysis was measured over a 5-minute interval by the change in absorbance at 405 nm using a Terumo Max® kinetic microplate reader, where less than 5% of the substrate added Was used. The concentration of added inhibitor that caused a 50% reduction in the initial rate of hydrolysis was determined by the IC 50 Value.
[0430]
(Factor Xa)
The catalytic activity of Factor Xa was determined by combining the chromogenic substrate S-2765 (N-benzyloxycarbonyl-D-arginine-L-glycine-L-arginine-p-nitroaniline) obtained from DiaPharma Group (Franklin, OH). Used and determined. All substrates were reconstituted with deionized water before use. The final concentration of S-2765 was 250 μM (approximately 5-fold Km). Purified human factor X was obtained from Enzyme Research Laboratories, Inc (South Bend, IN) and factor Xa (FXa) was described (Bock, PE, Craig, PA, Olson, PA). It was prepared by activating factor X as described in ST and Singh, P. Arch.Biochem.Biophys.273: 375-388 (1989)). The enzyme was diluted in HBSA prior to the assay to a final concentration of 0.25 nM.
[0431]
(Recombinant tissue plasminogen activator (rt-PA) assay)
The rt-PA catalytic activity was measured using Pefachrome t-PA (Pentapharma Ltd., CH 3 SO 2 -D-hexahydrotyrosine-glycyl-L-arginine-p-nitroaniline). This substrate was made in deionized water and subsequently diluted in HBSA prior to the assay to a final concentration of 500 μM (approximately 3-fold Km). Human rt-PA (Activase®) was purchased from Genentech Inc. Obtained from. The enzyme was reconstituted in deionized water and diluted in HBSA prior to the assay to a final concentration of 1.0 nM.
[0432]
(Plasmin assay)
Plasmin catalytic activity was determined using the chromogenic substrate S-2366 [L-pyroglutamyl-L-prolyl-L-arginine-p-nitroaniline hydrochloride] obtained from DiaPharma Group. This substrate was made in deionized water and subsequently diluted in HBSA prior to the assay to a final concentration of 300 μM (約 2.5-fold Km). Purified human plasmin was obtained from Enzyme Research Laboratories, Inc. The enzyme was diluted in HBSA prior to assay to a final concentration of 1.0 nM.
[0433]
(Activated protein C (aPC) assay)
The aPC catalytic activity was determined using a chromogenic substrate, Pefachrome PC (δ-carbobenzyloxy D-lysine-L prolyl-L-arginine-p-nitroaniline dihydrochloride, obtained from Pentapharma Ltd.). This substrate was made in deionized water and subsequently diluted in HBSA prior to the assay to a final concentration of 400 μM (approximately 3-fold Km). Purified human aPC was obtained from Hematological Technologies, Inc. This enzyme was diluted in HBSA to a final concentration of 1.0 nM prior to the assay.
[0434]
(Chymotrypsin assay)
The chromogenic substrate, S-2586 (methoxy-succinyl-L-arginine-L-prolyl-L-tyrosyl-p-nitroanilide), obtained from DiaPharma Group, was used to determine chymotrypsin catalytic activity. This substrate was made in deionized water and subsequently diluted in HBSA prior to the assay to a final concentration of 100 μM ((9-fold Km). Purified (3 × -crystallization; CDI) bovine pancreatic α-chymotrypsin was obtained from Worthington Biochemical Corp. The enzyme was reconstituted in deionized water and diluted in HBSA to a final concentration of 0.5 nM prior to the assay.
[0435]
(Trypsin assay)
Trypsin catalytic activity using S-2222 (benzoyl-L-isoleucine-L-glutamic acid- [γ-methyl ester] -L-arginine-p-nitroanilide), a chromogenic substrate, obtained from DiaPharma Group. It was determined. This substrate was made in deionized water and subsequently diluted in HBSA prior to the assay to a final concentration of 250 μM (approximately 4-fold Km). Purified (3x-crystallized; TRL3) bovine pancreatic alpha trypsin was obtained from Worthington Biochemical Corp. The enzyme was reconstituted in deionized water and diluted in HBSA to a final concentration of 0.5 nM prior to the assay.
[0436]
(Tables IA and IB)
Tables IA and IB show that for the compounds of the present invention, the K for specific enzymes listed above for the compounds of the invention shows a high degree of specificity for the inhibition of urokinase compared to other serine proteases. i Value or IC 50 Indicates the value.
[0437]
[Table 2]
[0438]
[Table 3]
(Example B)
(Evaluation of test compounds as inhibitors of angiogenesis in vivo)
A standard angiogenesis assay, the chicken CAM (chick embryo chorioallantoic membrane) model, is used to assess the ability of test compounds to inhibit angiogenesis. This model is an established model for assessing the activity of test compounds affecting new blood vessel formation.
[0439]
A filter disk saturated with a 0.5 μg / ml solution of basic fibroblast growth factor (bFGF) is placed on the CAM of a 10-day-old chick embryo to induce angiogenesis. Twenty-four hours later, 0-1 μg of test compound (contained in a total volume of 100 μl of sterile PBS) is injected intravenously into the embryo. After about 48 hours, the embryo is sacrificed and the filter disc and surrounding CAM tissue are excised for analysis. Angiogenesis is quantified by counting the number of vessel branch points in the confined area of the filter [Brooks, P .; C. Et al., Methods in Molecular Biology 120: 257-269 (1999)]. Angiogenic index is defined as the difference in the number of vascular branch points between the experimental group and untreated control embryos. Each experimental group contains 8-10 chicken embryos.
[0440]
(Example C)
(Evaluation of a test compound that inhibits the growth of human tumor cells in a chicken embryo model)
The activity of a test compound that inhibits the growth of human tumor cells in vivo is evaluated using a chicken embryo model. Single cell suspension of human fibrosarcoma cells (HT 1080) (4 × 10 4 in a total volume of 40 μl) 5 Cells (containing the cells) are described in Brooks et al. (“Brooks, PC et al.,“ Integrin α v β 3 Apply to 10-day-old chick embryos as described by Antagonists Promote Tumor Regression by Inducing Apotosis of Angiogenic Blood Vessels (Cell 79: 1157-1164 (1994)). Twenty-four hours later, embryos are injected intravenously with 0-10 μg of test compound. After this single dose of compound, control and treated embryos are incubated for a total of 7 days and then sacrificed. The tumor is excised, the surrounding CAM tissue is removed and weighed. Table 1 shows the wet weight of the tumor cut out in this example. Each experimental group contains 10-12 chicken embryos.
