Cyclic peptides for capture of interleukin-1 beta

Abstract

Compounds of Formula (I) or pharmaceutically acceptable salts thereof are provided, wherein X1, X2, X3, A1, A2, R1-R7, R8a, R8b, R9a, R9b, R10 and R11 are as described herein. The compounds of formula (I) can capture IL-1 beta and are expected to be used as therapeutic agents, for example, in the treatment of cardiovascular diseases and inflammatory disorders. The disclosure also provides pharmaceutical compositions comprising the compounds disclosed herein, or pharmaceutically acceptable salts thereof. The present disclosure also relates to methods of treating and preventing cardiovascular diseases and inflammatory disorders using the compounds or pharmaceutically acceptable salts thereof, as well as methods of preparing medicaments for this purpose. # imgabs0 #

Description

Technical Field

[0001] The present disclosure relates to certain cyclic peptides that capture interleukin-1β (IL-1β), pharmaceutical compositions comprising such peptides, and methods of using the compounds to treat, inhibit, or ameliorate one or more cardiovascular disease states that may benefit from capturing IL-1β, including atherosclerosis.

[0002] Reference to electronically submitted sequence listing

[0003] This application contains a sequence listing submitted electronically in XML format and is hereby incorporated by reference in its entirety. The XML file was created on May 25, 2023, is named 25545-WO-PCT_SL.XML, and is 916,839 bytes in size. Background Art

[0004] Atherosclerosis is a disease of the arteries characterized by the accumulation of cholesterol plaques on the inner wall of the arteries. The progression of atherosclerosis can lead to hardening or stenosis of the arteries and increase the risk of plaque rupture. These ruptures can release cholesterol globules and other substances into the blood, which may lead to the blockage of blood flow to the brain, heart or other organs. Medically, these are referred to as major adverse cardiac events (MACE).

[0005] Risk factors for the development and progression of atherosclerotic cardiovascular disease (ASCVD) include high cholesterol, high blood pressure, a diet high in saturated fat, smoking, obesity, diabetes, physical inactivity, and elevated levels of C-reactive protein (CRP), a marker of inflammation.

[0006] The first line of treatment to prevent the progression of ASCVD is a healthy diet and exercise, but compliance is often poor. Medical treatment for ASCVD focuses primarily on cholesterol-lowering drugs such as statins, cholesterol absorption inhibitors, and low-density lipoprotein (LDL) receptor inhibitors. These drugs are very effective in reducing the accumulation of fatty acid deposits and improving arterial health. Other prescription drugs used to treat ASCVD but not improve the disease state include blood thinners such as aspirin (used to prevent platelet aggregation in narrowed arteries) and blood pressure medications (used to reduce the risk and severity of heart attacks). For advanced cases of ASCVD, surgical options for more invasive interventions include angioplasty, stent placement, endarterectomy (surgical removal of plaque), and bypass surgery.

[0007] While cholesterol-lowering drugs have become an important standard of care for slowing the progression of atherosclerosis, clinical data support another key role of inflammation in the progression of untreated ASCVD. Biomarkers of inflammation, such as CRP, are associated with an increased risk of cardiovascular events, independent of cholesterol levels. The Canakinumab Anti-Inflammatory Thrombotic Outcomes Study (CANTOS) is the first clinical trial to show that reducing vascular inflammation without concurrent lipid reduction reduces the incidence of cardiovascular events. N Engl J Med 2017; 377: 1119-1131. Canakinumab is an anti-interleukin-1β (IL-1β) human monoclonal antibody approved for clinical treatment of rheumatic disorders. IL-1β is a proinflammatory cytokine that induces IL-6 and thereby elevates the downstream inflammatory biomarker high-sensitivity CRP (hsCRP). Therefore, CANTOS provides proof of concept that therapy targeting IL-1β can reduce the incidence of MACE in certain patients, a therapy that is a complement and potential addition to the standard of care for lowering LDL.

[0008] In addition to standard-of-care cholesterol-lowering drugs, additional nonsurgical treatment options are needed to slow the progression of atherosclerosis and reduce the risk of MACE. In addition, patients with inflammatory lesions may benefit from orally administered agents that block the same cytokine, IL-1β, as canakinumab. Summary of the invention

[0009] The present disclosure provides certain cyclic peptides that reduce inflammation by binding to IL-1β cytokines and preventing binding to IL-1 receptors, thereby leading to inhibition of downstream pro-inflammatory signaling. These cyclic peptides can be valuable pharmaceutically active compounds for the treatment of cardiovascular diseases and inflammatory lesions. In one aspect, the present disclosure provides compounds of formula (I) and pharmaceutically acceptable salts thereof

[0010]

[0011] The compound can capture IL-1β and thereby affect downstream pro-inflammatory signaling pathways that may be associated with cardiovascular disorders. Therefore, in another aspect, the present disclosure provides a method for treating cardiovascular disorders (e.g., atherosclerosis, vascular inflammation), the method comprising administering a therapeutically effective amount of a compound of the present disclosure to a subject in need thereof. In certain embodiments, the administration comprises oral administration of the compound.

[0012] In addition, the present disclosure provides methods for preparing the compounds of the present disclosure and pharmaceutical compositions comprising the compounds of the present disclosure and a pharmaceutically acceptable carrier. DETAILED DESCRIPTION OF THE INVENTION

[0014] Compounds of the Disclosure

[0015] In one embodiment, the present disclosure provides a compound having structural formula (I) as shown above, wherein:

[0016] R 1 is CH3C(O)NH-CH2CH2-O- or C 1 ;

[0017] C 1 yes:

[0018] (i) a 5- to 6-membered monocyclic aromatic or heteroaromatic group, wherein the heteroaromatic group contains 1 to 2 heteroatoms selected from N, O and S; or

[0019] (ii) a 5- to 6-membered monocyclic or bicyclic saturated cycloalkyl or heterocycloalkyl group containing 1 to 2 heteroatoms selected from N, O and S; or

[0020] (iii) a 5- to 6-membered monocyclic or bicyclic cycloalkyl group;

[0021] Among them C 1 is unsubstituted or substituted with 1 to 3 R C1 Substituent substituted, said R C1 The substituents are independently selected from halogen, C1-C3 alkyl, C1-C3 fluoroalkyl, carboxyl, C1-C3 alkoxy and C2-C3 acyl;

[0022] R 2 is H, C1-C3 alkyl, benzyl or phenyl-CH2CH2-;

[0023] R 3 is a 9- to 10-membered bicyclic aryl or heteroaryl group, wherein the heteroaryl group contains 1 to 2 heteroatoms selected from N, O and S;

[0024] Where R 3 is unsubstituted or substituted with 1 to 3 R 3a Substituent substituted, said R 3a The substituents are independently selected from halogen, C1-C3 alkyl, C1-C3 fluoroalkyl, hydroxyl and C1-C3 alkoxy;

[0025] R 4 It is C1-C3 alkyl, HO2C-(CH2) m -、H2NC(O)-(CH2) m -、(CH3)2NC(O)-(CH2) m - or tetrazolyl-(CH2) m -;

[0026] R 5It is amino, H2N(CH2) n -、H2NC(O)-(CH2) n -, CH3C(O)NH-, CH3C(O)NH(CH2) n -、C 5 or C 5 -CH2-,

[0027] C 5 yes:

[0028] (i) a 5- to 6-membered monocyclic aromatic or heteroaromatic group, wherein the heteroaromatic group contains 1 to 2 heteroatoms selected from N, O and S;

[0029] (ii) a 9- to 10-membered bicyclic aryl or heteroaryl group, wherein the bicyclic heteroaryl group contains 1 to 3 heteroatoms selected from N, O and S;

[0030] (iii) a 5- to 6-membered monocyclic or 9- to 10-membered heterocycloalkyl group, wherein the heterocycloalkyl group is saturated or partially unsaturated and contains 1 to 2 heteroatoms selected from N, O and S;

[0031] (iv) a 5- to 6-membered monocyclic cycloalkyl group;

[0032] (v) 2,3-dihydroindole;

[0033] Among them C 5 is unsubstituted or substituted with 1 to 3 R C5 Substituent substituted, said R C5 The substituents are independently selected from halogen, amino, hydroxy, C1-C3 alkyl, C1-C3 fluoroalkyl, C1-C3 alkoxy, H2N-(CH2) k -、H2NC(O)-(CH2) k -, H2C=CH-CH2O- and phenyl;

[0034] R 6 H, C1-C5 alkyl, H2N(CH2) p -, HOCH2-, (CH3)2NCH2-, H3CO-(CH2) q - or C 6 -CH2-;

[0035] C 6 is a 5- or 6-membered monocyclic saturated heterocycloalkyl group containing 1 to 2 heteroatoms selected from N, O and S; and wherein C 6 is unsubstituted or substituted with 1 to 3 R C6 Substituent substituted, said R C6 The substituents are independently selected from halogen, C1-C3 alkyl, C1-C3 fluoroalkyl, hydroxyl and C1-C3 alkoxy;

[0036] R 7 is H or C1-C3 alkyl;

[0037] R 8a H, C1-C5 alkyl, HOCH2-, H2N(CH2) r -、(CH3)3N + (CH2) r - or CH3C(O)NH(CH2) r -;

[0038] R 8b is H or C1-C3 alkyl;

[0039] R 9a is H or C1-C3 alkyl;

[0040] R 9b H, C1-C5 alkyl, C 9 -CH2- or C 9 -CH2CH2-;

[0041] C 9 yes:

[0042] (i) a 5- to 6-membered monocyclic aromatic or heteroaromatic group, wherein the heteroaromatic group contains 1 to 2 heteroatoms selected from N, O and S; or

[0043] (ii) a 5- to 6-membered monocyclic saturated cycloalkyl or heterocycloalkyl group, wherein the heterocycloalkyl group contains 1 to 2 heteroatoms selected from N, O and S;

[0044] Among them C 9 is unsubstituted or substituted with 1 to 3 R C9 Substituent substituted, said R C9 The substituents are independently selected from halogen, amino, hydroxy, cyano, C1-C3 alkyl, C1-C3 fluoroalkyl, C1-C3 alkoxy, H2N-(CH2) k -、H2NC(O)-(CH2) k -, H2NCH2CH2O-, CH3C(O)NH-CH2CH2O- and morpholinyl-CH2CH2O-;

[0045] R 10 is H, halogen or C1-C3 alkyl;

[0046] R 11 is H, halogen or C1-C3 alkyl;

[0047] Each occurrence of subscript k is independently 1 or 2;

[0048] Subscript m is 1 or 2;

[0049] Subscript n is 1, 2, 3, or 4;

[0050] The subscript p is 1, 2, 3, or 4;

[0051] Subscript q is 1 or 2;

[0052] The subscript r is 1, 2, 3, or 4;

[0053] X 1 , X 2 and X 3 are independently C(H) or N; and

[0054] A 1 and A 2 independently selected from HO2C-, H2NC(O)-, CH3C(O)N(H)-, H2NS(O)2-, CH3S(O)2N(H)-, tetrazolyl and 5-oxoxadiazolyl; or

[0055] or pharmaceutically acceptable salts thereof.

[0056] In another embodiment, the present disclosure provides a compound of formula (I), wherein:

[0057] R 5 It is amino, H2N(CH2) n -、H2NC(O)-(CH2) n -、C 5 or C 5 -CH2-,

[0058] C 5 yes:

[0059] (i) a 5- to 6-membered monocyclic aromatic or heteroaromatic group, wherein the heteroaromatic group contains 1 to 2 heteroatoms selected from N, O and S:

[0060] (ii) a 9- to 10-membered bicyclic aryl or heteroaryl group, wherein the bicyclic heteroaryl group contains 1 to 3 heteroatoms selected from N, O and S;

[0061] (iii) a 5- to 6-membered monocyclic or 9- to 10-membered heterocycloalkyl group, wherein the heterocycloalkyl group is saturated or partially unsaturated and contains 1 to 2 heteroatoms selected from N, O and S; or

[0062] (iv) 2,3-dihydroindole;

[0063] Among them C 5 is unsubstituted or substituted with 1 to 3 R C5 Substituent substituted, said R C5The substituents are independently selected from halogen, amino, hydroxy, C1-C3 alkyl, C1-C3 fluoroalkyl, C1-C3 alkoxy, H2N-(CH2) k -、H2NC(O)-(CH2) k -, H2C=CH-CH2O- and phenyl;

[0064] R 6 H, C2-C5 alkyl, H2N(CH2) p -, HOCH2-, (CH3)2NCH2-, H3CO-(CH2) q - or C 6 -CH2-;

[0065] R 8a is H, C1-C3 alkyl, HOCH2- or H2N(CH2) r -;

[0066] R 9a It is H;

[0067] R 9b is H, C1-C3 alkyl, C 9 -CH2- or C 9 -CH2CH2-;

[0068] R 10 It is H;

[0069] R 11 It is H;

[0070] subscript p is 1, 2, or 3; and

[0071] The subscript r is 2, 3, or 4.

[0072] In one embodiment, the compound of formula (I) has the structural formula (IA)

[0073]

[0074] In another embodiment, the present disclosure provides a compound of formula (I), wherein:

[0075] C 1 is phenyl, pyrimidinyl or piperazinyl, wherein C 1 is unsubstituted or substituted with 1 to 2 R C1 Substituent substitution;

[0076] R 3 is naphthyl or indolyl, where R 3 is unsubstituted or substituted with 1 to 2 R 3a Substituent substitution;

[0077] C5 is phenyl, pyridyl, pyrimidinyl, naphthyl, indolyl, 7-azaindolyl, indazolyl, 2,3-dihydroindolinyl, piperidinyl, tetrahydropyranyl or cyclohexyl, wherein C 5 is unsubstituted or substituted with 1 to 2 R C5 replace;

[0078] C 6 is tetrahydropyranyl or morpholinyl; wherein C 6 is unsubstituted or substituted with 1 to 2 R C6 replace; and

[0079] C 9a is H or methyl;

[0080] C 9b is phenyl, pyridyl, cyclohexyl, morpholinyl or piperidinyl, wherein C 9 is unsubstituted or substituted with 1 to 2 R C9 replace.

[0081] In another embodiment, the present disclosure provides a compound of formula (I), wherein:

[0082] X 1 and X 2 is C(H); and

[0083] R 1 It is a phenyl group substituted by a carboxyl group.

[0084] In another embodiment, the present disclosure provides a compound of formula (I), wherein:

[0085] X 1 and X 2 is C(H); and

[0086] R 1 It is CH3C(O)NH-CH2CH2-O-.

[0087] In another embodiment, the present disclosure provides compounds of formula (I), wherein R 2 It's H.

[0088] In another embodiment, the present disclosure provides compounds of formula (I), wherein R 3 It is an indolyl group substituted by a halogen.

[0089] In another embodiment, the present disclosure provides compounds of formula (I), wherein R 3 It is naphthyl.

[0090] In another embodiment, the present disclosure provides compounds of formula (I), wherein R 4It is HO2C-(CH2) m -.

[0091] In another embodiment, the present disclosure provides compounds of formula (I), wherein X 3 It is C(H).

[0092] In another embodiment, the present disclosure provides compounds of formula (I), wherein R 5 It is H2N(CH2) n -, indole, 7-azaindole, naphthyl or pyridinyl.

[0093] In another embodiment, the present disclosure provides compounds of formula (I), wherein R 5 It is H2N(CH2) n -, indole, naphthyl or pyridyl.

[0094] In another embodiment, the present disclosure provides compounds of formula (I), wherein R 6 is H, HOCH2-, C2-C5 alkyl or H2N(CH2) p -.

[0095] In another embodiment, the present disclosure provides compounds of formula (I), wherein R 6 Is C2-C5 alkyl or H2N(CH2) p -.

[0096] In another embodiment, the present disclosure provides compounds of formula (I), wherein R 7 It's H.

[0097] In another embodiment, the present disclosure provides a compound of formula (I), wherein:

[0098] R 8a is methyl or H2NCH2CH2-; and

[0099] R 8b It's H.

[0100] In another embodiment, the present disclosure provides compounds of formula (I), wherein R 9b yes

[0101]

[0102] In another embodiment, the present disclosure provides compounds of formula (I), wherein R 9b yes

[0103]

[0104] In one embodiment, the present disclosure provides a compound of formula (I), wherein A 1 Selected from HO2C-, H2NC(O)-, CH3C(O)N(H)-, H2NS(O)2-, CH3S(O)2N(H)-, tetrazolyl and 5-oxooxadiazolyl. Chemical technicians will recognize that such a part in an amino acid residue can be shown by the following substructure:

[0105]

[0106] In one embodiment, the present disclosure provides a compound of formula (I), wherein A 2 Selected from HO2C-, H2NC(O)-, CH3C(O)N(H)-, H2NS(O)2-, CH3S(O)2N(H)-, tetrazolyl and 5-oxooxadiazolyl. Chemical technicians will recognize that such a part in an amino acid residue can be shown by the following substructure:

[0107]

[0108] In another embodiment, the present disclosure provides a compound of formula (I), wherein:

[0109] R 10 is H; and

[0110] R 11 It is H, F or Cl.

[0111] In another embodiment, the present disclosure provides compounds of formula (I), wherein A 1 and A 2 All are HO2C- (i.e., carboxyl).

[0112] In certain embodiments, the present disclosure provides compounds of Formula (I), wherein:

[0113] R 1 is 4-CH3C(O)-piperazin-1-yl, CH3C(O)NH-CH2CH2-O-, 5-CO2H-pyrimidin-2-yl or 4-CO2H-phenyl;

[0114] R 2 is H, ethyl, benzyl or phenyl -CH2CH2-;

[0115] R 3 is naphthyl-1-yl, 4-fluoroindol-3-yl or 4-chloroindol-3-yl;

[0116] R 4 It is methyl, HO2C-(CH2) m- or H2NC(O)-(CH2) m -;

[0117] R 5 It is amino, H2N(CH2) n -, naphthyl-1-yl, indol-3-yl, 7-aza-indol-3-yl, indazol-1-yl, 2,3-dihydroindolin-1-yl, pyridin-3-yl, pyridin-4-yl, piperidin-4-yl, 3-aminomethylphenyl, 4-aminomethylphenyl, 3-aminophenyl, 4-aminophenyl, phenyl, pyridin-4-yl-CH2-, pyrimidin-5-yl, tetrahydropyran-4-yl-, H2NC(O)-(CH2)2-, 3-biphenyl, 3-CH2=CH-CH2O-phenyl, CH3C(O)NH-, CH3C(O)NH(CH2)3-, or cyclohexyl;

[0118] R 6 It is H, (CH3)2CHCH2-, (CH3)3CCH2-, H2N(CH2) p -, HOCH2-, H3CCH2CH2-, morpholin-4-yl-CH2-, tetrahydropyran-4-yl-CH2-, (CH3)2NCH2- or H3CO-(CH2) q -;

[0119] R 7 is H or methyl;

[0120] R 8a It is H, methyl, HOCH2-, H2N(CH2) r -, (CH3)3NCH2CH2- or CH3C(O)NH(CH2)4-;

[0121] R 8b is H or methyl;

[0122] R 9a is H or methyl;

[0123] R 9bis H, methyl, H3CCH2CH2CH2-, 4-HO-phenyl-CH2CH2-, phenyl-CH2CH2-, cyclohexyl-CH2CH2-, 5-NC-pyridin-3-yl-CH2CH2-, 4-F3C-phenyl-CH2-, 3-F3C-phenyl-CH2-, 2-F3C-phenyl-CH2-, morpholin-4-yl-CH2CH2O-phenyl-CH2-, 4-aminophenyl-CH2-, 4,4-difluorocyclohexyl-CH2-, 4-H2NCH2CH2O-phenyl-CH2-, 4-H2NCH2CH2O-pyridin-3-yl-CH2-, piperidin-4-yl-CH2-, or 4-CH3C(O)NH-CH2CH2O-phenyl-CH2-;

[0124] R 10 is H, fluorine or methyl;

[0125] R 11 is H, fluorine or chlorine;

[0126] A 1 and A 2 All are HO2C-; and

[0127] X 1 , X 2 and X 3 It is C(H).

[0128] In certain embodiments, the present disclosure provides compounds of Formula (I), wherein:

[0129] R 1 is 4-CH3C(O)-piperazin-1-yl, CH3C(O)NH-CH2CH2-O-, 5-CO2H-pyrimidin-2-yl or 4-CO2H-phenyl;

[0130] R 2 is H, ethyl, benzyl or phenyl -CH2CH2-;

[0131] R 3 is naphthyl-1-yl, 4-fluoroindol-3-yl or 4-chloroindol-3-yl;

[0132] R 4 It is methyl, HO2C-(CH2) m - or H2N(O)C-(CH2) m -;

[0133] R 5 It is amino, H2N(CH2) n-, naphthyl-1-yl, indol-3-yl, 7-aza-indol-3-yl, indazol-1-yl, 2,3-dihydroindolin-1-yl, pyridin-3-yl, pyridin-4-yl, piperidin-4-yl, 3-aminomethylphenyl, 4-aminomethylphenyl, 3-aminophenyl, 4-aminophenyl, phenyl, pyridin-4-yl-CH2-, pyrimidin-5-yl, tetrahydropyran-4-yl-, H2N(O)C-(CH2)2-, 3-biphenyl or 3-CH2=CH-CH2O-phenyl,

[0134] R 6 It is H, (CH3)2CHCH2-, H2N(CH2) p -, HOCH2-, H3CCH2CH2-, morpholin-4-yl-CH2-, tetrahydropyran-4-yl-CH2-, (CH3)2NCH2- or H3CO-(CH2) q -;

[0135] R 7 is H or methyl;

[0136] R 8a is H, methyl, HOCH2- or H2N(CH2) r -;

[0137] R 8b is H or methyl;

[0138] R 9a It is H;

[0139] R 9b is H, methyl, 4-HO-phenyl-CH2CH2-, phenyl-CH2CH2-, 5-NC-pyridin-3-yl-CH2CH2-, 4-F3C-phenyl-CH2-, 3-F3C-phenyl-CH2-, 2-F3C-phenyl-CH2-, morpholin-4-yl-CH2CH2O-phenyl-CH2-, 4-aminophenyl-CH2-, 4,4-difluorocyclohexyl-CH2-, 4-H2NCH2CH2O-phenyl-CH2-, 4-H2NCH2CH2O-pyridin-3-yl-CH2-, piperidin-4-yl-CH2-, or 4-CH3C(O)NH-CH2CH2O-phenyl-CH2-;

[0140] R 10 and R 11 are all H; and

[0141] A 1 and A 2 They are all HO2C-.

[0142] In certain embodiments, the disclosure provides a compound of formula (I), wherein the compound is selected from SEQ ID NOs: 1-215 shown in Table 1. In certain embodiments, the disclosure provides a compound of formula (I), wherein the compound is selected from SEQ ID NOs: 1-173 shown in Table 1.

[0143] In a specific embodiment, the present disclosure provides a compound of formula (I), wherein the compound is selected from (SEQ ID NOs 78, 71-72, 67, 65, 70, 69, 68, 66, 64, 77, 79, 209, 193, 215, 212 and 210, respectively, in order of appearance):

[0144]

[0145]

[0146]

[0147]

[0148] In certain embodiments, the present disclosure provides a compound of formula (I), wherein the compound is selected from SEQ ID NO: 78, 71-72, 67, 65, 70, 69, 68, 66, 64, 77 and 79.

[0149] While not being bound by any particular theory, applicants believe that the compounds of the present disclosure capture interleukin-1β, preventing signaling through the IL-1 receptor, and thereby reducing downstream markers IL-6 and CRP. Therefore, the compounds can be used to treat the inflammatory component of cardiovascular diseases such as ASCVD and heart failure with preserved ejection fraction (HFpEF). The compounds can also be used to treat inflammatory lesions such as hidradenitis suppurativa (acne inversa), inflammatory bowel disease, and osteoarthritis.

