Cyclic peptides for capturing interleukin-1 beta

Abstract

Compounds of formula (I) or pharmaceutically acceptable salts thereof are provided, wherein X 1 , X 2 , X 3 , X 4 , R 1 ~R 4 , R 5a , R 5b , R 6a , R 6b , R 7a , R 7b , R 8a , R 8b , R 9 ~R 17 and subscripts v and w are as described herein, and are capable of capturing IL-1β and are expected to have utility as therapeutic agents for treating, for example, cardiovascular disease. The present disclosure also provides pharmaceutical compositions comprising the compounds disclosed herein, or pharmaceutically acceptable salts thereof. The present disclosure also relates to methods for using the compounds, or pharmaceutically acceptable salts thereof, in the treatment and prevention of cardiovascular disease and inflammatory disorders, as well as methods for preparing medicaments for this purpose. [Formula 1] TIFF2025542068000189.tif75140

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Patent Application No. 63 / 496,260, filed April 14, 2023, the entire contents of which are incorporated herein by reference.

[0002] Reference to an electronically submitted sequence listing The contents of the Electronic Sequence Listing (25677-WO-PCT_SL.xml; size: 1,859,444 bytes; created April 4, 2024) are incorporated herein by reference in their entirety.

[0003] The present disclosure relates to certain cyclic peptides that sequester interleukin-1β (IL-1β), pharmaceutical compositions containing such peptides, and methods of using the compounds to treat, inhibit, or ameliorate one or more cardiovascular disease conditions that can benefit from sequestering IL-1β, including atherosclerosis. [Background technology]

[0004] Atherosclerosis is a disease of the arteries characterized by the buildup of cholesterol plaques on the inner walls of arteries. The progression of atherosclerosis can lead to hardening or narrowing of the arteries, increasing the risk of plaque rupture. These ruptures can release cholesterol globules and other substances into the bloodstream, leading to blockage of blood flow to the brain, heart, or other organs. Medically, these are known 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 for preventing the progression of ASCVD is a healthy diet and exercise, but compliance is generally poor. Pharmacological treatment of ASCVD has primarily focused on cholesterol-lowering medications such as statins, cholesterol absorption inhibitors, and low-density lipoprotein (LDL) receptor inhibitors. These medications are highly effective in reducing the accumulation of fatty acid deposits and improving arterial health. Other medications prescribed for ASCVD that do not improve disease symptoms include blood thinners such as aspirin to prevent platelets from clumping in small arteries, and blood pressure medications to reduce the risk and severity of heart attacks. Surgical options for more aggressive intervention in advanced cases of ASCVD include angioplasty, stent placement, endarterectomy (surgical removal of plaque), and bypass surgery.

[0007] Cholesterol-lowering drugs have served as an important standard of care for slowing the progression of atherosclerosis, but clinical data support an additional important role for inflammation in the progression of untreated ASCVD. Inflammatory biomarkers, such as CRP, are associated with increased risk of cardiovascular events, independent of cholesterol levels. The Canakinumab Anti-inflammatory Thrombosis Outcomes Study (CANTOS) was the first clinical trial to demonstrate that reducing vascular inflammation in the absence of concomitant lipid lowering 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 use in rheumatic disorders. IL-1β is a pro-inflammatory cytokine that induces IL-6, thereby elevating the downstream inflammatory biomarker high-sensitivity CRP (hsCRP). CANTOS therefore provides proof of concept that IL-1β-targeted therapy may reduce rates of MACE in certain patients in a manner that is complementary and potentially additive to LDL-lowering standard therapy.

[0008] Additional non-surgical treatment approaches beyond standard-of-care cholesterol-lowering medications to slow atherosclerosis progression and reduce the risk of MACE are needed. Furthermore, patients suffering from inflammatory disorders would benefit from an orally administered agent that blocks the same cytokine, IL-1β, as canakinumab. Summary of the Invention [Means for solving the problem]

[0009] The present disclosure provides certain cyclic peptides that reduce inflammation by binding to IL-1β cytokines, preventing their binding to IL-1 receptors and inhibiting downstream pro-inflammatory signaling. These cyclic peptides may be useful pharmaceutically active compounds for the treatment of cardiovascular diseases and inflammatory disorders. In one aspect, the present disclosure provides compounds of formula (I): [ka]

[0010] and pharmaceutically acceptable salts thereof.

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

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

[0013] Compounds of the Disclosure In one embodiment, the present disclosure provides a compound having formula (I) above, wherein: R 1 is R 1e -C(O)NH-CH2CH2-O-, C1-C4 alkyl, halo, or C 1 and R 1e -teeth, (a) C1-C4 alkyl, or (b)C Y1 (where C Y1 teeth, (i) C3-C6 cycloalkyl, (ii) phenyl, or (iii) a 5- to 6-membered monocyclic heteroaryl, the heteroaryl containing 1 to 2 heteroatoms independently selected from the group consisting of N, O, and S; where C Y1 is unsubstituted or has 1 to 3 R selected from the group consisting of C1-C3 alkyl, halo, and piperazinyl Y1 is substituted with a substituent, C 1 teeth, (i) a 5- to 6-membered monocyclic aryl or heteroaryl, wherein the heteroaryl contains 1 to 2 heteroatoms independently selected from the group consisting of N, O, and S; (ii) a 3- to 6-membered monocyclic or 5- to 8-membered bicyclic cycloalkyl; or (iii) a 5- to 6-membered monocyclic saturated heterocycloalkyl containing 1 to 2 heteroatoms independently selected from the group consisting of N, O, and S; where C 1 is unsubstituted or is substituted with 1 to 3 R independently selected from the group consisting of halo, C-C alkyl, C-C fluoroalkyl, carboxy, C-C alkoxy, C-C acyl, —C(O)NH and —C(O)N(CH) C1 is substituted with a substituent, R 2 teeth, (i) a 5- to 6-membered monocyclic aryl or heteroaryl, wherein the heteroaryl contains 1 to 2 heteroatoms independently selected from the group consisting of N, O, and S; and (ii) a 3- to 8-membered monocyclic or bicyclic cycloalkyl; where R 2 is unsubstituted or is substituted with 1 to 3 R independently selected from the group consisting of halo, amino, hydroxy, C1-C3 alkyl, C1-C3 fluoroalkyl, and C1-C3 alkoxy; 2a is substituted with a substituent, R 2bis H or hydroxy, R 3 is F or hydroxy, R 4 teeth, (i) naphthyl; or (ii) a 9- to 10-membered heteroaryl, the heteroaryl containing 1 to 2 heteroatoms independently selected from the group consisting of N, O, and S; where R 4 is unsubstituted or has 1 to 2 R independently selected from the group consisting of halo 4a is substituted with a substituent, R 5a is H, C1-C3 alkyl, H2N(CH2) m - or HOCH2-, R 5b is H, C1-C3 alkyl, H2N(CH2) m - or HOCH2-, Alternatively, R 5a and R 5b together with the carbon atoms to which they are attached form a C3-C6 cycloalkyl or a 4-6 membered saturated heterocycloalkyl containing one N atom; R 6a and R 6b each independently represents H, —(CH) n1 CH3, -(CH2) n2 -OH, or -(CH2) n2 CO2H, R 7a is H, C1-C3 alkyl, HOCH2-, H2N(CH2) p -, HO2CCH2-, H2NC(O)CH2-, CH3OCH2-, or PhCH2-, R 7b is H, C1-C3 alkyl, HOCH2-, H2N(CH2) p -, HO2CCH2-, H2NC(O)CH2-, CH3OCH2-, or PhCH2-, Alternatively, R 7a and R 7btogether with the carbon atoms to which they are attached form a 4- to 6-membered saturated heterocycloalkyl containing one N atom; R 8a is HO-(CH2) q -, CH3-O-(CH2) q -, CH3CH2-O-(CH2) q -, PhCH2-O-(CH2) q -, C 1~ C3 alkyl, C1-C3 fluoroalkyl, H2N-(CH2) r -, (CH3)3N-(CH2) r -, H2NC(NH)N(H)-(CH2) r -, H2NC(O)N(H)-(CH2) r -, HO2C-(CH2) r -, (CH3)SO2-(CH2) r -, C 8a , or C 8a- CH2-, where C 8a teeth, (i) a 5- to 6-membered monocyclic aryl or heteroaryl, wherein the heteroaryl contains 1 to 2 heteroatoms independently selected from the group consisting of N, O, and S; (ii) a 5- to 6-membered monocyclic saturated heterocycloalkyl, wherein the heterocycloalkyl contains 1 to 2 heteroatoms independently selected from the group consisting of N, O, and S; or (iii) C3-C6 cycloalkyl, where C 8a is unsubstituted or is selected from halo, amino, hydroxy, C1-C3 alkyl, C1-C3 fluoroalkyl, C1-C3 alkoxy, H2N-(CH2) s -, H2NC(O)-(CH2) s 1 to 3 R independently selected from the group consisting of -, H2C=CH-CHO-, and phenyl; C8a is substituted with a substituent, R 8b is H, methyl or hydroxy, R9 is HO-(CH2) t -, H2N-(CH2) u -, H2NC(NH)N(H)-(CH2) u -, H2NC(O)N(H)-(CH2) u - or C 9 and where C 9 is a 5-6 membered saturated heterocycloalkyl containing 1-2 heteroatoms independently selected from the group consisting of N, O and S; where C 9 is unsubstituted or is substituted with 1 to 2 R independently selected from the group consisting of halo, C1-C3 alkyl, C1-C3 fluoroalkyl, and C1-C3 alkoxy; C9 is partially substituted with R 10 is H or methyl, R 11 -H, -CH2-C 11 , or -CH2-C 11 -C a and where C 11 teeth, (i) phenyl; or (ii) a 5- to 6-membered monocyclic aryl or heteroaryl, wherein the heteroaryl contains 1 to 2 heteroatoms independently selected from the group consisting of N, O, and S; where C 11 is unsubstituted or is substituted with 1 to 3 R independently selected from the group consisting of halo, hydroxy, amino, C1-C3 alkyl, C1-C3 fluoroalkyl, carboxy, C1-C3 alkoxy, and C2-C3 acyl; C11 is substituted with a substituent, C a is a 5- to 6-membered monocyclic heteroaryl, the heteroaryl containing 1 to 2 heteroatoms independently selected from the group consisting of N, O, and S; where C ais unsubstituted or is substituted with 1 to 3 R independently selected from the group consisting of halo, hydroxy, amino, C1-C3 alkyl, C1-C3 fluoroalkyl, carboxy, C1-C3 alkoxy, and C2-C3 acyl; Ca is substituted with a substituent, R 12 is H or -CH2C 12 and where C 12 teeth, (i) phenyl, or (ii) a 5- to 6-membered monocyclic heteroaryl, the heteroaryl containing 1 to 2 heteroatoms independently selected from the group consisting of N, O, and S; where C 12 is unsubstituted or is substituted with 1 to 3 R independently selected from the group consisting of halo, hydroxy, amino, C1-C3 alkyl, C1-C3 fluoroalkyl, carboxy, C1-C3 alkoxy, and C2-C3 acyl; C12 is substituted with a substituent, R 13 is H or methyl, R 14 is a halo, R 15 is -OH or -NH2, R 16 is halo, hydroxy, C1-C3 alkyl, C1-C3 fluoroalkyl, or C1-C3 alkoxy; R 17 is H, hydroxy or methyl, each occurrence of subscript m is independently 1, 2, 3, or 4; The subscript n1 is 0, 1, 2, or 3, The subscript n2 is 0, 1, or 2, each occurrence of the subscript p is independently 2, 3, or 4; the subscript q is 0, 1, or 2; The subscript r is 0, 1, 2, or 3, each occurrence of the subscript s is independently 1 or 2; The subscript t is 0, 1, or 2, The subscript u is 0, 1, 2, or 3, The subscript v is 0, 1, or 2, The subscript w is 0, 1, or 2, X 1 and X 2 are independently C(H) or N, X 3 and X 4 are independently C(H), C(Cl), C(F) or N, or Pharmaceutically acceptable salts thereof are provided.

[0014] In some embodiments, the present disclosure provides a method for the preparation of a [ka]

[0015] but [ka]

[0016] The present invention provides a compound of formula (I) wherein

[0017] In certain embodiments, the present disclosure provides compounds of formula (I), wherein X 1 and X 2 is C(H), R 1 teeth, C 1 where C 1 is phenyl or bicyclo[1.1.1]pentanyl, where C 1 is substituted with carboxy, or R 1e -C(O)NH-CHCH-O- (where R 1e is C1-C4 alkyl) is.

[0018] In certain embodiments, the present disclosure provides R2b is H.

[0019] In certain embodiments, the present disclosure provides R 17 is H.

[0020] In one embodiment, the present disclosure provides a compound of formula (IA) [ka]

[0021] The present invention provides a compound of formula (I) having the formula:

[0022] In certain embodiments, the present disclosure provides a compound having formula (IA), wherein: R 1 is CH3C(O)NH-CH2CH2-O- or C 1 and C 1 teeth, (i) a 5- to 6-membered monocyclic aryl or heteroaryl, wherein the heteroaryl contains 1 to 2 heteroatoms independently selected from the group consisting of N, O, and S; (ii) a 3- to 6-membered monocyclic or 5- to 8-membered bicyclic cycloalkyl; or (iii) a 5- to 6-membered monocyclic saturated heterocycloalkyl containing 1 to 2 heteroatoms independently selected from the group consisting of N, O, and S; where C 1 is unsubstituted or is substituted with 1 to 3 R independently selected from the group consisting of halo, C1-C3 alkyl, C1-C3 fluoroalkyl, carboxy, C1-C3 alkoxy, and C2-C3 acyl. C1 is substituted with a substituent, R 2 teeth, (i) a 5- to 6-membered monocyclic aryl or heteroaryl, wherein the heteroaryl contains 1 to 2 heteroatoms independently selected from the group consisting of N, O, and S; and (ii) a 3- to 8-membered monocyclic or bicyclic cycloalkyl; where R 2 is unsubstituted or is substituted with 1 to 3 R independently selected from the group consisting of halo, amino, hydroxy, C1-C3 alkyl, C1-C3 fluoroalkyl, and C1-C3 alkoxy; 2a is substituted with a substituent, R 3 is F or hydroxy, R 4 teeth, (i) naphthyl; or (ii) a 9- to 10-membered heteroaryl, the heteroaryl containing 1 to 2 heteroatoms independently selected from the group consisting of N, O, and S; where R 4 is unsubstituted or has 1 to 2 R independently selected from the group consisting of halo 4a is substituted with a substituent, R 5a is H, C1-C3 alkyl, H2N(CH2) m - or HOCH2-, R 5b is H, C1-C3 alkyl, H2N(CH2) m - or HOCH2-, Alternatively, R 5a and R 5b together with the carbon atoms to which they are attached form a 4- to 6-membered saturated heterocycloalkyl containing one N atom; R 6a and R 6b each independently represents H, —(CH) n1 CH3, -(CH2) n2 -OH, or -(CH2) n2 CO2H, R 7a is H, C1-C3 alkyl, HOCH2-, H2N(CH2) p - or HO2CCH2-, R 7b is H, C1-C3 alkyl, HOCH2-, H2N(CH2)p - or HO2CCH2-, Alternatively, R 7a and R 7b together with the carbon atoms to which they are attached form a 4- to 6-membered saturated heterocycloalkyl containing one N atom; R 8a is HO-(CH2) q -, C1-C3 alkyl, H2N-(CH2) r -, H2NC(NH)N(H)-(CH2) r -, H2NC(O)N(H)-(CH2) r - or C 8a and where C 8a teeth, (i) a 5- to 6-membered monocyclic aryl or heteroaryl, wherein the heteroaryl contains 1 to 2 heteroatoms independently selected from the group consisting of N, O, and S; or (ii) a 5- to 6-membered monocyclic saturated heterocycloalkyl, wherein the heterocycloalkyl contains 1 to 2 heteroatoms independently selected from the group consisting of N, O, and S; where C 8a is unsubstituted or is selected from halo, amino, hydroxy, C1-C3 alkyl, C1-C3 fluoroalkyl, C1-C3 alkoxy, H2N-(CH2) s -, H2NC(O)-(CH2) s 1 to 3 R independently selected from the group consisting of -, H2C=CH-CHO-, and phenyl; C8a is substituted with a substituent, R 8b is H or CH3, R 9 is HO-(CH2) t -, H2N-(CH2) u -, H2NC(NH)N(H)-(CH2) u - or C 9 and where C 9is a 5-6 membered saturated heterocycloalkyl containing 1-2 heteroatoms independently selected from the group consisting of N, O and S; where C 9 is unsubstituted or is substituted with 1 to 2 R independently selected from the group consisting of halo, C1-C3 alkyl, C1-C3 fluoroalkyl, and C1-C3 alkoxy; C9 is partially substituted with R 10 is H or methyl, R 11 -H, -CH2-C 11 , or -CH2-C 11 -C a and C 11 teeth, (i) phenyl; or (ii) a 5- to 6-membered monocyclic aryl or heteroaryl, wherein the heteroaryl contains 1 to 2 heteroatoms independently selected from the group consisting of N, O, and S; where C 11 is unsubstituted or is substituted with 1 to 3 R independently selected from the group consisting of halo, hydroxy, amino, C1-C3 alkyl, C1-C3 fluoroalkyl, carboxy, C1-C3 alkoxy, and C2-C3 acyl; C11 is substituted with a substituent, C a is a 5- to 6-membered monocyclic heteroaryl, the heteroaryl containing 1 to 2 heteroatoms independently selected from the group consisting of N, O, and S; where C a is unsubstituted or is substituted with 1 to 3 R independently selected from the group consisting of halo, hydroxy, amino, C1-C3 alkyl, C1-C3 fluoroalkyl, carboxy, C1-C3 alkoxy, and C2-C3 acyl; Ca is substituted with a substituent, R 12 is H or -CH2C 12 and where C 12 teeth, (i) phenyl, or (ii) a 5- to 6-membered monocyclic heteroaryl, the heteroaryl containing 1 to 2 heteroatoms independently selected from the group consisting of N, O, and S; where C 12 is unsubstituted or is substituted with 1 to 3 R independently selected from the group consisting of halo, hydroxy, amino, C1-C3 alkyl, C1-C3 fluoroalkyl, carboxy, C1-C3 alkoxy, and C2-C3 acyl; C12 is substituted with a substituent, each occurrence of subscript m is independently 1, 2, 3, or 4; The subscript n1 is 0, 1, 2, or 3, The subscript n2 is 0, 1, or 2, each occurrence of the subscript p is independently 2, 3, or 4; the subscript q is 0, 1, or 2; The subscript r is 0, 1, 2, or 3, each occurrence of the subscript s is independently 1 or 2; The subscript t is 0, 1, or 2, The subscript u is 0, 1, 2, or 3, X 1 , X 2 , X 3 and X 4 are independently C(H) or N, or Pharmaceutically acceptable salts thereof are provided.