[Brief description of the drawings]
FIG.
FIG. 1 shows a reaction scheme for solution phase synthesis of intermediates useful in synthesizing compounds of the present invention. Compound 1-1 is N-α-Cbz-D-serine (Ot-butyl), and compound 1-2 is alanine methyl ester hydrochloride. In this figure, "i" to "iv" are defined as follows: i) Cbz-D-Ser (Ot-butyl)-by EDC, 1-hydroxybenzotriazole and acetonitrile, diisopropylethylamine. Yields Ala-OMe; (ii) ethanol / acetic acid / water (4: 1: 1), 10% Pd on carbon, 45 psi H 2 95% after workup for 2 hours; iii) acetonitrile, benzenesulfonyl chloride, diisopropylethylamine, 43% workup yield; and iv) methanol, 1.0M lithium hydroxide, on DOWEX ion exchange resin. , Elution with methanol / water, 95% yield after workup. See also Examples 60-62.
FIG. 2
FIG. 2 shows a reaction scheme for a solution phase synthesis route that can be used to prepare intermediates useful for preparing compounds of the present invention. In this figure, "i" to "iii" are defined as follows: i) isobutyl chloroformate, sodium carbonate, water, 99.5% post work-up yield; ii) alanine t-butyl. Hydroxybenzotriazole in ester hydrochloride, EDC, and acetonitrile; diisopropylethylamine, quantitative yield after workup; and iii) TFA, DCM, quantitative yield after workup. See also Examples 74-76.
FIG. 3
FIG. 3 shows a reaction scheme for the synthesis of intermediates that can be used in the preparation of the compounds of the present invention. In this figure, "i" to "xi" are defined as follows: i) CuCN, DMF, reflux (4 hours); ii) EtOAc, 10% aqueous NaCN; iii) N-bromosuccinimide, , 2'-azo-isobutyronitrile, CCl 4 Reflux (5 hours); iv) NaN 3 , DMF; v) triphenylphosphine, THF, water, 0 ° C, stirring (10 hours); vi) K 2 CO 3 , Boc 2 O, water, dioxane; vii) hydroxylamine HCl, NMM, MeOH; viii) 10% Pd / C, MeOH, 45 psi H 2 (10 hours); ix) 4M HCl in dioxane; x) CsCO in DMF 3 And iodopropane; and xi) 4M HCl, dioxane, 3 hours, room temperature.
FIG. 4
FIG. 4 shows a reaction scheme for the synthesis of intermediates that can be used to prepare compounds of the present invention. In this figure, “i” to “iv” are defined as follows: i) NaN 3 , DMF; ii) 10% Pd / C, EtOAc, 45 psi H 2 (11 hours); iii) hydroxylamine HCl, NMM, MeOH; and iv) 10% Pd / C, MeOH, 45 psi H. 2 (48 hours).
FIG. 5
FIG. 5 shows a reaction scheme for the synthesis of intermediates that can be used in the preparation of the compounds of the present invention. In this figure, “i” to “vii” are defined as follows: i) Cu (I) CN, DMF; ii) NBS, benzoyl peroxide, CCl 4 , 80 ° C (14 hours); iii) NaN 3 Iv, DMF, stirring (20 h); iv) hydroxylamine HCl, NMM, MeOH, stirring (3 days); v) CsCO 3 Iodopropane, DMF, 50 ° C. (20 h); vi) triphenylphosphine, THF, stirring (20 h); and vii) 3M NaOH to pH14.
FIG. 6
FIG. 2 Is -CH 2 OA 1 And A 1 Is -C (= O) R 6 Figure 9 shows a reaction scheme for the synthesis of compounds of the present invention where is used as an intermediate (compound of Example 9). In this figure, "i" is: i) pyridine, R 6 COCl is defined.
FIG. 7
FIG. 7 shows a reaction scheme for the synthesis of certain compounds of the invention. In this figure, “i” to “vi” are defined as follows: i) trifluoroacetic anhydride, stirred at 0 ° C. overnight; ice, CH 2 C1 2 , Na 2 SO 4 Ii) Pd / C (10%) in MeOH (overnight); iii) NN'-di-Boc-N "-trifluoromethanesulfonyl-guanidine, TEA, CH 2 Cl 2 , 6 hours; HC1, brine, Na 2 SO 2 Column chromatography (CH 2 Cl 2 / MeOH 99: 1); iv) potassium carbonate, H 2 O / MeOH (2:15), overnight; CH 2 Cl 2 / MeOH (9: 1), Na 2 SO 4 V) benzylsulfonyl-D-serine-L-alanine carboxylate, HATU, HOAT, DIEA, acetonitrile, overnight; EtOAc, HCl, NaHCO 3 , Brine; HPLC (CH 3 CN, H 2 O, 0.1% TFA); vi) CH 2 Cl 2 / TFA (1: 1), 90 min; HPLC (CH 3 CN, H 2 O, 0.1% TFA).
FIG. 8
FIG. 8 shows a reaction scheme for the synthesis of certain compounds of the present invention. In this figure, “i” to “v” are defined as follows: i) trifluoroacetic anhydride, stirred overnight; ice, CH 2 Cl 2 , Na 2 SO 4 Ii) Pd / C (10%) in MeOH (overnight); iii) NN'-Boc-N "-trifluoromethanesulfonyl-guanidine, TEA, CH 2 Cl 2 , 24 hours; HCl, brine, Na 2 SO 4 Column chromatography (CH 2 Cl 2 / MeOH 98: 2); iv) potassium carbonate, H 2 O / MeOH (1: 1), overnight; H 2 O, CH 2 Cl 2 / MeOH (9: 1), Na 2 SO 4 And v) benzylsulfonyl-D-serine-L-alanine carboxylate in HATU, HOAT, DIEA, acetonitrile, room temperature, overnight; EtOAc, HCl, NaHCO 3 , Brine; CH 2 Cl 2 / TFA (1: 1), room temperature, 2 hours, HPLC (CH 3 CN, H 2 O, 0.1% TFA).