[0150] definition

[0151] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0152] As used throughout the present disclosure, "a compound of the disclosure," "a compound of the present disclosure," and "compounds disclosed herein" are used interchangeably and should be understood to include the disclosed cyclic peptides and compounds of formula (I). References to compounds of formula (I) include compounds of other general formulae within the scope of formula (I), including, but not limited to, compounds of formula (IA). Compounds of formula (I) may form salts, which are also within the scope of the present disclosure. References herein to compounds of the present disclosure (or compounds of formula (I)) are understood to include references to their salts, unless otherwise indicated. The term "salts" as used herein refers to acidic salts formed with inorganic and / or organic acids, as well as basic salts formed with inorganic and / or organic bases. In addition, when the compound of formula (I) contains both a basic moiety (such as, but not limited to, an amino group, pyrrolidine or imidazole) and an acidic moiety (such as, but not limited to, a carboxylic acid), zwitterions ("inner salts") may be formed and are included in the term "salt" as used herein. In one embodiment, the salt is a pharmaceutically acceptable (i.e., non-toxic, physiologically acceptable) salt. In another embodiment, the salt is a non-pharmaceutically acceptable salt. For example, a salt of a compound of formula (I) may be formed by reacting a compound of formula (I) with an amount (such as an equivalent) of an acid or base in a medium (such as, a medium in which the salt is precipitated), or in an aqueous medium, followed by lyophilization.

[0153] "Acyl" refers to an alkyl-C(O)- group wherein alkyl is as defined below. The bond to the parent group is through the carbon atom of the carbonyl group.

[0154] "Alkyl", and other groups with the prefix "alk" (such as alkoxy, etc.) refer to carbon chains containing a specified number of carbon atoms, which can be straight or branched or a combination thereof. For example, C1-C6 alkyl refers to an alkyl group having 1 (i.e., methyl) to 6 carbon atoms (i.e., hexyl). In a particular embodiment, the straight chain alkyl group has 1-6 carbon atoms, and the branched chain alkyl group has 3-7 carbon atoms. Examples of alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, sec-butyl and tert-butyl, pentyl, hexyl, heptyl, octyl, nonyl, etc.

[0155] "Alkoxy" and "alkyl-O-" are used interchangeably and refer to an alkyl group attached to an oxygen.

[0156] "Amino" refers to a H2N- group. The bond to the parent group is through the nitrogen atom.

[0157] "Amino acid" means naturally occurring α-amino acids and stereoisomers thereof, as well as non-natural amino acids (such as β-amino acids and substituted amino acids) and stereoisomers thereof. In the sequences given for the peptides (compounds) according to the present disclosure, the amino acid residues have their conventional meanings. Thus, "G" is glycine, "W" is tryptophan, "A" is alanine, "S" is serine, and so on. It should be understood that the "D" isomers are represented by a "d" before the single letter code or amino acid name, so that, for example, dA is the D isomer of L-alanine. Amino acid residues not covered by the foregoing have the definitions provided in the table of the Examples section below.

[0158] As used herein, "aryl" refers to a monocyclic 6-membered or bicyclic 10-membered ring system in which at least one ring is aromatic and all ring atoms are carbon.

[0159] A "bicyclic ring system" means two linked rings. The rings may be fused, i.e., sharing two adjacent atoms, or "spiro," i.e., sharing only one atom.

[0160] "Carboxyl" refers to a HO2C- group. The bond to the parent group is through the carbon atom of the carbonyl component.

[0161] "Cycloalkyl" refers to a saturated cyclic hydrocarbon group. In certain embodiments, the cycloalkyl group has 3-12 carbon atoms, thereby forming 1-3 fused carbocyclic rings. Examples of cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, adamantyl, etc.

[0162] "Fluorinated alkyl" includes mono- and poly-fluorinated alkyl groups, up to perfluorinated alkyl groups, for example, fluoromethyl, 1,1-difluoroethyl, trifluoromethyl or 1,1,1,2,2-pentafluorobutyl.

[0163] Unless otherwise indicated, "halogen" or "halo" includes fluorine (fluoro), chlorine (chloro), bromine (bromo) and iodine (iodo). In one embodiment, halogen is fluorine (-F) or chlorine (-C).

[0164] "Heterocycloalkyl" refers to a non-aromatic monocyclic, bicyclic or tricyclic ring system containing about 3 to about 10 ring atoms, preferably about 5 to about 10 ring atoms, wherein one or more atoms in the ring system are elements other than carbon, such as nitrogen, oxygen or sulfur, alone or in combination. There are no adjacent oxygen and / or sulfur atoms in the ring system. In certain embodiments, heterocycloalkyl contains about 5 to about 6 ring atoms. The prefix aza, oxa or thia before the heterocyclyl root name refers to at least one nitrogen, oxygen or sulfur atom present as a ring atom, respectively. In certain embodiments, the nitrogen or sulfur atom of the heterocycloalkyl may be optionally oxidized to the corresponding N-oxide, S-oxide or S,S-dioxide. Non-limiting examples of suitable monocyclic heterocyclyl rings include piperidinyl, pyrrolidinyl, piperazinyl, morpholinyl, thiomorpholinyl, thiazolidinyl, 1,4-dioxane, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothienyl, etc.

[0165] "Heteroaryl" refers to aromatic monocyclic, bicyclic and tricyclic ring structures in which one or more atoms (i.e., heteroatoms) in the ring are elements other than carbon. The heteroatoms are typically O, S or N atoms. Examples of heteroaromatic groups include pyridyl, pyrimidyl, pyrrolyl, pyridazinyl, isoxazolyl, thiazolyl, oxazolyl, indolyl, benzoxazolyl, benzothiazolyl and imidazolyl.

[0166] When any variable (e.g., R C1 ) occurs more than once in any component or in Formula (I) or in other formulae herein, its definition at each occurrence is independent of its definition at every other occurrence. Combinations of substituents and / or variables are permissible only if such combinations result in stable compounds. In selecting compounds of the present disclosure, one of ordinary skill in the art will recognize that various substituents (e.g., R C9 ) should be selected in accordance with the well-known principles of chemical structure connectivity and stability. Unless explicitly stated to the contrary, substitution with a specified substituent is permitted on any atom in a ring (e.g., an aryl, heteroaryl ring, or saturated heteroaryl ring) as long as such ring substitution is chemically permitted and produces a stable compound. A "stable" compound is a compound that can be prepared and isolated, and whose structure and properties remain or are caused to remain substantially unchanged for a period of time sufficient to allow the compound to be used for the purposes described herein (e.g., therapeutic or prophylactic administration to a subject).

[0167] The term "substituted" should be considered to include multiple degrees of substitution of a specified substituent. Where multiple substituent moieties are disclosed or claimed, the substituted compound may be independently substituted with one or more of the disclosed or claimed substituent moieties in the singular or plural. Independently substituted means that the (two or more) substituents may be the same or different.

[0168] Unless otherwise explicitly depicted or described, variables depicted with "floating" bonds in a structural formula are allowed to occur on any available carbon atom in the ring to which the variable is attached. When a moiety in formula (I) or any embodiment thereof is noted as "optionally substituted", it means that formula (I) or its embodiments encompass compounds containing the noted substituent (or substituents) on that moiety as well as compounds not containing the noted substituent (or substituents) on that moiety.

[0169] The wavy lines used in this article The point of attachment to the rest of the compound is indicated.

[0170] Certain compounds described herein may exist as tautomers, which have different points of hydrogen attachment and are accompanied by one or more double bond shifts. For example, a ketone and its enol form are keto-enol tautomers. The compounds of the present disclosure encompass individual tautomers and mixtures thereof.

[0171] In the compounds of the present disclosure, the atoms may exhibit their natural isotopic abundance, or one or more of the atoms may be artificially enriched with a particular isotope having the same atomic number, but an atomic mass or mass number different from the atomic mass or mass number that predominantly occurs in nature. The present disclosure described and claimed herein is intended to include all suitable isotopic variants of the compounds of the present disclosure and embodiments thereof. For example, different isotopic forms of hydrogen (H) include protium ( 1 H) and deuterium ( 2 H, also denoted herein as D). Protium is the predominant isotope of hydrogen found in nature. Enrichment in deuterium may provide certain therapeutic advantages, such as increasing half-life in vivo or reducing dosage requirements, or may provide compounds that can be used as standards for characterization of biological samples. Isotopically enriched compounds of the present disclosure may be prepared without undue experimentation by conventional techniques well known to those skilled in the art, or by methods similar to those described in the schemes and examples herein, using appropriate isotopically enriched reagents and / or intermediates.

[0172] The term "pharmaceutically acceptable salt" refers to a salt prepared from a pharmaceutically acceptable non-toxic base or acid. When the compounds of the present disclosure are acidic (or have functional groups that may be anionic), their corresponding salts can be conveniently prepared from pharmaceutically acceptable non-toxic bases (including inorganic bases and organic bases). Examples of suitable inorganic cations include, but are not limited to, alkali metal ions such as Li + 、Na + and K + , alkaline earth metal cations such as Ca2 + and Mg2+ , and other cations such as Al 3+ and Zn + Examples of suitable organic cations include, but are not limited to, ammonium ions (i.e., NH4 + ) and substituted ammonium ions. Examples of suitable substituted ammonium ions are those derived from methylamine, ethylamine, diethylamine, triethylamine and ethylenediamine. When the compounds of the present disclosure are alkaline, their corresponding salts can be conveniently prepared from pharmaceutically acceptable non-toxic acids (including inorganic and organic acids). Examples of such acid addition salts include salts formed by hydrohalic acids (e.g., hydrochloric acid, hydrobromic acid, hydroiodic acid), formic acid, acetic acid, capric acid and citric acid. Salts containing acetate, formate, caprate, chloride or sodium salts are typically used with the compounds of the present disclosure. In certain embodiments, the salts of the compounds of the present disclosure can be formed by exchanges well known to those of ordinary skill in the art, such as by anion exchange, for example, replacing trifluoroacetate ions with chloride ions.

[0173] In addition, the compound of the present disclosure can exist in amorphous form and / or one or more crystalline forms, and therefore, all amorphous and crystalline forms of the compound of formula (I) and mixtures thereof (including embodiments) are intended to be included in the scope of the present disclosure. In addition, some compounds of the present disclosure can form solvates with water (that is, hydrates) or common organic solvents (such as, but not limited to acetic acid or acetonitrile). Such solvates and hydrates of the compound of the present invention, particularly pharmaceutically acceptable solvates and hydrates, together with non-solvated forms and anhydrous forms, are equally encompassed in the scope of the present disclosure.

[0174] Any pharmaceutically acceptable prodrug modifications of the compounds of this disclosure that result in conversion in vivo to compounds within the scope of this disclosure are also within the scope of this disclosure.

[0175] The present disclosure also relates to processes for preparing compounds of formula (I), which processes are described in the following examples and by which compounds of the present disclosure can be obtained.

[0176] "Treatment" and "treating" refer to all processes in which the progression of a disease or disorder described herein can be slowed, interrupted, stopped, controlled or stopped. These terms do not necessarily imply complete elimination of all disease or disorder symptoms.

[0177] As used herein, "Preventing" or "prophylaxis" means reducing the likelihood of contracting a disease or disorder described herein, or reducing the severity of a disease or disorder described herein.

[0178] The term "therapeutically effective (or efficacious) amount" and similar descriptions such as "therapeutically effective amount" or "effective dose" are intended to refer to the amount of a compound of the present disclosure that will elicit the biological or medical response of a tissue, system, animal or human that the researcher, veterinarian, physician or other clinician is seeking. In a preferred embodiment, the term "therapeutically effective amount" refers to the amount of a compound of the present disclosure that alleviates at least one clinical symptom in a human patient. The term "prophylactically effective (or efficacious) amount" and similar descriptions such as "effective preventive amount" are intended to refer to the amount of a compound of the present disclosure that will prevent or reduce the risk of the occurrence of a biological or medical event that the researcher, veterinarian, physician or other clinician is trying to prevent in a tissue, system, animal or human.

[0179] Dosages of Compounds of the Disclosure

[0180] The dosage regimen utilizing the compounds of the present disclosure is selected based on a variety of factors, including the type, race, age, weight, sex, and medical condition of the patient; the severity of the condition to be treated; the potency of the compound selected for administration; the route of administration; and the renal and liver function of the patient. For the purpose of determining the therapeutically effective or prophylactically effective dosage required to prevent, counteract, or prevent the progression of the condition, clinicians with ordinary skills may fully consider these factors. It should be understood that a specific daily dosage can be both a therapeutically effective dosage (e.g., for the treatment of an oncological condition) and a prophylactically effective dosage (e.g., for the prevention of an oncological condition).

[0181] Although individual needs vary, it is a technique in the art to determine the optimal range of the effective amount of the compound of the present disclosure. For example, in order to be administered to a person in a curative or preventive treatment of the illness and disorder identified herein, the typical dose of the compound of the present disclosure can be about 0.05 mg / kg / day to about 50 mg / kg / day. In certain embodiments, the compound of the present disclosure of about 5 mg / day to about 120 mg / day is administered to the patient, such as 10 mg / day, 20 mg / day, 30 mg / day, 40 mg / day, 50 mg / day, 60 mg / day, 70 mg / day, 80 mg / day, mg / day, 90 mg / day or 100 mg / day. In certain embodiments, the compound of the present disclosure of about 0.2 mg / kg to about 5 mg / kg is administered to the patient, such as 0.5 mg / kg, 0.75 mg / kg, 1.0 mg / kg, 1.25 mg / kg or 1.5 mg / kg. Such a dosage can be administered in a single dose or can be divided into multiple doses.

[0182] Pharmaceutical composition

[0183] The compounds of the present disclosure and their pharmaceutically acceptable salts can be administered to animals, preferably mammals, and particularly humans, as drugs themselves, in mixtures with each other, or in the form of pharmaceutical compositions. The term "subject" or "patient" includes animals, preferably mammals and particularly humans, that use the active agents of the present invention to prevent or treat medical conditions. Administration of drugs to subjects includes both self-administration and administration to patients by another person. The subject may need or desire to treat an existing disease or medical condition, or may need or desire preventive treatment to prevent a disease or medical condition or reduce the risk of its occurrence. The subject of "need" treatment of an existing condition or preventive treatment as used herein covers both the judgment of a medical professional on the need and the patient's desire for such treatment.

[0184] Therefore, the present disclosure also provides compounds of the present disclosure and pharmaceutically acceptable salts thereof for use as drugs, their use in regulating the activity of cytokine IL-1β, and in particular their use in treating and preventing the following diseases or disorders and their use in preparing drugs for these purposes. In certain embodiments, the compounds of the present disclosure and pharmaceutically acceptable salts thereof capture IL-1β.

[0185] In addition, the present disclosure provides pharmaceutical compositions comprising, as active ingredients, an effective dose of at least one compound of the present disclosure and / or a pharmaceutically acceptable salt thereof and conventional pharmaceutically acceptable carriers, i.e., one or more pharmaceutically acceptable carrier substances and / or additives.

[0186] Therefore, the present disclosure provides, for example, the compounds and pharmaceutically acceptable salts thereof for use as pharmaceutical compositions, the pharmaceutical compositions comprising, as active ingredients, an effective dose of the compounds of the present disclosure and / or pharmaceutically acceptable salts thereof and conventional pharmaceutically acceptable carriers, as well as the use of the compounds and / or pharmaceutically acceptable salts thereof in treating or preventing the following diseases or disorders (e.g., atherosclerosis), and their use in preparing medicaments for these purposes.

[0187] The pharmaceutical compositions according to the present disclosure can be administered orally, for example, in the form of pills, tablets, lacquered tablets, sugar-coated tablets, granules, hard and soft gelatin capsules, aqueous, alcoholic or oily solutions, syrups, emulsions or suspensions, or rectally, for example, in the form of suppositories. They can also be administered parenterally, for example, subcutaneously, intramuscularly or intravenously in the form of injection or infusion solutions.

[0188] Other suitable administration forms are, for example, transdermal or topical administration, for example in the form of ointments, tinctures, sprays or transdermal therapeutic systems, or, for example, microcapsules, implants or rods. The preferred administration form depends, for example, on the disease to be treated and its severity.

[0189] The present disclosure also provides pharmaceutical compositions comprising compounds of formula (I). Compounds of formula (I) can be used in combination with any suitable pharmaceutical carrier or excipient. Such pharmaceutical compositions comprise a therapeutically effective amount of one or more compounds of formula (I) and a pharmaceutically acceptable excipient and / or carrier. Specific pharmaceutical compositions will be suitable for the mode of administration. In particular aspects, the pharmaceutically acceptable carrier may be water or a buffer solution.

[0190] The excipients included in the pharmaceutical composition have different purposes, depending on, for example, the properties of the drug and the mode of administration. Examples of commonly used excipients include, but are not limited to: saline, buffered saline, dextrose, water for injection, glycerol, ethanol, and combinations thereof, stabilizers, solubilizers and surfactants, buffers and preservatives, tonicity agents, fillers, lubricants (such as talc or silicon dioxide, and fats such as vegetable stearin, magnesium stearate or stearic acid), emulsifiers, suspending agents or viscosity agents, inert diluents, fillers (such as cellulose, calcium hydrogen phosphate, vegetable fats and oils, lactose, sucrose, glucose, mannitol, sorbitol, calcium carbonate and magnesium stearate), disintegrants (such as cross-linked polyvinyl pyrrolidone, sodium starch glycolate, cross-linked sodium carboxymethyl cellulose), binders (such as starch, gelatin, cellulose, methylcellulose, The invention also includes but is not limited to cellulose or modified cellulose such as microcrystalline cellulose, hydroxypropyl cellulose, sugars such as sucrose and lactose, or sugar alcohols such as xylitol, sorbitol or maltitol, polyvinyl pyrrolidone and polyethylene glycol), wetting agents, antibacterial agents, chelating agents, coating agents (such as cellulose film coatings, synthetic polymers, shellac, zein, zein or other polysaccharides and gelatin), preservatives (including vitamin A, vitamin E, vitamin C, retinyl palmitate, selenium, cysteine, methionine, citric acid and sodium citrate, and synthetic preservatives (including methylparaben and propylparaben), sweeteners, aromas, flavoring agents, coloring agents, absorption enhancers, administration aids, and combinations thereof.

[0191] Carrier is the compound and material that improves and / or prolongs the delivery of active ingredient to the subject under the background of pharmaceutical composition.Using controlled release technology, carrier can be used to prolong the in vivo activity of drug or slow down the release of drug in subject.Carrier can also reduce drug metabolism and / or reduce the toxicity of drug in subject.Carrier can also be used to target drug delivery to specific cells or tissues in subject.Common carrier (hydrophilic and hydrophobic carrier both) include fat emulsion, lipid, PEGylated phospholipid, PEGylated liposome, PEGylated liposome wrapped with cyclic RGD peptide by PEG spacer, liposome and lipid sphere, microsphere (including those made of biodegradable polymer or albumin), polymer matrix, biocompatible polymer, protein-DNA complex, protein conjugate, erythrocyte, vesicle, nanoparticle and the side chain for hydrocarbon stapled.Aforesaid carrier can also be used to increase the cell membrane permeability of the compound of formula (I). In addition to their use in the pharmaceutical compositions of the present disclosure, the vectors may also be used in compositions for other uses, such as in vitro research uses (eg, for delivery to cultured cells) and / or in vivo research uses.

[0192] Pharmaceutical compositions suitable for oral administration can be presented as discrete units such as capsules or tablets; powders or granules; solutions, syrups or suspensions (in aqueous or non-aqueous liquids; or edible foams or whips; or emulsions). Suitable excipients for tablets or hard gelatin capsules include lactose, corn starch or its derivatives, stearic acid or its salts. Suitable excipients for use with soft gelatin capsules include, for example, vegetable oils, waxes, fats, semisolid or liquid polyols, etc. For the preparation of solutions and syrups, excipients that can be used include, for example, water, polyols and sugars. For the preparation of suspensions, oils (e.g., vegetable oils) can be used to provide oil-in-water or water-in-oil suspensions. Excipients that promote absorption from the gastrointestinal tract, for example, penetration enhancers, such as sodium caprate, can be included. In some cases, delayed release preparations may be advantageous, and compositions that can deliver the compounds of the present disclosure in a delayed or controlled release manner may also be prepared. Prolonged gastric residence poses problems for degradation by enzymes present in the stomach, and therefore enteric-coated capsules may also be prepared by standard techniques in the art, in which the active substance is released lower in the gastrointestinal tract.

[0193] Pharmaceutical compositions suitable for transdermal administration may be presented as discrete patches intended to remain in intimate contact with the epidermis of the recipient for a prolonged period of time. For example, the active ingredient may be delivered from the patch by iontophoresis as generally described in Pharmaceutical Research, 3(6):318 (1986).

[0194] The pharmaceutical composition suitable for topical application can be formulated as an ointment, cream, suspension, lotion, powder, solution, paste, gel, spray, aerosol or oil. When formulated in the form of an ointment, the active ingredient can be used together with a paraffinic or water-miscible ointment base. Alternatively, the active ingredient can be formulated in an ointment together with an oil-in-water cream base or a water-in-oil base. Pharmaceutical compositions suitable for topical application to the eye include eye drops, in which the active ingredient is dissolved or suspended in a suitable carrier (especially an aqueous solvent). Pharmaceutical compositions suitable for topical application in the oral cavity include lozenges, pastilles and mouthwashes.

[0195] Pharmaceutical compositions adapted for rectal administration may be presented as suppositories or as enemas.

[0196] Pharmaceutical compositions suitable for nasal administration (wherein the carrier is a solid) include coarse powders having a particle size, for example, in the range of 20 to 500 microns, which are administered by snorting, i.e., by holding the container containing the powder up close to the nose and rapidly inhaling through the nasal cavity. For administration as a nasal spray or as nose drops, suitable compositions (wherein the carrier is a liquid) include aqueous or oily solutions of the active ingredient.

[0197] Pharmaceutical compositions suitable for administration by inhalation include fine particle dusts or mists which can be generated by various types of metered dose pressurized aerosols, nebulizers or insufflators.

[0198] Pharmaceutical compositions adapted for vaginal administration may be presented as pessaries, tampons, creams, gels, pastes, foams or spray formulations.

[0199] Pharmaceutical compositions suitable for parenteral administration include aqueous and non-aqueous sterile injection solutions, which may contain antioxidants, buffers, bacteriostats, and solutes that make the preparations substantially isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions, which may include suspending agents and thickeners. Excipients that can be used for injection solutions include, for example, water for injection, alcohols, polyols, glycerol, and vegetable oils. The composition can be presented in unit dose or multiple dose containers (e.g., sealed ampoules and vials), and can be stored under freeze drying (lyophilization) conditions, requiring only the addition of a sterile liquid carrier, such as water or saline for injection, just before use. Ready-to-use injection solutions and suspensions can be prepared from sterile powders, granules, and tablets. The pharmaceutical composition may contain preservatives, solubilizers, stabilizers, wetting agents, emulsifiers, sweeteners, colorants, spices, salts (the substance of the present disclosure itself may be provided in the form of a pharmaceutically acceptable salt), buffers, coating agents, or antioxidants. In addition to the compounds of the disclosure, they may contain therapeutically active agents.

[0200] Methods of using the compounds of the present disclosure

[0201] The present application provides a method for IL-1 mediated cell signaling, comprising contacting a cell with a compound of the present disclosure or a pharmaceutically acceptable salt thereof. The inhibition of IL-1 mediated cell signaling can be assessed by detecting a decrease in the levels of downstream biomarkers IL-6 and CRP (e.g., hsCRP).

[0202] The present application also provides methods of using the compounds of the present disclosure (or pharmaceutically acceptable salts thereof) or pharmaceutical compositions containing such compounds to treat disease conditions, including but not limited to conditions associated with IL-1β.