[0023] In another embodiment, the disclosure provides a compound of formula (I) or (IA), wherein: C 1 is phenyl, pyrimidinyl or piperazinyl, and C 1 is unsubstituted or contains 1-2 R C1 is substituted with a substituent, R 2 is pyridyl or bicyclo[1.1.1]pentanyl, and R 2is unsubstituted or contains 1-2 R 2a is substituted with a substituent, R 4 is indolyl or naphthyl, and R 4 is unsubstituted or contains one R 4a is substituted with a substituent, C 8a is phenyl, pyridyl, pyrimidinyl, pyridazinyl, tetrahydropyranyl, piperidinyl, morpholinyl or piperazinyl, and C 8a is unsubstituted or contains 1-2 R C8a is replaced by C 9 is morpholinyl, and C 9 is unsubstituted or contains one R C9 is replaced by R 11 teeth, -CH2-C 11 (where C 11 is phenyl, pyridyl, pyrimidinyl or pyrazinyl, and C 11 is unsubstituted or contains one R C8 is replaced by ); -CH2-C 11 -C a (where C 11 is phenyl, and C 11 is unsubstituted or contains one R C11 and C a is pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, isoxazolyl or imidazolyl, where C a is unsubstituted or contains one R Ca substituted with ), and R 12 is -CH2C 12 (where C 12 is phenyl or pyridyl, where C 12 is unsubstituted or contains one R C12 (which is replaced by ).

[0024] In another embodiment, the disclosure provides a compound of formula (I) or (IA), wherein: X 1 and X 2 is C(H), R 1 is phenyl substituted with carboxy.

[0025] In another embodiment, the present disclosure provides R 2 is unsubstituted or 1 to 3 R 2a is a 5- to 6-membered monocyclic aryl or heteroaryl substituted with a substituent, 3 is C(H). For example, in one specific embodiment, R 2 is unsubstituted or substituted pyridyl.

[0026] In another embodiment, the present disclosure provides R 2 is an unsubstituted bicyclo[1.1.1]pentanyl.

[0027] In another embodiment, the present disclosure provides R 3 is fluoro.

[0028] In another embodiment, the present disclosure provides R 4 is 4-fluoroindolyl.

[0029] In another embodiment, the present disclosure provides R 5a and R 5b is methyl.

[0030] In another embodiment, the present disclosure provides a compound of formula (I) or (IA), wherein R 6a is H, -(CH2) n1 CH3, -(CH2) n2 -OH, or -(CH2) n2 CO2H, R 6b is H or methyl, The subscript n1 is 1, 2, or 3, The subscript n2 is 0, 1, or 2.

[0031] In another embodiment, the disclosure provides a compound of formula (I) or (IA), wherein: R 6a is -OH or -CH2CO2H, R 6b is H.

[0032] In another embodiment, the present disclosure provides X 4 is C(H).

[0033] In another embodiment, the present disclosure provides R 7a and R 7b is methyl.

[0034] In another embodiment, the present disclosure provides a compound of formula (I) or (IA), wherein R 8a is 1 to 3 R C8a phenyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, piperidinyl, tetrahydropyranyl, or morpholinyl, each of which may or may not be substituted by a substituent; R 8b is H.

[0035] In another embodiment, the present disclosure provides R 8a is an unsubstituted pyridyl, pyrimidinyl, or pyrazinyl.

[0036] In another embodiment, the present disclosure provides R 9 is H2N-(CH2) u - and the subscript u is 1 or 2.

[0037] In another embodiment, the present disclosure provides R 10 is H.

[0038] In another embodiment, the disclosure provides a compound of formula (I) or (IA), wherein: R 11 is H, R 12 is -CH2C 12 (where C 12 is phenyl or pyridyl, where C 12 is unsubstituted or contains one R C12 (which is replaced by ).

[0039] In another embodiment, the disclosure provides a compound of formula (I) or (IA), wherein: R 11 teeth, -CH2-C 11 (where C 11 is phenyl, pyridyl, pyrimidinyl or pyrazinyl, where C 11 is unsubstituted or contains one R C11 is replaced by ); or -CH2-C 11 -C a (where C 11 is phenyl, and C 11 is unsubstituted or contains one R C11 and C a is pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, isoxazolyl or imidazolyl, where C a is unsubstituted or contains one R Ca substituted with R 12 is H.

[0040] In another embodiment, the present disclosure provides a compound of formula (I), wherein: X 1 X 2 , X 3 and X 4 is C(H), R 1 is phenyl substituted with carboxy; R 2 is a 5-6 membered monocyclic aryl or heteroaryl that is unsubstituted or has 1-3 R 2a a 5- to 6-membered monocyclic aryl or heteroaryl substituted with a substituent; R 3 is fluoro, R 4 is 4-fluoroindolyl, R 5a and R 5b is methyl, R 6a is -OH or -CH2CO2H, R 6b is H, R 7a and R 7b is methyl, R 8a is unsubstituted or contains 1 to 3 R C8a phenyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, piperidinyl, tetrahydropyranyl, or morpholinyl, each of which is substituted by a substituent; R 8b is H, R 9 is H2N-(CH2) u - and R 10 is H, and The subscript u is either 1 or 2.

[0041] In a particular aspect of this embodiment, the disclosure provides compounds of formula (I) or (IA), wherein: R 11 is H, and R 12 is -CH2C 12 and C 12 is phenyl or pyridyl, where C 12 is unsubstituted or contains one R C12 is replaced by .

[0042] In another specific aspect of this embodiment, the present disclosure provides compounds of formula (I) or (IA), wherein: R 11 teeth, -CH2-C 11 (where C 11 is phenyl, pyridyl, pyrimidinyl or pyrazinyl, where C 11 is unsubstituted or contains one R C11 is replaced by ); or -CH2-C 11 -C a (where C 11 is phenyl, where C 11 is unsubstituted or contains one R C11 and C a is pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, isoxazolyl or imidazolyl, where C a is unsubstituted or contains one R Ca substituted with ), and R 12 is H.

[0043] In another embodiment, the disclosure provides a compound of formula (I) or (IA), wherein: R 1 is CH3C(O)NH-CH2CH2-O-, 5-CO2H-pyrimidin-2-yl, 4-CH3C(O)-piperazin-1-yl, 4-CO2H-cyclohex-4-yl, 4-CO2H-phenyl, or bicyclo[1.1.1]pentane-1-carboxylic acid; R 2 is pyridin-4-yl, pyridazin-4-yl, bicyclo[1.1.1]pentan-1-yl, or cyclobutyl; R 4 is 4-fluoroindol-3-yl, 4-chloroindol-3-yl, or naphth-1-yl; R 5a is CH3, HOCH2-, H2NCH2CH2CH2CH2-, or H2NCH2CH2-, R 5bis CH3, HOCH2-, or H2NCH2CH2-, Alternatively, R 5a and R 5b together with the carbon atoms to which they are attached form an azetidinyl ring, R 6a is -CH2CO2H, -OH, -H, -CO2H, -CH2OH, CH3, or -CH2CH3, R 6b is H or CH3-, R 7a is CH3, HOCH2-, H2NCH2CH2-, H2NCH2CH2CH2CH2-, or -CH2CO2H, R 7b is CH3, HOCH2-, H2NCH2CH2-, H2NCH2CH2CH2CH2-, or -CH2CO2H, Alternatively, R 7a and R 7b together with the carbon atoms to which they are attached form an azetidinyl or piperidinyl ring, R 8a is amino, hydroxy, methyl, HNC(NH)N(H)CHCH-, HNC(O)-N(H)CHCH-, HN CHCH-, phenyl, pyridin-4-yl, pyridin-3-yl, pyridin-2-yl, pyrimidin-5-yl, pyrimidin-2-yl, pyrazin-2-yl, pyridazin-3-yl, pyridazin-4-yl, piperidin-4-yl, tetrahydropyran-4-yl, or morpholin-4-yl; R 8b is H or CH3, R 9 is HO-, H2N-, H2NCH2-, H2NCH2CH2-, H2NCH2CH2CH2-, H2NC(NH)N(H)CH2CH2- or morpholin-4-yl, R 10 is H or methyl, R 11is -CH2Ph, -CH2-(4-bromophenyl), -CH2-(pyrimidin-5-yl), -CH2-4-(pyrimidin-5-yl)phenyl, -CH2-4-(2-aminopyrimidin-5-yl)phenyl, -CH2-4-(pyridin-4-yl)phenyl, CH2-4-(pyridin-3-yl)phenyl, -CH2-4-(5-aminopyrazin-2-yl)phenyl, -CH2-4-(2- -aminopyrimidin-5-yl)phenyl, -CH2-4-(2-methoxypyrimidin-5-yl)phenyl, -CH2-4-(pyridin-2-yl)phenyl, -CH2-4-[(3-methyl)-isoxazol-4-yl]phenyl, -CH2-4-[(1-methyl)imidazol-2-yl]phenyl, or -CH2-4-[(1-methyl)imidazol-4-yl]phenyl, and R 12 is H, —CH2Ph, —CH2-(4-Fphenyl), or —CH2-(4-pyridin-4-yl).

[0044] In some embodiments, the disclosure provides compounds of formula (I or (IA), wherein: R 1 is CH3C(O)NH-CH2CH2-O-, 5-CO2H-pyrimidin-2-yl, 4-CH3C(O)-piperazin-1-yl, 4-CO2H-cyclohex-4-yl, 4-CO2H-phenyl, or bicyclo[1.1.1]pentane-1-carboxylic acid; R 2 is pyridin-4-yl, pyridazin-4-yl, bicyclo[1.1.1]pentan-1-yl, or cyclobutyl; R 4 is 4-fluoroindol-3-yl, 4-chloroindol-3-yl, or naphth-1-yl; R 5a is CH3, HOCH2-, H2NCH2CH2CH2CH2-, or H2NCH2CH2-, R 5b is CH3, HOCH2-, or H2NCH2CH2-, Alternatively, R 5a and R5b together with the carbon atoms to which they are attached form an azetidinyl ring, R 6a is -CH2CO2H, -OH, -H, -CO2H, -CH2OH, CH3, or -CH2CH3, R 6b is H or CH3-, R 7a is CH3, HOCH2-, H2NCH2CH2-, H2NCH2CH2CH2CH2-, or -CH2CO2H, R 7b is CH3, HOCH2-, H2NCH2CH2-, H2NCH2CH2CH2CH2-, or -CH2CO2H, Alternatively, R 7a and R 7b together with the carbon atoms to which they are attached form an azetidinyl or piperidinyl ring, R 8a is amino, hydroxy, methyl, HNC(NH)N(H)CHCH-, HNC(O)-N(H)CHCH-, HN CHCH-, phenyl, pyridin-4-yl, pyridin-3-yl, pyridin-2-yl, pyrimidin-5-yl, pyrimidin-2-yl, pyrazin-2-yl, pyridazin-3-yl, pyridazin-4-yl, piperidin-4-yl, tetrahydropyran-4-yl, or morpholin-4-yl; R 8b is H or CH3, R 9 is HO-, H2N-, H2NCH2-, H2NCH2CH2-, H2NCH2CH2CH2-, H2NC(NH)N(H)CH2CH2- or morpholin-4-yl, R 10 is H or methyl, R 11is -CH2Ph, -CH2-(4-bromophenyl), -CH2-(pyrimidin-5-yl), -CH2-4-(pyrimidin-5-yl)phenyl, -CH2-4-(2-aminopyrimidin-5-yl)phenyl, -CH2-4-(pyridin-4-yl)phenyl, CH2-4-(pyridin-3-yl)phenyl, -CH2-4-(5-aminopyrazin-2-yl)phenyl, -CH2-4-(2- -aminopyrimidin-5-yl)phenyl, -CH2-4-(2-methoxypyrimidin-5-yl)phenyl, -CH2-4-(pyridin-2-yl)phenyl, -CH2-4-[(3-methyl)-isoxazol-4-yl]phenyl, -CH2-4-[(1-methyl)imidazol-2-yl]phenyl, or -CH2-4-[(1-methyl)imidazol-4-yl]phenyl, and R 12 is H, —CH2Ph, —CH2-(4-Fphenyl), or —CH2-(4-pyridin-4-yl).

[0045] In one embodiment, the present disclosure provides a compound of formula (IB): [ka]

[0046] The present invention provides a compound of formula (I) having the formula:

[0047] In specific embodiments, the present disclosure provides compounds of formula (IB), wherein: R 5a is methyl or HOCH2- R 6a is H, -OH or -CH2CO2H, R 8a teeth, (I C 8 (where C 8 is unsubstituted pyridyl, pyrimidinyl, or pyrazinyl); or (ii) CH3CH2-O- and R 8b is H, methyl, R11 is H or -CH2Ph, R 12 teeth, (i) H, or (ii)-CH2C 12 (where C 12 is phenyl or pyridyl, where C 12 is unsubstituted or substituted with one halo), and The subscript u is either 1 or 2.

[0048] In certain embodiments, the present disclosure provides a compound of formula (I), wherein the compound is selected from the group consisting of SEQ ID NOs: 1-213 and 215-385 as shown in Table 1.

[0049] In certain embodiments, the disclosure provides a compound of formula (I), wherein the compound is selected from the group consisting of (SEQ ID NO:22, SEQ ID NO:29, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:44, SEQ ID NO:48, SEQ ID NO:51, SEQ ID NO:67, SEQ ID NO:72, SEQ ID NOs:99, 101, 218, and SEQ ID NO:381, respectively, in order of appearance): TIFF2025542068000007.tif146140

[0050] TIFF2025542068000008.tif237155

[0051] TIFF2025542068000009.tif162156

[0052] TIFF2025542068000010.tif242152

[0053] TIFF2025542068000011.tif228139

[0054] Without being bound by any particular theory, the applicant believes that the compounds of the present disclosure capture interleukin-1β and prevent signal transduction through the IL-1 receptor, thereby reducing downstream markers IL-6 and CRP.Therefore, the compounds may be useful for treating inflammatory components of cardiovascular diseases, such as ASCVD and heart failure with preserved ejection fraction (HFpEF).The compounds may also be useful for treating inflammatory disorders, such as hidradenitis suppurativa (acne inversa), inflammatory bowel disease, and osteoarthritis.

[0055] definition Unless otherwise defined, 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.

[0056] Reference to compounds of structural formula (I) includes compounds of other general structural formulas and embodiments that fall within the scope of formula (I), including, but not limited to, compounds of formula (IA) or (IB).

[0057] As used throughout this disclosure, the terms "compounds of the present disclosure," "compounds 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). Compounds of Formula (I) can form salts that are also within the scope of this disclosure. Reference herein to compounds of the present disclosure (or compounds of Formula (I)) is understood to include reference to salts thereof, unless otherwise indicated. As used herein, the term "salt(s)" refers to acid salts formed with inorganic and / or organic acids, and basic salts formed with inorganic and / or organic bases. Furthermore, when compounds of Formula (I) contain 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 ("internal salts") may be formed and are included in the term "salt(s)" as used herein. In one embodiment, the salts are pharmaceutically acceptable (i.e., non-toxic, physiologically acceptable) salts. In another embodiment, the salt is other than a pharmaceutically acceptable salt. A salt of a compound of formula (I) can be formed, for example, by reacting a compound of formula (I) with an amount, such as an equivalent amount, of an acid or base in a medium such as one in which the salt precipitates or in an aqueous medium, followed by lyophilization.

[0058] "Acyl" means an alkyl-C(O)- group, where alkyl is as defined below. The bond to the parent group is through the carbon atom of the carbonyl group.

[0059] "Alkyl," as well as other groups having the prefix "alk," such as alkoxy, refer to carbon chains which may be linear or branched, or combinations thereof, containing the indicated number of carbon atoms. For example, C1-C6 alkyl refers to an alkyl group having from 1 (i.e., methyl) to 6 carbon atoms (i.e., hexyl). In certain embodiments, linear alkyl groups have from 1 to 6 carbon atoms, and branched alkyl groups have from 3 to 7 carbon atoms. Examples of alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, sec- and tert-butyl, pentyl, hexyl, heptyl, octyl, nonyl, and the like.

[0060] "Alkoxy" and "alkyl-O-" are used interchangeably and refer to an alkyl group attached to an oxygen.

[0061] "Amino" means a H2N- group. The bond to the parent group is through the nitrogen atom.