FIG. 9
FIG. 9 shows a reaction scheme for the synthesis of the compounds of the present invention. In this figure, “i” to “vi” are defined as follows: i) trifluoroacetic anhydride, 0 ° C., 1 hour; ii) KNO 3 -20 ° C, stirring overnight; CH 2 Cl 2 , Na 2 SO 4 Column chromatography; iii) HCl in MeOH, 0 ° C., SnC1 2 , Stirred for 30 minutes, NaHCO 3 , CH 2 Cl 2 , Na 2 SO 4 Iv) N 1 N'-di-Boc-N "-trifluoromethanesulfonyl-guanidine, TEA, CH 2 Cl 3 , Stirring, 24 hours; HCl (1M), brine, Na 2 SO 4 , Column chromatography; v) K 2 CO 3 , H 2 O / MeOH (1: 1), stirring overnight; H 2 O, CH 2 Cl 2 / MeOH (95: 5), Na 2 SO 4 And vi) benzylsulfonyl-D-serine-L-alanine carboxylate, HATU, HOAT, DIEA, AcN, stirring overnight; EtOAc, HCl (1M), NaHCO; 3 Aqueous solution, brine, Na 2 SO 4 HPLC.
FIG. 10A
FIG. 10A shows certain preferred compounds of the present invention.
FIG. 10B
FIG. 10B shows certain preferred compounds of the present invention.
FIG. 10C
FIG. 10C shows certain preferred compounds of the present invention.
FIG. 10D
FIG. 10D shows certain preferred compounds of the present invention.
FIG. 10E
FIG. 10E illustrates certain preferred compounds of the present invention.
FIG. 10F
FIG. 10F shows certain preferred compounds of the present invention.
FIG. 11
FIG. 11 shows a reaction scheme for the synthesis of the compounds of the present invention. In this figure, “i” to “xii” are defined as follows: i) CCl 4 , N-bromosuccinimide; N 2 , AIBN, stirring, flash chromatography; ii) DMF, NaN 3 , Stirred overnight, diethyl ether, water, brine, MgSO 4 (Iii) MeOH, TEA, 65 ° C., 4 hours; EtOAc, H 2 O, brine, MgSO 4 Iv) THF / water, Ph 3 P, stirring overnight; 1N HCl, H 2 O, DCM, pH about 9; v) dioxane / water, K 2 CO 3 , Boc 2 O, stir overnight; EtOAc, NaHCO 3 Aqueous solution, brine, Na 2 SO 4 , Flash column chromatography; vi) DMF, 2-iodopropane, CsCO 3 ; EtOAc, NaHCO 3 Aqueous solution, Na 2 SO 4 Flash column chromatography; vii) dioxane, 4N HCl in dioxane, solvent removal; viii) AcN, DIEA; Boc-alanine, EDC, HOBt, stirred overnight; EtOAc, NaHCO 3 Aqueous solution, brine, Na 2 SO 4 Ix) dioxane, 4N HCl in dioxane, solvent removal, x) AcN, DIEA, BnSO 2 -DSer (tBu) -OH, EDC, HOBt, stirred overnight; EtOAc, NaHCO 3 Aqueous solution, brine, Na 2 SO 4 RP-HPLC; xc) DCM, TFA; RP-HPLC; and xii) H 2 O, HOAc, Zn powder; RP-HPLC.
FIG.
FIG. 12 shows a reaction scheme for the synthesis of the compounds of the present invention. In this figure, “i” to “vi” are defined as follows: i) EDC, HOBt, DIEA and CH 3 CN; ii) TFA, CH 2 Cl 2 Iii) BNSO 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 Cl 2 .
FIG. 13A
FIG. 13A shows certain preferred compounds of the present invention.
FIG. 13B
FIG. 13B shows certain preferred compounds of the present invention.
FIG. 13C.
FIG. 13C shows certain preferred compounds of the present invention.
FIG. 14
FIG. 14 shows a reaction scheme for the synthesis of the compounds of the present invention. In this figure, “i” to “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, NaHCO 3 , CH 3 CN, H 2 O; v) TFA, CH 2 Cl 2 And vi) benzylsulfonyl-D-serine-L-alanine carboxylate, HATU, HOAT, DIEA, CH 3 CN.
FIG.
FIG. 15 shows a reaction scheme for the synthesis of the compounds of the present invention. In this figure, “i” to “iii” are defined as follows: i) thionyl chloride, MeOH; ii) Na / Hg (5%), CNNH 2 Reflux, 62% yield; and iii) benzylsulfonyl-D-serine-L-alanine carboxylate, HATU, HOAT, DIEA, CH 3 CN.
FIG.
FIG. 16 shows a reaction scheme for the synthesis of the compounds of the present invention. In this figure, “i” to “vi” are defined as follows: i) BH 3 , THF; ii) cl / dioxane, quantitative yield; iii) BOC-ON, THF; iv) TEA, CH 2 Cl 2 V) TFA / CH 2 Cl 2 And vi) benzylsulfonyl-D-serine-L-alanine carboxylate, HATU, HOAT, DIEA, CH 3 CN.
FIG.
FIG. 17 shows a reaction scheme for the synthesis of the compounds of the present invention. In this figure, “i” to “vi” are defined as follows: i) BH 3 , THF; ii) HCl, quantitative yield; iii) BOC-ON, THF; iv) TEA, CH 2 Cl 2 V) TFA, CH 2 Cl 2 And vi) benzylsulfonyl-D-serine-L-alanine carboxylate, HATU, HOAT, DIEA, CH 3 CN.