[0203] In certain embodiments, the present disclosure provides a method for treating cardiovascular disease, the method comprising administering to a subject in need of such treatment a therapeutically effective amount of a compound of the present disclosure (or a pharmaceutically acceptable salt thereof) or any of the aforementioned pharmaceutical compositions comprising such a compound. In certain embodiments, the cardiovascular disease is vascular inflammation. In certain embodiments, the cardiovascular disease is atherosclerosis. In certain embodiments, the cardiovascular disease is heart failure with normal ejection fraction (HFpEF). In other embodiments, the cardiovascular disease is heart failure with reduced ejection fraction (HFrEF).

[0204] In certain embodiments, the present disclosure provides methods for treating chronic kidney disease, comprising administering to a subject in need of such treatment a therapeutically effective amount of a compound of the present disclosure (or a pharmaceutically acceptable salt thereof) or any of the foregoing pharmaceutical compositions comprising such a compound.

[0205] In certain embodiments, the present disclosure provides a method of treating an inflammatory lesion, the method comprising administering to a subject in need of such treatment a therapeutically effective amount of a compound of the present disclosure (or a pharmaceutically acceptable salt thereof) or any of the aforementioned pharmaceutical compositions comprising such a compound. In certain embodiments, the inflammatory lesion is selected from hidradenitis suppurativa (acne inversa), inflammatory bowel disease, arthritis, and nonalcoholic steatohepatitis (NASH).

[0206] In certain embodiments, the inflammatory lesion is hidradenitis suppurativa (acne inversa).

[0207] In certain embodiments, the inflammatory disorder is inflammatory bowel disease, such as Crohn's disease or ulcerative colitis.

[0208] In certain embodiments, the inflammatory disorder is arthritis, e.g., osteoarthritis, rheumatoid arthritis, psoriatic arthritis, or gouty arthritis.

[0209] In other embodiments, the inflammatory disorder is nonalcoholic steatohepatitis (NASH).

[0210] Combination therapy

[0211] One or more additional pharmacologically active agents may be administered in combination with the compounds of the present disclosure. Additional one or more active agents are intended to represent one or more pharmaceutically active agents that are active in vivo, including prodrugs that are converted into pharmaceutically active forms after administration, which are different from the compounds of Formula I, and also include free acids, free bases, and pharmaceutically acceptable salts of the additional active agents. Typically, any suitable additional one or more active agents, including, but not limited to, antihypertensive agents, antiatherosclerotic agents (such as lipid regulating compounds), antidiabetic agents, and / or antiobesity agents, anti-inflammatory agents, may be used in any combination with the compounds of the present disclosure in a single dose formulation (fixed dose drug combination), or may be administered to a subject in one or more separate dose formulations that allow for concurrent or sequential administration of the active agents (co-administration of separate active agents).

[0212] Examples of additional active agents that can be used to treat cardiovascular disorders include, but are not limited to, angiotensin converting enzyme inhibitors (e.g., alacepril, benazepril, captopril, ciropril, cilazapril, delapril, enalapril, enalaprilat, fosinopril, imidapril, lisinopril, mavipril, perindopril, quinapril, ramipril, spirapril, temocapril or trandolapril), angiotensin II receptor antagonists (e.g., losartan, i.e., Valsartan (including combinations with sacubitril), candesartan, olmesartan, telmesartan, and any of these drugs used in combination with hydrochlorothiazide, such as ), sGC activators (e.g., riociguat and vericipamidol), PCSK9 inhibitors (e.g., evolocumab, alirocumab, MK-0616, and those disclosed in WO2019 / 246349), neutral endopeptidase inhibitors (e.g., thiomethyloxyphenylalanine and phospho-dipeptide), aldosterone antagonists, aldosterone synthase inhibitors, renin inhibitors, endothelin receptor antagonists, phosphodiesterase-5 inhibitors (e.g., sildenafil, tadalafil, and vardenafil), vasodilators, calcium channel blockers (e.g., amlodipine, nifedipine, verapamil, diltiazem, gallopamil, niludipine, nimodipine, , nicardipine), potassium channel activators (e.g., nicorandil, pinacidil, chromankalim, minoxidil, alprakalim, loprazolam), diuretics (e.g., hydrochlorothiazide), sympatholytics, beta-adrenergic blocking agents (e.g., propranolol, atenolol, bisoprolol, carvedilol, metoprolol or metoprolol tartrate), alpha-adrenergic blocking agents (e.g., doxazosin, prazosin or alpha methyldopa), central alpha-adrenergic agonists, peripheral vasodilators (e.g., hydralazine); lipid-lowering agents, for example, HMG-CoA reductase inhibitors such as simvastatin and lovastatin, which are in the form of lactone prodrugs as and and pharmaceutically acceptable salts of dihydroxyopenic acid HMG-CoA reductase inhibitors, such as atorvastatin (particularly as calcium salts sold), rosuvastatin (especially calcium salts sold), pravastatin (especially sodium salt), fluvastatin (especially sodium salt), crovastatin and pitavastatin; cholesterol absorption inhibitors such as ezetimibe and with any other lipid-lowering agent (such as the above-mentioned HMG-CoA reductase inhibitors) and in particular with simvastatin or ezetimibe in combination with atorvastatin calcium; nicotinic acid in immediate or controlled release form and / or with an HMG-CoA reductase inhibitor; nicotinic acid receptor agonists such as acipimox and acifuran, and nicotinic acid receptor partial agonists; metabolic altering agents, including insulin and insulin mimetics (e.g., degludec, glargine, lispro), dipeptidyl peptidase-IV (DPP-4) inhibitors (e.g., sitagliptin, alogliptin, linagliptin, vildagliptin); insulin sensitizers, including (i) P PARy agonists, such as glitazones (e.g., pioglitazone, miglitazone, lobeglitazone, rosiglitazone and balaglitazone) and other PPAR ligands, including (1) PPARα / γ dual agonists (e.g., sitaglitazar, muraglitazar, aleglitazar, soglitazar and naglitazar); (2) PPARα agonists such as fenofibric acid derivatives (e.g., gemfibrozil, clofibrate, ciprofibrate, fenofibrate, bezafibrate), (3) selective PPARγ modulators (SPPARγM's) (e.g., such as in WO 02 / 060388, WO 02 / 08188, WO 2004 / 019869, WO 2004 / 020409, WO 2004 / 020408 and WO 2004 / 066963); and (4) PPARγ partial agonists; (ii) biguanides, such as metformin and its pharmaceutically acceptable salts, especially metformin hydrochloride, and its extended release formulations, such as Glumetza TM 、Fortamet TM and GlucophageXR TMand (iii) protein tyrosine phosphatase-1B (PTP-1B) inhibitors; insulin or insulin analogs (e.g., insulin detemir, insulin glulisine, insulin degludec, insulin glargine, insulin lispro and inhaled formulations of each); leptin and leptin derivatives and agonists; amylin and amylin analogs (e.g., pramlintide); sulfonylurea and non-sulfonylurea insulin secretagogues (e.g., tolbutamide, glyburide, glipizide, glimepiride, mitiglinide, meglitinides, nateglinide and repaglinide); α-glucosidase inhibitors (e.g., acarbose, voglibose, and miglitol); glucagon receptor antagonists; incretin mimetics, such as GLP-1, GLP-1 analogs, derivatives, and mimetics; and GLP-1 receptor agonists (e.g., dulaglutide, semaglutide, albiglutide, exenatide, liraglutide, lixisenatide, tapoglutide, including intranasal, transdermal, and once-weekly formulations thereof); bile acid masking agents (e.g., colestilan, colestilide, colesevelam hydrochloride, colesti The invention also includes dialkylaminoalkyl derivatives of styrofoam, cholestyramine, and cross-linked dextran), acyl-CoA:cholesterol acyltransferase inhibitors (e.g., avasimibe); anti-obesity compounds; agents intended for use in inflammatory conditions, such as aspirin, nonsteroidal anti-inflammatory drugs or NSAIDs, glucocorticoids, and selective cyclooxygenase-2 or COX-2 inhibitors; glucokinase activators (GKA); inhibitors of 11β-hydroxysteroid dehydrogenase type 1 (e.g., such as those disclosed in U.S. Pat. No. 6,730,690); fructose 1,6-bisphosphatase inhibitors of acetyl-CoA carboxylase-1 or 2 (ACC1 or ACC2); AMP-activated protein kinase (AMPK) activators; other agonists of G protein-coupled receptors: (i) GPR-109, (ii) GPR-119, and (iii) GPR-40; SSTR3 antagonists (e.g., such as those disclosed in WO 2009 / 001836); neuromedin U receptor agonists (e.g., such as those disclosed in WO 2009 / 042053, including, but not limited to, neuromedin S (NMS)); SCD modulators; GPR-105 antagonists (e.g., such as those disclosed in WO 2009 / 000087); SGLT inhibitors (e.g., empagliflozin, dapagliflozin, canagliflozin, erpagliflozin, remogloflozin, topagliflozin and empagliflozin); inhibitors of acyl-CoA: diacylglycerol acyltransferase 1 and 2 (DGAT-1 and DGAT-2); inhibitors of fatty acid synthase;Inhibitors of acyl-CoA: monoacylglycerol acyltransferase 1 and 2 (MGAT-1 and MGAT-2); agonists of TGR5 receptor (also known as GPBAR1, BG37, GPCR19, GPR131 and M-BAR); ileal bile acid transporter inhibitors; PACAP, PACAP mimetics and PACAP receptor 3 agonists; PPAR agonists; protein tyrosine phosphatase-1B (PTP-1B) inhibitors; IL-1β antibodies (e.g., givolizumab and canakinumab); and bromocriptine mesylate and its rapid release formulations; or in combination with other drugs that are beneficial for treating the above-mentioned conditions or disorders (including chemically feasible free acid, free base and pharmaceutically acceptable salt forms of the above-mentioned active agents). ;

[0213] Examples of additional active agents that can be used to treat inflammatory lesions include, but are not limited to, steroidal and nonsteroidal anti-inflammatory agents, glucocorticoids, and therapeutic hormones. In a specific embodiment, in the treatment of hidradenitis suppurativa (acne inverse), the additional active agent can be an antibiotic, an injectable steroid, a therapeutic hormone, a TNF inhibitor (e.g., infliximab, adalimumab, etanercept, golimumab, certolizumab), an analgesic (e.g., codeine, hydrocodone, morphine, pregabalin, gabapentin, intralesional triamcinolone, corticosteroids, naproxen, ketoprofen, diclofenac, ibuprofen, acetaminophen). In other embodiments, in the treatment of inflammatory bowel disease, the additional active agent can be methotrexate, a TNF inhibitor, an oral sphingosine 1-phosphate receptor modulator (e.g., fingolimod, siponimod, ozanimod, pernaimod) or a selective JAK inhibitor (e.g., tofacitinib, baricitinib, upadacitinib). In certain embodiments, in the treatment of osteoarthritis, the additional active agent can be an analgesic (examples listed above). In other embodiments, in the treatment of gouty arthritis, the additional active agent can be colchicine, a nonsteroidal anti-inflammatory agent or a glucocorticoid.

[0214] Methods of preparing compounds of the present disclosure

[0215] The compounds described herein can be prepared according to the procedures of the following schemes and examples using appropriate materials, and are further illustrated by the following specific examples. The examples also include methods for testing such compounds in cell assays. However, the compounds shown in the examples should not be construed as forming the only species considered to be present disclosure.

[0216] The examples further illustrate the preparation details of the compounds of the present disclosure. It will be readily appreciated by those skilled in the art that known changes in the conditions and processes of the following preparation operations can be used to prepare these compounds. For example, in some cases, the order of the steps of executing the reaction scheme can be changed to promote the reaction or avoid undesirable reaction products. The starting materials and intermediates of the final compounds are purchased, prepared by known procedures, or as otherwise indicated. The examples are provided only for the purpose of further illustration and are not intended to limit the present disclosure.

[0217] NMR data were obtained in CDCl3, DMSO-d6 or methanol-d4 on a 300 MHz or 400 MHz instrument and chemical shifts are reported relative to a tetramethylsilane standard. Resonance signals are reported using the following abbreviations: s = singlet, d = doublet, t = triplet, q = quartet, dd = doublet of doublets, m = multiplets or overlaps of non-equivalent resonances. Coupling constants (J) are reported in Hertz (Hz).

[0218] Throughout the synthetic schemes and examples, unless otherwise indicated, abbreviations and acronyms may be used with the following meanings:

[0219] abbreviation

[0220]

[0221]

[0222]

[0223]

[0224]

[0225]

[0226] Intermediate synthesis:

[0227] Synthesis Scheme 1

[0228]

[0229] 1Inda

[0230]

[0231] (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(1H-indazol-1-yl)propanoic acid

[0232] Step 1: DMSO (150 mL), 1H-indazole (15 g, 127 mmol) and (S)-2-amino-3-hydroxypropionic acid (14.01 g, 133 mmol) and 0.2 M potassium phosphate buffer (600 mL, 127 mmol) at pH 8 were placed in a 2-L 4-neck round-bottom flask purged and maintained with inert nitrogen at 65°C. [(4-formyl-5-hydroxy-6-methylpyridin-3-yl)methoxy]phosphonic acid (PLP) (96 mg, 0.00284 eq.) in H2O (2 mL) and PfTrpB-0B2 (1.05 g, 10 wt %) in 0.2 M potassium phosphate buffer (2 mL) were added to the solution. The resulting solution was stirred at 65°C for 16 h. The reaction progress was monitored by HPLC-MS.

[0233] Step 2: Na2CO3 (21.55 g, 2.0 eq.) and Fmoc-OSu (37.68 g, 1.1 eq.) in THF (150 mL) were added to the resulting solution at 0°C. The solution was then stirred at 25°C for 2 h. The progress of the reaction was monitored by HPLC-MS. The solution was adjusted to pH 3 with hydrogen chloride (6 M). The solution was extracted with EtOAc (3 x 500 mL). The organic layers were combined and washed with H2O (2 x 1 L) and NaCl (1 x 1 L). The organic layer was dried over anhydrous sodium sulfate and concentrated under vacuum. The crude product was slurried with DCM (15 V) and the solid was collected by filtration. HPLC-MS: (ESI, m / z): 428 [M+H]+. 1 HNMR: (400MHz, DMSO-d6, ppm): δ13.08(s,1H),8.10(d,J=1.0Hz,1H),7.87(dt,J=7.7,1. 0Hz,2H),7.81-7.71(m,2H),7.71-7.65(m,1H),7.58(d,J=7.5Hz,1H),7.54(d,J=7.5Hz, 1H),7.41(dtd,J=6.0,4.4,4.3,3.2Hz,2H),7.40-7.33(m,1H),7.36-7.24(m,1H),7.29- 7.20(m,1H),7.16-7.08(m,1H),4.83-4.65(m,2H),4.61-4.51(m,1H),4.22-4.06(m,3H).

[0234] Synthesis Scheme 2

[0235]

[0236] 1Indolin

[0237]

[0238] (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(indolin-1-yl)propanoic acid

[0239] Step 1: Indoline (125 g, 1.05 mol, 1.00 equiv), DMSO (625 mL), 0.2 M potassium phosphate buffer (5625 mL) at pH 8 and PLP (778.32 mg, 2.94 mmol), L-serine (115.75 g, 1.10 mol, 1.05 equiv) and PfTrpB-4D11 (12.50 g) were added to a 10 L 3-neck round bottom flask at 25 ° C. The resulting solution was stirred at 65 ° C for 4 h under nitrogen. The reaction progress was monitored by HPLC-MS.

[0240] Step 2: Na2CO3 (129 g, 2.00 equiv.), Fmoc-OSu (306 g, 1.50 equiv.) and THF (2.5 L) were added to the above solution at 0 °C. The reaction mixture was stirred at 25 °C for 3 h under nitrogen. The solution was acidified to pH 3 with 6M HCl. The mother liquor was collected by filtration and then extracted with EtOAc (3 x 1 L). The organic layers were combined and washed with H2O (2 x 800 mL) and NaCl (1 x 800 mL), dried over Na2SO4 and concentrated under vacuum. The residue was purified by silica gel column chromatography, eluted with PE / EtOAc (3: 1) to provide a crude product. The crude product was then purified by preparative reverse phase HPLC (C18), eluted with 50% to 70% gradient of acetonitrile / water + 0.05% TFA for 30 min. The mixture was concentrated under vacuum at 20 °C to remove MeCN, then extracted with EtOAc (3x 1L), dried over Na2SO4 and concentrated under vacuum. The pure product was slurried in hexanes (1L) for 30 min. The solid was collected by filtration. This produces (2S)-3-(2,3-dihydroindole-1-yl)-2-[[(9H-fluorenes-9-ylmethoxy)carbonyl]amino]propionic acid. HPLC-MS: (ESI, m / z): 429[M+H]+. 1 H NMR: (400MHz, DMSO-d6, ppm) δ7.89 (d, J=7.5Hz, 2H), 7.74-7.67 (m, 2H), 7.41 (t, J=7.5Hz, 2H), 7.35-7.25 (m, 2H), 7 .00(ddd,J=17.4,7.5,1.3Hz,2H),6.61-6.48(m,2H),4.35-4.18(m,4H),3.47-3.28(m,4H),2.87(t,J=8.2Hz,2H).

[0241] Synthesis Scheme 3

[0242]

[0243] 3AzaPhe4AcPip

[0244]

[0245] (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(6-(4-acetylpiperazin-1-yl)pyridine- 3-amino)propionic acid

[0246] Step 1: To a mixture of 1-(piperazin-1-yl)ethane-1-one (21.85 g, 170 mmol) in DMF (150 mL) was added 5-bromo-2-fluoropyridine (15 g, 85 mmol) under argon at room temperature. The reactant was stirred at 100 ° C for 2 h, then diluted with 300 mL EtOAc and washed with H2O (3 x 80 mL), saturated aqueous NaCl solution (80 mL), dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with a gradient of 0%-100% EtOAc in PE. The fractions containing the desired product were combined and concentrated under reduced pressure to provide 1-(4-(5-bromopyridin-2-yl)piperazin-1-yl)ethane-1-one. C11H15BrN3O[M+H] + The MS ESI calculated values ​​were 284.03 and 286.03, and the found values ​​were 283.90 and 285.90.

[0247] Step 2: A mixture of nickel(II) chloride ethylene glycol dimethyl ether complex (0.696 g, 3.17 mmol) and 1,10-phenanthroline (0.571 g, 3.17 mmol) in DMA (2 mL) was heated at 50°C for 0.5 h. A mixture of 1-(4-(5-bromopyridin-2-yl)piperazin-1-yl)ethan-1-one (4.5 g, 15.84 mmol), (R)-tert-butyl 2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-bromopropanoate (7.78 g, 17.42 mmol) and tetrabutylammonium iodide (5.85 g, 15.84 mmol) in DMA (2 mL) was added at 25°C. Zinc powder (2.071 g, 31.7 mmol) was then added. The resulting mixture was stirred at 50°C for 1 h. The resulting mixture was poured into water (300 mL) and extracted with EtOAc (3 x 300 mL). The organic layer was washed with water (100 mL) and brine (2 x 80 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel, eluted with DCM-MeOH (10: 1) to provide (S)-2-((((9H-fluorenes-9-yl)methoxy)carbonyl)amino)-3-(6-(4-acetylpiperazine-1-yl)pyridin-3-yl)tert-butyl propionate. C33H39N4O5[M+H]+ The MS ESI calculated value was 571.28 and the found value was 571.40.

[0248] Step 3: To a mixture of (S)-tert-butyl 2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(6-(4-acetylpiperazin-1-yl)pyridin-3-yl)propanoate (7.3 g, 12.79 mmol) in DCM (70 mL) was added TFA (70 mL, 909 mmol) at room temperature under argon. The reaction was stirred at room temperature for 1 h and then concentrated under reduced pressure. The residue was dissolved in THF (20 mL) and the resulting mixture was purified by reverse phase Combi-Flash using the following conditions: column C18 silica gel column (330 g), 20-35 μm; mobile phase A: 5 mM TFA in water; mobile phase B: MeCN; (gradient: 0% B for 10 min, increase to 42.3% B in 35 min, 42.3% B for 3.2 min; increase to 95% B in 2 min, 95% B for 10 min); flow rate: 60 mL / min; detector: UV254 and 210 nm; RT: 35.32 min. The fractions containing the product were collected and concentrated under vacuum to provide (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(6-(4-acetylpiperazin-1-yl)pyridin-3-yl)propanoic acid. C29H31N4O5[M+H] + The MS ESI calculated value was 515.22 and the found value was 515.15. 1 H NMR (300 MHz, methanol-d4) δ 7.87-7.79 (m, 4H), 7.62-7.55 (m, 2H), 7.42-7.27 (m, 4H), 7.11-7.09 (m, 1H), 4.51-4.46 (m, 1H), 4.32-4.09 (m, 3H), 3.65-3.54 (m, 8H), 3.29-3.20 (m, 1H), 2.94-2.89 (m, 1H), 2.12 (s, 3H).

[0249] Synthesis Scheme 4

[0250]

[0251] 3AzaTyrEtNAc

[0252]

[0253] (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(6-(2-acetylaminoethoxy)pyridine-3- 4-(4-amino)propionic acid

[0254] Step 1: under argon at room temperature to N- (2-hydroxyethyl) acetamide (17.58g, 170mmol) in t-BuOH (150mL) mixture, add 5-bromo-2-fluoropyridine (15g, 85mmol), followed by potassium tert-butoxide (19.13g, 170mmol). The reactant was stirred at room temperature for 1h, then concentrated under reduced pressure. The residue was diluted with 500mL EtOAc, and washed with saturated NaHCO3 aqueous solution (3x 250mL), saturated NaCl aqueous solution (250mL), dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure to provide crude N- (2- ((5-bromopyridin-2-yl) oxygen base) ethyl) acetamide. C9H12BrN2O2[M+H] + The MS ESI calculated values ​​were 259.00 and 261.00, and the found values ​​were 258.90 and 261.90.

[0255] Step 2: A mixture of nickel(II) chloride ethylene glycol dimethyl ether complex (1.187 g, 5.40 mmol) and 1,10-phenanthroline (0.974 g, 5.40 mmol) in DMA (70 mL) was heated at 50°C for 0.5 h. A mixture of N-(2-((5-bromopyridin-2-yl)oxy)ethyl)acetamide (7 g, 27.0 mmol), (R)-tert-butyl 2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-bromopropanoate (13.26 g, 29.7 mmol) and tetrabutylammonium iodide (9.98 g, 27.0 mmol) in DMA (70 mL) was added at 25°C. Zinc powder (3.53 g, 54.0 mmol) was then added and the resulting mixture was stirred at 50°C for 1 h. The reaction mixture was diluted with 300mL EtOAc and washed with saturated NaHCO aqueous solution (3x 80mL), brine (80mL), dried over NaSO and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with a gradient of 0%-20% DCM in MeOH. The fractions containing the desired product were combined and concentrated under reduced pressure to provide (S)-2-((((9H-fluorenes-9-yl)methoxy)carbonyl)amino)-3-(6-(2-acetamidoethoxy)pyridin-3-yl)tert-butyl propionate. C31H36N3O6[M+H] + The MS ESI calculated value was 546.25 and the found value was 546.40.