[0062] "Amino acid" refers to naturally occurring α-amino acids and their stereoisomers, as well as non-naturally occurring amino acids (such as β-amino acids and substituted amino acids) and their stereoisomers. In the sequences shown for peptides (compounds) according to the present disclosure, amino acid residues have their conventional meanings. Thus, "G" is glycine, "W" is tryptophan, "A" is alanine, "S" is serine, etc. It is understood that "d" isomers are designated by a "d" before the single-letter code or amino acid name, e.g., dA is the d isomer of l-alanine. Amino acid residues not encompassed by the foregoing have the definitions provided in the tables in the Examples section below.

[0063] "Aryl," as used herein, 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.

[0064] A "bicyclic ring system" refers to two linked rings. The rings may be fused, i.e., share two adjacent atoms, or "spirocyclic," i.e., share only a single atom.

[0065] "Carboxy" refers to a HO2C- group. The bond to the parent group is through the carbon atom of the carbonyl moiety.

[0066] "Cycloalkyl" refers to a saturated cyclic hydrocarbon radical. In certain embodiments, a cycloalkyl group has 3 to 12 carbon atoms, forming 1 to 3 carbon rings. The rings may be fused or "spirocyclic," i.e., sharing only a single atom, or "bridged," i.e., sharing three or more atoms, with two bridgehead atoms connected by a bridge containing at least one atom. Examples of cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, adamantyl, bicyclo[1.1.1]pentanyl, and the like.

[0067] "Fluoroalkyl" includes mono- and multiply fluoro-substituted alkyl groups, up to perfluoro-substituted alkyl, such as fluoromethyl, 1,1-difluoroethyl, trifluoromethyl, or 1,1,1,2,2-pentafluorobutyl.

[0068] "Halogen" or "halo," unless otherwise specified, includes fluorine (fluoro), chlorine (chloro), bromine (bromo), and iodine (iodo). In one embodiment, halo is fluoro (-F) or chloro (-Cl).

[0069] "Heterocycloalkyl" means 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, in which one or more atoms of the ring system, alone or in combination, is an element other than carbon, e.g., nitrogen, oxygen, or sulfur. The rings of bicyclic and tricyclic rings may be fused or "spirocyclic," i.e., sharing only a single atom, or "bridged," i.e., sharing three or more atoms, with two bridgehead atoms being connected by a bridge containing at least one atom. There are no adjacent oxygen and / or sulfur atoms in the ring system. In some embodiments, a heterocycloalkyl contains about 5 to about 6 ring atoms. The prefix aza, oxa, or thia before the heterocyclyl root name means that at least a nitrogen, oxygen, or sulfur atom, respectively, is present as a ring atom. In some embodiments, the nitrogen or sulfur atom of a heterocycloalkyl can be oxidized to the corresponding N-oxide, S-oxide, or S,S-dioxide. Non-limiting examples of suitable monocyclic heterocyclyl rings include piperidyl, pyrrolidinyl, piperazinyl, morpholinyl, thiomorpholinyl, thiazolidinyl, 1,4-dioxanyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothiophenyl, and the like.

[0070] "Heteroaryl" refers to aromatic monocyclic, bicyclic, and tricyclic ring systems in which one or more atoms in the ring, the heteroatom(s), are elements other than carbon. The heteroatoms are typically O, S, or N atoms. Examples of heteroaromatic groups include pyridinyl, pyrimidinyl, pyrrolyl, pyridazinyl, isoxazolyl, thiazolyl, oxazolyl, indolyl, benzoxazolyl, benzothiazolyl, and imidazolyl.

[0071] Any component or any variable (e.g., R C1When R occurs more than one time, its definition on 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 this disclosure, those skilled in the art will appreciate the various substituents, e.g., R C9 It will be recognized that the substituents should be selected in accordance with well-known principles of connectivity and stability of chemical structures. Unless otherwise specified, substitution by the designated substituents is permissible for any atom in the ring (e.g., an aryl, heteroaryl ring, or saturated heteroaryl ring) so long as such ring substitution is chemically permissible and results in a stable compound. A "stable" compound is one that can be prepared and isolated and whose structure and properties remain or remain essentially unchanged for a period of time sufficient to permit use of the compound for the purposes described herein (e.g., therapeutic or prophylactic administration to a subject).

[0072] The term "substituted" is deemed to include multiple degrees of substitution by a specified substituent. Where multiple substituent moieties are disclosed or claimed, the substituted compound may be singly or multiply independently substituted with one or more of the disclosed or claimed substituent moieties. Independently substituted means that the (two or more) substituents may be the same or different.

[0073] Unless otherwise expressly shown or stated, variables depicted in structural formulas having "floating" bonds are allowed on any available carbon atom in the ring to which the variable is attached. When a moiety is described in Formula (I) or any embodiment thereof as "optionally substituted," it means that Formula (I) or any embodiment thereof encompasses compounds that contain the substituent(s) described on the moiety as well as compounds that do not contain the substituent(s) described on the moiety.

[0074] As used herein, a wavy line [ka]

[0075] indicates the point of attachment to the rest of the compound.

[0076] Some of the compounds described herein may exist as tautomers with different attachment points of hydrogen accompanied by one or more double bond shifts. For example, a ketone and its enol form are keto-enol tautomers. Individual tautomers as well as mixtures thereof are encompassed by the compounds of the present disclosure.

[0077] In the compounds of the present disclosure, atoms may exhibit their natural isotopic abundance, or one or more of the atoms may be artificially enriched in a particular isotope having the same atomic number but an atomic mass or mass number different from that predominantly found in nature. The present disclosure as described and claimed herein is meant to include all suitable isotopic variations 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 referred to herein as D). Protium is the predominant hydrogen isotope found in nature. Enrichment with deuterium may provide certain therapeutic advantages, such as increased in vivo half-life or reduced dosage requirements, or may result in compounds useful as standards for characterization of biological samples. Isotopically enriched compounds of the present disclosure can be prepared without undue experimentation by conventional techniques well known to those skilled in the art, or by processes similar to those described in the schemes and examples herein using appropriate isotopically enriched reagents and / or intermediates.

[0078] The term "pharmaceutically acceptable salts" refers to salts prepared from pharmaceutically acceptable non-toxic bases or acids. When a compound of the present disclosure is acidic (or has a functional group that may be anionic), its corresponding salt can be conveniently prepared from pharmaceutically acceptable non-toxic bases, including inorganic bases and organic bases. Examples of suitable inorganic cations include Li + , Na + , and K. + Alkali metal ions such as Ca2 + , and Mg2 + Alkaline earth metal cations such as Al 3+ and Zn + Examples of suitable organic cations include, but are not limited to, ammonium ion (i.e., NH4 + ) and substituted ammonium ions. Examples of suitable substituted ammonium ions are those derived from methylamine, ethylamine, diethylamine, triethylamine, and ethylenediamine. When a compound of the present disclosure is basic, its corresponding salt can be conveniently prepared from pharmaceutically acceptable non-toxic acids, including inorganic and organic acids. Examples of such acid addition salts include salts formed from 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 typical for use with the compounds of the present disclosure. In some embodiments, salts of the compounds of the present disclosure can be formed by anion exchange, such as by replacing a trifluoroacetate ion with a chloride ion, as is well known to those skilled in the art.

[0079] Additionally, the compounds of the present disclosure may exist in amorphous form and / or one or more crystalline forms, and therefore, all amorphous and crystalline forms of the compounds of Formula (I), including the Examples, and mixtures thereof, are intended to be included within the scope of the present disclosure. Additionally, some of the compounds of the present disclosure may form solvates with water (i.e., hydrates) or common organic solvents, such as, but not limited to, acetic acid or acetonitrile. Such solvates and hydrates of the compounds, particularly pharmaceutically acceptable solvates and hydrates, are also included within the scope of the present disclosure, along with unsolvated and anhydrous forms.

[0080] Any pharmaceutically acceptable prodrug modification of a compound of the present disclosure that results in conversion in vivo to a compound within the scope of the present disclosure is also within the scope of the present disclosure.

[0081] The present disclosure also relates to methods for preparing compounds of formula (I), which are described in the Examples below and from which compounds of the present disclosure can be obtained.

[0082] "Treatment" and "treating" refer to any process that may slow, interrupt, halt, control, or stop the progression of a disease or disorder described herein. These terms do not necessarily indicate the complete elimination of all disease or disorder symptoms.

[0083] As used herein, "preventing" or "prevention" refers to reducing the likelihood of contracting a disease or disorder described herein or reducing the severity of a disease or disorder described herein.

[0084] The term "therapeutically effective (or effective) amount" and similar descriptions such as "therapeutically effective amount" or "effective dose" are intended to mean an amount of a compound of the present disclosure that elicits a biological or medical response in a tissue, system, animal, or human that is sought by a researcher, veterinarian, physician, or other clinician. In preferred embodiments, the term "therapeutically effective amount" refers to an amount of a compound of the present disclosure that alleviates at least one clinical symptom in a human patient. The term "prophylactically effective (or effective) amount" and similar descriptions such as "prophylactically effective amount" are intended to mean an amount of a compound of the present disclosure that prevents or reduces the risk of occurrence of a biological or medical event sought to be prevented in a tissue, system, animal, or human by a researcher, veterinarian, physician, or other clinician.

[0085] Dosage of Compounds of the Disclosure The dosage regimen utilizing the compounds of the present disclosure is selected according to various factors, including the type, species, age, weight, sex and condition of the patient; the severity of the symptoms to be treated; the efficacy of the compound selected for administration; the route of administration; the renal and hepatic function of the patient.Consideration of these factors is well within the understanding of those skilled in the art to determine the therapeutically or prophylactically effective dosage required to prevent, counter or stop the progression of symptoms.It is understood that a specific daily dosage can be both a therapeutically effective amount for the treatment of, for example, oncological conditions, and a prophylactically effective amount for the prevention of, for example, oncological conditions.

[0086] While individual needs vary, determining optimal ranges for effective amounts of the compounds of the present disclosure is within the skill of one in the art. For administration to humans in the curative or prophylactic treatment of the conditions and disorders identified herein, for example, a typical dosage of a compound of the present disclosure can be about 0.05 mg / kg / day to about 50 mg / kg / day. In some embodiments, a patient is administered about 5 mg / day to about 120 mg / day, e.g., 10 mg / day, 20 mg / day, 30 mg / day, 40 mg / day, 50 mg / day, 60 mg / day, 70 mg / day, 80 mg / day, 90 mg / day, or 100 mg / day of a compound of the present disclosure. In certain embodiments, a patient is administered about 0.2 mg / kg to about 5 mg / kg, e.g., 0.5 mg / kg, 0.75 mg / kg, 1.0 mg / kg, 1.25 mg / kg, or 1.5 mg / kg of a compound of the present disclosure. Such doses may be administered in a single dose or may be divided into multiple doses.

[0087] Pharmaceutical Composition The compounds of the present disclosure and their pharmaceutically acceptable salts can be administered to animals, preferably mammals, particularly humans, as pharmaceuticals by themselves, mixed with each other, or in the form of pharmaceutical compositions.The term "subject" or "patient" includes animals, preferably mammals, particularly humans, that use the active agent for the prevention or treatment of medical conditions.Administration of a drug to a subject includes both self-administration and administration to a patient by another person.A subject may need or desire treatment for an existing disease or medical condition, or may need or desire preventive treatment to prevent or reduce the risk of developing a disease or medical condition.As used herein, a subject "in need" of treatment or preventive treatment of an existing condition includes both the determination of need by a medical professional and the patient's desire for such treatment.

[0088] Therefore, the present disclosure also provides compounds of the present disclosure and pharmaceutically acceptable salts thereof for use as pharmaceuticals, their use for modulating the activity of cytokine IL-1β, particularly in the treatment and prevention of the diseases or disorders described below, and their use for preparing medicaments for these purposes. In certain embodiments, the compounds of the present disclosure and pharmaceutically acceptable salts thereof capture IL-1β.

[0089] Furthermore, the present disclosure provides pharmaceutical compositions comprising as an active ingredient an effective amount of at least one compound of the present disclosure and / or a pharmaceutically acceptable salt thereof, and a conventional pharmaceutically acceptable carrier, i.e., one or more pharmaceutically acceptable carrier substances and / or excipients.

[0090] Thus, the present disclosure provides, for example, compounds of the present disclosure and pharmaceutically acceptable salts thereof for use as pharmaceutical compositions comprising an effective amount of the compounds of the present disclosure and / or pharmaceutically acceptable salts thereof and a conventional pharmaceutically acceptable carrier as active ingredients, as well as the use of the compounds and / or pharmaceutically acceptable salts thereof in the treatment or prevention of diseases or disorders described below, such as atherosclerosis, and their use for preparing medicaments for these purposes.

[0091] The pharmaceutical compositions according to the present disclosure can be administered orally, for example, in the form of pills, tablets, lacquered tablets, dragees, granules, hard and soft gelatin capsules, aqueous, alcoholic or oily solutions, syrups, emulsions or suspensions, or rectally, for example, in the form of suppositories. Administration can also be parenterally, for example, subcutaneously, intramuscularly or intravenously, in the form of solutions for injection or infusion.

[0092] 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.

[0093] 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 pharmaceutically acceptable excipient(s) and / or carrier(s). A particular pharmaceutical composition is suited to the mode of administration. In certain embodiments, the pharmaceutically acceptable carrier may be water or a buffer solution.

[0094] The excipients contained in pharmaceutical compositions have different purposes, for example, depending on the nature of drug and the mode of administration.The examples of commonly used excipients include, but are not limited to, saline, buffered saline, dextrose, water for infection, glycerol, ethanol and their combinations, stabilizers, solubilizers and surfactants, buffers and preservatives, isotonicity agents, bulking agents, lubricants (for example, talc or silica, and fats (such as vegetable stearin, magnesium stearate or stearic acid)), emulsifiers, suspending agents or viscosity agents, inert diluents, fillers (such as cellulose, dibasic calcium phosphate, vegetable oils, lactose, sucrose, glucose, mannitol, sorbitol, calcium carbonate, magnesium stearate, etc.), disintegrants (such as cross-linked polyvinylpyrrolidone, sodium starch glycolate, cross-linked sodium carboxymethylcellulose, etc.), binders (such as starch, gelatin, cellulose, modified celluloses such as methylcellulose or microcrystalline cellulose, hydroxypropyl cellulose, sugars such as sucrose and lactose, or sugar alcohols such as xylitol, sorbitol or maltitol, polyvinylpyrrolidone and polyethylene glycol), humectants, antimicrobial agents, chelating agents, coatings (e.g., cellulose film coatings, synthetic polymers, shellac, corn protein zein or other polysaccharides, and gelatin), preservatives (including vitamin A, vitamins, vitamin C, retinyl palmitate and selenium, cysteine, methionine, citric acid and sodium citrate, and synthetic preservatives including methylparaben and propylparaben), sweeteners, fragrances, flavorings, colorants, absorption enhancers, administration aids, and combinations thereof.

[0095] Carriers are compounds and substances that improve and / or prolong the delivery of active ingredients to a subject in the context of a pharmaceutical composition. Carriers can help extend the in vivo activity of a drug in a subject or delay the release of a drug using controlled-release technology. Carriers can also reduce drug metabolism and / or reduce drug toxicity in a subject. Carriers can also be used to target the delivery of a drug to specific cells or tissues in a subject. Common carriers (both hydrophilic and hydrophobic) include lipid emulsions, lipids, PEGylated phospholipids, PEGylated liposomes, PEGylated liposomes coated with cyclic RGD peptide via a PEG spacer, liposomes and liposomes, microspheres (including those made of biodegradable polymers or albumin), polymer matrices, biocompatible polymers, protein-DNA complexes, protein conjugates, red blood cells, vesicles, nanoparticles, and hydrocarbon stapling side chains. The aforementioned carriers 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, carriers may also be used in compositions for other uses, such as in vitro (e.g., for delivery to cultured cells) and / or in vivo research uses.

[0096] Pharmaceutical compositions suitable for oral administration may be provided as discrete units such as capsules or tablets; as powders or granules; as solutions, syrups, or suspensions (in aqueous or non-aqueous liquids; or as edible foams or whips, or as emulsions). Suitable excipients for tablets or hard gelatin capsules include lactose, corn starch or its derivatives, and stearic acid or its salts. Suitable excipients for use with soft gelatin capsules include, for example, vegetable oils, waxes, fats, semi-solid 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, such as vegetable oils, can be used to provide oil-in-water or oil-in-water suspensions. Excipients that promote absorption from the gastrointestinal tract, such as permeation enhancers such as sodium caprate, can be included. In certain circumstances, delayed-release preparations may be advantageous, and compositions that can deliver the compounds of the present disclosure in a delayed-release or controlled-release manner can also be prepared. Prolonged gastric retention poses the problem of degradation by enzymes present in the stomach, therefore enteric coated capsules can also be prepared by standard techniques in the art in which the active substance for release is lowered in the gastrointestinal tract.

[0097] Pharmaceutical compositions adapted 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).

[0098] Pharmaceutical compositions suitable for topical administration can be formulated as ointment, cream, suspension, lotion, powder, solution, paste, gel, spray, aerosol or oil.When formulated into ointment, active ingredient can be used with either paraffinic ointment base or water-miscible ointment base.Alternatively, active ingredient can be formulated into cream containing oil-in-water cream base or water-in-oil base.Pharmaceutical compositions suitable for topical administration to the eye include eye drops, in which active ingredient is dissolved or suspended in suitable carrier, especially aqueous solvent.Pharmaceutical compositions suitable for topical administration to the mouth include lozenges, troches and mouthwash.

[0099] Pharmaceutical compositions adapted for rectal administration may be presented as suppositories or as enemas.