Claims (87)

  1. A compound of the following formula and pharmaceutically acceptable salts thereof:
    here,
    (A) X is -S (O) 2- , -N (R ')-S (O) 2 -,-(C = O)-, -OC (= O)-, -NH-C (= O)-, -P- (O) (R ')-, and a direct bond, wherein R' is independently hydrogen, alkyl of 1 to about 4 carbon atoms, about 6 to An aryl of about 14 carbon atoms, or an aralkyl of about 7 to about 16 carbon atoms, provided that when X is -P (O) (R ')-, then R' is not hydrogen;
    (B) R 1 is selected from the group consisting of:
    (1) alkyl of 1 to about 12 carbon atoms, unsubstituted or substituted with one or two substituents selected from the group consisting of Y 1 and Y 2 ;
    (2) alkyl of 1 to about 3 carbon atoms substituted with cycloalkyl of about 3 to about 8 carbon atoms, wherein the cycloalkyl is unsubstituted or Y 1 , Y 2 , and Y 3 Alkyl substituted with 1 to 3 substituents selected from the group consisting of
    (3) a cycloalkyl of 3 to about 15 carbon atoms, 1 to the cycloalkyl is unsubstituted or Y 1, Y 2, and which is selected from the group consisting of Y 3 on the ring A cycloalkyl, mono-, di-, or tri-substituted with three substituents;
    (4) 4 to about 10 ring atom heterocycloalkyl having a ring atom selected from carbon and heteroatoms, wherein the heteroatom comprises oxygen, nitrogen, and S (O) i Selected from the group, wherein i is 0, 1 or 2, and said heterocycloalkyl is unsubstituted or selected from the group consisting of Y 1 , Y 2 , and Y 3 on said ring Heterocycloalkyl, mono-, di- or tri-substituted with 1 to 3 substituents,
    (5) 4 to about 10 ring atom heterocyclo having a ring atom selected from carbon and heteroatoms, wherein said heteroatom is selected from the group consisting of oxygen, nitrogen, and S (O) i. Wherein i is 0, 1, or 2, and the heterocyclo is
    Where the said
    Is a 5-7 membered heterocyclic ring having 3-6 ring carbon atoms, wherein, V is, -CH 2 -, - O - , - S (= O) -, - S (O) 2 - or -S-, the heterocycle may be unsubstituted or substituted with 1-3 substituents selected from Y 1, Y 2, and the group consisting of Y 3 on the ring carbons one A substituted, disubstituted or trisubstituted heterocyclo,
    (6) alkenyl of from 2 to about 6 carbon atoms, unsubstituted or substituted with cycloalkyl of from about 3 to about 8 carbon atoms, wherein the cycloalkyl is unsubstituted or Y on the ring. Alkenyl, mono-, di- or tri-substituted with 1 to 3 substituents selected from the group consisting of 1 , Y 2 and Y 3 ;
    (7) aryl of from about 6 to about 14 carbon atoms, wherein the aryl is unsubstituted or 1-3 substituents selected from the group consisting of Y 1 , Y 2 and Y 3 Aryl, mono-, di-, or tri-substituted with
    (8) about 5 to about 14 ring atom heteroaryl having a ring atom selected from carbon and heteroatoms, wherein said heteroatom is selected from oxygen, nitrogen, and sulfur; Heteroaryl is a heteroaryl, which is unsubstituted or mono-, di- or tri-substituted with one to three substituents selected from the group consisting of Y 1 , Y 2 and Y 3 .
    (9) an aralkyl of about 7 to about 15 carbon atoms, wherein the aralkyl is unsubstituted or substituted on the alkyl chain with hydroxy or halogen, and the aralkyl is unsubstituted, Or an aralkyl, which is mono-, di- or trisubstituted on the aryl ring with 1 to 3 substituents selected from the group consisting of Y 1 , Y 2 and Y 3 .
    (10) a heteroaralkyl of about 5 to about 14 ring atoms having a ring atom selected from carbon and a heteroatom, wherein the heteroatom is selected from oxygen, nitrogen, and sulfur; Is unsubstituted or substituted on the alkyl chain with hydroxy or halogen, and the heteroaralkyl is unsubstituted on the ring or Y 1 , Y 2 , and Y on the ring monosubstituted with 1-3 substituents selected from the group consisting of 3, disubstituted, or trisubstituted, heteroaralkyl,
    (11) A aralkenyl of about 8 to about 16 carbon atoms, said aralkenyl 1, is unsubstituted, or is selected from the group consisting of Y 1, Y 2, and Y 3 on the aryl ring Aralkenyl, mono-, di- or trisubstituted with three substituents,
    (12) about 5 to about 14 ring atom heteroaralkenyl having a ring atom selected from carbon and heteroatoms, wherein said heteroatom is selected from oxygen, nitrogen, and sulfur; The heteroaralkenyl is unsubstituted or mono-substituted, di-substituted, or substituted on the ring carbon with 1 to 3 substituents selected from the group consisting of Y 1 , Y 2 , and Y 3. Trisubstituted heteroaralkenyl,
    (17) a fused carbocyclic alkyl of about 9 to about 15 carbon atoms,
    (18) difluoromethyl or perfluoroalkyl of 1 to about 12 carbon atoms,
    (19) perfluoroaryl of about 6 to about 14 carbon atoms,
    (20) perfluoroaralkyl of about 7 to about 15 carbon atoms, and (21) hydrogen when X is a direct bond;
    Wherein each Y 1 , Y 2 , and Y 3 are independently selected and are (i) halogen, cyano, nitro, tetrazolyl, guanidino, amidino, methylguanidino, —CF 3 , —CF 2 CF 3 , -CH (CF 3) 2, -C (OH) (CF 3) 2, -OCF 3, -OCF 2 H, -OCF 2 CF 3, -OC (O) NH 2, -OC (O) NHZ 1 , -OC (O) NZ 1 Z 2, -NHC (O) Z 1, -NHC (O) NH 2, -NHC (O) NZ 1, -NHC (O) NZ 1 Z 2, -C (O) OH, -C (O) OZ 1 , -C (O) NH 2, -C (O) NHZ 1, -C (O) NZ 1 Z 2, -P (O) 3 H 2, -P (O) 3 (Z 1) 2, -S (O) 3 H, -S (O) m Z 1, -Z 1, -OZ 1, -OH -NH 2, -NHZ 1, -NZ 1 Z 2, -C (= NH) NH 2, -C (= NOH) NH 2, -N- morpholino, and -S (O) m (CF 2 ) q CF 3 wherein m is 0, 1 or 2; q is an integer from 0 to 5; and Z 1 and Z 2 are independently from 1 to about 12 carbon atoms. Selected from the group consisting of alkyl, 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. that, or (ii) Y 1 and Y 2, together -O [C (Z 3) ( Z 4)] r O- or -O [C (Z 3) ( Z 4)] r + 1 - become so is selected, wherein, r is a integer of from 1 to 4, and Z 3 and Z 4 Independently, 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 about 5 Selected from the group consisting of heteroaralkyls of up to about 14 ring atoms;
    (C) R 2 is, -CH 3, -C 2 H 5 , - (CH 2) 2 OH, - (CH 2) 2 OA 1, -CH (R 5) OH, -CH (R 5) OA 1 , and it is selected from the group consisting of -CH 2 NH-X'-R 6 , wherein, A 1 is, -C (= O) oR 6 , -C (= O) R 6 or -C (= O) be NR 5 R 6; X 'is, -S (O) 2 -, - S (O) 2 -N (R ") -, - (C = O) -, - C (= O) -O- , -C (= O) -NH-, -P (O) (R ")-, and a direct bond, wherein R" is hydrogen, alkyl of 1 to about 4 carbon atoms, aryl of about 6 to about 14 carbon atoms or aralkyl of about 7 to about 16 carbon atoms, with the proviso, X 'is -P (O) (R ") - in the case of, R" is not hydrogen; R 5 Is a group consisting of Is et al selected:
    (1) alkyl having 1 to about 4 carbon atoms, which is unsubstituted or substituted with 1 to 2 substituents selected from the group consisting of Y 1 and Y 2 ;
    (2) alkyl of 1 to about 3 carbon atoms substituted with cycloalkyl of 3 to about 6 carbon atoms, wherein the cycloalkyl is unsubstituted or represents Y 1 , Y 2 , and monosubstituted with one to three substituents selected from the group consisting of Y 3, is disubstituted or trisubstituted, alkyl,
    (3) cycloalkyl of 3 to about 6 carbon atoms, wherein the cycloalkyl is unsubstituted or 1 to 3 on the ring selected from the group consisting of Y 1 , Y 2 , and Y 3 A cycloalkyl, mono-, di-, or tri-substituted with three substituents;
    (4) 4 to about 6 ring atom heterocycloalkyl having a ring atom selected from carbon and heteroatoms, wherein the heteroatom consists of oxygen, nitrogen, and S (O) i Selected from the group, wherein i is 0, 1 or 2, and said heterocycloalkyl is unsubstituted or selected from the group consisting of Y 1 , Y 2 , and Y 3 on said ring Heterocycloalkyl, mono-, di- or tri-substituted with 1 to 3 substituents,
    (5) 4 to about 6 ring atom heterocyclo having a ring atom selected from carbon and heteroatom, wherein the heteroatom is selected from the group consisting of oxygen, nitrogen, and S (O) i. Wherein i is 0, 1, or 2, and the heterocyclo is
    Where the said
    Is a 5-7 membered heterocyclic ring having 3-6 ring carbon atoms, wherein, V is, -CH 2 -, - O - , - S (= O) -, - S (O) 2 - or -S-, the heterocycle is unsubstituted or substituted, with 1-3 substituents selected from the group consisting of Y 1, Y 2, and Y 3 on one had the ring carbon Mono-, di- or trisubstituted heterocyclo,
    (6) alkenyl of 2 to about 6 carbon atoms, unsubstituted or substituted with cycloalkyl of about 3 to about 6 carbon atoms, wherein the cycloalkyl is unsubstituted or Y on the ring. Alkenyl, mono-, di- or tri-substituted with 1 to 3 substituents selected from the group consisting of 1 , Y 2 and Y 3 ;
    (7) phenyl, which is unsubstituted or mono-, di- or tri-substituted by 1 to 3 substituents selected from the group consisting of Y 1 , Y 2 and Y 3 , Phenyl,
    (8) about 5 to about 6 ring atom heteroaryl having a ring atom selected from carbon and heteroatoms, wherein the heteroatom is selected from oxygen, nitrogen, and sulfur; Heteroaryl is a heteroaryl, which is unsubstituted or mono-, di- or tri-substituted with one to three substituents selected from the group consisting of Y 1 , Y 2 and Y 3 .
    (9) alkyl of 1 to about 4 carbon atoms substituted with phenyl, wherein the phenyl is unsubstituted or selected from the group consisting of Y 1 , Y 2 , and Y 3 on the phenyl ring Alkyl, mono-, di- or tri-substituted with one to three substituents
    (10) a heteroaralkyl of about 5 to about 6 ring atoms having a ring atom selected from carbon and heteroatoms, wherein the heteroatoms are selected from oxygen, nitrogen, and sulfur; An aralkyl is unsubstituted or substituted on the alkyl chain with hydroxy or halogen, and the heteroaralkyl is unsubstituted on the ring or Y 1 , Y 2 , and Heteroaralkyl, mono-substituted, di-substituted or tri-substituted with 1 to 3 substituents selected from the group consisting of Y 3 ,
    (11) an aralkenyl of about 8 to about 12 carbon atoms, wherein the aralkenyl is unsubstituted or selected from the group consisting of Y 1 , Y 2 , and Y 3 on the aryl ring Aralkenyl, mono-, di- or trisubstituted with three substituents,
    (12) about 5 to about 6 ring atom heteroaralkenyl having a ring atom selected from carbon and heteroatoms, wherein the heteroatom is selected from oxygen, nitrogen, and sulfur; The heteroaralkenyl is unsubstituted or mono-substituted, di-substituted, or substituted on the ring carbon with 1 to 3 substituents selected from the group consisting of Y 1 , Y 2 , and Y 3. Trisubstituted heteroaralkenyl, and (13) hydrogen; and R 6 is selected from the group consisting of:
    (1) alkyl of 1 to about 12 carbon atoms, unsubstituted or substituted with one or two substituents selected from the group consisting of Y 1 and Y 2 ;
    (2) alkyl of 1 to about 3 carbon atoms substituted with cycloalkyl of 3 to about 8 carbon atoms, wherein the cycloalkyl is unsubstituted or represents Y 1 , Y 2 , and monosubstituted with one to three substituents selected from the group consisting of Y 3, is disubstituted or trisubstituted, alkyl,
    (3) a cycloalkyl of 3 to about 15 carbon atoms, 1 to the cycloalkyl is unsubstituted or Y 1, Y 2, and which is selected from the group consisting of Y 3 on the ring A cycloalkyl, mono-, di-, or tri-substituted with three substituents;
    (4) 4 to about 10 ring atom heterocycloalkyl having a ring atom selected from carbon and heteroatoms, wherein the heteroatom comprises oxygen, nitrogen, and S (O) i Selected from the group, wherein i is 0, 1 or 2, and said heterocycloalkyl is unsubstituted or selected from the group consisting of Y 1 , Y 2 , and Y 3 on said ring Heterocycloalkyl, mono-, di- or tri-substituted with 1 to 3 substituents,
    (5) 4 to about 10 ring atom heterocyclo having a ring atom selected from carbon and heteroatoms, wherein said heteroatom is selected from the group consisting of oxygen, nitrogen, and S (O) i. Wherein i is 0, 1, or 2, and the heterocyclo is
    Where the said
    Is a 5-7 membered heterocyclic ring having 3-6 ring carbon atoms, wherein, V is, -CH 2 -, - O - , - S (= O) -, - S (O) 2 - or -S-, the heterocycle is unsubstituted or substituted, with 1-3 substituents selected from the group consisting of Y 1, Y 2, and Y 3 on one had the ring carbon Mono-, di- or trisubstituted heterocyclo,
    (6) aryl of about 6 to about 14 carbon atoms, wherein said aryl is unsubstituted or 1-3 substituents selected from the group consisting of Y 1 , Y 2 and Y 3 Aryl, mono-, di-, or tri-substituted with
    (7) about 5 to about 14 ring atoms heteroaryl having a ring atom selected from carbon and heteroatoms, wherein the heteroatoms are selected from oxygen, nitrogen, and sulfur; Heteroaryl is a heteroaryl, which is unsubstituted or mono-, di- or tri-substituted with one to three substituents selected from the group consisting of Y 1 , Y 2 and Y 3 .