[0256] Step 3: To a mixture of (S)-tert-butyl 2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(6-(2-acetamidoethoxy)pyridin-3-yl)propanoate (8 g, 14.66 mmol) in DCM (80 mL) was added TFA (80 mL, 1038 mmol) at room temperature under argon. The reaction was stirred at room temperature for 1 h and then concentrated under reduced pressure. The residue was purified by RP flash column chromatography using the following conditions: Column: C18 silica gel column (330 g), 20-35 μm; Mobile phase A: 5 mM NH4HCO3 aqueous solution; Mobile phase B: MeCN; (Gradient: 0% B for 5 min, increase to 45% B in 20 min, 45% B for 10 min; increase to 95% B in 15 min, 95% B for 10 min); Flow rate: 60 mL / min; Detector: UV 254 and 210 nm; RT: 35.32 min. The fractions containing the product were collected and concentrated under reduced pressure to give (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(6-(2-acetamidoethoxy)pyridin-3-yl)propanoic acid. C27H28N3O6[M+H] + The MS ESI calculated value was 490.19 and the found value was 490.10. 1 H NMR (400 MHz, methanol-d4) δ 8.02-8.00 (m, 1H), 7.79-7.77 (m, 2H), 7.67-7.64 (m, 1H), 7.61-7.57 (m, 2H), 7.40-7.36 (m, 2H), 7.31-7.27 (m, 2H), 6.82-6.80 (m, 1H), 4.42-4.39 (m, 1H), 4.33-4.22 (m, 4H), 4.16-4.13 (m, 1H), 3.53-3.51 (m, 2H), 3.20-3.15 (m, 1H), 2.93-2.87 (m, 1H), 1.92 (s, 3H).

[0257] Synthesis Scheme 5

[0258]

[0259] 3Pal4Ph4CO2H

[0260]

[0261] (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(6-(4-(tert-butoxycarbonyl)phenyl)pyrrolidone pyridin-3-yl)propionic acid

[0262] Step 1: To a stirred solution of NiCl2-ethylene glycol dimethyl ether (0.918 g, 4.18 mmol) in DMA (100 mL) was added 1,10-phenanthroline (0.905 g, 4.18 mmol) at room temperature under nitrogen atmosphere. The resulting solution was stirred at 50 °C for 1 h. 2-Chloro-5-iodopyridine (5 g, 20.88 mmol), (R)-tert-butyl 2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-iodopropanoate (12.36 g, 25.06 mmol), TBAI (8.01 g, 20.88 mmol) and Zn (2.73 g, 41.8 mmol) were added to the above mixture at room temperature and the resulting mixture was stirred at 25 °C for 2 h. The reaction was quenched with H2O (200 mL) and extracted with EtOAc (2 x 500 mL). The combined organic layers were washed with brine (3 x 200 mL), dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography eluting with 0-30% EtOAc in PE to yield (S)-tert-butyl 2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(6-chloropyridin-3-yl)propanoate. 27 H 28 ClN2O4[M+H] + The MS ESI calculated value was 479.17 and the found value was 479.20.

[0263] Step 2: To a stirred solution of (S)-tert-butyl 2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(6-chloropyridin-3-yl)propanoate (5 g, 10.48 mmol) in DCM (5 mL) was added TFA (10 mL) at room temperature. The solution was stirred at 25 °C for 1 h. The solvent was concentrated under reduced pressure and the residue was purified by RP-flash using the following conditions: Column: Flash C18 (330 g); Mobile phase A: water (0.1% TFA), Mobile phase B: MeCN; (Gradient: 5% B for 5 min, increased to 30% B in 15 min, 30% B for 5 min; increased to 95% B in 20 min, 95% B for 10 min); Flow rate: 90 mL / min; Detector: UV 210 nm; RT = 40 min. The fractions containing the product were collected and evaporated in vacuo to yield (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(6-chloropyridin-3-yl)propanoic acid. 23 H 20 ClN2O4[M+H] + The MS ESI calculated value was 423.10 and the found value was 423.10.

[0264] Step 3: To a stirred solution of (S)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(6-chloropyridin-3-yl)propanoic acid (3 g, 7.09 mmol) in THF (25 mL) and water (5 mL) at 25 °C under nitrogen was added (4-(tert-butoxycarbonyl)phenyl)boronic acid (1.890 g, 8.51 mmol) and K3PO4 (7.53 g, 35.5 mmol). The resulting solution was stirred at 25 °C for 10 min. Pd(dtbpf)Cl2 (0.694 g, 1.064 mmol) was added to the solution, and the mixture was then stirred at 60 °C for 16 h. The reaction was cooled to room temperature and quenched with H2O (200 mL), and extracted with EtOAc (2 x 500 mL). The combined organic layers were washed with brine (3 x 200 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by RP-flash using the following conditions: Column: Flash C18 (330 g); Mobile phase A: Water (0.1% TFA), Mobile phase B: MeCN; (Gradient: 5% B for 5 min, increasing to 60% B in 15 min, 60% B for 15 min; increasing to 95% B in 10 min, 95% B for 10 min); Flow rate: 90 mL / min; Detector: UV 210 nm; RT = 55 min. The fractions containing the product were collected and evaporated in vacuo to yield (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(6-(4-(tert-butoxycarbonyl)phenyl)pyridin-3-yl)propanoic acid. C 34 H 33 N2O6[M+H] + The MS ESI calculated value was 565.23 and the found value was 565.15; 1 H NMR (400MHz, methanol-d4) δ8.66(d,J=1.9Hz,1H),8.12-8.07(m,3H),7.99-7.92(m,3H),7.77(d,J=7.5Hz,2H),7.59-7.56(m,2H),7.37-7.33 (m,2H),7.30-7.22(m,2H),4.60-4.56(m,1H),4.29-4.27(m,2H),4.14-4.10(m,1H),3.45-3.41(m,1H),3.14-3.10(m,1H),1.62(s,9H).

[0265] Synthesis Scheme 6

[0266]

[0267] AbuM

[0268]

[0269] (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-morpholinobutanoic acid

[0270] Step 1: To a stirred mixture of ((benzyloxy)carbonyl)-L-homoserine (2.5 g, 9.87 mmol) and NaHCO3 (2.488 g, 29.6 mmol) in DMF (50 mL) at 0°C under nitrogen atmosphere was added benzyl bromide (2.026 g, 11.85 mmol). The mixture was stirred at 25°C for 16 h, then quenched with water (200 mL) and extracted with EtOAc (3 x 200 mL). The combined organic layers were washed with brine (3 x 100 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography eluting with 0-45% EtOAc in PE to provide ((benzyloxy)carbonyl)-L-homoserine benzyl ester. C 19 H 21 NO5Na[M+Na] + The MS ESI calculated value was 366.14 and the found value was 366.15.

[0271] Step 2: Under N2 atmosphere in a two-necked 500mL flask, I2 (22.17g, 87mmol) was added to a solution of PPh3 (17.19g, 65.5mmol) and 1H-imidazole (4.46g, 65.5mmol) in anhydrous CH2Cl2 (100mL) at 0°C. The reactant was warmed to 25°C and stirred for 15min. A solution of ((benzyloxy)carbonyl)-L-homoserine benzyl ester (15g, 43.7mmol) in CH2Cl2 (100mL) was added over 10 minutes, and then stirred at room temperature for 2h. The mixture was washed with saturated Na2S2O3 solution (3x 200mL) and subsequently washed with brine (3x 50mL). The organic layer was dried over Na2SO4, the filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography eluting with 0-40% EtOAc in PE to provide (S)-benzyl 2-(((benzyloxy)carbonyl)amino)-4-iodobutanoate. 19 H 21 INO4[M+H] + The MS ESI calculated value was 454.04 and the found value was 454.05. 1H NMR (400MHz, CDCl3) δ7.45-7.31(m,10H),5.37-5.35(m,1H),5.19-5.15(m,2H),5.11 (s,2H),4.48-4.41(m,1H),3.15-3.10(m,2H),2.46-2.44(m,1H),2.25-2.20(m,1H).

[0272] Step 3: To a stirred mixture of (S)-benzyl 2-(((benzyloxy)carbonyl)amino)-4-iodobutyrate (13.6 g, 30.0 mmol) dissolved in THF (140 mL) was added morpholine (20.91 g, 240 mmol) at room temperature. The resulting mixture was stirred at 50 °C for 16 h. The reaction mixture was concentrated under reduced pressure and the residue was purified by silica gel column chromatography eluting with 0-80% EtOAc in PE, and the fractions containing the product were collected and concentrated under reduced pressure to provide benzyl (S)-2-(((benzyloxy)carbonyl)amino)-4-morpholinobutyrate. 23 H 29 N2O5[M+H] + The MS ESI calculated value was 413.21 and the found value was 413.35. 1 H NMR (400MHz, CDCl3) δ7.36-7.34(m,10H),6.67(s,1H),5.24-5.02(m,4H),4 .52-4.47(m,1H),3.67-3.52(m,4H),2.38-2.51(m,6H),2.15-1.81(m,2H).

[0273] Step 4: (S)-2-(((benzyloxy)carbonyl)amino)-4-morpholinobutyric acid benzyl ester (6.5 g, 15.76 mmol) was dissolved in EtOH (65 mL) and the resulting mixture was evacuated and a nitrogen atmosphere was applied at ambient temperature. Pd-C (2.6 g, dried, 20 wt %) was then added under a nitrogen atmosphere. The suspension was degassed under vacuum and purged with H2 several times, and the reaction solution was stirred at 25 ° C under 1 atmosphere of H2 for 5 h. The suspension was filtered and the filtrate was concentrated under reduced pressure to produce crude (S)-2-amino-4-morpholinobutyric acid. CH 17 N2O3[M+H] + The MS ESI calculated value was 189.12 and the found value was 189.25.

[0274] Step 5: To a solution of (S)-2-amino-4-morpholinobutyric acid (2.97 g, 15.76 mmol) in THF (25 mL) and water (25 mL) was added NaHCO3 (6.63 g, 79 mmol) and Fmoc-OSu (4.85 g, 14.20 mmol) at 25°C. The reaction mixture was stirred at 25°C for 16 h. After completion, the pH of the solution was adjusted to 3 with 1 M HCl. The mixture was purified by reverse phase flash chromatography using the following conditions: C18 column (120 g); mobile phase A: water (0.05% TFA), mobile phase B: MeCN; (gradient: 5% B for 5 min, increased to 38% B in 30 min, 38% B for 2.6 min; increased to 95% B in 2 min, 95% B for 5 min); flow rate: 60 mL / min; detector: UV 210 nm; RT = 31 min. The fractions containing the product were collected and concentrated under reduced pressure to give (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-morpholinobutanoic acid. 23 H 27 N2O5[MH] - The MSESI calculated value is 411.19 and the measured value is 411.30. 1 H NMR (300MHz, methanol-d4) δ7.81-7.79(m,2H),7.68-7.65(m,2H),7.42-7.37(m,2H),7.33-7.29(m,2H),4.48-4. 45(m,2H),4.42-4.23(m,2H),4.01-3.77(m,4H),3.33-3.30(m,2H),3.29-3.14(m,4H),2.45-2.05(m,2H).

[0275] Synthesis Scheme 7

[0276]

[0277] Bip4CO2H

[0278]

[0279] (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4'-(tert-butoxycarbonyl)-[1,1'- phenyl]-4-yl)propanoic acid

[0280] Argon was bubbled through a mixture of (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-bromophenyl)propanoic acid (6 g, 12.87 mmol), (4-(tert-butoxycarbonyl)phenyl)boronic acid (4.29 g, 19.30 mmol) and K3PO4 (8.19 g, 38.6 mmol) in THF (40 mL) for 10 min, followed by the addition of [1,1'-bis(di-tert-butylphosphino)ferrocene]dichloropalladium(II) (0.839 g, 1.287 mmol). After the resulting mixture was stirred at 50° C. for 16 h, it was diluted with EtOAc (300 mL) and washed with saturated aqueous NaHCO3 solution (3× 80 mL), brine (2× 40 mL), dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with a gradient of 0%-50% EtOAc in PE. The fractions containing the product were collected and rotary evaporated in vacuo. The residue was re-purified by combi-Flash with the following conditions: Column: Column: C18 gel column (330 g), 20-35 μm; Mobile phase A: 0.5% TFA in water; Mobile phase B: MeCN; (Gradient: 0% B for 10 min, increasing to 62.3% B in 25 min, 62.3% B for 6.2 min; increasing to 95% B in 2 min, 95% B for 10 min); Flow rate: 90 mL / min; Detector: UV 254 and 210 nm; RT: 32.32 min. The fractions containing the product were collected and concentrated under reduced pressure to afford (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4'-(tert-butoxycarbonyl)-[1,1'-biphenyl]-4-yl)propanoic acid. 35 H 34 NO6[M+1] + The MS ESI calculated value was 564.23 and the found value was 564.15. 1 H NMR (300 MHz, methanol-d4) δ 7.97-7.95 (m, 2H), 7.78-7.76 (m, 2H), 7.61-7.53 (m, 6H), 7.38-7.21 (m, 2H), 4.51-4.11 (m, 4H), 3.32-3.25 (m, 1H), 3.03-2.95 (m, 1H), 1.61 (s, 9H).

[0281] Synthesis Scheme 8

[0282]

[0283] h3Pal5CN

[0284]

[0285] (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-(5-cyanopyridin-3-yl)butanoic acid

[0286] Step 1: To a stirred solution of (((9H-fluoren-9-yl)methoxy)carbonyl)-L-homoserine (15 g, 43.9 mmol) in DCM (200 mL) was added (Z)-tert-butyl N,N'-diisopropylaminocarbamidate (35.2 g, 176 mmol) at room temperature. The solution was stirred at 30 °C for 4 h. The solid was filtered off and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with 0-32% EtOAc in PE to provide (((9H-fluoren-9-yl)methoxy)carbonyl)-L-homoserine tert-butyl ester. 23 H 28 NO5[M+H] + The MS ESI calculated value was 398.19 and the found value was 398.15. 1 H NMR (300MHz, CDCl3) δ7.81-7.74(m,2H),7.64-7.56(m,2H),7.45-7.37(m,2H),7.37-7.28(m,2H),5.61(d,J=7.7Hz, 1H), 4.54-4.32 (m, 3H), 4.22 (t, J = 6.9Hz, 1H), 3.76-3.52 (m, 2H), 2.25-2.07 (m, 1H), 1.71-1.55 (m, 1H), 1.48 (s, 9H).

[0287] Step 2: To a stirred solution of PPh3 (12.67 g, 48.3 mmol) and 1H-imidazole (4.38 g, 64.4 mmol) in DCM (200 mL) was added I2 (12.26 g, 48.3 mmol) at room temperature under nitrogen atmosphere. The mixture was stirred at room temperature for 10 min. (((9H-fluoren-9-yl)methoxy)carbonyl)-L-homoserine tert-butyl ester (12.8 g, 32.2 mmol) was added to the mixture and stirred at 25 °C for 2 h. The solvent was concentrated under reduced pressure and the residue was purified by silica gel column chromatography eluting with 0-18% EtOAc in PE to provide (S)-tert-butyl 2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-iodobutyrate. C 23 H 26 INO4Na[M+Na] + The MS ESI calculated value was 530.09 and the found value was 530.20. 1HNMR (300MHz, CDCl3) δ7.83-7.71(m,2H),7.65-7.56(m,2H),7.47-7.36(m,2H),7.36-7.28(m,2H),5.35(d,J=8.3H z,1H),4.52-4.35(m,2H),4.34-4.16(m,2H),3.21-3.04(m,2H),2.52-2.32(m,1H),2.28-2.07(m,1H),1.48(s,9H).

[0288] Step 3: To a stirred solution of 1,10-phenanthroline (1.083 g, 6.01 mmol) in DMA (300 mL) was added NiCl2-ethylene glycol dimethyl ether (1.321 g, 6.01 mmol) at room temperature under argon atmosphere. The mixture was stirred at 50 °C for 1 h and then cooled to room temperature. Then (S)-tert-butyl 2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-iodobutyrate (15.25 g, 30.1 mmol), 5-bromonicotinonitrile (5.5 g, 30.1 mmol), TBAI (15.25 g, 30.1 mmol) and zinc (3.93 g, 60.1 mmol) were added to the mixture at room temperature and stirred at 35 °C for 2.5 h. The mixture was cooled to room temperature and diluted with water (500 mL) and EtOAc (800 mL). The solid was filtered off and the organic layer was separated. The organic layer was washed with brine (3 x 150 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography eluting with 0-40% EtOAc in PE to provide (S)-tert-butyl 2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-(5-cyanopyridin-3-yl)butanoate. 29 H 30 N3O4[M+H] + The MS ESI calculated value was 484.22 and the found value was 484.20. 1 H NMR (300MHz, CDCl3) δ8.75(s,1H),8.66(s,1H),7.87-7.71(m,3H),7.61(d,J=7.4Hz,2H),7.47-7.28(m,4H),5.40(d,J= 7.8Hz,1H),4.55-4.37(m,2H),4.36-4.18(m,2H),2.85-2.59(m,2H),2.26-2.08(m,1H),2.02-1.82(m,1H),1.49(s,9H).

[0289] Step 4: To a stirred solution of (S)-tert-butyl 2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-(5-cyanopyridin-3-yl)butanoate (7.5 g, 15.51 mmol) in DCM (50 mL) was added TFA (100 mL) at room temperature. The solution was stirred at 25 °C for 3 h and then concentrated under reduced pressure. The residue was purified by RP-flash using the following conditions: C18 column, 330 g, 5%-5% in 10 min, 5%-55% in 40 min, 55%-55% in 10 min of MeCN in water (0.05% TFA) to produce (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-(5-cyanopyridin-3-yl)butanoic acid. 25 H 22 N3O4[M+H] + The MS ESI calculated value was 428.15 and the found value was 428.05. 1 H NMR (400MHz, DMSO-d6) δ12.78(br,1H),8.87(s,1H),8.75(s,1H),8.18(s,1H),7.92-7.90(m,2H),7.76-7. 73(m,3H),7.44-7.33(m,4H),4.39-4.20(m,3H),3.94-3.89(m,1H),2.79-2.67(m,2H),2.09-1.89(m,2H).

[0290] Synthesis Scheme 9

[0291]

[0292] Phe4AcPip

[0293]

[0294] (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-(4-acetylpiperazin-1-yl)phenyl) Propionic acid

[0295] Step 1: To a stirred solution of (S)-3-(4-bromophenyl)-2-((tert-butoxycarbonyl)amino)propanoic acid (6 g, 17.43 mmol) in toluene (180 mL) was added XPhos Pd G2 (2.057 g, 2.61 mmol) at 25 °C under nitrogen. The resulting solution was stirred at 100 °C for 10 min. 1-(Piperazin-1-yl)ethane-1-one (2.234 g, 17.43 mmol) and Cs2CO3 (5.04 g, 26.1 mmol) were added and the resulting solution was stirred at 110 °C for 2 h. The reaction was cooled to room temperature and quenched with H2O (500 mL) and extracted with EtOAc (2x500 mL). The combined organic layers were washed with brine (3x 200 mL), dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography eluting with 0-60% EtOAc in PE to give (S)-3-(4-(4-acetylpiperazin-1-yl)phenyl)-2-((tert-butoxycarbonyl)amino)propanoic acid. 20 H 30 N3O5[M+H] + The MS ESI calculated value was 392.21 and the found value was 392.25. 1 H NMR (400 MHz, methanol-d4) δ 7.12 (d, J = 8 Hz, 2H), 6.89 (d, J = 8 Hz, 2H), 4.16-4.13 (m, 1H), 3.72-3.65 (m, 4H), 3.14-3.04 (m, 4H), 2.93-2.81 (m, 2H), 2.13 (s, 3H), 1.38-1.29 (m, 9H).

[0296] Step 2: To a stirred solution of (S)-3-(4-(4-acetylpiperazin-1-yl)phenyl)-2-((tert-butoxycarbonyl)amino)propanoic acid (10 g, 25.5 mmol) in DCM (30 mL) was added TFA (30 mL) at room temperature. The solution was stirred at 25 °C for 1 h and then concentrated under reduced pressure. The crude (S)-3-(4-(4-acetylpiperazin-1-yl)phenyl)-2-aminopropanoic acid was used directly in the next step without any further purification. 15 H 22 N3O3[M+H] + The MS ESI calculated value was 292.16 and the found value was 292.20.

[0297] Step 3: To a stirred solution of (S)-3-(4-(4-acetylpiperazin-1-yl)phenyl)-2-(carboxylamino)propanoic acid (7 g, 20.87 mmol) in THF (25 mL) and water (25 mL) was added Fmoc-OSu (6.34 g, 18.79 mmol) at 25°C under nitrogen. And then NaHCO3 (8.77 g, 104 mmol) was added. The resulting mixture was stirred at 25°C for 2h. The pH was adjusted to 5 with 1N HCl and extracted with EtOAc (2 x 200 mL). The combined organic layers were washed with brine (3 x 100 mL), dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by reverse phase flash chromatography using the following conditions: Column: C18 silica gel (330 g); Mobile phase A: water (0.1% TFA), Mobile phase B: MeCN; (Gradient: 5% B for 5 min, increase to 55% B in 15 min, 55% B for 5 min; increase to 95% B in 20 min, 95% B for 5 min); Flow rate: 90 mL / min; Detector: UV 210 nm; RT = 45 min. The fractions containing the product were collected and concentrated in vacuo to yield (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-(4-acetylpiperazin-1-yl)phenyl)propanoic acid. 30 H 32 N3O5[M+H] + The MS ESI calculated value was 514.23 and the found value was 514.30. 1 H NMR (400 MHz, methanol-d4) δ 7.78-7.76 (m, 2H), 7.61-7.52 (m, 2H), 7.40-7.37 (m, 2H), 7.32-7.22 (m, 4H), 7.08-6.98 (m, 2H), 4.47-4.43 (m, 1H), 4.33-4.31 (m, 1H), 4.14-4.02 (m, 2H), 3.68-3.63 (m, 4H), 3.23-3.08 (m, 5H), 2.91-2.85 (m, 1H), 2.11 (s, 3H).

[0298] Synthesis Scheme 10

[0299]

[0300] Phe4Pyrim5CO2H

[0301]

[0302] (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-(5-(tert-butoxycarbonyl)pyrimidine-2- 1-(4-(4-amino)phenyl)propionic acid

[0303] Step 1: To a mixture of 2-chloropyrimidine-5-carboxylic acid (10 g, 63.1 mmol) in t-BuOH (100 mL) was added DMAP (0.771 g, 6.31 mmol) and Boc2O (16.52 g, 76 mmol) at room temperature under argon. The resulting mixture was stirred at 50 °C for 16 h, then it was cooled to room temperature and concentrated under vacuum. The residue was purified by silica gel column chromatography, eluting with 0-60% EtOAc in PE to give tert-butyl 2-chloropyrimidine-5-carboxylate. C9H 12 ClN2O2[M+H] + The MS ESI calculated value was 215.05 and the found value was 214.95.

[0304] Step 2: To a mixture of tert-butyl 2-chloropyrimidine-5-carboxylate (4 g, 18.64 mmol) in 1,4-dioxane (40 mL) and water (8 mL) was added (S)-3-(4-dihydroxyborylphenyl)-2-((tert-butoxycarbonyl)amino)propionic acid (8.64 g, 28.0 mmol), PdCl2(dppf) (1.364 g, 1.864 mmol) and K2CO3 (7.73 g, 55.9 mmol) at room temperature under argon. The resulting mixture was stirred at 80 ° C for 3 h, then the pH was adjusted to 4 with 1N HCl and extracted with EtOAc (2x300 mL). The combined organic layers were washed with brine (3x 50 mL), dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with 0-100% EtOAc in PE to give (S)-2-((tert-butoxycarbonyl)amino)-3-(4-(5-(tert-butoxycarbonyl)pyrimidin-2-yl)phenyl)propanoic acid. 23 H 30 N3O6[M+H] + The MS ESI calculated value was 444.21 and the found value was 444.35. 1 HNMR (400MHz, CDCl3) δ 9.27 (s, 2H), 8.42 (d, J = 7.8Hz, 2H), 7.36 (d, J = 7.9Hz, 2H), 4.69-4.68 (m, 1H), 3.29-3.24 (m, 2H), 1.63 (s, 9H), 1.44 (s, 9H).