[0100] Pharmaceutical compositions adapted for nasal administration wherein the carrier is a solid include a coarse powder having a particle size in the range, for example, 20 to 500 microns, administered in the manner in which snuff is taken, i.e. by rapid inhalation through the nasal passage from a container of the powder held close to the nose. Compositions wherein the carrier is a liquid, suitable for administration as nasal drops or as nasal sprays, include aqueous or oily solutions of the active ingredient.

[0101] Pharmaceutical compositions adapted for administration by inhalation include fine particle dusts or mists, which may be generated by means of various types of metered dose pressurized aerosols, nebulizers or insufflators.

[0102] Pharmaceutical compositions adapted for vaginal administration may be presented as pessaries, tampons, creams, gels, pastes, foams or spray formulations.

[0103] Pharmaceutical compositions suitable for parenteral administration include aqueous and non-aqueous sterile injection solutions, which may contain antioxidants, buffers, bacteriostats, and solutes that render the formulation substantially isotonic with the blood of the intended recipient, as well as aqueous and non-aqueous sterile suspensions, which may contain suspending agents and thickening agents. Excipients that may be used for injection solutions include, for example, water for injection, alcohols, polyols, glycerin, and vegetable oils. The compositions may be presented in unit-dose or multi-dose containers, such as sealed ampoules and vials, and may be stored in a freeze-dried (lyophilized) state, requiring only the addition of a sterile liquid carrier, such as water for injection or physiological saline, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules, and tablets. Pharmaceutical compositions may contain preservatives, solubilizers, stabilizers, wetting agents, emulsifiers, sweeteners, colorants, odorants, salts (the substances of the present disclosure themselves may be provided in the form of pharmaceutically acceptable salts), buffers, coating agents, or antioxidants. They may also contain therapeutically active agents in addition to the compounds of the present disclosure.

[0104] Methods of Use of the Disclosed Compounds The present application provides a method for inhibiting IL-1-mediated cell signaling, comprising contacting a cell with a compound of the present disclosure or a pharmaceutically acceptable salt thereof. 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).

[0105] The present application also provides methods of treating disease conditions, including but not limited to those in which IL-1β is involved, using the compounds of the present disclosure (or pharmaceutically acceptable salts thereof) or pharmaceutical compositions containing such compounds.

[0106] In some embodiments, the present disclosure provides a method of treating cardiovascular disease, comprising administering 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 to a subject in need of such treatment. In some embodiments, the cardiovascular disease is vascular inflammation. In some embodiments, the cardiovascular disease is atherosclerosis. In some embodiments, the cardiovascular disease is heart failure with preserved ejection fraction (HFpEF). In other embodiments, the cardiovascular disease is heart failure with reduced ejection fraction (HFrEF).

[0107] In some embodiments, the present disclosure provides a method of treating chronic kidney disease, comprising administering 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 to a subject in need of such treatment.

[0108] In some embodiments, the present disclosure provides a method of treating an inflammatory disorder, comprising administering a therapeutically effective amount of a compound of the present disclosure (or a pharmaceutically acceptable salt thereof) or any of the aforementioned pharmaceutical compositions containing such a compound to a subject in need of such treatment. In certain embodiments, the inflammatory disorder is selected from the group consisting of hidradenitis suppurativa (acne inversa), inflammatory bowel disease, arthritis, and nonalcoholic steatohepatitis (NASH).

[0109] In some embodiments, the inflammatory disorder is hidradenitis suppurativa (also acne inversa).

[0110] In certain embodiments, the inflammatory disorder is inflammatory bowel disease, such as Crohn's disease or ulcerative colitis.

[0111] In some embodiments, the inflammatory disorder is arthritis, eg, osteoarthritis, rheumatoid arthritis, psoriatic arthritis, or gouty arthritis.

[0112] In another embodiment, the inflammatory disorder is non-alcoholic steatohepatitis (NASH).

[0113] Combination therapy One or more additional pharmacologically active agents can be administered in combination with the compounds of the present disclosure. The term "one or more additional active agents" is intended to mean one or more pharmaceutically active agents that are active in the body, including prodrugs that are different from the compounds of Formula I and are converted into pharmaceutically active forms after administration, and also includes the free acid, free base, and pharmaceutically acceptable salts of the additional active agents. Generally, any suitable one or more additional active agents, including but not limited to antihypertensive agents, antiatherosclerotic agents such as lipid-modifying compounds, antidiabetic and / or antiobesity agents, and anti-inflammatory agents, can be used in any combination with the compounds of the present disclosure in a single dosage form (fixed-dose drug combination), or can be administered to a subject in one or more separate dosage forms that allow for simultaneous or sequential administration of the active agents (co-administration of separate active agents).

[0114] Examples of additional active agents that may be used in the treatment of cardiovascular disorders include, but are not limited to, angiotensin converting enzyme inhibitors (e.g., alacepril, benazepril, captopril, ceronapril, cilazapril, delapril, enalapril, enalaprilat, fosinopril, imidapril, lisinopril, moveltipril, perindopril, quinapril, ramipril, spirapril, temocapril, or trandolapril), angiotensin II receptor antagonists (e.g., losartan, supra), i.e., COZAAR®, valsartan (including in combination with sacubitril), candesartan, olmesartan, telmesartan, and any of these drugs used in combination with hydrochlorothiazide, such as HYZAAR®; sGC activators (e.g., riociguat and vericiguat), PCSK9 inhibitors (e.g., evolocumab, alirocumab, MK-0616, and those disclosed in WO 2019 / 246349), neutral endopeptides thidase inhibitors (e.g., thiorphan and phosphoramidon), 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., nicoran, 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 commercially available in the lactone prodrug form ZOCOR® and MEVACOR® and function as inhibitors after administration, and pharmaceutically acceptable salts of dihydroxy open-acid HMG-CoA reductase inhibitors such as atorvastatin (particularly the calcium salt sold under LIPITOR®), rosuvastatin (particularly the calcium salt sold under CRESTOR®), pravastatin (particularly the sodium salt sold under PRAVACHOL®), fluvastatin (particularly the sodium salt sold under LESCOL®), sodium salt sold under the trademark [registered trademark]), crivastatin and pitavastatin; cholesterol absorption inhibitors such as ezetimibe (ZETIA®) and ezetimibe in combination with any other lipid-lowering agent such as the HMG-CoA reductase inhibitors mentioned above, in particular simvastatin (VYTORIN®) or atorvastatin calcium; immediate-release or controlled-release niacin and / or niacin with HMG-CoA reductase inhibitors; niacin receptor agonists such as acipimox and acifran, and niacin receptor partial agonists; insulin and insulin mimetics (e.g., insulin degludec metabolic modifiers, including degludec, insulin glargine, insulin lispro, dipeptidyl peptidase-IV (DPP-4) inhibitors (e.g., sitagliptin, alogliptin, omarigliptin, linagliptin, vildagliptin); insulin sensitizers, for example, (i) PPARy agonists, for example, glitazones (e.g., pioglitazone, mitoglitazone, lobeglitazone, rosiglitazone, and balaglitazone), and other PPAR ligands, for example, (i) PPARα / γ dual agonists (e.g., tiglitazar, muraglitazar, aleglitazar, sodelglitazar, and naveglitazar);(2) PPARα agonists, such as fenofibic acid derivatives (e.g., gemfibrozil, clofibrate, ciprofibrate, fenofibrate, bezafibrate), (3) selective PPARγ modulators (SPPARγ M's), (e.g., those disclosed in WO 02 / 060388, WO 02 / 08188, WO 2004 / 019869, WO 2004 / 020409, WO 2004 / 020408, and WO 2004 / 066963); and (4) PPAR gamma partial agonists; ii) biguanides, such as metformin and its pharmaceutically acceptable salts, particularly metformin hydrochloride, and sustained-release formulations thereof, such as Glumetza™, Fortamet™, and GlucophageXR™; (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 inhalable formulations of each); leptin and leptin derivatives and agonists; amylin and amylin analogs (e.g., pramlintide); sulfonylureas and non-sulfonylurea insulin secretagogues (e.g., tolbutamide, glyburide, glipizide, glimepiride, mitigrin, etc.); linides, meglitinides, nateglinide, and repaglinide; alpha-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, taspoglutide (including nasal, transdermal, and once-weekly formulations thereof)); bile acid sequestrants (e.g., colestilan, colestimide, colesebalam hydrochloride, colestipol, cholestyramine, and dialkylaminoalkyl derivatives of cross-linked dextran), acyl-CoA:cholesterol acyltransferase inhibitors (e.g., avasimibe); anti-obesity compounds;Drugs 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 (GKAs); inhibitors of 11β-hydroxysteroid dehydrogenase type 1 (such as those disclosed in U.S. Pat. No. 6,730,690); inhibitors of fructose 1,6-bisphosphatase, (such as those disclosed in U.S. Pat. No. 6,054,587; U.S. Pat. No. 6,110,903; U.S. Pat. No. 6,284,748; U.S. Pat. No. 6, 399,782; and U.S. Pat. No. 6,489,476); 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 (such as those disclosed in WO 2009 / 001836); neuromedin U receptor agonists (such as, but not limited to, neuromedin S (NMS)), such as those disclosed in Publication No. 2009 / 042053; SCD modulators; GPR-105 antagonists (such as those disclosed in WO 2009 / 000087); SGLT inhibitors (such as empagliflozin, dapagliflozin, canagliflozin, ertugliflozin, remogroflozin, tofogliflozin, and ipragliflozin); inhibitors of acyl-coenzyme A: inhibitors of diacylglycerol acyltransferase 1 and 2 (DGAT-1 and DGAT-2) fatty acid synthase; acyl-coenzyme A: inhibitors of monoacylglycerol acyltransferase inhibitors of glycerol acyltransferase 1 and 2 (MGAT-1 and MGAT-2); agonists of the 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., gevokizumab and canakinumab); and bromocriptine mesylate and its rapid-release formulations;or, where chemically possible, in combination with other drugs useful in treating the above-mentioned conditions or disorders, including the free acid, free base and pharmaceutically acceptable salt forms of the above-mentioned active agents;

[0115] Examples of additional active agents that can be used in the treatment of inflammatory disorders include, but are not limited to, steroidal and nonsteroidal anti-inflammatory agents, glucocorticoids, and therapeutic hormones. In certain embodiments, in the treatment of hidradenitis suppurativa (acne inversa), 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, ponesimod), or a selective JAK inhibitor (e.g., tofacitinib, baricitinib, upadacitinib). In some 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 drug, or a glucocorticoid.

[0116] Methods of Preparing the Compounds of the Disclosure 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 cellular assays. However, the compounds shown in the examples should not be construed as forming the only genus that is considered in this disclosure.

[0117] The examples further illustrate the details of preparing the compounds of the present disclosure. Those skilled in the art will readily understand that known variations of the conditions and processes of the following preparative procedures can be used to prepare these compounds. For example, in some cases, the order of carrying out the steps of the reaction schemes can be varied to facilitate the reaction or to avoid undesired reaction products. Starting materials and intermediates of the final compounds are purchased, prepared from known procedures, or otherwise exemplified. The examples are provided for the purpose of further illustration only and are not intended to limit the present disclosure.

[0118] NMR data were obtained on 300 MHz or 400 MHz instruments in CDCl3, DMSO-d6, or methanol-d4, with chemical shifts reported relative to a tetramethylsilane standard. Resonance signals are reported by the following abbreviations: s = singlet, d = doublet, t = triplet, q = quartet, dd = doublet of doublets, m = multiplet or overlap of unequal resonances. Coupling constants (J) are reported in Hertz (Hz).

[0119] Throughout the synthetic schemes and examples, the following abbreviations and acronyms may be used, unless otherwise indicated: [Table 1]

[0120] TIFF2025542068000014.tif250165

[0121] TIFF2025542068000015.tif250165

[0122] TIFF2025542068000016.tif253166

[0123] TIFF2025542068000017.tif254166

[0124] TIFF2025542068000018.tif171110

[0125] TIFF2025542068000019.tif251166

[0126] TIFF2025542068000020.tif248166

[0127] TIFF2025542068000021.tif15165

[0128] Intermediate synthesis: The following schemes describe suitable syntheses of certain protected amino acid precursors used to prepare compounds of the present disclosure.

[0129] Synthetic Scheme 1 [ka]

[0130] Precursor to 3Pal4CO2H [ka]

[0131] (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(6-(tert-butoxycarbonyl)pyridin-3-yl)propanoic acid Step 1: To a stirred solution of NiCl-glyme (710 mg, 3.23 mmol) in DMA (160 mL) was added 1,10-phenanthroline (700 mg, 3.23 mmol) under a nitrogen atmosphere at 25° C. The resulting mixture was stirred at 50° C. for 1 h, and then tert-butyl 5-bromopicolinate (4.17 g, 16.15 mmol), benzyl (R)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-iodopropanoate (8.51 g, 16.15 mmol), TBAI (6.19 g, 16.15 mmol), and Zn (2.11 g, 32.3 mmol) were added at room temperature. The resulting mixture was stirred at 25° C. for 2 h, after which it was quenched with HO (200 mL) and extracted with EA (2×500 mL). The combined organic layers were washed with brine (3 x 200 mL), dried over anhydrous NaSO, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography eluting with 0-20% EA in PE to give tert-butyl (S)-5-(2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(benzyloxy)-3-oxopropyl)picolinate (7.5 g, 12.96 mmol, 80% yield) as an off-white solid. 35 H 35 N2O6[M+H] + MS ESI calculated for 579.24, found 579.40.

[0132] Step 2: To a stirred solution of tert-butyl (S)-5-(2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(benzyloxy)-3-oxopropyl)picolinate (7.5 g, 12.96 mmol) in EA (75 mL) was added Pd—C (1.379 g, 12.96 mmol, dry, 10% wt) at room temperature under nitrogen atmosphere. The resulting mixture was degassed three times with hydrogen and stirred at 25° C. for 4 h. The solid was filtered off and the filtrate was concentrated under reduced pressure. The residue was purified by RP-flash using the following conditions: Column: Flash C 18(330 g); Mobile phase A: water (0.1% TFA), Mobile phase B: ACN; (Gradient: 5 min hold at 5% B, hold to 30% B within 15 min, hold at 30% B for 5 min; up to 95% B within 20 min, hold at 95% B for 10 min); Flow rate: 90 mL / min; Detector: UV 210 nm; RT = 40 min. Product-containing fractions were collected and evaporated in vacuo to give (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(6-(tert-butoxycarbonyl)pyridin-3-yl)propanoic acid (5.4625 g, 11.18 mmol, 86% yield) as a yellow solid. C 28 H 29 N2O6[M+H] + MS ESI calculated for 489.19, found 489.20; 1 H NMR(300 MHz,CD3OD)δ 8.57(s,1H),8.03-7.91(m,1H),7.86-7.74(m,3H)7.54-7.53(m,2H),7.36-7.28(m,4H),4.5 1-4.40(m,1H),4.32-4.13(m,3H),3.39-3.33(m,1H),3.06-3.05(m,1H),1.65-1.59(m,9H). Synthetic Scheme 2 [ka]

[0133] 3 Precursor to PyridaAla [ka]

[0134] (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(pyridazin-3-yl)propanoic acid Step 1: To a mixture of NiCl-glyme (0.713 g, 3.24 mmol) in DMA (20 mL) was added 1,10-phenanthroline (0.703 g, 3.24 mmol) at room temperature. The resulting mixture was stirred at 50° C. for 1 hour, then cooled to room temperature. A mixture of 3-bromopyridazine (5.16 g, 32.4 mmol), tert-butyl (R)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-iodopropanoate (8.0 g, 16.22 mmol), and TBAI (5.99 g, 16.22 mmol) was poured into 120 mL of DMA, and then zinc (2.120 g, 32.4 mmol) was added at room temperature. The resulting mixture was stirred at room temperature for 5 hours. The reaction mixture was diluted with water (200 mL) and extracted with EA (3×200 mL). The combined organic layers were washed with brine (3×100 mL), dried over NaSO, and filtered. The filtrate was concentrated in vacuo. The residue was purified by silica gel column chromatography eluting with EA / PE (4 / 1) to give tert-butyl (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(pyridazin-3-yl)propanoate (3.6 g, 8.08 mmol, 50% yield) as a yellow solid. 26 H 28 N3O4[M+H] + MS ESI calculated for 446.20, found 446.30.