    (8) an aralkyl of about 7 to about 15 carbon atoms, wherein the aralkyl is unsubstituted or substituted on the alkyl chain with hydroxy or halogen, and the aralkyl is unsubstituted, Or an aralkyl, which is mono-, di- or trisubstituted on the aryl ring with 1 to 3 substituents selected from the group consisting of Y 1 , Y 2 and Y 3 .
    (9) about 5 to about 14 ring atom heteroaralkyl having a ring atom selected from carbon and heteroatom, wherein the heteroatom is selected from oxygen, nitrogen, and sulfur; An aralkyl is unsubstituted or substituted on the alkyl chain with hydroxy or halogen, and the heteroaralkyl is unsubstituted on the ring or Y 1 , Y 2 , and monosubstituted with 1-3 substituents selected from the group consisting of Y 3, disubstituted, or trisubstituted, heteroaralkyl, and (10) hydrogen, provided that, a 1 is -C (= O) oR When 6 , R 6 is not hydrogen;
    (D) R 3 is selected from H or methyl, or R 3 and R 4a and R 4b are selected together as described in (f);
    (E) (i) R 4a is a configuration of the S, and H, -CH 2 -S-CH 3 , -CH 2 OH, -CH 2 CN, 1~ about 3 lower alkyl carbon atoms, - CH 2 C≡CH, —CH 2 CH = CH 2 and —CH = CH 2 , and R 4b is hydrogen;
    (Ii) R 4a and R 4b are independently lower alkyl of 1-3 carbon atoms;
    (Iii) R 4a and R 4b are selected together and are- (CH 2 ) k- , where k is 5 or 6, providing a spirocycloalkyl; or (iv) R 3 And R 4a and R 4b are selected together as described in (f);
    (F) Alternatively, R 3 and R 4a are selected together to be in the configuration of S to form prolyl, pipecolyl, azetidine-2-carbonyl, 4-hydroxyprolyl, 3-hydroxyprolyl, 4- Aminopuroriru, 4 - (- CH 2 NH 2) - prolyl, 3,4 Metanopuroriru, and a group selected from the group consisting of 3,4 Dehidoropuroriru provided to P2, and R 4b is hydrogen;
    (G) R 7 is hydrogen or alkyl of 1 to about 4 carbon atoms; and (h) E is QT;
    Here, (i) Q is, -C (R 13 R 14) t -, phenyl substituted with R 8 and R 9, substituted by R 8 or R 8 and R 9, 1 to 2 hetero 5-membered or 6-membered heterocyclic ring having atoms, and substituted with R 8 and R 9, is selected from one or two of the group consisting of heterocyclic ring 9-membered or 10-membered having heteroatoms Wherein the heteroatom is selected from nitrogen and sulfur; and (ii) T is —C (= NR 10 ) NHR 11 , —NH—C (= NR 10 ) NHR 11 , and —NHR 15 Selected from the group consisting of:
    Wherein R 8 and R 9 are independently 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, R 10 and R 11 are independently hydrogen, hydroxy, alkoxy of 1 to about 3 carbon atoms, and 3 to about 16 carbon atoms. Is an alkyl of 1 to about 3 carbon atoms, or —C (= O) R 12 , wherein R 10 and R 11 are not both hydroxy or both alkoxy; R 12 is hydrogen , alkyl of 1 to about 6 carbon atoms, 1 to about 6 carbon atoms alkoxy or (CF 2), a j CF 3, where, j is 0, 1, 2 or 3; R 13 And each R 14 is independently selected from the group consisting of lower alkyl of hydrogen and from 1 to about 3 carbon atoms; R 15 is hydrogen, alkyl of 1 to about 6 carbon atoms, and - (CF 2 H ) selected from the group consisting of CF 3 , wherein h is 0, 1, 2, or 3 and t is an integer from 0 to 6,
    Compounds and pharmaceutically acceptable salts thereof.
  2. 2. The compound according to claim 1, wherein E is:
    A compound selected from the group consisting of:
  3. 2. The compound according to claim 1, wherein E is:
    A compound selected from the group consisting of:
  4. 4. The compound according to claim 3, wherein X is -S (O) 2- or -OC (= O)-.
  5. R 2 is -CH 3 or -CH (R 5) OH, The compound according to claim 4.
  6. R 5 is hydrogen, alkyl substituted with unsubstituted alkyl or Y 1, A compound according to claim 5.
  7. R 2 is -CH 2 OH or -CH (CH 3) OH, The compound according to claim 6.
  8. R 2 is -CH (CH 3) OH, and has a configuration of R, the compounds according to claim 7.
  9. R 3 is hydrogen, A compound according to claim 7.
  10. R 4 is methyl or propargyl, and R 4b is hydrogen, A compound according to claim 9.
  11. R 3 and R 4a are selected together and consist of prolyl, pipecolyl, azetidine-2-carbonyl, 4-hydroxyprolyl, 3-hydroxyprolyl, 3,4-methanoprolyl, and 3,4-dehydroprolyl providing a group selected from the group in P 2, and R 4b is hydrogen, a compound according to claim 7.
  12. R 3 and R 4a are selected together to form a group selected from the group consisting of prolyl, 4-cis-hydroxyprolyl, 3,4-dehydroprolyl, 3,4-methanoprolyl, and azetidine-2-carbonyl providing a P 2, and R 4b is hydrogen, a compound according to claim 11.
  13. The compound according to claim 5, wherein R 5 is H.
  14. 4. The compound of claim 3, wherein X is selected from the group consisting of -S (O) 2- , -OC (= O)-, -NH-C (= O)-, and a direct bond.