[0305] Step 3: To a mixture of (S)-2-((tert-butoxycarbonyl)amino)-3-(4-(5-(tert-butoxycarbonyl)pyrimidin-2-yl)phenyl)propanoic acid (7 g, 15.78 mmol) in DCM (70 mL) was added TFA (14 mL, 182 mmol) at room temperature under argon. After stirring at room temperature for 1 h, the reaction was concentrated to give (S)-2-amino-3-(4-(5-(tert-butoxycarbonyl)pyrimidin-2-yl)phenyl)propanoic acid. 18 H 22 N3O4[M+H] + The MS ESI calculated value was 344.15 and the found value was 344.20.

[0306] Step 4: To a mixture of (S)-2-amino-3-(4-(5-(tert-butoxycarbonyl)pyrimidin-2-yl)phenyl)propanoic acid (4 g, 11.65 mmol) in THF (40 mL) and H2O (40 mL) was added NaHCO3 (4.89 g, 58.2 mmol) and Fmoc-OSu (3.54 g, 10.48 mmol) under argon at room temperature. After stirring at room temperature for 1 h, the pH was adjusted to 4 with 1N HCl and the solution was extracted with EtOAc (2 x 300 mL). The combined organic layers were washed with brine (150 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by RP-flash using the following conditions: column: C18 column (330 g); mobile phase A: water (0.05% TFA); mobile phase B: MeCN; (gradient: 0% B for 5 min, increase to 73% B in 30 min, 73% B for 10 min; increase to 95% B in 4 min, 95% B for 10 min); flow rate: 60 mL / min; detector: UV 254 and 210 nm; RT: 35.32 min to produce (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-(5-(tert-butoxycarbonyl)pyrimidin-2-yl)phenyl)propanoic acid. 33 H 32 N3O6[M+H] + The MS ESI calculated value was 566.22 and the found value was 566.35. 1 H NMR(300MHz,DMSO-d6)δ12.85(s,1H),9.24(s,2H),8.37(d,J=8.0Hz,2H),7.88-7.79(m,3H ),7.66-7.14(m,8H),4.30-4.14(m,4H),3.22-3.16(m,1H),3.05-2.82(m,1H),1.59(s,9H).

[0307] Synthesis Scheme 11

[0308]

[0309]

[0310] Prot4NHB

[0311]

[0312] (2S,4R)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-4-(benzylamino)pyrrolidine-2-carboxylic acid

[0313] Step 1: TEA (14.26 mL, 102 mmol) and NsCl (10.89 g, 49.1 mmol) were added to a mixture of (2S, 4R)-4-aminopyrrolidine-1,2-dicarboxylic acid 1-(tert-butyl) 2-methyl ester (10 g, 40.9 mmol) in DCM (100 mL) at room temperature. The reactant was stirred at room temperature for 4 h. The resulting solution was quenched with water (150 mL) and extracted with EtOAc (3x300 mL). The organic layers were combined, washed with brine (2x 150 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated in vacuo. The residue was purified by silica gel chromatography, eluting with a gradient of 0% to 45% EtOAc in PE to provide (2S, 4R)-4-((4-nitrophenyl)sulfonylamino)pyrrolidine-1,2-dicarboxylic acid 1-(tert-butyl) 2-methyl ester. C 17 H 24 N3O8S[M+H-Boc] + The MS ESI calculated value was 330.12 and the found value was 330.10. 1 HNMR(400MHz, CDCl3)δ8.18-8.16(m,1H),7.91-7.87(m,1H),7.79-7.75(m,2H),4.40-4.29(m,1H),4 .18-4.15(m,1H),3.72(s,3H),3.66-3.62(m,1H),3.26-3.19(m,1H),2.29-2.15(m,2H),1.38(s,9H).

[0314] Step 2: To a mixture of 1-(tert-butyl)2-methyl (2S,4R)-4-((4-nitrophenyl)sulfonylamino)pyrrolidine-1,2-dicarboxylate (17 g, 39.6 mmol) in DCM (60 mL) was added TFA (30 mL, 389 mmol) at ambient temperature. The reaction was stirred at ambient temperature for 2 h and then concentrated under vacuum to provide methyl (2S,4R)-4-((4-nitrophenyl)sulfonylamino)pyrrolidine-2-carboxylate.12 H 16 N3O6S[M+H] + The MS ESI calculated value was 330.07 and the found value was 330.10.

[0315] Step 3: NaHCO (16.52 g, 197 mmol) and Alloc-OSu (5.46 mL, 35.4 mmol) were added to a solution of (2S, 4R)-4-((4-nitrophenyl)sulfonylamino)pyrrolidine-2-formic acid methyl ester (18.5 g, 39.3 mmol) in THF (150 mL) and water (150 mL) at ambient temperature. The reactant was stirred at ambient temperature for 4 h, then diluted with water (100 mL), and extracted with EtOAc (3 x 250 mL). The organic layers were combined, washed with brine (2 x 150 mL), dried over anhydrous Na SO, and filtered. The filtrate was concentrated in vacuo. The residue was purified by silica gel chromatography eluting with a gradient of 0% to 55% EtOAc in PE to afford 1-allyl 2-methyl (2S,4R)-4-((4-nitrophenyl)sulfonylamino)pyrrolidine-1,2-dicarboxylate. 16 H 20 N3O8S[M+H] + The MS ESI calculated value was 414.09 and the found value was 413.95. 1 H NMR (300MHz, CDCl3) δ8.18-8.15(m,1H),7.91-7.88(m,1H),7.82-7.77(m,2H),5.84-5.70(m,1H),5.30-5.16(m,2H) ,4.58-4.52(m,2H),4.50-4.48(m,1H),4.47-4.15(m,1H),3.78-3.73(m,4H),3.32-3.28(m,1H),2.30-2.19(m,2H).

[0316] Step 4: (2S, 4R) -4- ((4- nitrophenyl) sulfonylamino) pyrrolidine -1,2- dicarboxylic acid 1-allyl 2-methyl ester (8.1g, 19.59mmol) in DMF (100mL) solution was added (bromomethyl) benzene (4.02g, 23.51mmol) and K2CO3 (8.12g, 58.8mmol) at ambient temperature. The reactant was stirred at ambient temperature for 4h, then quenched with water (100mL), and extracted with EtOAc (3x 250mL). The organic layers were combined, washed with brine (4x 200mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under vacuum. The residue was purified by silica gel chromatography eluting with a gradient of 0% to 55% EtOAc in PE to afford 1-allyl 2-methyl (2S,4R)-4-((N-benzyl-4-nitrophenyl)sulfonylamino)pyrrolidine-1,2-dicarboxylate. 23 H 26 N3O8S[M+H] + The MS ESI calculated value was 504.14 and the found value was 504.20. 1 H NMR (300MHz, CDCl3) δ7.88-7.84(m,1H),7.68-7.67(m,2H),7.66-7.55(m,1H),7.29-7.23(m,5H) ,5.85-5.75(m,1H),5.25-5.18(m,2H),4.19-4.15(m,6H),3.73-3.25(m,5H),2.23-2.15(m,2H).

[0317] Step 5: To a solution of 1-allyl 2-methyl (2S, 4R)-4-((N-benzyl-4-nitrophenyl)sulfonylamino)pyrrolidine-1,2-dicarboxylate (9.3 g, 18.47 mmol) in DMF (40 mL) was added 4-methoxythiophenol (3.11 g, 22.16 mmol) and K2CO3 (7.66 g, 55.4 mmol) at ambient temperature. The reaction was stirred at ambient temperature for 1 h, then diluted with water (100 mL) and extracted with EtOAc (3 x 150 mL). The organic layers were combined, washed with brine (2 x 100 mL), dried over anhydrous Na2SO4, and filtered. The residue was purified by RP-flash using the following conditions: column: Flash C18 (330 g); mobile phase A: water (0.05% TFA), mobile phase B: MeCN; (gradient: 5% B for 5 min, increase to 32% B in 15 min, 32% B for 6 min; increase to 95% B in 5 min, 95% B for 5 min); flow rate: 90 mL / min; detector: UV 210 nm; RT = 36 min. The fractions containing the product were collected and concentrated in vacuo to yield 1-allyl 2-methyl (2S, 4R)-4-(benzylamino)pyrrolidine-1,2-dicarboxylate. 17 H 23 N2O4[M+H] + The MS ESI calculated value was 319.16 and the found value was 319.10. 1 HNMR (300MHz, CDCl3) δ7.39-7.34(m,5H),5.90-5.85(m,1H),5.32-5.17(m,2H),4.60-4.36(m,3H),3.96-3.57(m,8H),2.07-1.99(m,2H).

[0318] Step 6: add Boc2O (4.16g, 19.08mmol) and TEA (5.32mL, 38.2mmol) to the solution of (2S, 4R)-4-(benzylamino) pyrrolidine-1,2-dicarboxylic acid 1-allyl 2-methyl ester (4.5g, 12.72mmol) in DCM (40mL) at ambient temperature. The reactant is stirred at ambient temperature for 4h, then quenched with water (30mL), and extracted with EtOAc (3x 200mL). The organic layers are merged, washed with brine (2x 100mL), dried over anhydrous Na2SO4, and filtered. The filtrate is concentrated under vacuum. The residue was purified by silica gel chromatography eluting with a gradient of 0% to 40% EtOAc in PE to afford 1-allyl 2-methyl (2S,4R)-4-(benzyl(tert-butoxycarbonyl)amino)pyrrolidine-1,2-dicarboxylate. 22H 31 N2O6[M+H] + The MS ESI calculated value was 419.21 and the found value was 419.15. 1 HNMR(300MHz, CDCl3)δ7.34-7.15(m,5H),5.90-5.85(m,1H),5.25-5.18(m,2H),4.58 -4.32(m,6H),3.74-3.44(m,5H),2.49-2.38(m,1H),2.12-1.99(m,1H),1.42(s,9H).

[0319] Step 7: To a mixture of (2S,4R)-4-(benzyl(tert-butoxycarbonyl)amino)pyrrolidine-1,2-dicarboxylic acid 1-allyl 2-methyl ester (3 g, 5.73 mmol) in THF (15 mL) was added LiOH (11.47 mL, 11.47 mmol, 1 M in water) at 0°C. The reaction was stirred at ambient temperature for 2 h, then acidified to pH 3-4 with aqueous HCl and extracted with EtOAc (3x150 mL). The organic layers were combined, washed with brine (2x 80 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under vacuum to provide (2S,4R)-1-((allyloxy)carbonyl)-4-(benzyl(tert-butoxycarbonyl)amino)pyrrolidine-2-carboxylic acid. C 21 H 29 N2O6[M+H] + The MS ESI calculated value was 405.19 and the found value was 405.30. 1 HNMR(300MHz, CDCl3)δ7.35-7.14(m,5H),5.89-5.75(m,1H),5.14-5.19(m, 2H),4.61-4.30(m,6H),3.82-3.32(m,2H),2.47-2.18(m,2H),1.42(s,9H).

[0320] Step 8: To a mixture of (2S,4R)-1-((allyloxy)carbonyl)-4-(benzyl(tert-butoxycarbonyl)amino)pyrrolidine-2-carboxylic acid (2.6 g, 5.46 mmol) in DCM (150 mL) was added AcOH (0.751 mL, 13.11 mmol) and Pd(Ph3P)4 (0.126 g, 0.109 mmol) at ambient temperature. And then Bu3SnH (1.749 g, 6.01 mmol) was added to the reaction. After the reaction was stirred at ambient temperature for 4 h, it was concentrated under reduced pressure to provide (2S,4R)-4-(benzyl(tert-butoxycarbonyl)amino)pyrrolidine-2-carboxylic acid. 17 H25 N2O4[M+H] + The MS ESI calculated value was 321.17 and the found value was 321.15.

[0321] Step 9: To a mixture of (2S, 4R)-4-(benzyl(tert-butoxycarbonyl)amino)pyrrolidine-2-carboxylic acid (3.2 g, 4.99 mmol) in THF (50 mL) and water (50 mL) was added NaHCO3 (2.098 g, 24.97 mmol) to a pH of 8-9. And then Fmoc-OSu (1.516 g, 4.49 mmol) was added to the reaction. The resulting mixture was stirred at ambient temperature for 4 h, then acidified to pH 3-4 with aqueous HCl solution, and extracted with EtOAc (3 x 200 mL). The organic layers were combined, washed with brine (2 x 100 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under vacuum to provide a crude product. The residue was purified by RP-flash using the following conditions: column: Flash C18 (330 g); mobile phase A: water (0.05% TFA), mobile phase B: MeCN; (gradient: 5% B for 5 min, increase to 70% B in 25 min, 70% B for 8 min; increase to 95% B in 2 min, 95% B for 5 min); flow rate: 90 mL / min; detector: UV 210 nm; RT = 45 min. The fractions containing the product were collected and concentrated in vacuo to give (2S,4R)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-4-(benzyl(tert-butoxycarbonyl)amino)pyrrolidine-2-carboxylic acid. 32 H 35 N2O6[M+H] + The MS ESI calculated value was 543.24 and the found value was 543.20. 1 HNMR(400MHz,DMSO-d6)δ12.93(s,2H),7.88(d,J=7.6Hz,2H),7.61-7.57(m,2H),7.43-7.18(m ,9H),4.43-4.15(m,8H),3.53-3.25(m,2H),2.51-2.50(m,1H),2.03-1.92(m,1H),1.37(s,2H).

[0322] Synthesis Scheme 12

[0323]

[0324] Prot4NHCH2CH2Ph

[0325]

[0326] (2S,4R)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-4-(phenethylamino)pyrrolidine-2-carboxylic acid

[0327] Step 1: To a mixture of (2S,4R)-4-((4-nitrophenyl)sulfonylamino)pyrrolidine-1,2-dicarboxylic acid 1-allyl 2-methyl ester (7.2 g, 17.42 mmol), DIAD (5.08 ml, 26.1 mmol), and PPh3 (6.85 g, 26.1 mmol) in THF (70 mL) was added 2-phenylethane-1-ol (3.19 g, 26.1 mmol) at ambient temperature. The reactants were stirred at ambient temperature for 4 h, then quenched with water (100 mL), and extracted with EtOAc (3 x 250 mL). The organic layers were combined, washed with brine (4 x 200 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography eluting with a gradient of 0% to 55% EtOAc in PE to afford 1-allyl 2-methyl (2S,4R)-4-((4-nitro-N-phenethylphenyl)sulfonamido)pyrrolidine-1,2-dicarboxylate. 24 H 28 N3O8S[M+H] + The MS ESI calculated value was 518.15 and the found value was 518.20.

[0328] Step 2: To a mixture of 1-allyl 2-methyl (2S,4R)-4-((4-nitro-N-phenethylphenyl)sulfonylamino)pyrrolidine-1,2-dicarboxylate (7.2 g, 13.91 mmol) in DMF (80 mL) was added 4-methoxythiophenol (1.950 g, 13.91 mmol) and K2CO3 (1.923 g, 13.91 mmol) at ambient temperature. The reaction was stirred at ambient temperature for 1 h, then diluted with water (200 mL) and extracted with EtOAc (3 x 150 mL). The organic layers were combined, washed with brine (2 x 500 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by RP-flash using the following conditions: column: Flash C18 (330 g); mobile phase A: water (0.05% TFA), mobile phase B: MeCN; (gradient: 5% B for 5 min, increase to 32% B in 15 min, 32% B for 6 min; increase to 95% B in 5 min, 95% B for 5 min); flow rate: 90 mL / min; detector: UV 210 nm; RT = 36 min. The fractions containing the product were collected and concentrated in vacuo to yield 1-allyl 2-methyl (2S, 4R)-4-(phenethylamino)pyrrolidine-1,2-dicarboxylate. C 18 H 25N2O4[M+H] + The MS ESI calculated value was 333.17 and the found value was 333.25.

[0329] Step 3: add Boc2O (3.93g, 18.04mmol) and TEA (3.35mL, 24.07mmol) to the mixture of (2S, 4R)-4-(phenethylamino)pyrrolidine-1,2-dicarboxylic acid 1-allyl 2-methyl ester (4g, 12.03mmol) in DCM (40mL) at ambient temperature. The reactant is stirred at ambient temperature for 4h, then quenched with water (30mL), and extracted with EtOAc (3x200mL). The organic layers are merged, washed with saline (2x 100mL), dried over anhydrous Na2SO4, and filtered. The filtrate is concentrated under reduced pressure. The residue was purified by silica gel chromatography eluting with a gradient of 0% to 40% EtOAc in PE to afford 1-allyl 2-methyl (2S,4R)-4-((tert-butoxycarbonyl)(phenethyl)amino)pyrrolidine-1,2-dicarboxylate. 23 H 33 N2O6[M+H] + The MS ESI calculated value was 433.23 and the found value was 433.25.

[0330] Step 4: To a mixture of 1-allyl 2-methyl (2S,4R)-4-((tert-butoxycarbonyl)(phenethyl)amino)pyrrolidine-1,2-dicarboxylate (3.84 g, 8.88 mmol) in THF (30 mL) was added LiOH (17.76 mL, 17.76 mmol, 1 M in water) at 0°C. The reaction was stirred at ambient temperature for 2 h, then acidified to pH 3-4 with aqueous HCl and extracted with EtOAc (3 x 150 mL). The organic layers were combined, washed with brine (2 x 80 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure and the crude product was used directly in the next step without any further purification. C 22 H 31 N2O6[M+H] + The MS ESI calculated value was 419.21 and the found value was 419.35.

[0331] Step 5: To a mixture of (2S,4R)-1-((allyloxy)carbonyl)-4-((tert-butoxycarbonyl)(phenethyl)amino)pyrrolidine-2-carboxylic acid (7 g, 16.73 mmol) in DCM (150 mL) was added AcOH (2.411 g, 40.1 mmol) and Pd(Ph3P)4 (3.87 g, 3.35 mmol) at ambient temperature. And then Bu3SnH (5.36 g, 18.40 mmol) was added to the reaction. The reaction was stirred at ambient temperature for 4 h and then concentrated under reduced pressure. The crude product was used directly in the next step without any further purification. C 18 H 27 N2O4[M+H] + The MS ESI calculated value was 335.19 and the found value was 335.25.

[0332] Step 6: To a mixture of (2S,4R)-4-((tert-butoxycarbonyl)(phenethyl)amino)pyrrolidine-2-carboxylic acid (5.4 g, 16.15 mmol) in THF (50 mL) and water (50 mL) was added NaHCO (8.56 g, 81 mmol) and then Fmoc-OSu (4.89 g, 14.54 mmol) at room temperature. After the reaction was stirred at ambient temperature for 4 h, it was acidified to pH 3-4 with aqueous HCl and extracted with EtOAc (3 x 400 mL). The organic layers were combined, washed with brine (2 x 200 mL), dried over anhydrous NaSO and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by RP-flash using the following conditions: column: Flash C18 (330 g); mobile phase A: water (0.05% TFA), mobile phase B: MeCN; (gradient: 5% B for 5 min, increase to 70% B in 25 min, 70% B for 8 min; increase to 95% B in 2 min, 95% B for 5 min); flow rate: 90 mL / min; detector: UV 210 nm; RT = 45 min. The fractions containing the product were collected and concentrated in vacuo to give (2S,4R)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-4-((tert-butoxycarbonyl)(phenethyl)amino)pyrrolidine-2-carboxylic acid. 33 H 37 N2O6Na[M+Na] + The MS ESI calculated value was 579.26 and the found value was 579.25. 1H NMR (300MHz, methanol-d4) δ7.78-7.76(m,2H),7.63-7.57(m,2H),7.44-7.24(m,6H),7.20-7.17(m,3H),4.51-4.28(m,4H),4.25 -4.15(m,1H),3.60-3.41(m,1H),3.39-3.31(m,3H),2.82-2.77(m,2H),2.45-2.44(m,1H),1.98-1.94(m,1H),1.46(s,9H).

[0333] Synthesis Scheme 13

[0334]

[0335] Prot4NHEt

[0336]

[0337] (2S,4R)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-4-(ethylamino)pyrrolidine-2-carboxylic acid

[0338] Step 1: Add iodoethane (4.35g, 27.9mmol) and KCO (9.63g, 69.7mmol) to a mixture of (2S, 4R)-4-((4-nitrophenyl)sulfonylamino)pyrrolidine-1,2-dicarboxylic acid 1-allyl 2-methyl ester (9.6g, 23.22mmol) in DMF (100mL) at room temperature. The reactant is stirred at room temperature for 12h, then quenched with water (80mL), and extracted with EtOAc (3x200mL). The organic layers are combined, washed with brine (4x 150mL), dried over anhydrous NaSO, and filtered. The filtrate is concentrated under reduced pressure. The residue was purified by silica gel chromatography eluting with a gradient of 0% to 55% EtOAc in PE to afford 1-allyl 2-methyl (2S,4R)-4-((N-ethyl-4-nitrophenyl)sulfonylamino)pyrrolidine-1,2-dicarboxylate. 18 H 24 N3O8S[M+H] + The MS ESI calculated value was 442.12 and the found value was 442.05. 1 H NMR (300MHz, CDCl3) δ8.02-8.01(m,1H),7.73-7.62(m,3H),5.95-5.78(m,1H),5.29-5.18(m,2H) ,4.58-4.43(m,4H),3.76-3.73(m,4H),3.38-3.29(m,3H),2.29-2.26(m,2H),1.26-1.20(m,3H).

[0339] Step 2: To a stirred mixture of (2S,4R)-4-((N-ethyl-4-nitrophenyl)sulfonylamino)pyrrolidine-1,2-dicarboxylic acid 1-allyl 2-methyl ester (5.5 g, 12.46 mmol) and K2CO3 (5.17 g, 37.4 mmol) in DMF (80 mL) was added 4-methoxythiophenol (3.49 g, 24.92 mmol) at room temperature. After the mixture was stirred at room temperature for 1 h, it was quenched with water (200 mL) and extracted with EtOAc (3 x 200 mL). The combined organic layers were washed with brine (3 x 100 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography eluting with 0-3% MeOH in DCM to provide 1-allyl 2-methyl (2S,4R)-4-(ethylamino)pyrrolidine-1,2-dicarboxylate. 12 H 21 N2O4[M+H] + The MS ESI calculated value was 257.14 and the found value was 257.15. 1 H NMR (300MHz, CDCl3) δ6.02-5.76(m,1H),5.40-5.11(m,2H),4.68-4.38(m,3H),3.91-3.77(m,1H),3.77-3.67( m,3H),3.61-3.42(m,1H),3.42-3.21(m,1H),2.68(q,J=7.2Hz,2H),2.28-2.00(m,2H),1.14(t,J=7.1Hz,3H).

[0340] Step 3: To a stirred solution of (2S,4R)-4-(ethylamino)pyrrolidine-1,2-dicarboxylic acid 1-allyl 2-methyl ester (2.9 g, 11.31 mmol) and TEA (3.15 mL, 22.63 mmol) in DCM (50 mL) was added Boc2O (3.94 mL, 16.97 mmol) at room temperature. The resulting solution was stirred at 25 °C for 16 h and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with 0-30% EtOAc in PE to provide (2S,4R)-4-((tert-butoxycarbonyl)(ethyl)amino)pyrrolidine-1,2-dicarboxylic acid 1-allyl 2-methyl ester. 17 H 29 N2O6[M+H] + The MS ESI calculated value was 357.19 and the found value was 357.30. 1H NMR (400MHz, CDCl3) δ6.01-5.79(m,1H),5.37-5.14(m,2H),4.70-4.40(m,4H),3.85-3.75(m,1H),3.73(d,J=7.8Hz, 3H), 3.49-3.36 (m, 1H), 3.28-3.06 (m, 2H), 2.53-2.38 (m, 1H), 2.22-2.07 (m, 1H), 1.46 (s, 9H), 1.11 (t, J = 7.0Hz, 3H).