[0135] Step 2: To a solution of tert-butyl (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(pyridazin-3-yl)propanoate (3.6 g, 8.08 mmol) in DCM (10 mL) was added TFA (20 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 hours, after which it was concentrated in vacuo. The residue was purified by RP-flash eluting with 0-50% MeCN in water (0.05% TFA) to give (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(pyridazin-3-yl)propanoic acid (2.5 g, 6.42 mmol, 79% yield) as a pale yellow solid. C 22 H 20N3O4[M+H] + MS ESI calculated for 390.14, found 390.10. 1 H NMR(300 MHz,CD3OD)δ 9.17(s,1H),7.90-7.77(m,4H),7.61-7.59(m,2H),7.41-7.36(m,2H),7.31-7.26(m,2H),4.74-4.70(m,1H),4.29(d,J=6.0 Hz,2H),4.18-4.14(m,1H),3.67-3.61(m,1H),3.42-3.37(m,1H). Synthetic Scheme 3 [ka]

[0136] Precursor to aMeDab [ka]

[0137] (S)-2-((((9H-Fluoren-9-yl)methoxy)carbonyl)amino)-4-((tert-butoxycarbonyl)amino)-2-methylbutanoic acid Step 1: To a stirred solution of (S)-2-amino-2-methylpent-4-enoic acid (2 g, 15.48 mmol) in dioxane (20 mL) and water (20 mL) was added DIEA (8.11 mL, 46.5 mmol) and Fmoc-OSu (5.22 g, 15.48 mmol) at room temperature. The resulting solution was stirred at 25° C. for 16 h. The pH of the solution was adjusted to 6N. HCl to give 3, which was purified by RP flash under the following conditions: 5% to 5% in 5 min, 5% to 60% in 30 min, 98% to 98% in 5 min, MeCN (0.05% TFA) in water, RT = 35 min to give (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2-methylpent-4-enoic acid (4.6 g, 12.44 mmol, 80% yield) as an off-white solid. 21 H 20 NO4[MH] + MS ESI calculated for 350.15, found 350.00.1 H NMR(300 MHz,CDCl3)δ 9.67(s,1H),7.82-7.68(m,2H),7.58(d,J=7.4 Hz,2H),7.45-7.19(m,4H),5.83-5.53(m,1H),5.52-5.33(m,1H),5.16-5.12(m,2H),4.64-4.27(m,2H),4.21(t,J=6.6 Hz,1H),2.94-2.48(m,2H). Step 2: To a stirred solution of (S)-2-((((9H-Fluoren-9-yl)methoxy)carbonyl)amino)-2-methylpent-4-enoic acid (4.6 g, 13.09 mmol) in acetone (80 mL), 4-methylmorpholine (2.91 g, 14.40 mmol, 50% in water) and OsO (3.33 g, 1.309 mmol, 10% in water) were added at 25 °C. The resulting solution was stirred at 25 °C for 4 h. Sodium periodate (10.27 g, 14.40 mmol, 30% in water) was then added, and the resulting mixture was stirred at 25 °C for 16 h. The mixture was diluted with water (100 mL) and extracted with EA (3 × 200 mL). The combined organic layers were washed with brine (2 × 100 mL) and dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure to give (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2-methyl-4-oxobutanoic acid (4.9 g, 11.79 mmol, 90% yield) as a black solid. 20 H 18 NO5[MH] - MS ESI calculated for 352.13, found 352.10.

[0138] Step 3: To a stirred solution of (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2-methyl-4-oxobutanoic acid (1 g, 2.83 mmol) in toluene (50 mL), TFA (0.968 g, 8.49 mmol) and tert-butyl carbamate (1.989 g, 16.98 mmol) were added at room temperature. The resulting solution was stirred at 25° C. for 2 hours. The solvent was concentrated under reduced pressure to give (S,Z)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((tert-butoxycarbonyl)imino)-2-methylbutanoic acid (1.3 g, 2.011 mmol, 71% yield) as a black solid. 25 H 27 N2O6[MH] - MS ESI calculated for 451.19, found 451.00.

[0139] Step 4: To a stirred solution of (S,Z)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((tert-butoxycarbonyl)imino)-2-methylbutanoic acid (5 g, 7.73 mmol) and dimethyl(phenyl)silane (5.27 g, 38.7 mmol) in toluene (50 mL) was added tris(pentafluorophenyl)borane (0.396 g, 0.773 mmol) at room temperature. The resulting solution was stirred at 25° C. for 16 hours. The solvent was concentrated under reduced pressure, and the residue was purified under the following conditions: 18 Purification by RP-flash column, 330 g, 5% to 5% in 5 min, 5% to 50% in 30 min, 98% to 98% in 5 min, MeCN (0.025% TFA) in water, RT = 35 min, afforded (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((tert-butoxycarbonyl)amino)-2-methylbutanoic acid (2.5183 g, 5.26 mmol, 68% yield) as an off-white solid. 25 H 29 N2O6[MH] - MS ESI calculated for 453.21, found 453.00. 1H NMR(400 MHz,DMSO-d6)δ 12.49(s,1H),7.90(d,J=6.8 Hz,2H),7.73(d,J=7.6 Hz,2H),7.54(s,1H),7.44-7.40(m,2H),7.36-7.32(m,2H),6.77(s,1H),4 .25-4.22(m,3H),2.93-2.91(m,2H),1.97-1.81(m,2H),1.37-1.34(m,9H). Synthetic Scheme 4 [ka]

[0140] Precursor to daMeDab [ka]

[0141] (R)-2-((((9H-Fluoren-9-yl)methoxy)carbonyl)amino)-4-((tert-butoxycarbonyl)amino)-2-methylbutanoic acid Step 1: To a stirred solution of (R)-2-amino-2-methylpent-4-enoic acid (1.3 g, 10.07 mmol) and DIEA (5.27 mL, 30.2 mmol) in dioxane (20 mL) and water (20 mL), Fmoc-OSu (3.73 g, 11.07 mmol) was added at room temperature. The resulting solution was stirred at 25° C. for 16 hours. The pH was adjusted to 3 with 1 N HCl, and the solution was purified by RP flash using the following conditions: 18 Column, 330 g, 5% to 5% in 5 min, 5% to 50% in 30 min, 98% to 98% in 5 min, MeCN in water (0.05% TFA), RT = 30 min, afforded (R)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2-methylpent-4-enoic acid (3 g, 8.11 mmol, 81% yield) as an off-white solid. 21 H 21 NO4Na[M+Na] + MS ESI calculated for 374.15, found 374.05. 1H NMR(400 MHz,CDCl3)δ 7.79(d,J=7.5 Hz,2H),7.61(d,J=7.5 Hz,2H),7.48-7.29(m,4H),5.71(s,1H),5.48(s,1H),5.20-5.16(m,2H),4.44-4.40(m,2H),4.25(t,J=6.7 Hz,1H),2.82-2.71(m,2H). 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 NMO (2.200 g, 9.39 mmol, 50% in water) in acetone (60 mL) was added OsO (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 (2.009 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 EA (3 × 200 mL). The combined organic layers were washed with brine (2 × 100 mL) and dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure to give (R)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2-methyl-4-oxobutanoic acid (3.4 g, 7.70 mmol, 90% yield) as a black solid. 20 H 20 NO5[M+H] + MS ESI calculated for 354.13, found 354.00.

[0142] 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), tert-butyl carbamate (4.77 g, 40.8 mmol) and TFA (2.323 g, 20.38 mmol) were added at room temperature. The resulting solution was stirred at 25° C. for 2 hours. 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 (3.1 g, 5.48 mmol, 81% yield) as a black solid. 25 H 27 N2O6[MH] + MS ESI calculated for 451.19, found 450.90.

[0143] 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 (1.867 g, 13.70 mmol) in toluene (120 mL) was added tris(pentafluorophenyl)borane (0.281 g, 0.548 mmol) at room temperature. The solution was stirred at 25° C. for 16 hours. The solvent was concentrated under reduced pressure, and the residue was purified under the following conditions: 330 g C 18 Purification by column, 5% to 5% in 5 min, 5% to 55% in 30 min, 98% to 98% in 5 min, MeCN in water (0.05% TFA), RP-flash at RT = 35 min gave (R)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((tert-butoxycarbonyl)amino)-2-methylbutanoic acid (2.2528 g, 4.81 mmol, 88% yield) as an off-white solid. 25 H 31 N2O6[M+H] + MS ESI calculated for 455.21, found 455.10; 1H NMR(400 MHz,DMSO-d6)δ 12.48(s,1H),7.90(d,J=7.2 Hz,2H),7.73(d,J=7.2 Hz,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). Synthetic Scheme 5 [ka]

[0144] Precursor to dProc3Bn [ka]

[0145] (2R,3S)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-3-benzylpyrrolidine-2-carboxylic acid Step 1: To a mixture of ethyl (R)-N-(but-3-en-1-yl)-N-(1-phenylethyl)glycinate (10.48 g, 40.1 mmol) in THF (100 mL) was added LDA (20.05 mL, 40.1 mmol, 2 M in THF) under argon at −78° C. After the mixture was stirred at −78° C. for 30 minutes, dry ZnBr (120 mL, 120 mmol, 1 N in THF) was added to the mixture at −78° C. The resulting mixture was slowly warmed to ambient temperature and stirred at ambient temperature for 4 hours. Iodobenzene (10.63 g, 52.1 mmol), Pd2(dba)3 (1.102 g, 1.203 mmol), and tri-o-tolylphosphine (1.587 g, 5.21 mmol) were added sequentially, and the reaction was stirred at ambient temperature for 16 h. The resulting mixture was quenched with saturated NH4Cl (100 mL) and extracted with EA (3 × 200 mL). The organic layers were combined, washed with brine (2 × 130 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel chromatography eluting with a gradient of 0–15% EA in PE to afford ethyl (2R,3S)-3-benzyl-1-((R)-1-phenylethyl)pyrrolidine-2-carboxylate (8.6 g, 25.5 mmol, 64% yield) as an orange semisolid. 22 H 28 NO2[M+H] + MS ESI calculated for 338.20, found 338.30.

[0146] Step 2: To a stirred solution of ethyl (2R,3S)-3-benzyl-1-((R)-1-phenylethyl)pyrrolidine-2-carboxylate (10 g, 29.6 mmol) in EtOH (100 mL) was added Pd / C (4 g, 37.6 mmol, dry, 10% by weight) under nitrogen at 25° C. The resulting mixture was stirred at 25° C. for 10 minutes, then degassed under vacuum and purged with H several times. The resulting mixture was stirred at 60° C. under 2 atmospheres of H for 6 hours. After filtration, the filtrate was concentrated in vacuo to afford (2R,3S)-3-benzylpyrrolidine-2-carboxylate (6 g, 25.7 mmol, 87% yield) as a colorless oil. 14 H 20 NO2[M+H]+ MS ESI calculated for 234.14, found 234.20.

[0147] Step 3: To a stirred solution of ethyl (2R,3S)-3-benzylpyrrolidine-2-carboxylate (6 g, 25.7 mmol) in THF (60 mL) was added LiOH (51.4 mL, 51.4 mmol, 1N in water) at room temperature. The solution was stirred at 25° C. for 12 hours, and the pH value of the solution was adjusted to 7 with 1N HCl. The solution was used directly in the next step without further purification. 12 H 16 NO2[M+H] + MS ESI calculated for 206.11, found 206.15.

[0148] Step 4: To a stirred solution of ((2R,3S)-3-benzylpyrrolidin-2-yl)(11-oxidanyl)methanone (5 g, 24.48 mmol) in THF (50 mL) and water (50 mL) was added NaHCO (10.28 g, 122 mmol) at 25 °C under nitrogen atmosphere. The resulting mixture was stirred at 25 °C for 10 min. Fmoc-OSu (7.43 g, 22.03 mmol) was added to the mixture and stirred at 25 °C for 2 h. The pH value of the solution was adjusted to 3 with 1 N HCl. The aqueous layer was extracted with EA (2 × 500 mL). The combined organic layers were washed with brine (3 × 50 mL), dried over anhydrous NaSO, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by RP-flash using the following conditions: Column: Flash C 18 (330 g); Mobile phase A: water (0.1% TFA), Mobile phase B: ACN; (Gradient: 5 min hold at 5% B, hold to 30% B within 15 min, hold at 30% B for 5 min; up to 95% B within 20 min, hold at 95% B for 10 min); Flow rate: 90 mL / min; Detector: UV 210 nm; RT = 40 min. Product-containing fractions were collected and rotary evaporated in vacuo to give (2R,3S)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-3-benzylpyrrolidine-2-carboxylic acid (8.2727 g, 19.35 mmol, 79% yield) as an off-white solid. C 27 H 26 NO4[M+H] +MS ESI calculated for 428.18, found 428.10. 1 H NMR(400 MHz,CD3OD)δ 7.79-7.77(m,2H),7.65-7.60(m,2H),7.40-7.36(m,2H),7.33-7.18(m,7H),4.42-4.18(m,4H),3.68-3.6 4(m,1H),3.31-3.30(m,1H),3.05-3.02(m,1H),2.78-2.65(m,1H),2.37-2.34(m,1H),1.80-1.77(m,2H). Synthetic Scheme 6 [ka]

[0149] Precursor to dProc3Bn4Br [ka]

[0150] (2R,3S)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-3-(4-bromobenzyl)pyrrolidine-2-carboxylic acid Step 1: To a solution of ethyl (R)-N-(but-3-en-1-yl)-N-(1-phenylethyl)glycinate (5 g, 19.13 mmol) in THF (30 mL) was added LDA (9.57 mL, 19.13 mmol, 2 M in THF) at −78° C. under argon. The solution was stirred at −78° C. for 30 minutes. To the solution was then added dry ZnBr (57.4 mL, 57.4 mmol, 1 N in THF) at −78° C. The reaction was allowed to warm slowly to ambient temperature and stirred at ambient temperature for 4 hours. Then, 1-bromo-4-iodobenzene (7.04 g, 24.87 mmol), Pd2(dba)3 (0.526 g, 0.574 mmol), and tri-o-tolylphosphine (0.757 g, 2.487 mmol) were added sequentially, and the reaction was stirred at ambient temperature for 16 h. The resulting solution was quenched with saturated NH4Cl (50 mL) and extracted with EA (3 × 150 mL). The organic layers were combined, washed with brine (2 × 80 mL), dried over anhydrous Na2SO4, and filtered. After concentration, the residue was purified by silica gel chromatography eluting with a gradient of 0% to 11% EA in PE to afford ethyl (2R,3S)-3-(4-bromobenzyl)-1-((R)-1-phenylethyl)pyrrolidine-2-carboxylate (3.6 g, 8.65 mmol, 45% yield) as an orange semisolid. C 22 H 27 BrNO2[M+H] + MS ESI calculated values: 416.11, 418.11, found values: 416.05, 418.05.

[0151] Step 2: To a mixture of ethyl (2R,3S)-3-(4-bromobenzyl)-1-((R)-1-phenylethyl)pyrrolidine-2-carboxylate (3.7 g, 8.97 mmol) in DCM (220 mL) was added CAN (14.76 g, 26.9 mmol, dissolved in 44 mL of water) at 0 °C for 3 min. The resulting mixture was stirred at ambient temperature for 6 h. The reaction was quenched with 60 mL of saturated NaHCO and extracted with EA (3 × 250 mL). The organic layers were combined, washed with brine (2 × 150 mL), dried over anhydrous NaSO, and filtered. The filtrate was concentrated in vacuo and purified by RP-flash using the following conditions: Column: Flash C18 (330 g); Mobile phase A: water (0.05% TFA), Mobile phase B: ACN; (Gradient: 5 min hold at 2% B, hold to 32% B within 15 min, hold at 32% B for 4 min; hold to 98% B within 10 min, hold at 98% B for 3 min); Flow rate: 35 mL / min; Detector: UV 220 nm; RT = 38 min. Product-containing fractions were collected and concentrated in vacuo to give ethyl (2R,3S)-3-(4-bromobenzyl)pyrrolidine-2-carboxylate (1.1 g, 3.52 mmol, 39% yield) as a brown semi-solid. 14 H 19 BrNO2[M+H] + MS ESI calculated values: 312.05, 314.05, found values: 312.10, 314.10.

[0152] Step 3: To a mixture of ethyl (2R,3S)-3-(4-bromobenzyl)pyrrolidine-2-carboxylate (3.3 g, 10.57 mmol) in THF (22 mL) was added LiOH (21.14 mL, 21.14 mmol, 1 M in water) at 0 °C. After the reaction was stirred at ambient temperature for 4 h, it was acidified to pH 3-4 with aqueous HCl and concentrated in vacuo to afford the crude product (2R,3S)-3-(4-bromobenzyl)pyrrolidine-2-carboxylic acid (3.5 g, 9.85 mmol, 93% yield) as a white solid. 12 H 15 BrNO2[M+H] + MS ESI calculated values: 284.02, 286.02, found values: 284.05, 286.05.

[0153] Step 4: To a mixture of (2R,3S)-3-(4-bromobenzyl)pyrrolidine-2-carboxylic acid (3.5 g, 9.85 mmol) in THF (30 mL) and water (30 mL) was added NaHCO3 (4.14 g, 49.3 mmol) and Fmoc-OSu (2.99 g, 8.87 mmol) at ambient temperature. The reaction was stirred at ambient temperature for 4 h. The resulting solution was acidified to pH 3-4 with aqueous HCl and extracted with EA (3 x 100 mL). The organic layers were combined, washed with brine (2 x 100 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated in vacuo. The residue was purified by Rp-flash using the following conditions: Column: C 18 (330 g); Mobile phase A: water (0.05% TFA), Mobile phase B: ACN; (Gradient: 5 min hold at 5% B, hold to 78% B within 28 min, hold at 78% B for 8.5 min; up to 95% B within 2 min, hold at 95% B for 10 min); Flow rate: 90 mL / min; Detector: UV 210 nm; RT = 53.5 min. Product-containing fractions were collected and concentrated in vacuo to give the crude product. The crude product was separated by Prep-SFC using the following conditions: Column: CHIRALPAK IH, 3 × 25 cm, 5 μm; Mobile phase A: CO2, Mobile phase B: IPA:ACN = 1:1 (0.1% 2M NH3-MeOH); Flow rate: 70 mL / min; Gradient: isocratic 35% B; Column temperature (°C): 35; Back pressure (bar): 100; Wavelength: 220 nm; A slower peak was obtained at 8.39 min. The collected fractions were combined and concentrated in vacuo. The residue was lyophilized to give (2R,3S)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-3-(4-bromobenzyl)pyrrolidine-2-carboxylic acid (3.0099 g, 5.74 mmol, 58% yield) as an off-white solid. 27 H 25 BrNO4[M+H] + MS ESI calculated values ​​were 506.09 and 508.09, and experimental values ​​were 506.05 and 508.05. 1H NMR(400 MHz,CD3OD)δ 7.78-7.76(m,2H),7.64-7.60(m,2H),7.45-7.36(m,4H),7.32-7.28(m,2H),7.18-7.14(m,2H),4.39-4.17(m,4H), 3.66-3.63(m,1H),3.32-3.28(m,1H),3.00-2.95(m,1H),2.76-2.62(m,1H),2.38-2.35(m,1H),1.82-1.77(m,2H). Synthetic Scheme 7 [ka]

[0154] Precursor to dProc3CH2Pyrim [ka]

[0155] (2R,3S)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-3-(pyrimidin-5-ylmethyl)pyrrolidine-2-carboxylic acid Step 1: To a solution of ethyl (R)-N-(but-3-en-1-yl)-N-(1-phenylethyl)glycinate (2.61 g, 10 mmol) in THF (10 mL) was added LDA (6 mL, 12.00 mmol, 2 M in THF) under argon at −78° C. After the reaction solution was stirred at −20° C. for 30 minutes, a solution of ZnBr in THF (15 mL, 30.0 mmol) was added to the reaction at −78° C. The resulting mixture was allowed to warm and stirred at ambient temperature for 4 hours, after which I (2.66 g, 10.50 mmol) in THF (10 mL) was added. The reaction mixture was stirred at ambient temperature for 16 hours, then quenched with saturated NH4Cl (100 mL) and extracted with EtOAc (2×100 mL). The combined organic layers were washed with brine (2 x 100 mL), dried over Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by Rp-flash using the following conditions: Column: C 18Gel column (330 g); Mobile phase A: water (5 mM NH4HCO3); Mobile phase B: MeCN; (Gradient: 5 min hold at 0% B, hold to 36% B within 20 min, hold at 36% B for 10 min; up to 95% B within 5 min, hold at 95% B for 10 min); Flow rate: 80 mL / min; Detector: UV 254 and 210 nm; RT: 35.32 min. Product-containing fractions were collected and rotary evaporated in vacuo to give ethyl (2R,3S)-3-(iodomethyl)-1-((R)-1-phenylethyl)pyrrolidine-2-carboxylate (3.5 g, 9.04 mmol, 90% yield) as a colorless oil. C 16 H 23 INO2[M+H] + MS ESI calculated for 388.08, found 388.15.