  15. 15. The compound according to claim 14, wherein X is -S (O) 2- or -OC (= O)-.
  16. R 1 is phenyl, benzyl, 2-phenylethyl, isobutyl, n- butyl, 3-phenylpropyl, 4-chlorobenzyl, is selected from the group consisting of 3-chlorobenzyl and 2-fluorobenzyl, to claim 15 A compound as described.
  17. R 1 -X- is phenyl -S (O) 2 -, benzyl -S (O) 2 -, 2- phenylethyl -S (O) 2 -, 3- phenylpropyl -S (O) 2 -; n -Butyl-S (O) 2- , benzyl-OC (= O)-, isobutyl-OC (= O)-, 4-chlorobenzyl-S (O) 2- , 3-chlorobenzyl-S (O) 2 -, and 2-fluorobenzyl -S (O) 2 - is selected from the group consisting of a compound according to claim 16.
  18. R 2 is selected to provide a D- seryl group P3, compounds of claim 17.
  19. The compound according to claim 18, wherein E is 4-amidinophenyl, 4-guanidinophenyl or 5- (2-amidino-thienyl).
  20. A compound according to claim 19, wherein, (i) R 3 and R 4a are selected together, prolyl, azetidine-2-carbonyl, 3,4 Metanopuroriru, and 3,4-dehydro or providing a group selected from the group consisting of prolyl the P 2, or (ii) R 3 is hydrogen and R 4a is methyl and R 4b is hydrogen, compound.
  21. 18. The compound according to claim 17, wherein E is 4-amidinophenyl, 4-guanidinophenyl or 5- (2-amidino-thienyl).
  22. A compound according to claim 21, wherein, (i) R 3 and R 4a are selected together, prolyl, azetidine-2-carbonyl, 3,4 Metanopuroriru and 3,4 Dehidoropuroriru a group selected from the group consisting of providing a P 2 or (ii) R 3 is hydrogen and R 4a is methyl and R 4b is hydrogen, compound.
  23. A compound according to claim 17, wherein, (i) R 3 and R 4a are selected together, prolyl, azetidine-2-carbonyl, 3,4 Metanopuroriru and 3,4 Dehidoropuroriru a group selected from the group consisting of providing a P 2 or (ii) R 3 is hydrogen and R 4a is methyl and R 4b is hydrogen, compound.
  24. R 2 is selected from the group consisting of -CH 2 NH (X ') ( R 6) , and -CH (R 5) OH, a compound of claim 1.
  25. R 5 is hydrogen, unsubstituted alkyl, selected from the group consisting of alkyl substituted alkyl and 1-4 carbon atoms in Y 1, alkyl of the 1 to 4 carbon atoms, unsubstituted phenyl or Y 1, Y monosubstituted with 1-3 substituents selected from the group consisting of 2 and Y 3, an alkyl substituted with disubstituted, or trisubstituted phenyl, a compound according to claim 24.
  26. R 2 is D-seryl, (R, R) D-arothreonyl, D-2-aminobutyryl, N-β-methyloxycarbonyl-D-2,3-diaminopropionyl, N-β- (2-phenylethylcarbonyl 26) is selected to provide P3 with a group selected from the group consisting of -D-2,3-diaminopropionyl, and N-β-benzyloxycarbonyl-D-2,3-diaminopropionyl. The compound according to the above.
  27. 27. The compound according to claim 26, wherein P3 is D-seryl or (R, R) D-arothreonyl.
  28. The compound according to claim 1, wherein R 3 is hydrogen.
  29. The compound of claim 1, wherein R 4a is methyl, vinyl, allyl or propargyl, and R 4b is hydrogen.
  30. R 3 and R 4a are selected together and consist of prolyl, pipecolyl, azetidine-2-carbonyl, 4-hydroxyprolyl, 3-hydroxyprolyl, 3,4-methanoprolyl, and 3,4-dehydroprolyl 2. A compound according to claim 1, wherein a group selected from the group is provided for P2 and R4b is hydrogen.
  31. R 3 and R 4a are selected together to form a group selected from the group consisting of prolyl, 4-cis-hydroxyprolyl, 3,4-dehydroprolyl, 3,4-methanoprolyl, and azetidine-2-carbonyl To P2 and R 4b is hydrogen.
  32. R 2 is, D- seryl and (R, R) D- Arosureoniru a group selected from the group consisting of is selected to provide the P3, compounds of claim 31.
  33. R 2 is selected D- seryl and (R, R) a group selected from the group consisting of -D- Arosureoniru to provide the P3, R 3 is hydrogen, R 4a is methyl and The compound of claim 1, wherein R 4b is hydrogen.
  34. R 3 and R 4a is selected together to provide prolyl, azetidine-2-carbonyl, 3,4 Metanopuroriru, and a group selected from the group consisting of 3,4 Dehidoropuroriru to P2, and R 2. The compound according to claim 1, wherein 4b is hydrogen.
  35. R 2 is -CH 2 OH, and have a configuration of R, the compounds according to claim 7.
  36. R 2 is, - (CH 2) 2 OA 1 , or -CH (R 5) is OA 1, A compound according to claim 1.
  37. R 2 is -CH (R 5) OA 1, A compound according to claim 36.
  38. R 5 is hydrogen, unsubstituted alkyl, selected from the group consisting of alkyl substituted alkyl, and from 1 to about 4 carbon atoms in Y 1, alkyl of the 1 to about 4 carbon atoms, unsubstituted phenyl or Y 1, Y 2 and monosubstituted with one to three substituents selected from the group consisting of Y 3, is alkyl substituted with disubstituted, or trisubstituted phenyl, a compound according to claim 37.
  39. R 2 is selected to provide a group selected from the group consisting of carboxylic acid esters of acyl and D- seryl to P3, compounds of claim 37.
  40. R 3 is hydrogen, A compound according to claim 39.
  41. 41. The compound of claim 40, wherein R4a is methyl, vinyl, allyl or propargyl, and R4b is hydrogen.
  42. R 3 and R 4a are selected together to form a group consisting of prolyl, pipecolyl, azetidine-2-carbonyl, 4-hydroxyprolyl, 3-hydroxyprolyl, 3,4-methanoprolyl and 3,4-dehydroprolyl 40. The compound of claim 39, wherein P2 provides a group selected from and R4b is hydrogen.
  43. 2. The compound of claim 1, wherein R2 is selected to provide P3 with a group selected from the group consisting of acyl and carboxylic esters of D-seryl.