[0341] Step 4: To a stirred solution of 1-allyl 2-methyl (2S,4R)-4-((tert-butoxycarbonyl)(ethyl)amino)pyrrolidine-1,2-dicarboxylate (3.8 g, 10.66 mmol) in THF (50 mL) was added LiOH (32 mL, 32.0 mmol, 1 N in water) at room temperature. After the solution was stirred at 25 °C for 2 h, the pH of the solution was adjusted to 3 with 1N HCl and then extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with brine (100 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to give (2S,4R)-1-((allyloxy)carbonyl)-4-((tert-butoxycarbonyl)(ethyl)amino)pyrrolidine-2-carboxylic acid. C 16 H 27 N2O6[M+H] + The MS ESI calculated value was 343.18 and the found value was 343.30.

[0342] Step 5: To a stirred solution of (2S,4R)-1-((allyloxy)carbonyl)-4-((tert-butoxycarbonyl)(ethyl)amino)pyrrolidine-2-carboxylic acid (3.4 g, 9.93 mmol), AcOH (1.431 g, 23.83 mmol) and Pd(Ph3P)4 (2.295 g, 1.986 mmol) in DCM (150 mL) was added Bu3SnH (3.18 g, 10.92 mmol) dropwise at room temperature. The resulting solution was stirred at 25 °C for 1 h and then concentrated under reduced pressure to give (2S,4R)-4-((tert-butoxycarbonyl)(ethyl)amino)pyrrolidine-2-carboxylic acid. 12 H 23 N2O4[M+H] + The MS ESI calculated value was 259.16 and the found value was 259.30.

[0343] Step 6: To a stirred solution of (2S,4R)-4-((tert-butoxycarbonyl)(ethyl)amino)pyrrolidine-2-carboxylic acid (2.5 g, 9.68 mmol) and Na2CO3 (3.08 g, 29.0 mmol) in THF (60 mL) and water (60 mL) was added Fmoc-OSu (3.26 g, 9.68 mmol) at room temperature. After the resulting mixture was stirred at 25°C for 16 h, the pH was adjusted to 3 with 1N HCl and then extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with brine (2 x 50 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure and the residue was purified by RP-flash using the following conditions: 330 g C18 column, 5%-5% in 5 min, 5%-65% in 40 min of MeCN in water (0.05% TFA) to yield (2S,4R)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-4-((tert-butoxycarbonyl)(ethyl)amino)pyrrolidine-2-carboxylic acid. 27 H 33 N2O6[M+H] + The MS ESI calculated value was 481.23 and the found value was 481.10. 1 H NMR (400MHz, DMSO-d6) δ12.98(br,1H),7.90(d,J=7.6Hz,2H),7.68-7.64(m,2H),7.44-7.41(m,2H),7.35-7.31(m,2H),4.43-4.18(m ,5H),3.60-3.56(m,1H),3.29-3.22(m,1H),3.16-3.12(m,2H),2.42-2.38(m,1H),2.08-2.01(m,1H),1.40(s,9H),1.05-1.00(m,3H).

[0344] Synthesis Scheme 14

[0345]

[0346] PyrimAla4AcPip

[0347]

[0348] (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(2-(4-acetylpiperazin-1-yl)pyrimidine- 5-amino)propionic acid

[0349] Step 1: To a mixture of 5-bromo-2-fluoropyrimidine (7 g, 39.6 mmol) in DMF (70 ml) was added 1-(piperazin-1-yl)ethane-1-one (10.14 g, 79 mmol) and K2CO3 (10.93 g, 79 mmol) at room temperature. The reaction mixture was stirred at 100 °C for 2 h and then cooled back to room temperature. The reaction mixture was extracted with 500 mL of EtOAc, washed with H2O (3 x 100 mL) and brine (80 mL), dried over Na2SO4, filtered and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with a gradient of 0%-80% EtOAc in PE. The fractions containing the desired product were combined and concentrated under reduced pressure to provide 4-(5-bromopyrimidin-2-yl)piperazine-1-carbaldehyde. C 10 H 14 BrN4O[M+H] + The MS ESI calculated values ​​were 285.03, 287.03, and the measured values ​​were 284.95, 286.95. 1 H NMR (300MHz, CDCl3) δ8.32(s,2H),3.90-3.74(m,4H),3.74-3.64(m,2H),3.58-3.48(m,2H),2.15(s,3H).

[0350] Step 2: A mixture of nickel(II) chloride ethylene glycol dimethyl ether complex (0.693 g, 3.16 mmol) and 1,10-phenanthroline (0.569 g, 3.16 mmol) in DMA (50 mL) was heated at 50°C for 0.5 h. A solution of 1-(4-(5-bromopyrimidin-2-yl)piperazin-1-yl)ethan-1-one (4.5 g, 15.78 mmol), (R)-tert-butyl 2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-iodopropanoate (11.68 g, 23.67 mmol) and TBAI (5.83 g, 15.78 mmol) in DMA (50 mL) was added at 25°C. Zn (2.064 g, 31.6 mmol) was then added. After the resulting mixture was stirred at 30°C for 24 h, the reaction mixture was filtered and washed with DCM. The organic phase was concentrated under reduced pressure. The residue was purified by RP-flash using the following conditions: column: C18 column (330 g); mobile phase A: water (0.05% TFA); mobile phase B: MeCN; (gradient: 0% B for 5 min, increasing to 82% B in 30 min, 82% B for 6 min; increasing to 95% B in 2 min, 95% B for 3 min); flow rate: 60 mL / min; detector: UV 254 and 210 nm; RT: 35 min to produce (S)-tert-butyl 2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(2-(4-acetylpiperazin-1-yl)pyrimidin-5-yl)propanoate. C 32 H 38 N5O5[M+H] + The MS ESI calculated value was 572.28 and the found value was 572.30. 1 H NMR (300MHz, CDCl3) δ8.19 (s, 2H), 7.77 (d, J = 7.6Hz, 2H), 7.63-7.52 (m, 2H), 7.46-7.26 (m, 4H), 4.48-4.42 (m, 2H), 4.35-4.30 (m, 1H), 4.2 2-4.17(m,1H),3.89-3.81(m,4H),3.71-3.68(m,2H),3.56-3.52(m,2H),3.08-3.02(m,1H),2.92-2.87(m,1H),2.17(s,3H),1.48(s,9H).

[0351] Step 3: To a mixture of (S)-tert-butyl 2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(2-(4-acetylpiperazin-1-yl)pyrimidin-5-yl)propanoate (4.6 g, 8.05 mmol) in DCM (40 mL) was added TFA (80 mL, 1038 mmol) at room temperature under argon. The reaction was stirred at room temperature for 3 h and then concentrated under reduced pressure. The residue was purified by RP-flash using the following conditions: column: C18 gel column (330 g); mobile phase A: water (0.05% TFA); mobile phase B: MeCN; (gradient: 0% B for 5 min, increase to 72% B in 30 min, 72% B for 6 min; increase to 95% B in 2 min, 95% B for 10 min); flow rate: 60 mL / min; detector: UV 254 and 210 nm; RT: 35 min to produce (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(2-(4-acetylpiperazin-1-yl)pyrimidin-5-yl)propanoic acid. 28 H 30 N5O5[M+H] + The MS ESI calculated value was 516.22 and the found value was 516.35. 1 H NMR (300MHz, DMSO-d6) δ8.31(s,2H),7.89(d,J=7.5Hz,2H),7.81(d,J=8.6Hz,1H),7.74-7.62(m,2H),7.59-7.22(m ,4H),4.29-4.09(m,4H),3.74-3.60(m,4H),3.47-3.46(m,4H),2.98-2.92(m,1H),2.76-2.68(m,1H),2.03(s,3H).

[0352] Synthesis Scheme 15

[0353]

[0354] wxya

[0355]

[0356] (2S,3S)-2-Amino-3-(4-fluoro-1H-indol-3-yl)butanoic acid

[0357] Into a 1-L 3-neck round-bottom flask purged and maintained with inert nitrogen, 4-fluoro-1H-indole (10 g, 1.00 equiv.), L-threonine (10.6 g, 1.20 equiv.), DMSO (100 mL), potassium phosphate buffer (0.2 M, 300 mL, pH = 7.4) were placed. The reaction mixture was heated to 65 ° C, and then PfTrpB-7E6 (2.5 g, 25 wt%) and 3-hydroxy-2-methyl-5-([phosphonooxy]methyl)-4-pyridinecarboxaldehyde (0.078 g, 0.004 equiv.) were added. The resulting solution was stirred at 65 ° C overnight. The mixture was then cooled to room temperature and used directly in the next step.

[0358] THF (100mL), sodium carbonate (23.56g, 3.0 equivalents) and 2,5-dioxopyrrolidin-1-yl 9H-fluorene-9-ylmethyl ester (29.96g, 1.20 equivalents) were added to the above reaction mixture at 0°C. The resulting solution was stirred overnight at room temperature. The pH was adjusted to 4 with 3M HCl and the solid precipitate was filtered out. The resulting solution was extracted with EtOAc (3x 500mL). The organic fractions were combined and washed with brine (1L), dried over anhydrous sodium sulfate and concentrated under vacuum. The mixture was applied to a silica gel column with MeOH:DCM=1:5. HPLC-MS:(ES,m / z):[M+1]:459. 1 H NMR (300MHz, DMSO-d6) δ12.60(s,1H),11.15(s,1H),7.87(d,J=7.6Hz,2H),7.76-7.49(m,3H),7.47-7.34(m,2H),7.34-7.16(m,4H),7.03(td,J=7 .9,5.0Hz,1H),6.73(dd,J=11.8,7.7Hz,1H),4.36(t,J=8.5Hz,1H),4.31 -4.02(m,3H),3.51(q,J=7.4Hz,1H),1.31(d,J=7.0Hz,4H),0.78(s,1H).

[0359] Synthesis Scheme 16

[0360]

[0361] sbW4Cl

[0362]

[0363] (2S,3S)-2-Amino-3-(4-chloro-1H-indol-3-yl)butanoic acid

[0364] Into a 1-L 3-neck round-bottom flask purged and maintained with inert nitrogen, 4-chloro-1H-indole (10 g, 1.00 equivalent), L-threonine (14.09 g, 1.8 equivalent), DMSO (100 mL), potassium phosphate buffer (0.2 M, 300 mL, pH = 7.4) were placed, and the reaction mixture was heated to 65 ° C., and then PfTrpB-7E6 (7.5 g, 25 wt%) and 3-hydroxy-2-methyl-5-([phosphonooxy]methyl)-4-pyridinecarboxaldehyde (174 mg, 0.01 equivalent) were added. The resulting solution was stirred at 65 ° C. for 36 h. The mixture was then cooled to room temperature and used directly in the next step.

[0365] THF (100 mL), sodium carbonate (20.9 g, 3.0 equiv.) and 2,5-dioxopyrrolidin-1-yl 9H-fluorene-9-ylmethyl carbonate (31.0 g, 1.40 equiv.) were added to the above reaction mixture at 0°C. The resulting solution was stirred at room temperature overnight. The pH was adjusted to 4 with 3M HCl and the solid precipitate was filtered out. The resulting solution was extracted with EtOAc (3 x 500 mL). The organic fractions were combined and washed with brine (1 L), dried over anhydrous sodium sulfate and concentrated under vacuum. HPLC-MS: (ES, m / z): [M+1]: 475

[0366] Synthesis Scheme 17

[0367]

[0368] sbMe1Nal

[0369]

[0370] (2S,3S)-2-Amino-3-(naphthalen-1-yl)butanoic acid

[0371] Step 1: To a solution of (2S,3R)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-hydroxybutanoic acid (250 g, 1.00 equiv) in DMF (1.5 L) was added benzyl bromide (250 g, 2.00 equiv) dropwise at 20 °C. Then, cesium carbonate (477 g, 2.00 equiv) was added and the solution was stirred at 20 °C for 3 h. The reactant was poured into ice H2O (3 L) and extracted with EtOAc (500 mL x 3). The organic phase was washed with 3% LiCl solution (500 mL x 2) and brine (500 mL), dried over sodium sulfate and concentrated under vacuum at 40 °C. The crude product was triturated with methyl tert-butyl ether:PE=6:1. 1H NMR (400MHz, CDCl3): δ7.77(d,J=7.6Hz,1H),7.40(d,J=8.0Hz,1H),7.31-7.36(m,10H),5.6 5-5.71m,1H),5.13-5.31(m,3H),4.39-4.43(m,3H),4.22-4.25(m,1H),1.25(d,J=6.4Hz,3H)

[0372] Step 2: To a 3-neck round bottom flask under inert nitrogen was added benzyl (2S,3R)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-hydroxybutanoate (125 g, 1.00 equiv) and DCE (750 mL). The reactants were cooled to 0 °C, followed by the addition of NIS (195 g, 3.00 equiv) and PPh3 (228 g, 3.00 equiv). The temperature was raised to 50 °C and the reaction mixture was stirred for 3 h. The reactants were poured into ice H2O (500 mL) and extracted with DCM (500 mL x 2). The organic phase was dried over sodium sulfate and concentrated under vacuum at 40 °C. The residue was purified by silica gel column chromatography (PE / EtOAc=1 / 0 to 0 / 1). 1 H NMR (400MHz, CDCl3): δ7.78(d,J=7.6Hz,2H),7.68(d,J=7.2Hz,2H),7.33-7.43(m,9H),5.2 7-5.68(m,1H),5.21-5.23(m,2H),4.39-4.52(m,3H),4.25-4.38(m,1H),1.91-1.95(m,3H).

[0373] Step 3: Into a 3-neck round bottom flask was placed 2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-iodobutyrate, 1-iodonaphthalene (42.2 g, 1.20 equiv), TBAI (76.7 g, 1.50 equiv), Zn (19.0 g, 2.10 equiv) and DMA (750 mL). Into a second 3-neck round bottom flask at 25°C was placed picolinamidine.2HCl (42.2 g, 1.20 equiv), NiCl2.ethylene glycol dimethyl ether (7.61 g, 0.25 equiv) and DMA (750 mL). Under argon, the contents of the second flask were added to the first flask. The resulting mixture was then stirred at 25°C for 12 h. The reaction was poured into ice H2O (3 L) and extracted with EtOAc (1 L x 2). The organic phase was dried over sodium sulfate and concentrated under vacuum at 40° C. The crude product was purified by reverse phase HPLC (MeCN:H 2 O). HPLC-MS: [M+23]: 564. 1H NMR(400MHz, CDCl3)δ:8.17-8.24(m,1H),8.15-8.17(m,1H),7.77-7.87(m,2H),7.76-7.77(m,4H),7.30-7.41(m,10H ),5.30-5.38(m,1H),4.96-5.04(m,3H),4.85-4.87(m,1H),4.30-4.34(m,1H),4.18-4.26(m,4H),1.43-1.45(m,3H).

[0374] Step 4: 143 g of benzyl (2S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(naphthalen-1-yl)butanoate were isolated by SFC. The organic phase was concentrated under vacuum at 35°C.

[0375] Peak 1: benzyl (2S,3R)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(naphthalen-1-yl)butanoate. 1 HNMR (400MHz, DMSO-d6): δ8.11-8.12(m,2H),8.10-8.11(m,1H),7.88-7.9 0(m,2H),7.54-7.88(m,1H),7.44-7.53(m,2H),7.42-7.44(m,4H),7.33-7 .42(m,3H),7.27-7.33(m,6H),7.08-7.09(m,2H),4.91-4.94(m,1H),4.79 -4.82(m,1H),4.58(t,J=8.0Hz),4.17-4.25(m,4H),1.39(d,J=6.8Hz,3H). Peak 2: benzyl (2S,3S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(naphthalen-1-yl)butanoate. 1 H NMR (400MHz, DMSO-d6): δ7.92-8.15(m,1H),7.86-7.92(m,1H),7.84-7.86(m,1H),7.57-7.84(m,2H),7.56-7.57(m,1H),7.41-7.54(m,4H) ,7.30-7.38(m,4H),7.27-7.30(m,7H),5.08-5.14(m,2H),4.65(t,J=8.0Hz),4.23-4.26(m,1H),4.05-4.18(m,3H),1.30(d,J=6.8Hz,3H).

[0376] Step 5: To a 3-neck round bottom flask was added benzyl (2S,3S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(naphthalen-1-yl)butanoate (40.0 g, 1.00 equiv) and THF (200 mL). 10% wet Pd / C (7.00 g) was added and the reaction was purged with H2 3 times and stirred at 25°C under H2 (15 psi) for 12 h. The reaction was filtered through a pad of celite and concentrated under vacuum at 35°C. The crude product was ground with PE at 25°C for 1 h. After filtration, the filter cake was dissolved in MeCN (100 mL) and concentrated under vacuum at 35°C to remove residual solvent. HPLC-MS: [M+23]: 474. 1 H NMR (400MHz, DMSO-d6) δ12.78 (s, 1H), 8.23 ​​(d, J = 7.6Hz, 1H), 7.86-7.88 ( m,1H),7.80-7.86(m,2H),7.61-7.80(m,2H),7.55-7.59(m,4H),7.481-7. 55(m,1H),7.40-7.48(m,3H),7.27-7.29(m,2H),4.28-4.60(m,1H),4.24- 4.28(m,1H),4.17-4.24(m,2H),4.04-4.15(m,1H),1.36(d,J=6.8Hz,3H).

[0377] Synthesis Scheme 18

[0378]

[0379]

[0380] TyrEtNAc

[0381]

[0382] L-Tyrosine O-ethylacetamide or (S)-3-(4-(2-acetylaminoethoxy)phenyl)-2-aminopropionic acid

[0383] Step 1: To a stirred solution of (tert-butoxycarbonyl)-L-tyrosine methyl ester (10.0 g, 33.9 mmol), (2-bromoethyl) benzyl carbamate (26.2 g, 102 mmol) and TBAB (5.46 g, 16.93 mmol) in DMF (150 mL) was added potassium carbonate (14.04 g, 102 mmol) at room temperature. The mixture was then stirred at 50 °C for 24 h. The mixture was cooled to room temperature, quenched with water (250 mL), and extracted with EtOAc (2 x 500 mL). The combined organic layers were washed with brine (3 x 150 mL), dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography, eluting with 0-30% EtOAc in PE. C 25 H 32 N2O7[M+Na] + The MS ESI calculated value was 495.22, and the found value was 495.10; 1 HNMR (300MHz, CDCl3) δ7.38-7.32(m,5H),7.04(d,J=8.4Hz,2H),6.81(d,J=8.4Hz,2H),5.31(br,1H),5.21(s,2H),4. 97(br,1H),4.56-4.53(m,1H),4.03(t,J=5.0Hz,2H),3.72(s,3H),3.64-3.58(m,2H),3.06-3.01(m,2H),1.43(s,9H).

[0384] Step 2: To a stirred solution of (S)-methyl 3-(4-(2-(((benzyloxy)carbonyl)amino)ethoxy)phenyl)-2-((tert-butoxycarbonyl)amino)propanoate (16.0 g, 33.9 mmol) and acetic anhydride (6.39 mL, 67.7 mmol) in THF (200 mL) was added Pd / C (3.60 g, 33.9 mmol, dried, 10 wt %) at room temperature under nitrogen atmosphere. The mixture was degassed 3 times with hydrogen and stirred at 20 °C for 4 h. DIPEA (17.74 mL, 102 mmol) was added to the mixture and stirred at 20 °C for 1 h. The mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with 0-3% MeOH in DCM. 19 H 28 N2O6[M+Na] + The MS ESI calculated value was 403.19 and the found value was 403.10; 1H NMR(300MHz, CDCl3)δ7.05(d,J=8.4Hz,2H),6.85-6.80(m,2H),5.99(br,1H),5.32(br,1H),4.99-4.97(m ,1H),4.02(t,J=5.0Hz,2H),3.72(s,3H),3.69-3.63(m,2H),3.10-2.89(m,2H),2.02(s,3H),1.42(s,9H).

[0385] Step 3: To a stirred solution of (S)-methyl 3-(4-(2-acetamidoethoxy)phenyl)-2-((tert-butoxycarbonyl)amino)propanoate (12.5 g, 32.9 mmol) in THF (100 mL) was added lithium hydroxide (65.7 mL, 65.7 mmol, 1 N in water) at room temperature. The solution was stirred at 20 °C for 2 h. The pH of the solution was adjusted to 3 with 1 N HCl. The aqueous layer was extracted with EtOAc (2 x 250 mL). The combined organic layers were washed with brine (150 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure.

[0386] MS ESI[M+H] + :367.10.

[0387] Step 4: To a stirred solution of (S)-3-(4-(2-acetylaminoethoxy)phenyl)-2-((tert-butoxycarbonyl)amino)propanoic acid (12.5 g, 30.7 mmol) in THF (20 mL) was added 4N HCl in dioxane (200 mL) at room temperature. The solution was stirred at 20 °C for 1 h. The solvent was concentrated under reduced pressure. MS ESI [M+H] + :267.05.

[0388] Step 5: To a stirred mixture of (S)-3-(4-(2-acetylaminoethoxy)phenyl)-2-aminopropanoic acid hydrochloride (9.50 g, 25.1 mmol) and NaHCO3 (10.54 g, 126 mmol) in THF (100 mL) and water (100 mL) was added Fmoc-OSu (7.62 g, 22.59 mmol) at room temperature. The mixture was stirred at 20 °C for 1 h. The pH of the solution was adjusted to 3 with 1N HCl. The aqueous phase was extracted with EtOAc (2 x 500 mL). The combined organic layers were washed with brine (150 mL), dried over anhydrous sodium bicarbonate and filtered. The filtrate was concentrated under reduced pressure and the residue was recrystallized from EtOAc (200 mL). The solid was collected by filtration and dried under vacuum. MS ESI [M+H] + :489.05; 1H NMR (300MHz, methanol-d4) δ7.79(d,J=7.6Hz,2H),7.62-7.57(m,2H),7.42-7.26(m,4H),7.17-7.14(m,2H),6.83(d,J=8.4Hz,2H),4.41 -4.31(m,2H),4.29-4.10(m,2H),3.96(t,J=4.8Hz,2H),3.51(t,J=5.4Hz,2H),3.19-3.13(m,1H),2.92-2.84(m,1H),1.94(s,3H).

[0389] Synthesis Scheme 19

[0390]

[0391]

[0392] TyrEtMor

[0393]

[0394] (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-(2-morpholinoethoxy)phenyl)propanoic acid

[0395] Step 1: To a solution of (tert-butoxycarbonyl)-L-tyrosine tert-butyl ester (400 mg, 1.185 mmol) in DMF (4.742 ml) at 0°C was added cesium carbonate (1 g, 3.08 mmol). The reaction was stirred for 15 min and 2-morpholinoethyl 4-methylbenzenesulfonate (474 ​​mg, 1.660 mmol) was added and stirred at 0°C for 2.5 hours. The reaction was diluted with EtOAc and washed with 10% aqueous LiCl solution. The organic layer was washed with 1M HCl, saturated aqueous NaHCO3, and aqueous NaCl solution, then dried over sodium sulfate, filtered, and concentrated. The crude material was purified by column chromatography using a RediSep Rf Gold silica gel 24g pre-packed column on Isco, eluting with a gradient of 0-100% EtOAc in hexanes over 30 min to produce the product. Molecular weight: 450.576; MS ESI [M+H] + :451.3.

[0396] Step 2: Dissolve (S)-tert-butyl 2-((tert-butoxycarbonyl)amino)-3-(4-(2-morpholinoethoxy)phenyl)propanoate (525 mg, 1.165 mmol) in DCM (3 mL) at ambient temperature. Add TFA (3 mL) and allow to stir at ambient temperature for 1.5 hours. The reaction was concentrated under reduced pressure and used crude in the next step. Molecular weight: 294.351, MS ESI [M+H] +The actual measured value is: 295.1.