[0156] Step 2: Argon gas was bubbled through a mixture of ethyl (2R,3S)-3-(iodomethyl)-1-((R)-1-phenylethyl)pyrrolidine-2-carboxylate (2 g, 5.16 mmol), 5-iodopyrimidine (1.383 g, 6.71 mmol), and TBAI (1.908 g, 5.16 mmol) in DMA (10 mL), which was designated as solution A. Argon gas was bubbled through a mixture of NiCl-glyme (0.227 g, 1.033 mmol) and 1,10-phenanthroline (0.186 g, 1.033 mmol) in DMA (10 mL), which was stirred at 50 °C for 0.5 h, which was designated as solution B. Solution A was injected into solution B, followed by the addition of Zn (0.675 g, 10.33 mmol). The reaction solution was degassed with a syringe containing argon for 10 minutes while stirring, and then stirred at 30° C. The reaction was stirred at 30° C. for 16 hours. The reaction mixture was purified by Rp-flash under the following conditions: Column: Flash C 18Column (120 g); Mobile phase A: water (0.1% TFA), Mobile phase B: ACN; (Gradient: 5 min hold at 5% B, hold to 76% B within 30 min, hold at 76% B for 8.6 min; up to 95% B within 2 min, hold at 95% B for 10 min); Flow rate: 60 mL / min; Detector: UV 210 nm; RT = 34 min. Product-containing fractions were collected and rotary evaporated in vacuo to give ethyl (2R,3S)-1-((R)-1-phenylethyl)-3-(pyrimidin-5-ylmethyl)pyrrolidine-2-carboxylate (760 mg, 2.239 mmol, 43% yield) as a yellow solid. 20 H 26 N3O2[M+H] + MS ESI calculated for 340.20, found 340.25.

[0157] Step 3: Ethyl (2R,3S)-1-((R)-1-phenylethyl)-3-(pyrimidin-5-ylmethyl)pyrrolidine-2-carboxylate (710 mg, 2.092 mmol) was dissolved in THF (20 mL), then evacuated and an argon atmosphere was applied at ambient temperature. Pd / C (100 mg, 0.094 mmol, dry, 10% by weight) was then added under an argon atmosphere. The suspension was degassed under vacuum and purged with H2 several times. The reaction solution was stirred under 2 atmospheres of H2 at 50°C for 1.5 hours. The suspension was filtered, and the filtrate was concentrated under reduced pressure to give ethyl (2R,3S)-3-(pyrimidin-5-ylmethyl)pyrrolidine-2-carboxylate (492 mg, 2.092 mmol, 100% yield) as a pale yellow solid. C 12 H 18 N3O2[M+H] + MS ESI calculated for 236.14, found 236.15.

[0158] Step 4: To a stirred mixture of ethyl (2R,3S)-3-(pyrimidin-5-ylmethyl)pyrrolidine-2-carboxylate (0.492 g, 2.092 mmol) in THF (10 mL) was added a solution of LiOH (4.18 mL, 4.18 mmol, 1N in water) at ambient temperature. The resulting mixture was stirred at ambient temperature for 2 hours and then concentrated under reduced pressure to give crude (11-oxidanyl)((2R,3S)-3-(pyrimidin-5-ylmethyl)pyrrolidin-2-yl)methanone (0.431 g, 2.092 mmol, 100% yield) as a yellow oil. 10 H 14 N3O2[M+H] + MS ESI calculated for 208.11, found 208.15.

[0159] Step 5: To a solution of (11-oxidanyl)((2R,3S)-3-(pyrimidin-5-ylmethyl)pyrrolidin-2-yl)methanone (431 mg, 2.092 mmol) in THF (5 mL) and water (5 mL) was added NaHCO (879 mg, 10.46 mmol) and Fmoc-OSu (635 mg, 1.883 mmol) at ambient temperature. The reaction mixture was stirred at room temperature for 16 h, and then 1 M HCl was added to adjust the pH to 5. The resulting solution was diluted with EA (100 mL) and washed with brine (3 × 30 mL). The organic layer was dried over anhydrous NaSO and filtered. The filtrate was concentrated, and the residue was purified by column chromatography using Flash C 18(330 g) was purified by mobile phase A: water (0.1% TFA), mobile phase B: ACN; (Gradient: 5 min hold at 5% B, hold to 60% B within 30 min, hold at 60% B for 2.6 min; hold to 95% B within 2 min, hold at 95% B for 5 min); flow rate: 80 mL / min; detector: UV 210 nm; RT = 31 min. Product-containing fractions were collected and rotary evaporated in vacuo to give the crude product. The crude product was separated by Prep-SFC using the following conditions: Column: CHIRALPAK IH, 3 × 25 cm, 5 μm; Mobile phase A: CO2, Mobile phase B: MeOH (0.1% 2M NH3-MeOH); Flow rate: 60 mL / min; Gradient: Isocratic 30% B; Column temperature (°C): 35; Back pressure (bar): 100; Wavelength: 220 nm; RT1 (min): 5.72; RT2 (min): 7.18; Sample solvent: MeOH (0.1% 2M NH3-MeOH); Injection volume: 1.5 mL; Number of runs: 20. The fractions of the second peak (RT2: 7.18 min) were collected, concentrated in vacuo, and then lyophilized overnight to give (2R,3S)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-3-(pyrimidin-5-ylmethyl)pyrrolidine-2-carboxylic acid (450 mg, 1.048 mmol, 50% yield) as a white solid. 25 H 24 N3O4[M+H] + MS ESI calculated for 430.18, found 430.20. 1 H NMR(400 MHz,CD3OD)δ 9.03(s,1H),8.76-8.75(m,2H),7.80-7.79(m,2H),7.77-7.63(m,2H),7.41-7.35(m,2H),7.32-7.29(m,2H),4.40-4.33(m,3H),4. 32-4.17(m,1H),3.67-3.66(m,1H),3.37-3.35(m,1H),3.06-2.96(m,1H),2.86-2.72(m,1H),2.67-2.64(m,1H),1.94-1.90(m,2H). Synthetic Scheme 8 [ka]

[0160] Precursor to dProc4Bn4Br [ka]

[0161] (2R,4R)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-4-(4-bromobenzyl)pyrrolidine-2-carboxylic acid Step 1: To a stirred solution of di-tert-butyl (2S,4R)-4-(4-bromobenzyl)pyrrolidine-1,2-dicarboxylate (4 g, 9.08 mmol) in DCM (40 mL) was added TFA (8 mL) at 0° C. The solution was stirred at 25° C. for 40 min. The solution was concentrated under reduced pressure and the residue was purified in the following conditions: 330 g, C 18 Purification by RP-flash column, 5% to 5% in 5 min, 5% to 35% in 20 min, 98% to 98% in 5 min, MeCN (0.05% TFA) in water, RT = 25 min. The collected material contained the desired product. The pH of the solution was adjusted to 10 with saturated NaHCO3. The aqueous layer was extracted with EA (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 tert-butyl (2S,4R)-4-(4-bromobenzyl)pyrrolidine-2-carboxylate (2.4 g, 6.70 mmol, 73.8% yield) as a yellow solid. C 16 H 23 NO2[M+H] + MS ESI calculated values: 340.08, 342.08, found values: 340.10, 342.10.

[0162] Step 2: To a stirred solution of tert-butyl (2S,4R)-4-(4-bromobenzyl)pyrrolidine-2-carboxylate (2.4 g, 7.05 mmol) and TEA (1.071 g, 10.58 mmol) in DCM (40 mL) was added NCS (1.036 g, 7.76 mmol) under a nitrogen atmosphere at 0 °C. The solution was 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 0-20% EA in PE to afford (R)-3-(4-bromobenzyl)-3,4-dihydro-2H-pyrrole-5-carboxylate (2.2 g, 6.18 mmol, 88% yield) as a pale yellow oil. 16 H 21 BrNO2[M+H] + MS ESI calculated values: 338.07, 340.07, found values: 338.00, 340.00. 1 H NMR(300 MHz,CDCl3)δ 7.39-7.31(m,2H),7.00-6.92(m,2H),4.06(dd,J=17.7,6.9 Hz,1H),3.74(dd,J=16.2,3.6 Hz,1H),2.84-2.81(m,1H),2.68-2.41(m,4H),1.48(s,9H). Step 3: To a stirred solution of tert-butyl (R)-3-(4-bromobenzyl)-3,4-dihydro-2H-pyrrole-5-carboxylate (2.2 g, 6.50 mmol) in MeOH (40 mL) and AcOH (10 mL) was added NaBH (0.492 g, 13.01 mmol) at −45° C. under a nitrogen atmosphere. The solution was stirred at −45° C. for 2 hours. The solution was quenched with water (2 mL). The solution was purified under the following conditions: 330 g, C 18Purification by RP flash using a column, 5% to 5% in 5 min, 5% to 35% in 20 min, 98% to 98% in 5 min, MeCN (0.05% TFA) in water, RT = 25 min. The collected material contained the desired product. The pH of the solution was adjusted to 10 with saturated NaHCO3. The aqueous layer was extracted with EA (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 the crude product (1.6 g, 75:25 in SFC). The crude product was separated by SFC using the following conditions: Column: Chiral Art Amylose-SA, 3 x 25 cm, 5 μm; Mobile phase A: CO2, Mobile phase B: MeOH (0.1% 2M NH3-MeOH); Flow rate: 100 mL / min; Gradient: Isocratic 15% B; Column temperature (°C): 35; Back pressure (bar): 100; Wavelength: 220 nm; RT1 (min): 3.13; RT2 (min): 3.8; Sample solvent: MeOH (0.1% 2M NH3-MeOH); Injection volume: 1 mL; Number of runs: 20. The 3.8 min fraction was collected and concentrated under reduced pressure to give tert-butyl (2R,4R)-4-(4-bromobenzyl)pyrrolidine-2-carboxylate (0.95 g, 2.65 mmol, 40.8% yield) as a yellow oil. 16 H 23 BrNO3[M+H] + MS ESI calculated values: 340.08, 342.08, found values: 340.00, 342.00.

[0163] Step 4: To a stirred solution of tert-butyl (2R,4R)-4-(4-bromobenzyl)pyrrolidine-2-carboxylate (950 mg, 2.79 mmol) in DCM (10 mL) was added TFA (20 mL) at room temperature. The solution was stirred at 25° C. for 2 hours. The solvent was concentrated under reduced pressure to give (2R,4R)-4-(4-bromobenzyl)pyrrolidine-2-carboxylic acid (0.8 g, 2.53 mmol, 91% yield) as a pale yellow oil. 12 H 15 BrNO2[M+H] + MS ESI calculated values: 284.02, 286.02, experimental values: 284.00, 286.00. Step 5: To a stirred solution of (2R,4R)-4-(4-bromobenzyl)pyrrolidine-2-carboxylic acid (0.8 g, 2.53 mmol) in THF (20 mL) and water (20.00 mL), NaHCO (0.639 g, 7.60 mmol) and Fmoc-OSu (0.855 g, 2.53 mmol) were added at room temperature. The mixture was stirred at 25° C. for 16 hours. The pH was adjusted to 3 with 1 N HCl. The aqueous layer was extracted with EA (3×200 mL). The combined organic layers were washed with brine (100 mL) and dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure, and the residue was purified under the following conditions: 120 g C 18 Purification by RP flash using a column, 5% to 5% in 5 min, 5% to 60% in 30 min, 98% to 98% in 5 min, MeCN (0.05% TFA) in water, RT = 35 min, afforded (2R,4R)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-4-(4-bromobenzyl)pyrrolidine-2-carboxylic acid (1.1603 g, 2.268 mmol, 90% yield) as an off-white solid. 27 H 25 BrNO4[M+H] + MS ESI calculated values: 506.09, 508.09, found values: 506.00, 508.00. 1 H NMR(400 MHz,CD3OD)δ 7.77-7.26(m,10H),7.14-7.09(m,2H),4.46-4.14(m,4H),3.69-3.31(m,1H),3.15-2.97(m,1H),2.72-2.46(m,4H),1.74-1.58(m,1H). Synthetic Scheme 9 [ka]

[0164] Precursor to dProc4Bn4F [ka]

[0165] (2R,4R)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-4-(4-fluorobenzyl)pyrrolidine-2-carboxylic acid Step 1: To a mixture of (4-fluorobenzyl)triphenylphosphonium chloride (17.82 g, 43.8 mmol) in THF (30 mL) was added potassium 2-methylpropan-2-olate (4.92 g, 43.8 mmol) under argon. The reaction was stirred at room temperature for 1 hour. To the mixture was added a solution of di-tert-butyl (R)-4-oxopyrrolidine-1,2-dicarboxylate (5 g, 17.52 mmol) in THF (20 mL). The reaction was stirred at room temperature for 2 hours. The resulting solution was quenched with water (50 mL) and extracted with ethyl acetate (3 x 300 mL). The organic layers were combined, washed with brine (2 x 50 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated in vacuo, and the residue was purified by silica gel chromatography, eluting with a gradient of ethyl acetate:petroleum ether—0:1 to 1:5—to afford di-tert-butyl (R,E)-4-(4-fluorobenzylidene)pyrrolidine-1,2-dicarboxylate (4.8 g, 12.72 mmol, 73% yield) as a colorless oil. 21 H 29 FNO4[M+H] + MS ESI calculated for 378.21, found 378.20.

[0166] Step 2: To a mixture of di-tert-butyl (R,E)-4-(4-fluorobenzylidene)pyrrolidine-1,2-dicarboxylate (5 g, 13.25 mmol) in MeOH (50 mL) was added Raney Ni (1.2 g, 20.45 mmol) at room temperature under argon. The suspension was degassed under vacuum and purged with H2 several times, and the reaction solution was stirred under 2 atmospheres of H2 at room temperature for 6 hours. LCMS showed that the major component was the product. The resulting solution was filtered. The filtrate was concentrated in vacuo to give di-tert-butyl (2R)-4-(4-fluorobenzyl)pyrrolidine-1,2-dicarboxylate (4.5 g, 11.86 mmol, 90% yield) as a colorless oil. C 21 H 31 FNO4[M+H] +MS ESI calculated for 380.22, found 380.20.

[0167] Step 3: To a stirred solution of di-tert-butyl (2R)-4-(4-fluorobenzyl)pyrrolidine-1,2-dicarboxylate (4.5 g, 11.86 mmol) in TFA (10 mL) and DCM (25 mL) was added at room temperature. The solution was stirred at room temperature for 1 hour. 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: ACN; (Gradient: 5% B, 5 min hold, to 42% B within 15 min, 5 min hold at 42% B; 5 min hold at 95% B within 5 min, 5 min hold at 95% B); Flow rate: 90 mL / min; Detector: UV 210 nm; RT = 26 min. Product-containing fractions were collected and rotary evaporated in vacuo to give tert-butyl (2R)-4-(4-fluorobenzyl)pyrrolidine-2-carboxylate (2.8 g, 10.02 mmol, 85% yield) as a pale yellow oil. 16 H 23 FNO2[M+H] + MS ESI calculated for 280.17, found 280.25.