  44. R 3 is hydrogen, R 4a is methyl and R 4b is hydrogen, A compound according to claim 43.
  45. R 3 and R 4a is selected together to provide prolyl, azetidine-2-carbonyl, 3,4 Metanopuroriru, and a group selected from the group consisting of 3,4 Dehidoropuroriru to P2, and R 44. The compound of claim 43, wherein 4b is hydrogen.
  46. 2. The compound of claim 1, wherein R2 is selected to provide P3 with a group selected from the group consisting of acyl and carboxylic esters of D-seryl.
  47. The compound according to claim 1, wherein the compound is selected from the group consisting of the compounds shown in FIGS. 10A to 10F, compound AX, compound AY and compound AZ.
  48. The compound of claim 1, wherein the compound is selected from the group consisting of the compounds shown in FIGS.
  49. 49. The compound according to claim 48, wherein the compound is selected from the group consisting of compound BG, compound BJ, and compound BK.
  50. 49. The compound of claim 48, which is compound BQ.
  51. Q is phenyl substituted with R 8 and R 9, or pyridyl substituted with R 8 and R 9, compounds of claim 1.
  52. T is, -C (= NR 10) is NHR 11 or -NH-C (= NR 10) NHR 11, compound of claim 51.
  53. 53. The compound of claim 52, wherein T is at position 4 of Q.
  54. Q is phenyl substituted with R 8 and R 9, A compound according to claim 53.
  55. T is -NH-C (= NR 10) NHR 11, compound of claim 54.
  56. Q is a pyridyl substituted with R 8 and R 9, A compound according to claim 53.
  57. T is -NH-C (= NR 10) NHR 11, compound of claim 56.
  58. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a therapeutically effective amount of a compound of claim 1.
  59. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a therapeutically acceptable amount of the compound of claim 6.
  60. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a therapeutically acceptable amount of the compound of claim 17.
  61. 28. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a therapeutically acceptable amount of the compound of claim 27.
  62. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a therapeutically acceptable amount of the compound of claim 30.
  63. 40. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a therapeutically acceptable amount of the compound of claim 39.
  64. 42. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a therapeutically acceptable amount of a compound of claim 41.
  65. 43. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a therapeutically acceptable amount of the compound of claim 42.
  66. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a therapeutically effective amount of a compound according to claim 44.
  67. 46. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a therapeutically effective amount of a compound according to claim 45.
  68. 48. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a therapeutically effective amount of a compound of claim 47.
  69. 49. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a therapeutically effective amount of a compound according to claim 48.
  70. 50. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a therapeutically effective amount of a compound according to claim 49.
  71. A method of treating a condition that is ameliorated by inhibiting or decreasing urokinase activity in a mammal in need thereof, comprising administering to said mammal a therapeutically effective amount of a compound of claim 1. A method comprising the step of:
  72. A method of treating a condition that is ameliorated by inhibiting or decreasing urokinase activity in a mammal in need of such treatment, comprising administering to said mammal a therapeutically effective amount of a compound of claim 6. A method comprising the step of:
  73. 18. A method of treating a condition that is ameliorated by inhibiting or decreasing urokinase activity in a mammal in need thereof, comprising administering to the mammal a therapeutically effective amount of a compound of claim 17. A method comprising the step of:
  74. 28. A method of treating a condition that is ameliorated by inhibiting or decreasing urokinase activity in a mammal in need thereof, comprising administering to said mammal a therapeutically effective amount of a compound of claim 27. A method comprising the step of:
  75. 33. A method of treating a condition that is ameliorated by inhibiting or decreasing urokinase activity in a mammal in need of such treatment, comprising administering to said mammal a therapeutically effective amount of a compound of claim 32. A method comprising the step of:
  76. A method of treating a condition that is reversed by inhibiting or decreasing urokinase activity in a mammal in need thereof, comprising administering to said mammal a therapeutically effective amount of a compound of claim 39. A method comprising the step of:
  77. 47. A method of treating a condition that is ameliorated by inhibiting or decreasing urokinase activity in a mammal in need thereof, comprising administering to said mammal a therapeutically effective amount of a compound of claim 41. A method comprising the step of:
  78. 48. A method of treating a condition that is ameliorated by inhibiting or decreasing urokinase activity in a mammal in need thereof, comprising administering to said mammal a therapeutically effective amount of a compound of claim 42. A method comprising the step of:
  79. 47. A method of treating a condition that is ameliorated by inhibiting or decreasing urokinase activity in a mammal in need thereof, comprising administering to said mammal a therapeutically effective amount of a compound of claim 44. A method comprising the step of:
  80. 47. A method of treating a condition that is ameliorated by inhibiting or decreasing urokinase activity in a mammal in need thereof, comprising administering to said mammal a therapeutically effective amount of a compound of claim 45. A method comprising the step of:
  81. 48. A method of treating a condition that is ameliorated by inhibiting or decreasing urokinase activity in a mammal in need thereof, comprising administering to said mammal a therapeutically effective amount of a compound of claim 47. A method comprising the step of:
  82. 49. A method of treating a condition that is ameliorated by inhibiting or decreasing urokinase activity in a mammal in need of said treatment, comprising administering to said mammal a therapeutically effective amount of a compound of claim 48. A method comprising the step of:
  83. 50. A method of treating a condition that is ameliorated by inhibiting or decreasing urokinase activity in a mammal in need thereof, comprising administering to said mammal a therapeutically effective amount of a compound of claim 49. A method comprising the step of:
  84. 48. A method of reducing or inhibiting angiogenesis in a mammal in need of treatment, said method comprising administering to said mammal a therapeutically effective amount of a compound of claim 47.
  85. 85. The method of claim 84, wherein said angiogenesis is associated with a pathological condition.
  86. 49. A method of reducing or inhibiting angiogenesis in a mammal in need of treatment, said method comprising administering to said mammal a therapeutically effective amount of a compound of claim 48.
  87. 87. The method of claim 86, wherein said angiogenesis is associated with a pathological condition.
JP2002519486A 2000-08-11 2001-08-10 Non-covalent inhibitors of urokinase and angiogenesis Pending JP2004506648A (en)

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EP20000126874 EP1182207B1 (en) 2000-08-11 2000-12-07 Non-covalent inhibitors of urokinase and blood vessel formation
US09/733,645 US6586405B2 (en) 2000-08-11 2000-12-07 Non-covalent inhibitors of urokinase and blood vessel formation
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