[0397] Step 3: (S)-2-amino-3-(4-(2-morpholinoethoxy)phenyl)propanoic acid (354 mg, 1.203 mmol) was dissolved in tetrahydrofuran (3 mL) and water (3 mL) at ambient temperature. The reactants were cooled to 0°C. Sodium carbonate (382 mg, 3.61 mmol) was added and the solution was stirred at 0°C for 5 min. (9H-fluorene-9-yl)methyl (2,5-dioxopyrrolidin-1-yl)carbonate (406 mg, 1.203 mmol) was added and stirred for 2 h. THF was removed under reduced pressure. The aqueous solution was acidified to pH 3 with 1N HCl. The reactants were extracted with EtOAc, washed with water, dried over sodium sulfate, filtered, and concentrated. The crude material was purified by reverse phase column chromatography using a 50 g C18 prepacked column on Isco, eluting with a gradient of 10-100% TFA conditioned MeCN in TFA conditioned water over 30 min to give the product in fractions. The fractions were combined, frozen in a -78°C dry ice / acetone bath, and lyophilized to afford the product. Molecular weight: 516.94. MS ESI [M+H] + The actual measured value is: 517.2.

[0398] Synthesis Scheme 20

[0399]

[0400] daMeDab

[0401]

[0402] (R)-2,4-Diamino-2-methylbutyraldehyde

[0403] Step 1: To a stirred solution of (R)-2-amino-2-methylpent-4-enoic acid (1.3 g, 10.07 mmol) and DIPEA (5.27 mL, 30.2 mmol) in dioxane (20 mL) and water (20 mL) was added Fmoc-OSu (3.73 g, 11.07 mmol) at room temperature. The resulting solution was stirred at 25 °C for 16 h. The pH was adjusted to 3 with 1 N HCl and the solution was purified by reverse phase flash column chromatography using the following conditions: C18 column, 330 g, 5%-5% in 5 min, 5%-50% in 30 min, 98%-98% in 5 min of MeCN in water (0.05% TFA) to yield (R)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2-methylpent-4-enoic acid. 21 H 21 NO4Na[M+Na] +The MS ESI calculated value was 374.15 and the found value was 374.05. 1 H NMR (400MHz, CDCl3) δ7.79(d,J=7.5Hz,2H),7.61(d,J=7.5Hz,2H),7.48-7.29(m,4H),5.71(s,1 H), 5.48 (s, 1H), 5.20-5.16 (m, 2H), 4.44-4.40 (m, 2H), 4.25 (t, J = 6.7Hz, 1H), 2.82-2.71 (m, 2H).

[0404] Step 2: To a stirred solution of (R)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2-methylpent-4-enoic acid (3 g, 8.54 mmol) and N-methylmorpholine N-oxide (2.2 g, 9.39 mmol, 50% in water) in acetone (60 mL) was added OsO4 (2.170 g, 0.854 mmol, 10% in water) at room temperature. The resulting solution was stirred at 25 °C for 4 h. Sodium periodate (2.00 g, 9.39 mmol) in water (20 mL) was added to the solution and the resulting mixture was stirred at 25 °C for 16 h. The reaction mixture was diluted with water (50 mL) and extracted with EtOAc (3 x 200 mL). The combined organic layers were washed with brine (2 x 100 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was then concentrated under reduced pressure to give (R)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2-methyl-4-oxobutanoic acid. 20 H 20 NO5[M+H] + The MS ESI calculated value was 354.13, the found value was 354.00.

[0405] Step 3: To a stirred solution of (R)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2-methyl-4-oxobutanoic acid (3 g, 6.79 mmol) in toluene (120 mL) was added tert-butyl carbamate (4.77 g, 40.8 mmol) and TFA (2.32 g, 20.38 mmol) at room temperature. The resulting solution was stirred at 25 °C for 2 h. The solution was concentrated under reduced pressure to give (R,Z)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((tert-butoxycarbonyl)imino)-2-methylbutanoic acid. 25 H 27 N2O6[MH] + The MS ESI calculated value was 451.19 and the found value was 450.90.

[0406] Step 4: To a stirred solution of (R,Z)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((tert-butoxycarbonyl)imino)-2-methylbutanoic acid (3.1 g, 5.48 mmol) and dimethyl(phenyl)silane (1.87 g, 13.7 mmol) in toluene (120 mL) was added tri(pentafluorophenyl)borane (0.281 g, 0.548 mmol) at room temperature. The solution was stirred at 25 °C for 16 h. The solvent was concentrated under reduced pressure and the residue was purified by reverse phase flash chromatography using the following conditions: 330 g C18 column, 5%-5% in 5 min, 5%-55% in 30 min, 98%-98% MeCN in water (0.05% TFA) in 5 min, RT = 35 min to yield (R)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((tert-butoxycarbonyl)amino)-2-methylbutanoic acid. 25 H 31 N2O6[M+H] + The MS ESI calculated value was 455.21, and the found value was 455.10; 1 H NMR (400MHz, DMSO-d6) δ12.48(s,1H),7.90(d,J=7.2Hz,2H),7.73(d,J=7.2Hz,2H),7.54(s,1H),7.44-7.41(m,2H),7.36- 7.32(m,2H),6.78(s,1H),4.25-4.23(m,3H),2.92-2.91(m,2H),1.99-1.93(m,1H),1.87-1.81(m,1H),1.37-1.34(m,9H).

[0407] Synthesis Scheme 21

[0408]

[0409] dDabMe3

[0410]

[0411] [(R)-3-Amino-3-carboxypropyl]-trimethylammonium

[0412] Step 1: To a mixture of (R)-4-amino-2-((tert-butoxycarbonyl)amino)butanoic acid (7 g, 32.1 mmol) in MeOH (30 mL) was added CH3I (10.03 mL, 160 mmol) and KHCO3 (16.05 g, 160 mmol) at ambient temperature. The reactants were warmed to 35 °C for 12 h. The reaction mixture was then filtered and the filtrate was concentrated under reduced pressure to provide a crude product. The residue was dissolved in ethanol and then filtered again. The filtrate was concentrated under reduced pressure to provide (R)-3-((tert-butoxycarbonyl)amino)-3-carboxy-N,N,N-trimethylpropane-1-ammonium iodide. C 12 H 25 IN2O4[MI] + The MS ESI calculated value was 261.18 and the found value was 261.30.

[0413] Step 2: To a mixture of (R)-3-((tert-butoxycarbonyl)amino)-3-carboxy-N,N,N-trimethylpropane-1-ammonium iodide (15 g, 23.18 mmol) in DCM (60 mL) was added TFA (30 mL, 389 mmol) at ambient temperature. The reaction was stirred at ambient temperature for 1 h and then concentrated under reduced pressure to provide (R)-3-amino-3-carboxy-N,N,N-trimethylpropane-1-ammonium 2,2,2-trifluoroacetate. C9H 17 F3N2O4[M-CF3COO] + The MS ESI calculated value was 161.13 and the found value was 161.30.

[0414] Step 3: To a mixture of (R)-3-amino-3-carboxy-N,N,N-trimethylpropane-1-ammonium 2,2,2-trifluoroacetate (11.5 g, 20.97 mmol) in THF (40 mL) and water (40 mL) was added NaHCO3 (8.81 g, 105 mmol) and Fmoc-OSu (6.37 g, 18.87 mmol). The reaction was stirred at ambient temperature for 2 h, then acidified to pH 3-4 with aqueous HCl and filtered to provide a crude product. The crude product was purified by reverse phase column chromatography using the following conditions: 330 g C18 column; mobile phase A: water (0.05% TFA), mobile phase B: MeCN; (gradient: 0% B for 5 min, increased to 33% B in 19 min, 38% B for 7 min; increased to 95% B in 5 min, 95% B for 5 min); flow rate: 80 mL / min. The fractions containing the product were collected and lyophilized to provide (R)-3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-carboxy-N,N,N-trimethylpropane-1-ammonium 2,2,2-trifluoroacetate.24 H 27 F3N2O6[M-CF3COO] + The MS ESI calculated value was 383.20 and the found value was 383.25. 1 H NMR(300MHz, DMSO-d6)δ7.91(d,J=7.2Hz,2H),7.79-7.71(m,3H),7.46-7.41(m,2H),7.37-7.32(m,2H),4.42-4.22( m,3H),4.07-4.01(m,1H),3.99-3.41(m,1H),3.33-3.28(m,1H),3.07(s,9H),2.21-2.16(m,1H),2.08-2.04(m,1H). 19 F-NMR (282MHz, DMSO-d6) δ73.931.

[0415] Synthesis Scheme 22

[0416]

[0417] FqV

[0418]

[0419] 3-[4-[(S)-2-amino-2-carboxyethyl]phenyl]bicyclo[1.1.1]pentane-1-carboxylic acid

[0420] Step 1: To a stirred mixture of (S)-2-((tert-butoxycarbonyl)amino)-3-(4-iodophenyl)propanoic acid (3.91 g, 10 mmol) in DMF (40 mL) at 0 ° C under an argon atmosphere, 3-bromoprop-1-ene (3.63 g, 30.0 mmol) and NaHCO (0.840 g, 10.00 mmol) were added. The resulting mixture was stirred at 40 ° C for 16 h. The reaction mixture was cooled to room temperature and quenched with water (200 mL) and extracted with EtOAc (2x200 mL). The combined organic layers were washed with brine (3x 100 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography (eluting with 0-40% EtOAc in PE), and the fractions containing the product were collected and evaporated under reduced pressure to afford (S)-2-((tert-butoxycarbonyl)amino)-3-(4-iodophenyl)propionic acid allyl ester. 27 H 25 INO4[M+H] + The MS ESI calculated value was 554.08 and the found value was 554.20.

[0421] Step 2: To a stirred solution of 3-(methoxycarbonyl)bicyclo[1.1.1]pentane-1-carboxylic acid (8 g, 47.0 mmol) in DCM (100 mL) was added (Z)-tert-butyl N,N'-diisopropylaminocarbamidate (37.7 g, 188 mmol) at room temperature. The resulting solution was stirred at 40 °C for 2 h. The mixture was then cooled to room temperature. The solid was filtered off and the filtrate was concentrated under reduced pressure to produce 1-(tert-butyl) 3-methyl bicyclo[1.1.1]pentane-1,3-dicarboxylate. 1 H NMR (400MHz, DMSO-d6) δ3.61 (s, 3H), 2.17 (s, 6H), 1.39 (s, 9H).

[0422] Step 3: To a stirred solution of 1-(tert-butyl) 3-methyl bicyclo[1.1.1]pentane-1,3-dicarboxylate (15 g, 39.8 mmol) in THF (150 mL) was added LiOH (119 mL, 119 mmol, 1 N in water) at room temperature. The resulting solution was stirred at 25 °C for 5 h. The pH of the solution was adjusted to 3 with 1N HCl and then extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with brine (2 x 50 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was then concentrated under reduced pressure to produce 3-(tert-butyloxycarbonyl)bicyclo[1.1.1]pentane-1-carboxylic acid. 11 H 15 O4[MH] - The MS ESI calculated value was 211.10 and the found value was 211.10.

[0423] Step 4: To a stirred solution of 3-(tert-butoxycarbonyl)bicyclo[1.1.1]pentane-1-carboxylic acid (10 g, 28.3 mmol), 2-hydroxyisoindoline-1,3-dione (6.00 g, 36.7 mmol) and DMAP (0.345 g, 2.83 mmol) in DCM (100 mL) was added N,N'-dicyclohexylcarbodiimide (6.42 g, 31.1 mmol) at room temperature. The resulting mixture was stirred at 25 °C for 16 h. The solid was filtered off and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with 0-40% EtOAc in PE to provide 1-(tert-butyl) 3-(1,3-dioxoisoindolin-2-yl)bicyclo[1.1.1]pentane-1,3-dicarboxylate. 1 H NMR (400MHz, CDCl3) δ7.89 (dd, J = 5.5, 3.1 Hz, 2H), 7.79 (dd, J = 5.5, 3.1 Hz, 2H), 2.50 (s, 6H), 1.47 (s, 9H).

[0424] Step 5: To a stirred solution of NiBr2·3H2O (0.791 g, 2.90 mmol) in DMA (80 mL) was added 4,4'-di-tert-butyl-2,2'-bipyridine (0.973 g, 3.63 mmol) at room temperature under nitrogen atmosphere. After the resulting mixture was stirred at 50°C for 30 min, it was cooled to room temperature. 1-(tert-butyl)3-(1,3-dioxoisoindolin-2-yl)bicyclo[1.1.1]pentane-1,3-dicarboxylate (4.8 g, 12.09 mmol), (S)-allyl 2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-iodophenyl)propanoate (6.69 g, 12.09 mmol), trimethylsilyl chloride (0.131 g, 1.209 mmol) and zinc (3.95 g, 60.4 mmol) were added to the above mixture at room temperature. The resulting mixture was stirred at 25 °C for 2 h. The reaction was quenched with brine (150 mL) and extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with brine (3 x 100 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was then concentrated under reduced pressure and the residue was purified by silica gel column chromatography eluting with 0-40% EtOAc in PE to afford (S)-tert-butyl 3-(4-(2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(allyloxy)-3-oxopropyl)phenyl)bicyclo[1.1.1]pentane-1-carboxylate. 37 H 40 NO6[M+H] + The MS ESI calculated value was 594.28 and the found value was 594.30.

[0425] Step 6: To a stirred solution of (S)-tert-butyl 3-(4-(2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(allyloxy)-3-oxopropyl)phenyl)bicyclo[1.1.1]pentane-1-carboxylate (2.3 g, 1.55 mmol) and phenylsilane (0.335 g, 3.10 mmol) in THF (30 mL) was added Pd(Ph3P)4 (0.090 g, 0.077 mmol) at room temperature under nitrogen atmosphere. The resulting mixture was stirred at room temperature for 1 h, then concentrated under reduced pressure and the residue was purified by reverse phase column chromatography using the following conditions: C18 column, 330 g, 5%-5% in 5 min, 5%-70% in 40 min of MeCN in water (0.05% TFA) to yield (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-(3-(tert-butoxycarbonyl)bicyclo[1.1.1]pentan-1-yl)phenyl)propanoic acid. 34 H 34NO6[MH] - The MS ESI calculated value was 552.25 and the found value was 552.30. 1 H NMR (400MHz, methanol-d4) δ7.80-7.77(m,2H),7.67-7.65(m,2H),7.48-7.23(m,4H),7.19-7.17(m,2H),7.10-7.07(m,2H),4. 45-4.44(m,1H),4.42-4.41(m,1H),4.34-4.29(m,2H),3.24-3.18(m,1H),2.94-2.86(m,1H),2.12(s,6H),1.45(s,9H).

[0426] Synthesis Scheme 23

[0427]

[0428] F4PcCA

[0429]

[0430] (1R,4s)-4-(4-((S)-2-amino-2-carboxyethyl)phenyl)cyclohexane-1-carboxylic acid

[0431] Step 1: (S)-2-((tert-butoxycarbonyl)amino)-3-(4-iodophenyl)methyl propionate (15.5g, 38.2mmol) and Pd(Ph3P)4(2.21g, 1.91mmol) were added to a mixture of tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-ene-1-formate (29.5g, 96mmol) in THF (30mL) at room temperature. The reactant was warmed to 60°C for 4h. The resulting solution was quenched with water (100mL) and extracted with EtOAc (3x 300mL). The organic layers were combined, washed with brine (2x 200mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to provide a crude product. The residue was purified by silica gel chromatography eluting with a gradient of ethyl acetate in petroleum ether (0% to 25%) to provide tert-butyl 4'-((S)-2-((tert-butoxycarbonyl)amino)-3-methoxy-3-oxopropyl)-2,3,4,5-tetrahydro-[1,1'-biphenyl]-4-carboxylate. 26 H 38 NO6Na[M+Na] + The MS ESI calculated value was 482.26 and the found value was 482.10.

[0432] Step 2: To a mixture of tert-butyl 4'-((S)-2-((tert-butoxycarbonyl)mino)-3-methoxy-3-oxopropyl)-2,3,4,5-tetrahydro-[1,1'-biphenyl]-4-carboxylate (16 g, 34.8 mmol) in methanol (160 mL) was added Pd-C (10% on carbon, wetted with about 55% water, 5.3 g, 4.98 mmol) at room temperature. The solution was degassed with H2 three times and stirred at room temperature under H2 atmosphere (1.5 atm) for 1 h. The resulting solution was filtered and the filtrate was concentrated under reduced pressure to provide tert-butyl (S)-4-(4-(2-((tert-butoxycarbonyl)amino)-3-methoxy-3-oxopropyl)phenyl)cyclohexane-1-carboxylate. C 26 H 41 NO6[M+H] + The MS ESI calculated value was 462.28 and the found value was 462.30.

[0433] Step 3: To a stirred solution of (S)-tert-butyl 4-(4-(2-((tert-butoxycarbonyl)amino)-3-methoxy-3-oxopropyl)phenyl)cyclohexane-1-carboxylate (15 g, 32.5 mmol) in THF (300 mL) was added LiOH (65.0 mL, 65.0 mmol) at room temperature. The solution was stirred at 20 °C for 1 h. The pH of the solution was adjusted to 3 with 1N HCl. The reaction was concentrated under reduced pressure to give (S)-2-((tert-butoxycarbonyl)amino)-3-(4-(4-(tert-butoxycarbonyl)cyclohexyl)phenyl)propanoic acid. 25 H 38 NO6Na[M+Na] + The MS ESI calculated value was 470.26 and the found value was 470.30.

[0434] Step 4: Separate (S)-2-((tert-butoxycarbonyl)amino)-3-(4-(4-(tert-butoxycarbonyl)cyclohexyl)phenyl)propanoic acid (14 g, 31.3 mmol) using preparative supercritical fluid chromatography using the following conditions: column: CHIRALART Cellulose-SB, 3 x 25 cm, 5 μm; mobile phase A: CO2, mobile phase B: MeOH (0.1% 2M NH3-MEOH); flow rate: 80 mL / min; gradient: 10% B; 220 nm; retention time of peak 1: 7.45; retention time of peak 2: 8.38; to provide the first eluting peak (S)-2-((tert-butoxycarbonyl)amino)-3-(4-((1s,4R)-4-(tert-butoxycarbonyl)cyclohexyl)phenyl)propanoic acid. C 25 H 38 NO6Na[M+Na] +The MS ESI calculated value was 470.26 and the found value was 470.30. 1H NMR (300 MHz, CDCl3) δ7.11 (d, J=3.0 Hz, 4H), 4.27 (s, 1H), 3.18-3.12 (m, 1H), 2.91 (s, 1H), 2.60 (s, 1H), 2.49 (s, 1H), 2.20 (d, J=10.9 Hz, 2H), 1.78-1.52 (m, 6H), 1.49 (d, J=0.7 Hz, 9H), 1.36 (s, 9H).

[0435] and the second eluting peak (S)-2-((tert-butoxycarbonyl)amino)-3-(4-((1r,4S)-4-(tert-butoxycarbonyl)cyclohexyl)phenyl)propanoic acid. 25 H 38 NO6Na[M+Na] + The MS ESI calculated value is 470.26, and the measured value is 470.30. 1HNMR (300 MHz, CDCl3) δ7.14-7.06 (m, 4H), 4.25 (s, 1H), 3.14 (d, J = 12.3 Hz, 1H), 2.88 (s, 1H), 2.49-2.41 (m, 1H), 2.29-2.15 (m, 1H), 2.05 (d, J = 12.2 Hz, 2H), 1.90 (d, J = 12.1 Hz, 2H), 1.69-1.47 (m, 2H), 1.47 (s, 11H), 1.34 (s, 9H).

[0436] Step 5: To a solution of (S)-2-((tert-butoxycarbonyl)amino)-3-(4-((1s,4R)-4-(tert-butoxycarbonyl)cyclohexyl)phenyl)propanoic acid (8.3 g, 18.54 mmol) in THF (80 mL) was added HCl (9.27 mL, 18.54 mmol) portionwise at room temperature. The reaction was concentrated under reduced pressure to afford (S)-2-amino-3-(4-((1s,4R)-4-(tert-butoxycarbonyl)cyclohexyl)phenyl)propanoic acid. 20 H 30 NO4[M+H] + The MS ESI calculated value was 348.21 and the found value was 348.25.

[0437] Step 6: To a stirred solution of (S)-2-amino-3-(4-((1s,4R)-4-(tert-butoxycarbonyl)cyclohexyl)phenyl)propanoic acid (6 g, 17.27 mmol) and NaHCO (7.25 g, 86 mmol) in THF (60 mL) and water (60.0 mL) was added Fmoc-OSu (5.24 g, 15.54 mmol). The mixture was stirred at room temperature for 1 h. The pH of the solution was adjusted to 3 with 1N HCl. The aqueous phase was extracted with EtOAc (2 x 200 mL). The combined organic layers were washed with brine (150 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by reverse phase column chromatography to give (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-((1s,4R)-4-(tert-butoxycarbonyl)cyclohexyl)phenyl)propanoic acid. 35 H 41 NO6[M+H] + The MS ESI calculated value was 570.28 and the found value was 570.15. 1 HNMR (300MHz, methanol-d4) δ7.81(d,J=7.5Hz,2H),7.61(d,J=7.5Hz,2H),7.43-7.27(m,4H),7.16(d,J=7.8Hz,2H),7.07(d,J=7.9Hz,2H),4.45-4.3 3(m,2H),4.22-4.06(m,2H),3.26-3.14(m,1H),2.96-2.88(m,1H),2.5 9(s,1H),2.48(s,1H),2.15(s,2H),1.61(d,J=5.5Hz,6H),1.49(s,9H).

[0438] Synthesis Scheme 24

[0439]

[0440] F4PtCC

[0441]

[0442] (1S,4r)-4-(4-((S)-2-amino-2-carboxyethyl)phenyl)cyclohexane-1-carboxylic acid

[0443] Step 1: To a mixture of (S)-2-((tert-butoxycarbonyl)amino)-3-(4-((1r,4S)-4-(tert-butoxycarbonyl)cyclohexyl)phenyl)propanoic acid (3 g, 6.70 mmol) in THF (30 mL) was added hydrogen chloride (30 mL, 60.0 mmol) portionwise at room temperature. The reaction was concentrated under reduced pressure to provide (S)-2-amino-3-(4-((1r,4S)-4-(tert-butoxycarbonyl)cyclohexyl)phenyl)propanoic acid. 20 H 31 NO4[M+H] + The MS ESI calculated value was 348.21 and the found value was 348.25.

[0444] Step 2: To a stirred solution of (S)-2-amino-3-(4-((1r,4S)-4-(tert-butoxycarbonyl)cyclohexyl)phenyl)propanoic acid (2 g, 5.76 mmol) (1.748 g, 5.18 mmol) and NaHCO3 (2.418 g, 28.8 mmol) in THF (20 mL) and water (20 mL) was added Fmoc-OSu (1.748 g, 5.18 mmol) at room temperature. The mixture was stirred at 20 °C for 1 h. The pH of the solution was adjusted to 3 with 1N HCl. The aqueous phase was extracted with EtOAc (2 x 200 mL). The combined organic layers were washed with brine (3 x 50 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by reverse phase column chromatography to give (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-((1r,4S)-4-(tert-butoxycarbonyl)cyclohexyl)phenyl)propanoic acid. 35 H 41 NO6[M+H] + The MS ESI calculated value was 570.28 and the found value was 570.35. 1 H NMR (300MHz, methanol-d4) δ7.81(d,J=7.6Hz,2H),7.61(d,J=7.5Hz,2H),7.43-7.38(m,2 H),7.34-7.27(m,2H),7.17(d,J=8.0Hz,2H),7.08(d,J=7.8Hz,2H),4.47-4.43(m, 1H),4.35-4.30(m,1H),4.21-4.02(m,2H),3.24-3.18(m,1H),2.94-2.87(m,1H),2 .40(s,1H),2.17(d,J=11.9Hz,1H),1.96(s,3H),1.82-1.55(m,2H),1.47(s,13H).