[0168] Step 4: tert-Butyl (2R)-4-(4-fluorobenzyl)pyrrolidine-2-carboxylate (2.8 g, 10.02 mmol) was separated by Prep-SFC using the following conditions: Column: Lux Cellulose-4, 4.6 × 50 mm, 3 μm; Mobile Phase A: Hex (0.1% NH H O), Mobile Phase B: MeOH prep; Flow Rate: 1.0 mL / min; Gradient: 30% B; Column Temperature: 25 °C; Back Pressure: 100 bar; 190 nm; RT1: 3.42 min; RT2: 4.16 min. The fractions of the first peak (RT1: 3.42 min) were collected and rotary evaporated in vacuo to give tert-butyl (2R,4S)-4-(4-fluorobenzyl)pyrrolidine-2-carboxylate (300 mg, 1.074 mmol, 11% yield) as a yellow oil. 16 H 23 FNO2[M+H] +MS ESI calculated for 280.17, found 280.25. Fractions of the second peak (RT2: 4.16 min) were collected and rotary evaporated in vacuo to give tert-butyl (2R,4R)-4-(4-fluorobenzyl)pyrrolidine-2-carboxylate (2.1 g, 7.52 mmol, 75% yield) as a yellow oil. 16 H 23 FNO2[M+H] + MS ESI calculated for 280.17, found 280.25. 1 H NMR (400 MHz, methanol-d₄) δ 7.25–7.21 (m, 2H), 7.05–7.01 (m, 2H), 4.35–4.31 (m, 1H), 3.43–3.39 (m, 1H), 3.08–3.03 (m, 1H), 2.78–2.77 (m, 2H), 2.76–2.72 (m, 1H), 2.48–2.47 (m, 1H), 1.77–1.74 (m, 1H), 1.52 (s, 9H). Step 5: A solution of tert-butyl (2R,4R)-4-(4-fluorobenzyl)pyrrolidine-2-carboxylate (2.1 g, 7.52 mmol) in CHCl (10 mL) and TFA (10.00 mL) was stirred at 25° C. for 3 hours. The reaction progress was monitored by LCMS. The reaction mixture was concentrated in vacuo to give crude (2R,4R)-4-(4-fluorobenzyl)pyrrolidine-2-carboxylic acid (1.678 g, approximately 7.52 mmol, 100% yield) as a yellow solid. 12 H 15 FNO2[M+H] + MS ESI calculated for 224.11, found 224.15.

[0169] Step 6: To a solution of (2R,4R)-4-(4-fluorobenzyl)pyrrolidine-2-carboxylic acid (1.678 g, 7.52 mmol) in THF (10 mL) and water (10.00 mL), sodium bicarbonate (3.95 g, 47.0 mmol) and N-(9-fluorenylmethoxycarbonyloxy)succinimide (2.86 g, 8.47 mmol) were added at room temperature. The reaction mixture was stirred at room temperature for 16 hours and then extracted with ethyl acetate (3 × 200 mL). The combined organic layer was washed with brine (3 × 100 mL), dried over anhydrous NaSO, and filtered. The filtrate was concentrated and purified by column: Flash C18 (120 g). Mobile phase A: water (0.1% TFA), mobile phase B: ACN; (Gradient: 5 min hold at 5% B, hold to 65% B within 25 min, hold at 65% B for 2.6 min; hold to 95% B within 2 min, hold at 95% B for 5 min); Flow rate: 70 mL / min; Detector: UV 210 nm; RT = 26 min. Product-containing fractions were collected and rotary evaporated in vacuo to give (2R,4R)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-4-(4-fluorobenzyl)pyrrolidine-2-carboxylic acid (2.6866 g, 6.03 mmol, 80% yield) as a white solid. 27 H 25 FNO4[M+H] + MS ESI calculated for 446.18, found 446.25. 1 H NMR (300MHz, methanol-d4)δ 7.78-7.71(m,2H),7.63-7.53(m,2H),7.39-7.26(m,4H),7.21-7.14(m,2H),7.07-6.97(m,2H),4.39-4.37(m,1H), 4.32-4.14(m,3H),3.68-3.32(m,1H),3.22-2.96(m,1H),2.71-2.37(m,2H),2.43-2.36(m,2H),1.81-1.62(m,1H). Synthetic Scheme 10 [ka]

[0170] Precursor to dProc4CH24Pal [ka]

[0171] (2R,4R)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-4-(pyridin-4-ylmethyl)pyrrolidine-2-carboxylic acid Step 1: To a mixture of diethyl phosphonate (8.29 g, 60.0 mmol) in THF (100 mL) was added sodium hydride (3.00 g, 125 mmol) at 0 °C under argon. The reaction was stirred at 0 °C for 30 minutes, and then 4-(chloromethyl)pyridine hydrochloride (8.20 g, 50 mmol) was added to the mixture at 0 °C. The resulting mixture was stirred at ambient temperature for 2 hours, after which it was quenched with water (150 mL) and extracted with ethyl acetate (3 × 300 mL). The organic layers were combined, washed with brine (2 × 180 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography eluting with a gradient of dichloromethane:methanol—1:0 to 15:1—to afford diethyl (pyridin-4-ylmethyl)phosphonate (5.5 g, 23.99 mmol, 48% yield) as an orange oil. C 10 H 17 NO3P[M+H] + MS ESI calculated for 230.09, found 230.15.

[0172] Step 2: To a mixture of diethyl (pyridin-4-ylmethyl)phosphonate (2.4 g, 10.47 mmol) in THF (20 mL) was added NaH (60% in mineral oil) (0.523 g, 13.09 mmol) under argon at 0° C. The mixture was stirred at 0° C. for 30 minutes, and then tert-butyl (R)-2-(((tert-butyldimethylsilyl)oxy)methyl)-4-oxopyrrolidine-1-carboxylate (3.11 g, 9.42 mmol) was added under argon at 0° C. The resulting mixture was stirred at ambient temperature under argon for 1 hour, after which it was quenched with water (40 mL) and extracted with ethyl acetate (3×100 mL). The organic layers were combined, washed with brine (2×50 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography eluting with a gradient of ethyl acetate:petroleum ether—0:1 to 1:3—to afford tert-butyl (R,E)-2-(((tert-butyldimethylsilyl)oxy)methyl)-4-(pyridin-4-ylmethylene)pyrrolidine-1-carboxylate (2.8 g, 6.92 mmol, 66% yield) as an orange semi-solid. 22 H 37 N2O3Si[M+H] + MS ESI calculated for 405.25, found 405.30.

[0173] Step 3: tert-Butyl (R,E)-2-(((tert-butyldimethylsilyl)oxy)methyl)-4-(pyridin-4-ylmethylene)pyrrolidine-1-carboxylate (7.58 g, 18.73 mmol) (combined with other batches) was dissolved in 2-propanol (80 mL), and the resulting mixture was evacuated and a nitrogen atmosphere was applied at ambient temperature. Raney Ni (30 g, 511 mmol, washed several times with 2-propanol) was then added under a nitrogen atmosphere. The suspension was degassed under vacuum and purged several times with H. The resulting mixture was stirred under 1 atmosphere of H at 65° C. for 12 hours, after which it was filtered through diatomaceous earth and washed with 2-propanol (2×150 mL). The filter was concentrated in vacuo to give the racemic product. The racemic product was separated by prep-SFC-HPLC using the following conditions: Column: CHIRAL ART Cellulose-SC, 5 x 25 cm, 5 μm; Mobile phase A: CO₂, Mobile phase B: IPA (0.5% 2M NH₃-MeOH); Flow rate: 250 mL / min; Gradient: Isocratic 44% B; Column temperature (°C): 35; Back pressure (bar): 100; Wavelength: 220 nm; Sample solvent: MeOH:DCM = 2:1; Injection volume: 3 mL. A fast peak was obtained at 7.3 min. The collected fractions were combined and concentrated under vacuum. The residue was lyophilized to give tert-butyl (2R,4S)-2-(((tert-butyldimethylsilyl)oxy)methyl)-4-(pyridin-4-ylmethyl)pyrrolidine-1-carboxylate (420 mg, 1.03 mmol, 6% yield) as a pale yellow semi-solid. 22 H 39 NO3Si[M+H] + MS ESI calculated for 407.27, found 407.30.

[0174] A slower peak was obtained at 8.6 minutes. The collected fractions were combined and concentrated in vacuo. The residue was lyophilized to give tert-butyl (2R,4R)-2-(((tert-butyldimethylsilyl)oxy)methyl)-4-(pyridin-4-ylmethyl)pyrrolidine-1-carboxylate (3.4 g, 8.36 mmol, 45% yield) as a pale yellow semi-solid. 22 H 39 NO3Si[M+H]+ MS ESI calculated for 407.27, found 407.70.

[0175] Step 4: To a mixture of tert-butyl (2R,4R)-2-(((tert-butyldimethylsilyl)oxy)methyl)-4-(pyridin-4-ylmethyl)pyrrolidine-1-carboxylate (3.4 g, 8.36 mmol) in THF (16.72 mL) was added TBAF (1 M in THF, 16.72 mL, 16.72 mmol). The reaction was stirred at ambient temperature for 2 hours and then concentrated in vacuo to give the crude product. The residue was purified by RP Flash under the following conditions: Column: Flash C18 (330 g); Mobile phase A: water (0.05% TFA), Mobile phase B: ACN; (Gradient: 2% B, 3 min hold, to 44% B within 15 min, 4.5 min hold at 44% B; 95% B within 5 min, 5 min hold at 95% B); Flow rate: 80 mL / min; Detector: UV 210 nm; RT = 32.5 min. Product-containing fractions were collected and concentrated in vacuo to give tert-butyl (2R,4R)-2-(hydroxymethyl)-4-(pyridin-4-ylmethyl)pyrrolidine-1-carboxylate (2.2 g, 7.52 mmol, 90% yield) as a pale orange oil. 16 H 25 N2O3[M+H] + MS ESI calculated for 293.18, found 293.20.

[0176] Step 5: To a mixture of tert-butyl (2R,4R)-2-(hydroxymethyl)-4-(pyridin-4-ylmethyl)pyrrolidine-1-carboxylate (2.2 g, 7.52 mmol) in DMF (20 mL) was added PDC (14.15 g, 37.6 mmol). The reaction was stirred at ambient temperature for 12 h. After completion, the pH value of the solution was adjusted to 3 with 1 N HCl. The aqueous layer was extracted with EA (2 × 250 mL). The combined organic layers were washed with brine (2 × 25 mL), dried over anhydrous NaSO, 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: ACN; (Gradient: 5 min hold at 2% B, hold to 33% B within 12 min, hold at 33% B for 8 min; hold to 95% B within 5 min, hold at 95% B for 5 min); Flow rate: 80 mL / min; Detector: UV 210 nm; RT = 35 min. Product-containing fractions were collected and concentrated in vacuo to give (2R,4R)-1-(tert-butoxycarbonyl)-4-(pyridin-4-ylmethyl)pyrrolidine-2-carboxylic acid (2 g, 6.53 mmol, 87% yield) as a black semi-solid. 16 H 23 N2O4[M+H] + MS ESI calculated for 307.16, found 307.15.

[0177] Step 6: To a mixture of (2R,4R)-1-(tert-butoxycarbonyl)-4-(pyridin-4-ylmethyl)pyrrolidine-2-carboxylic acid (2.0 g, 6.53 mmol) in CHCl (10 mL) was added TFA (10 mL, 130 mmol). The reaction was stirred at ambient temperature for 1 hour. The reaction was concentrated in vacuo to give the crude product (2R,4R)-4-(pyridin-4-ylmethyl)pyrrolidine-2-carboxylic acid (2.3 g, 5.58 mmol, 85% yield) as a black semi-solid. 11 H 15 N2O2[M+H] + MS ESI calculated for 207.11, found 207.20.

[0178] Step 7: To a mixture of (2R,4R)-4-(pyridin-4-ylmethyl)pyrrolidine-2-carboxylic acid (2.3 g, 5.58 mmol) in THF (10 mL) and water (10.00 mL) was added sodium bicarbonate (2.342 g, 27.9 mmol) and N-(9-fluorenylmethoxycarbonyloxy)succinimide (1.693 g, 5.02 mmol). The reaction was stirred at ambient temperature for 4 hours and then concentrated in vacuo. 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: ACN; (Gradient: 5 min hold at 5% B, hold to 35% B within 20 min, hold at 35% B for 5.8 min; hold to 95% B within 5 min, hold at 95% B for 5 min); Flow rate: 80 mL / min; Detector: UV 210 nm; RT = 40.8 min. Product-containing fractions were collected and concentrated in vacuo to give (2R,4R)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-4-(pyridin-4-ylmethyl)pyrrolidine-2-carboxylic acid (2.02 g, 4.71 mmol, 85% yield) as an off-white solid. C 26 H 25 N2O4[M+H] + MS ESI calculated value of 429.17, found value of 429.20. 1 H NMR (400MHz, methanol-d4)δ 8.78-8.74(m,2H),7.98-7.93(m,2H),7.79-7.74(m,2H),7.64-7.57(m,2H),7.40-7.27(m,4H),4.45-4.43(m,1H), 4.35-4.16(m,3H),3.75-3.73(m,1H),3.51-3.48(m,1H),3.23-3.05(m,3H),2.60-2.55(m,2H),1.82-1.65(m,1H). Synthetic Scheme 11 [ka]

[0179] Precursor to F4bcpA [ka]

[0180] (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-(3-(tert-butoxycarbonyl)bicyclo[1.1.1]pentan-1-yl)phenyl)propanoic acid 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) was added 3-bromoprop-1-ene (3.63 g, 30.0 mmol) and NaHCO (0.840 g, 10.00 mmol) under an argon atmosphere at 0° C. The resulting mixture was stirred at 40° C. for 16 h. The reaction mixture was cooled to room temperature, quenched with water (200 mL), and extracted with EA (2×200 mL). The combined organic layers were washed with brine (3×100 mL), dried over NaSO, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluting with 0-40% EA in PE). The product-containing fractions were collected and rotary evaporated in vacuo to give allyl (S)-2-((tert-butoxycarbonyl)amino)-3-(4-iodophenyl)propanoate (4 g, 9.27 mmol, 93% yield) as an off-white solid. 27 H 25 INO4[M+H] + MS ESI calculated for 554.08, found 554.20.

[0181] 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 tert-butyl (Z)-N,N'-diisopropylcarbamimidate (37.7 g, 188 mmol) at room temperature. The resulting solution was stirred at 40 °C for 2 h. The mixture was cooled to room temperature. The solid was filtered off, and the filtrate was concentrated under reduced pressure to give 1-(tert-butyl)3-methyl bicyclo[1.1.1]pentane-1,3-dicarboxylate (15 g, 39.8 mmol, 85% yield) as a pale yellow oil. 1H NMR(400 MHz,DMSO-d6)δ 3.61(s,3H),2.17(s,6H),1.39(s,9H). Step 3: To a stirred solution of 1-(tert-butyl) 3-methylbicyclo[1.1.1]pentane-1,3-dicarboxylate (15 g, 39.8 mmol) in THF (150 mL) was added LiOH (119 mL, 119 mmol, 1N in water) at room temperature. The resulting solution was stirred at 25° C. for 5 hours. The pH of the solution was adjusted to 3 with 1N HCl and then extracted with EA (3×100 mL). The combined organic layers were washed with brine (2×50 mL) and dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure to give 3-(tert-butoxycarbonyl)bicyclo[1.1.1]pentane-1-carboxylic acid (10 g, 28.3 mmol, 71% yield) as a pale yellow solid. 11 H 15 O4[MH] - MS ESI calculated for 211.10, found 211.10.

[0182] 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 DCC (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% EA in PE to give 1-(tert-butyl) 3-(1,3-dioxoisoindolin-2-yl)bicyclo[1.1.1]pentane-1,3-dicarboxylate (4.8 g, 12.09 mmol, 43% yield) as an off-white solid. 1 H NMR(400 MHz,CDCl3)δ 7.89(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). Step 5: To a stirred solution of NiBr2.3H2O (0.791 g, 2.90 mmol) in DMA (80 mL) was added dtbbpy (0.973 g, 3.63 mmol) at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at 50 °C for 30 min and then 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), allyl (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-iodophenyl)propanoate (6.69 g, 12.09 mmol), TMSCl (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 EA (3 × 100 mL). The combined organic layers were washed with brine (3 × 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 0-40% EA in PE to give tert-butyl (S)-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, 1.550 mmol, 13% yield) as an off-white solid. 37 H 40 NO6[M+H] + MS ESI calculated for 594.28, found 594.30.

[0183] Step 6: To a stirred solution of tert-butyl (S)-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, 1.550 mmol) and phenylsilane (0.335 g, 3.10 mmol) in THF (30 mL) was added Pd(PhP) (0.090 g, 0.077 mmol) at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at room temperature for 1 h. It was then concentrated under reduced pressure, and the residue was purified by RP-flash using the following conditions: C 18 Column, 330 g, 5% to 5% in 5 min, 5% to 70% in 40 min, MeCN in water (0.05% TFA), (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-(3-(tert-butoxycarbonyl)bicyclo[1.1.1]pentan-1-yl)phenyl)propanoic acid (522.8 mg, 0.897 mmol, 58% yield) was obtained as a pale yellow solid. 34 H 34 NO6[MH] - MS ESI calculated for 552.25, found 552.30. 1 H NMR(400 MHz,CD3OD)δ 7.80-7.77(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). Synthetic Scheme 12 [ka]

[0184] Precursor to F4pcCCA [ka]

[0185] (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-((1s,4R)-4-(tert-butoxycarbonyl)cyclohexyl)phenyl)propanoic acid Step 1: To a mixture of tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-ene-1-carboxylate (29.5 g, 96 mmol) in THF (30 mL) was added methyl (S)-2-((tert-butoxycarbonyl)amino)-3-(4-iodophenyl)propanoate (15.5 g, 38.2 mmol) and Pd(PhP) (2.210 g, 1.912 mmol) at room temperature. The reaction was warmed to 60 °C for 4 h. The resulting solution was quenched with water (100 mL) and extracted with ethyl acetate (3 × 300 mL). The organic layers were combined, washed with brine (2 × 200 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated in vacuo to give the crude product. The residue was purified by silica gel chromatography eluting with a gradient of ethyl acetate:petroleum ether—0:1 to 1:4 to give tert-butyl 4′-((S)-2-((tert-butoxycarbonyl)amino)-3-methoxy-3-oxopropyl)-2,3,4,5-tetrahydro-[1,1′-biphenyl]-4-carboxylate (16 g, 34.8 mmol, 91% yield) as a colorless solid. 26 H 38 NO6Na[M+Na] + MS ESI calculated for 482.26, found 482.10.