[0445] Preparation of final compound:

[0446] A. General Procedure for the Synthesis of Linear Peptide Precursors

[0447] The peptides in Table 1 were synthesized using standard solid phase synthesis using Fmoc / tBu chemistry as exemplified in Chan, WC; White, PD "Fmoc Solid-Phase Synthesis: a Practical Approach", Oxford University Press, Oxford, 2000; Steward, J.; Young, J. "Solid Phase Peptide Synthesis", Pierce Chemical Company, Rockford, 1984.; Benoiton, NL "Chemistry of Peptide Synthesis", CRC Press, New York, 2006; and Lloyd-Williams, P.; Albericio, F.; Giralt, E. "Chemical Approaches to the Synthesis of Peptides and Proteins", CRC Press, New York, 1997.

[0448] During the peptide chain extension process, the α-amino group of each amino acid is protected by a 9H-fluoren-9-ylmethoxycarbonyl group (Fmoc). In order to avoid any side reactions during the chain extension step, any reactive amino acid side chains also carry acid-labile protecting groups, effectively masking the reactive groups until they are removed by treatment with strong acid. After each coupling step, the Fmoc group of the N-terminal amino acid is removed with piperidine or 4-methylpiperidine, and the resin is thoroughly washed in preparation for the subsequent coupling of Fmoc-protected amino acid derivatives.

[0449] The side chain protecting groups used were: tert-butyl (tBu) for S, hY, Bip4CO2H, Phe4COOH, F4pcCCA, F4ptCCA, F4bcpA; tert-butoxy-carbonyl (Boc) for Dap, dDab, dK, dOrn, K, Prot4NH2, dDap, daMeDab; and β-methylpentyl ester (OMpe) for D.

[0450] Fmoc-protected amino acids are commonly available from suppliers such as Sigma-Aldrich, Novabiochem, Chem-Impex, and Combi-Block.

[0451] B. Synthetic Procedure for Preparing Cyclic Peptides

[0452] Synthesis Scheme 25

[0453]

[0454] Solid Phase Peptide Synthesis Protocol A

[0455] In LibertyBlue from CEM Corporation TM Peptides were synthesized using standard solid phase synthesis on a synthesizer using Fmoc / tBu chemistry as outlined above in Scheme 25. N,N'-diisopropylcarbodiimide (DIC) and ethyl cyano(oxime)acetate (Oxyma) were used as coupling agents to form an amide bond between the free amino terminus of the resin-bound protected peptide and the carboxylic acid of the Fmoc-protected amino acid.

[0456] The synthesis was performed using 2-chlorotrityl resin loaded with H-Gly (200-400 mesh, 0.79 mmol / g loading, 1% cross-linked polystyrene, Novabiochem). All amino acids were dissolved in DMF (N,N-dimethylformamide) at a concentration of 0.2 M. The amino acids were activated with an equimolar amount of Oxyma solution (0.5 M in DMF) and a 2-fold molar excess of DIC solution (1.0 M in DMF). Alternatively, the amino acids were dissolved in DMF (N,N-dimethylformamide) at a concentration of 0.125 M. The amino acids were activated with an equimolar amount of pure Oxyma solution (0.125 M in DMF; containing 0.05 M DIEA) and a 2-fold molar excess of DIC solution (0.25 M in DMF). The reactions were usually performed on a 25 μmol scale.

[0457] Each synthesis cycle includes: Fmoc-amino acid deprotection with 20% piperidine in DMF (90°C microwave assisted heating, 2 min) and coupling with Fmoc-protected amino acid / DIC / Oxyma (5, 5 and 10 equivalents, respectively; 90°C microwave assisted heating, 2 min or 4 min) (may be repeated 2 times for difficult couplings). The cycle of Fmoc deprotection and Fmoc-protected amino acid coupling was repeated using the desired monomer until a complete linear peptide was formed.

[0458] Solid Phase Peptide Synthesis Scheme B

[0459] Alternatively, in Peptides were synthesized using standard solid phase synthesis using Fmoc / tBu chemistry on a Syro II peptide synthesizer as outlined in Scheme 25 above. HATU and DIPEA were used as coupling agents to form an amide bond between the free amino terminus of the resin-bound protected peptide and the carboxylic acid of the Fmoc-protected amino acid. Synthesis was performed using 2-chlorotrityl resin loaded with H-Gly (200-400 mesh, 0.79 mmol / g loading, 1% cross-linked polystyrene, Novabiochem). All amino acids were dissolved in 1:1 DMF:NMP at a concentration of 0.2 M. Reactions were typically performed on a 12 μmol scale.

[0460] Each synthesis cycle included: (1) coupling with Fmoc-protected amino acid / HATU / DIPEA (4, 4 and 8 equivalents, respectively; room temperature; 15 min) (repeated twice). The mixture was filtered and the peptidyl resin was washed with DMF (2 x 1 mL); (2) Fmoc deprotection (repeated 3 times): 20% 4-methylpiperidine in DMF (1 mL; room temperature; 3 min). The mixture was filtered and the peptidyl resin was washed with DMF (4 x 1 mL). The cycle of Fmoc deprotection and Fmoc-protected amino acid coupling was repeated using the desired monomer until a complete linear peptide was formed.

[0461] Selective cleavage and macrolactamization of protected peptides

[0462] To cleave the protected linear peptide from the solid support, the peptidyl resin (~16 mg) was treated with 25% hexafluoroisopropanol (HFIP) in DCM at room temperature for 20 min, filtered, and the solvent removed under reduced pressure. The resulting residue was dissolved in DMF (5 mL). HATU (0.44 eq.) and DIPEA (2.5 eq.) were added. The mixture was stirred at room temperature for 5 min. An additional 0.66 eq. of HATU was then added. After completion of macrolactamization (monitored by UPLC-MS), the solvent was removed under reduced pressure.

[0463] Final sidechain deprotection

[0464] A solution of TFA / HO / TIS (90 / 8 / 2, v / v / v, 1 mL) was added to the crude protected cyclic peptide. The mixture was stirred at room temperature for 10 min. Cold ether (15 mL) was added to the solution. The peptide was precipitated by centrifugation (3200 rpm, -10 ° C). The precipitate was washed with ether (2 x 10 mL) and dried overnight under vacuum to produce a crude deprotected cyclic peptide as a solid.

[0465] HPLC purification

[0466] Using a Waters MS-Directed AutoPurification HPLC / MS system, the HPLC-MS was carried out on a Waters X-BridgePrep C18 OBD Prep column ( 5pm, column size 19*100mm) by preparative reverse phase high performance liquid chromatography (RP-HPLC). Mobile phase: (A) 0.16% TFA in HPLC water and (B) 0.16% TFA in HPLC acetonitrile; flow rate: 25mL / min; UV wavelength λ=215nm; gradient: 25-50% B over 5min. Alternatively, purification was performed on a Waters CSH-C18 column (19x250mm, 5uM) using Agilent with a 1290infinityII preparative LC system and LC-MSD XT mass spectrometer. Mobile phase: (A) 0.1% formic acid in HPLC water and (B) 0.1% formic acid in HPLC acetonitrile; flow rate: 25mL / min; UV wavelength λ=215nm; gradient: 20% B over 2.5min, 55% B over 2.5-20min. The UV absorbing fractions containing the target m / z ion were collected and the fractions containing the product were confirmed by LC / MS.

[0467] The identity and purity of the final compound were confirmed by UPLC-MS, which was measured by a reversed-phase WatersAcquity UPLC-MS system. Column: Waters XSelect CSH C18 column ( 2.5 μm, column size 2.1*50 mm). Mobile phase: (A) 0.05% TFA in HPLC water and (B) 0.05% TFA in HPLC acetonitrile; injection volume: 1 μL; flow rate: 1 mL / min; UV wavelength λ=215 nm; gradient: 5-100% B in 5 min. The pooled fractions containing the pure peptide were lyophilized to obtain the final cyclized product as a powder.

[0468] Biological assays:

[0469] Procedure for IL-6 assay in MRC5 cells

[0470] Inhibition of IL-1β-induced IL-6 secretion was evaluated in MRC5 cells. 2X EC was prepared in EMEM (ATCC 30-2003) containing 0.025% BSA (Sigma A9576), 1X penicillin / streptomycin (Gibco 15070-063), 1X NEAA (Gibco 11140-050), 1X GlutaMax (Gibco 35050-061) and 1X sodium pyruvate (Gibco 11360-070). 80 Concentration of recombinant human IL-1β (BioLegend 579404). 20 μL of IL-1β was added to a 384-well collagen-coated plate (Corning 354664) and pre-incubated for 1 hour at ambient temperature with 200 nL of compound dispensed using an ECHO 555 liquid handler. Human lung fibroblast MRC5 cells (ATCC CCL-171) were added at a density of 3000 cells / 20 μL per well. Cells were prepared by passage three times in growth medium EMEM (ATCC30-2003) containing 10% fetal bovine serum (Gibco 16140-071), 1X penicillin / streptomycin (Gibco 15070-063), 1X NEAA (Gibco 11140-050), 1X GlutaMax (Gibco 35050-061) and 1X sodium pyruvate (Gibco 11360-070) in collagen-coated T175 flasks (Greiner 661950) and harvested in seeding medium after 5 minutes of digestion with 0.25% trypsin-EDTA (Gibco 25200-056). 384-well collagen-coated plates containing a final volume of 40 μL were incubated overnight at 37° C., 5% CO 2 . 5 μL of conditioned medium was transferred to a 384-well AlphaLISA plate (PerkinElmer 6005350) and IL-6 was detected using a human AlphaLISA IL-6 kit (PerkinElmer AL223F) according to the manufacturer's protocol. 20 μL of the acceptor bead / biotinylated antibody mixture was added to a 384-well AlphaLISA plate and incubated at ambient temperature for 1 hour. The donor bead mixture was protected from light and 25 μL was added to the plate and incubated at ambient temperature for 30 minutes. The AlphaLISA plate was read on an EnVision multimode plate reader (PerkinElmer model 2104) using the AlphaScreen setting (laser excitation wavelength of 680 nm and emission wavelength of 570 nm). Dose response curves and IC were analyzed using a 4-parameter logistic equation in Spotfire software (Tibco, Palo Alto, CA). 50 value.

[0471] The amino acid sequences, biological activities (MRC IC 50 ), calculated monoisotopic mass, molecular formula, calculated molecular weight, and mass spectral data ([M+H]+ or [M+2H] / 2+).

[0472] Table 1

[0473]

[0474]

[0475]

[0476]

[0477]

[0478]

[0479]

[0480]

[0481]

[0482]

[0483]

[0484]

[0485]

[0486]

[0487]

[0488]

[0489]

[0490]

Claims

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof in: R 1 is CH3C(O)NH-CH2CH2-O- or C 1 ; C 1 yes: (i) a 5- to 6-membered monocyclic aromatic or heteroaromatic group, wherein the heteroaromatic group contains 1 to 2 heteroatoms selected from N, O and S; or (ii) a 5- to 6-membered monocyclic or bicyclic saturated cycloalkyl or heterocycloalkyl group containing 1 to 2 heteroatoms selected from N, O and S; or (iii) a 5- to 6-membered monocyclic or bicyclic cycloalkyl group; Among them C 1 is unsubstituted or substituted with 1 to 3 R C1 Substituent substituted, said R C1 The substituents are independently selected from halogen, C1-C3 alkyl, C1-C3 fluoroalkyl, carboxyl, C1-C3 alkoxy and C2-C3 acyl; R 2 is H, C1-C3 alkyl, benzyl or phenyl-CH2CH2-; R 3 is a 9- to 10-membered bicyclic aryl or heteroaryl group, wherein the heteroaryl group contains 1 to 2 heteroatoms selected from N, O and S; Where R 3 is unsubstituted or substituted with 1 to 3 R 3a Substituent substituted, said R 3a The substituents are independently selected from halogen, C1-C3 alkyl, C1-C3 fluoroalkyl, hydroxyl and C1-C3 alkoxy; R 4 It is C1-C3 alkyl, HO2C-(CH2) m -、H2NC(O)-(CH2) m -、(CH3)2NC(O)-(CH2) m - or tetrazolyl-(CH2) m -; R 5 It is amino, H2N(CH2) n -、H2NC(O)-(CH2) n -, CH3C(O)NH-, CH3C(O)NH(CH2) n -、C 5 or C 5 -CH2-, C 5 yes: (ii) a 5- to 6-membered monocyclic aromatic or heteroaromatic group, wherein the heteroaromatic group contains 1 to 2 heteroatoms selected from N, O and S; (ii) a 9- to 10-membered bicyclic aryl or heteroaryl group, wherein the bicyclic heteroaryl group contains 1 to 3 heteroatoms selected from N, O and S; (iii) a 5- to 6-membered monocyclic or 9- to 10-membered heterocycloalkyl group, wherein the heterocycloalkyl group is saturated or partially unsaturated and contains 1 to 2 heteroatoms selected from N, O and S; (iv) a 5- to 6-membered monocyclic cycloalkyl group; (v) 2,3-dihydroindole; Among them C 5 is unsubstituted or substituted with 1 to 3 R C5 Substituent substituted, said R C5 The substituents are independently selected from halogen, amino, hydroxy, C1-C3 alkyl, C1-C3 fluoroalkyl, C1-C3 alkoxy, H2N-(CH2) k -、H2NC(O)-(CH2) k -, H2C=CH-CH2O- and phenyl; R 6 H, C1-C5 alkyl, H2N(CH2) p -, HOCH2-, (CH3)2NCH2-, H3CO-(CH2) q - or C 6 -CH2-; C 6 is a 5- or 6-membered monocyclic saturated heterocycloalkyl group containing 1 to 2 heteroatoms selected from N, O and S; and wherein C 6 is unsubstituted or substituted with 1 to 3 R C6 Substituent substituted, said R C6 The substituents are independently selected from halogen, C1-C3 alkyl, C1-C3 fluoroalkyl, hydroxyl and C1-C3 alkoxy; R 7 is H or C1-C3 alkyl; R 8a H, C1-C5 alkyl, HOCH2-, H2N(CH2) r -、(CH3)3N + (CH2) r - or CH3C(O)NH(CH2) r -; R 8b is H or C1-C3 alkyl; R 9a is H or C1-C3 alkyl; R 9b H, C1-C5 alkyl, C 9 -CH2- or C 9 -CH2CH2-; C 9 yes: (i) a 5- to 6-membered monocyclic aromatic or heteroaromatic group, wherein the heteroaromatic group contains 1 to 2 heteroatoms selected from N, O and S; or (ii) a 5- to 6-membered monocyclic saturated cycloalkyl or heterocycloalkyl group, wherein the heterocycloalkyl group contains 1 to 2 heteroatoms selected from N, O and S; Among them C 9 is unsubstituted or substituted with 1 to 3 R C9 Substituent substituted, said R C9 The substituents are independently selected from halogen, amino, hydroxy, cyano, C1-C3 alkyl, C1-C3 fluoroalkyl, C1-C3 alkoxy, H2N-(CH2) k -、H2NC(O)-(CH2) k -, H2NCH2CH2O-, CH3C(O)NH-CH2CH2O- and morpholinyl-CH2CH2O-; R 10 is H, halogen or C1-C3 alkyl; R 11 is H, halogen or C1-C3 alkyl; Each occurrence of subscript k is independently 1 or 2; Subscript m is 1 or 2; Subscript n is 1, 2, 3, or 4; The subscript p is 1, 2, 3, or 4; Subscript q is 1 or 2; The subscript r is 1, 2, 3, or 4; X 1 , X 2 and X 3 are independently C(H) or N; and A 1 and A 2 Independently selected from HO2C-, H2NC(O)-, CH3C(O)N(H)-, H2NS(O)2-, CH3S(O)2N(H)-, tetrazolyl and 5-oxooxadiazolyl.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein: R 5 It is amino, H2N(CH2) n -、H2NC(O)-(CH2) n -、C 5 or C 5 -CH2-, C 5 yes: (i) a 5- to 6-membered monocyclic aromatic or heteroaromatic group, wherein the heteroaromatic group contains 1 to 2 heteroatoms selected from N, O and S: (ii) a 9- to 10-membered bicyclic aryl or heteroaryl group, wherein the bicyclic heteroaryl group contains 1 to 3 heteroatoms selected from N, O and S; (iii) a 5- to 6-membered monocyclic or 9- to 10-membered heterocycloalkyl group, wherein the heterocycloalkyl group is saturated or partially unsaturated and contains 1 to 2 heteroatoms selected from N, O and S; or (iv) 2,3-dihydroindole; Among them C 5 is unsubstituted or substituted with 1 to 3 R C5 Substituent substituted, said R C5 The substituents are independently selected from halogen, amino, hydroxy, C1-C3 alkyl, C1-C3 fluoroalkyl, C1-C3 alkoxy, H2N-(CH2) k -、H2NC(O)-(CH2) k -, H2C=CH-CH2O- and phenyl; R 6 H, C2-C5 alkyl, H2N(CH2) p -, HOCH2-, (CH3)2NCH2-, H3CO-(CH2) q - or C 6 -CH2-; R 8a is H, C1-C3 alkyl, HOCH2- or H2N(CH2) r -; R 9a It is H; R 9b is H, C1-C3 alkyl, C 9 -CH2- or C 9 -CH2CH2-; R 10 It is H; R 11 It is H; subscript p is 1, 2, or 3; and The subscript r is 2, 3, or 4.

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein: C1 is phenyl, pyrimidinyl or piperazinyl, wherein C 1 is unsubstituted or substituted with 1 to 2 R C1 Substituent substitution; R 3 is naphthyl or indolyl, where R 3 is unsubstituted or substituted with 1 to 2 R 3a Substituent substitution; C 5 is phenyl, pyridyl, pyrimidinyl, naphthyl, indolyl, 7-azaindolyl, indazolyl, 2,3-dihydroindolinyl, piperidinyl, tetrahydropyranyl or cyclohexyl, wherein C 5 is unsubstituted or substituted with 1 to 2 R C5 replace; C 6 is tetrahydropyranyl or morpholinyl; wherein C 6 is unsubstituted or substituted with 1 to 2 R C6 replace; and C 9a is H or methyl; C 9b is phenyl, pyridyl, cyclohexyl, morpholinyl or piperidinyl, wherein C 9 is unsubstituted or substituted with 1 to 2 R C9 replace.

4. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein X1 and X2 are C(H); and R1 is a phenyl group substituted with a carboxyl group.

5. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein X1 and X2 are C(H); and R1 is CH3C(O)NH-CH2CH2-O-.

6. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R 2 It's H.

7. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R 3 It is an indolyl group substituted by a halogen.

8. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R 3 It is naphthyl.

9. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R 4 It is HO2C-(CH2) m -.

10. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein X 3 It is C(H).

11. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R 5 It is H2N(CH2) n -, indole, 7-azaindole, naphthyl or pyridinyl.

12. The compound according to claim 5 or a pharmaceutically acceptable salt thereof, wherein R 5 It is H2N(CH2) n -, indole, naphthyl or pyridyl.

13. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R 6 is H, HOCH2-, C2-C5 alkyl or H2N(CH2) p -.

14. The compound according to claim 13 or a pharmaceutically acceptable salt thereof, wherein R 6 Is C2-C5 alkyl or H2N(CH2) p -.

15. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein R 7 It's H.

16. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein: R 8a is methyl or H2NCH2CH2-; and R 8b It's H.

17. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein R 9b yes 18. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein R 9b yes 19. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein R 10 is H; and R 11 It is H, F or Cl.

20. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein A 1 and A 2 They are all HO2C-.

21. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein: R 1 is 4-CH3C(O)-piperazin-1-yl, CH3C(O)NH-CH2CH2-O-, 5-CO2H-pyrimidin-2-yl or 4-CO2H-phenyl; R 2 is H, ethyl, benzyl or phenyl -CH2CH2-; R 3 is naphthyl-1-yl, 4-fluoroindol-3-yl or 4-chloroindol-3-yl; R 4 It is methyl, HO2C-(CH2) m - or H2NC(O)-(CH2) m -; R 5 It is amino, H2N(CH2) n -, naphthyl-1-yl, indol-3-yl, 7-aza-indol-3-yl, indazol-1-yl, 2,3-dihydroindolin-1-yl, pyridin-3-yl, pyridin-4-yl, piperidin-4-yl, 3-aminomethylphenyl, 4-aminomethylphenyl, 3-aminophenyl, 4-aminophenyl, phenyl, pyridin-4-yl-CH2-, pyrimidin-5-yl, tetrahydropyran-4-yl-, H2NC(O)-(CH2)2-, 3-biphenyl, 3-CH2=CH-CH2O-phenyl, CH3C(O)NH-, CH3C(O)NH(CH2)3-, or cyclohexyl; R 6 It is H, (CH3)2CHCH2-, (CH3)3CCH2-, H2N(CH2) p -, HOCH2-, H3CCH2CH2-, morpholin-4-yl-CH2-, tetrahydropyran-4-yl-CH2-, (CH3)2NCH2- or H3CO-(CH2) q -; R 7 is H or methyl; R 8a It is H, methyl, HOCH2-, H2N(CH2) r -, (CH3)3NCH2CH2- or CH3C(O)NH(CH2)4-; R 8b is H or methyl; R 9a is H or methyl; R 9b is H, methyl, H3CCH2CH2CH2-, 4-HO-phenyl-CH2CH2-, phenyl-CH2CH2-, cyclohexyl-CH2CH2-, 5-NC-pyridin-3-yl-CH2CH2-, 4-F3C-phenyl-CH2-, 3-F3C-phenyl-CH2-, 2-F3C-phenyl-CH2-, morpholin-4-yl-CH2CH2O-phenyl-CH2-, 4-aminophenyl-CH2-, 4,4-difluorocyclohexyl-CH2-, 4-H2NCH2CH2O-phenyl-CH2-, 4-H2NCH2CH2O-pyridin-3-yl-CH2-, piperidin-4-yl-CH2-, or 4-CH3C(O)NH-CH2CH2O-phenyl-CH2-; R 10 is H, fluorine or methyl; R 11 is H, fluorine or chlorine; A 1 and A 2 All are HO2C-; and X 1 , X 2 and X 3 It is C(H).

22. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound is selected from SEQ ID NO: 1-215.

23. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound is selected from (SEQ ID NOs 78, 71-72, 67, 65, 70, 69, 68, 66, 64, 77, 79, 209, 193, 215, 212 and 210, respectively, in order of appearance):

24. A pharmaceutical composition comprising the compound according to any one of claims 1 to 23 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

25. A method for treating atherosclerosis, the method comprising administering to a subject in need thereof a therapeutically effective amount of a compound according to any one of claims 1 to 23 or a pharmaceutically acceptable salt thereof.

26. A method for treating vascular inflammation, the method comprising administering to a subject in need thereof a therapeutically effective amount of a compound according to any one of claims 1 to 23 or a pharmaceutically acceptable salt thereof.

27. A method for treating inflammatory lesions, the method comprising administering to a subject in need thereof a therapeutically effective amount of a compound according to any one of claims 1 to 23 or a pharmaceutically acceptable salt thereof.

28. The method of claim 25, 26 or 27, wherein an effective amount of the compound or a pharmaceutically acceptable salt thereof is orally administered to the subject.

29. A compound according to any one of claims 1 to 23 or a pharmaceutically acceptable salt thereof for use in therapy.

30. Use of the compound according to any one of claims 1 to 23 or a pharmaceutically acceptable salt thereof for treating atherosclerosis.

31. Use of a compound according to any one of claims 1 to 23 or a pharmaceutically acceptable salt thereof for treating vascular inflammation.

32. Use of the compound according to any one of claims 1 to 23 or a pharmaceutically acceptable salt thereof for treating inflammatory lesions.

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