[0186] Step 2: A mixture of tert-butyl 4'-((S)-2-((tert-butoxycarbonyl)amino)-3-methoxy-3-oxopropyl)-2,3,4,5-tetrahydro-[1,1'-biphenyl]-4-carboxylate (16 g, 34.8 mmol) in methanol (160 mL) was degassed with Pd-C (10% on carbon, ca. 55% water, 5.3 g, 4.98 mmol) three times with H at room temperature and stirred under an atmosphere of H (1.5 atm) at room temperature for 1 h. The resulting solution was filtered. The filtrate was concentrated in vacuo to give tert-butyl (S)-4-(4-(2-((tert-butoxycarbonyl)amino)-3-methoxy-3-oxopropyl)phenyl)cyclohexane-1-carboxylate (15 g, 29.2 mmol, 84% yield) as a colorless semi-solid. 26 H 41 NO6[M+H] + MS ESI calculated for 462.28, found 462.30.

[0187] Step 3: To a stirred solution of tert-butyl (S)-4-(4-(2-((tert-butoxycarbonyl)amino)-3-methoxy-3-oxopropyl)phenyl)cyclohexane-1-carboxylate (15 g, 32.5 mmol) in THF (300 mL), LiOH (65.0 mL, 65.0 mmol) was added at room temperature. The solution was stirred at 20° C. for 1 hour. The pH value of the solution was adjusted to 3 with 1 N HCl. The reaction was concentrated under reduced pressure to give (S)-2-((tert-butoxycarbonyl)amino)-3-(4-(4-(tert-butoxycarbonyl)cyclohexyl)phenyl)propanoic acid (14 g, 31.3 mmol, 96% yield) as a white solid. C 25 H 38 NO6Na[M+Na] + MS ESI calculated for 470.26, found 470.30.

[0188] Step 4: (S)-2-((tert-butoxycarbonyl)amino)-3-(4-(4-(tert-butoxycarbonyl)cyclohexyl)phenyl)propanoic acid (14 g, 31.3 mmol) was separated by Prep-SFC using the following conditions: Column: CHIRAL ART Cellulose-SB, 3 x 25 cm, 5 µm; Mobile phase A: CO, Mobile phase B: MeOH (0.1% 2M NH-MEOH); Flow rate: 80 mL / min; Gradient: 10% B; 220 nm; RT1: 7.45; RT2: 8.38; Injection volume: 1.3 ml; Number of runs: 131; (S)-2-((tert-butoxycarbonyl)amino)-3-(4-((1s,4R)-4-(tert-butoxycarbonyl)cyclohexyl)phenyl)propanoic acid (8.3 g, 18.54 mmol, 59.3% yield) of the fast peak was obtained as a white solid; C 25 H 38 NO6Na[M+Na] + MS ESI calculated for 470.26, found 470.30. 1H NMR (300 MHz, chloroform-d) δ 7.11 (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), and a late peak of (S)-2-((tert-butoxycarbonyl)amino)-3-(4-((1r,4S)-4-(tert-butoxycarbonyl)cyclohexyl)phenyl)propanoic acid (3 g, 6.70 mmol, 21.43% yield) was obtained as a white solid; C 25 H 38 NO6Na [M+Na] +MS ESI calculated for 470.26, found 470.30. 1H NMR (300 MHz, chloroform-d) δ 7.14-7.06 (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). Step 5: To a mixture 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 hydrogen chloride (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 (6 g, 17.27 mmol, 93% yield) as a white solid. 20 H 30 NO4[M+H] + MS ESI calculated for 348.21, found 348.25.

[0189] 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), n-(9-fluorenylmethoxycarbonyloxy)succinimide (5.24 g, 15.54 mmol) was added at room temperature. The mixture was stirred at room temperature for 1 hour. The pH value of the solution was adjusted to 3 with 1 N HCl. The aqueous layer was extracted with acetic ether (2 × 200 mL). The combined organic layers were washed with brine (150 mL), dried over anhydrous NaSO, and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by RP flash to give (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-((1s,4R)-4-(tert-butoxycarbonyl)cyclohexyl)phenyl)propanoic acid (7.7 g, 13.52 mmol, 78% yield) as an off-white solid. 35 H 41 NO6[M+H] + MS ESI calculated for 570.28, found 570.15. 1 H NMR (300MHz, methanol-d4) δ 7.81(d,J=7.5 Hz,2H),7.61(d,J=7.5 Hz,2H),7.43-7.27(m,4H),7.16(d,J=7.8 Hz,2H),7.07(d,J=7.9 Hz,2H),4.45-4.33(m,2H),4.22-4.06(m,2H),3.26-3.14(m,1H),2.96-2.88(m,1H),2.59(s,1H),2.48(s,1H),2.15(s,2H),1.61(d,J=5.5 Hz,6H),1.49(s,9H). Synthetic Scheme 13 [ka]

[0190] Precursor to F4ptCCA (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-((1r,4S)-4-(tert-butoxycarbonyl)cyclohexyl)phenyl)propanoic acid 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 afford (S)-2-amino-3-(4-((1r,4S)-4-(tert-butoxycarbonyl)cyclohexyl)phenyl)propanoic acid (2 g, 5.76 mmol, 86% yield) as a white solid. 20 H 31 NO4[M+H] + MS ESI calculated for 348.21, found 348.25.

[0191] 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 NaHCO (2.418 g, 28.8 mmol) in THF (20 mL) and water (20 mL), N-(9-fluorenylmethoxycarbonyloxy)succinimide (1.748 g, 5.18 mmol) was added at room temperature. The mixture was stirred at 20 °C for 1 hour. The pH value of the solution was adjusted to 3 with 1 N HCl. The aqueous layer was extracted with acetic ether (2 × 200 mL). The combined organic layers were washed with brine (3 × 50 mL), dried over anhydrous NaSO, and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by RP flash to give (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-((1r,4S)-4-(tert-butoxycarbonyl)cyclohexyl)phenyl)propanoic acid (1.89 g, 3.32 mmol, 57.6% yield) as an off-white solid. 35 H 41 NO6[M+H] +MS ESI calculated for 570.28, found 570.35. 1 H NMR (300MHz, methanol-d4) δ 7.81(d,J=7.6 Hz,2H),7.61(d,J=7.5 Hz,2H),7.43-7.38(m,2H),7.34-7.27(m,2H),7.17(d,J=8.0 Hz,2H),7.08(d,J=7.8 Hz,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=11.9 Hz,1H),1.96(s,3H),1.82-1.55(m,2H),1.47(s,13H). Synthetic Scheme 14 [ka]

[0192] Precursor to PyrimAla4Ph4CO2H [ka]

[0193] (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(2-(4-(tert-butoxycarbonyl)phenyl)pyrimidin-5-yl)propanoic acid Step 1: To a stirred solution of NiBr2-glyme (0.951 g, 2.432 mmol) in DMA (100 mL) was added 1,10-phenanthroline (0.527 g, 2.432 mmol) under a nitrogen atmosphere at 25° C. The resulting solution was stirred at 50° C. for 1 hour, then cooled to room temperature, and to it was added 2-chloro-5-iodopyrimidine (5.85 g, 24.32 mmol), tert-butyl (R)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-iodopropanoate (6 g, 12.16 mmol), TBAI (4.66 g, 12.16 mmol), and zinc (1.590 g, 24.32 mmol) at room temperature. The resulting mixture was stirred at 25 °C for 2 h, then quenched with HO (200 mL) and extracted with EA (2 × 500 mL). The combined organic layers were washed with brine (3 × 200 mL), dried over anhydrous NaSO, and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with 0–20% EA in PE to afford tert-butyl (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(2-chloropyrimidin-5-yl)propanoate (4 g, 8.35 mmol, 70% yield) as an off-white solid. 26 H 27 ClN3O4[M+H] + MS ESI calculated for 480.16, found 480.25.

[0194] Step 2: To a stirred solution of tert-butyl (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(2-chloropyrimidin-5-yl)propanoate (6 g, 12.50 mmol) in DCM (15 mL) was added TFA (30 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: Column: Flash C 18(330 g); Mobile phase A: water (0.1% TFA), Mobile phase B: ACN; (Gradient: 5 min hold at 5% B, hold to 30% B within 15 min, hold at 30% B for 5 min; up to 95% B within 20 min, hold at 95% B for 10 min); Flow rate: 90 mL / min; Detector: UV 210 nm; RT = 40 min. Product-containing fractions were collected and evaporated in vacuo to give (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(2-chloropyrimidin-5-yl)propanoic acid (2.56 g, 6.04 mmol, 51% yield) as a yellow oil. C 22 H 19 ClN3O4[M+H] + MS ESI calculated for 424.10, found 424.15.

[0195] Step 3: To a stirred mixture of (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(2-chloropyrimidin-5-yl)propanoic acid (2.56 g, 6.04 mmol), (4-(tert-butoxycarbonyl)phenyl)boronic acid (1.609 g, 7.25 mmol) and K3PO4 (6.41 g, 30.2 mmol) in water (20 mL) and dioxane (20 mL) was added Pd(dtbpf)Cl2 (0.590 g, 0.906 mmol) at room temperature. The resulting mixture was stirred at 80 °C for 2 h, then cooled to room temperature and concentrated under reduced pressure. The residue was purified by RP-flash using the following conditions: Column: Flash C 18 (330 g); Mobile phase A: water (0.1% TFA), Mobile phase B: ACN; (Gradient: 5 min hold at 5% B, hold to 30% B within 15 min, hold at 30% B for 5 min; up to 95% B within 20 min, hold at 95% B for 10 min); Flow rate: 90 mL / min; Detector: UV 210 nm; RT = 40 min. Product-containing fractions were collected and evaporated in vacuo to give (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(2-(4-(tert-butoxycarbonyl)phenyl)pyrimidin-5-yl)propanoic acid (2.5629 g, 4.53 mmol, 75% yield) as a yellow oil. C 33 H 32 N3O6[M+H]+ MS ESI calculated for 566.22, found 566.40. 1 H NMR(300 MHz,CD3OD)δ 8.75(s,2H),8.42(d,J=8.3 Hz,2H),8.02(d,J=8.2 Hz,2H),7.75(d,J=7.5 Hz,2H),7.66-7.45(m,2H),7.42-7.15(m,4H),4.62-4.44(m,1H),4.38-4.20(m,2H),4.12(t,J=7.0 Hz,1H),3.35-3.33(m,1H),3.11-2.97(m,1H),1.62(s,9H). Synthetic Scheme 15 [ka]

[0196] Precursor to Trp4Az [ka]

[0197] (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(1H-pyrrolo[3,2-b]pyridin-3-yl)propanoic acid To a solution of (S)-2-amino-3-(1H-pyrrolo[3,2-b]pyridin-3-yl)propanoic acid (250 mg, 1.22 mmol) and NaCO (232 mg, 2.2 mmol) in THF (4 mL) and water (2 mL) was added Fmoc-OSu (431 mg, 1.28 mmol) at room temperature. The mixture was stirred for 16 hours, then adjusted to pH 3 with 1N HCl and concentrated in vacuo. The resulting aqueous suspension was washed with EtOAc (3 × 100 mL) and then extracted with 20% i-PrOH / DCM (3 × 100 mL). The combined organic extracts were dried over Na2SO4, filtered, and concentrated in vacuo to give (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(1H-pyrrolo[3,2-b]pyridin-3-yl)propanoic acid (470 mg, 1.10 mmol, 90% yield) as a white solid. MS-ESI [M+H]+ =428.4.1H NMR(500 MHz,DMSO-d6)δ 11.14(br s,1H),8.30(dd,J=4.6,1.3 Hz,1H),7.88(d,J=7.6 Hz,2H),7.75(dd,J=8.1,1.3 Hz,1H),7.63(t,J=7.9 Hz,2H),7.47(d,J=2.3 Hz,1H),7.40(td,J=7.3,3.4 Hz,2H),7.32-7.24(m,2H),7.12(dd,J=8.1,4.6 Hz,1H),4.23(td,J=8.1,4.3 Hz,1H),4.19-4.15(m,3H),3.30(dd,J=14.5,4.0 Hz,1H),3.09(dd,J=14.6,8.6 Hz,1H). Synthetic Scheme 16 [ka]

[0198] Precursor to 3AzaPhe4AcPip [ka]

[0199] (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(6-(4-acetylpiperazin-1-yl)pyridin-3-yl)propanoic acid Step 1: To a mixture of 1-(piperazin-1-yl)ethan-1-one (21.85 g, 170 mmol) in DMF (150 mL) was added 5-bromo-2-fluoropyridine (15 g, 85 mmol) at room temperature under argon. The reaction was stirred at 100° C. for 2 hours, then diluted with 300 mL of EtOAc, washed with HO (3×80 mL), saturated aqueous NaCl (80 mL), dried over NaSO, and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with a gradient of 0% to 100% EtOAc in PE. Fractions containing the desired product were combined and concentrated under reduced pressure to give 1-(4-(5-bromopyridin-2-yl)piperazin-1-yl)ethan-1-one. C11 H 15 BrNO[M+H] + MS ESI calculated values ​​for 284.03 and 286.03, found values ​​283.90 and 285.90.

[0200] 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), tert-butyl (R)-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. Then, zinc powder (2.071 g, 31.7 mmol) was added. The resulting mixture was stirred at 50° C. for 1 hour. The resulting mixture was poured into water (300 mL) and extracted with EtOAc (3×300 mL). The organic layer was washed with water (100 mL) and brine (2×80 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with DCM-MeOH (10:1) to give tert-butyl (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(6-(4-acetylpiperazin-1-yl)pyridin-3-yl)propanoate. C 33 H 39 N4O5[M+H] + MS ESI calculated for 571.28, found 571.40.

[0201] Step 3: To a mixture of tert-butyl (S)-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 aqueous solution. TFA; Mobile phase B: MeCN; (Gradient: 0% B, 10 min hold, to 42.3% B within 35 min, 42.3% B, 3.2 min hold; to 95% B within 2 min, 10 min hold 95% B); Flow rate: 60 mL / min; Detector: UV 254 and 210 nm; RT: 35.32 min. Product-containing fractions were collected and concentrated under vacuum to give (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(6-(4-acetylpiperazin-1-yl)pyridin-3-yl)propanoic acid. 29 H 31 N4O5[M+H] + MS ESI calculated value of 515.22, found value of 515.15. 1 H NMR (300MHz, 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,1 H),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). Synthetic Scheme 17 [ka]

[0202] Precursor to 3AzaTyrEtNAc [ka]

[0203] (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(6-(2-acetamidoethoxy)pyridin-3-yl)propanoic acid Step 1: To a mixture of N-(2-hydroxyethyl)acetamide (17.58 g, 170 mmol) in t-BuOH (150 mL) was added 5-bromo-2-fluoropyridine (15 g, 85 mmol) followed by potassium tert-butoxide (19.13 g, 170 mmol) at room temperature under argon. The reaction was stirred at room temperature for 1 hour and then concentrated under reduced pressure. The residue was diluted with 500 mL of EtOAc and washed with saturated aqueous NaHCO3 (3 x 250 mL), saturated aqueous NaCl (250 mL), dried over Na2SO4, and filtered. The filtrate was concentrated under reduced pressure to give crude N-(2-((5-bromopyridin-2-yl)oxy)ethyl)acetamide. CH 12 BrN2O2[M+H] + MS ESI calculated for 259.00 and 261.00, found 258.90 and 261.90.

[0204] 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), tert-butyl (R)-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 300 mL of EtOAc, washed with saturated aqueous NaHCO3 (3 x 80 mL), brine (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% to 20% DCM in MeOH. Fractions containing the desired product were combined and concentrated under reduced pressure to give tert-butyl (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(6-(2-acetamidoethoxy)pyridin-3-yl)propanoate. C 31 H 36 N3O6[M+H] + MS ESI calculated for 546.25, found 546.40.

[0205] Step 3: To a mixture of tert-butyl (S)-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 aqueous solution. NH4HCO3; Mobile phase B: MeCN; (Gradient: 5 min hold at 0% B, hold to 45% B within 20 min, 10 min hold at 45% B; up to 95% B within 15 min, 10 min hold at 95% B); Flow rate: 60 mL / min; Detector: UV 254 and 210 nm; RT: 35.32 min. Product-containing fractions 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. C 27 H 28 N3O6[M+H] + MS ESI calculated for 490.19, found 490.10. 1 H NMR (400MHz, 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,1) H),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). Synthetic Scheme 18 [ka]

[0206] Precursor to 3Pal4Ph4CO2H [ka]

[0207] (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(6-(4-(tert-butoxycarbonyl)phenyl)pyridin-3-yl)propanoic acid Step 1: To a stirred solution of NiCl-glyme (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 a nitrogen atmosphere. The resulting solution was stirred at 50 °C for 1 hour. 2-Chloro-5-iodopyridine (5 g, 20.88 mmol), tert-butyl (R)-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 hours. The reaction was quenched with HO (200 mL) and extracted with EtOAc (2 × 500 mL). The combined organic layers were washed with brine (3 × 200 mL), dried over anhydrous NaSO, 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 give tert-butyl (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(6-chloropyridin-3-yl)propanoate. 27 H 28 ClN2O4[M+H] + MS ESI calculated for 479.17, found 479.20.

[0208] Step 2: To a stirred solution of tert-butyl (S)-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 C 18 (330 g); Mobile phase A: water (0.1% TFA), Mobile phase B: ACN; (Gradient: 5 min hold at 5% B, hold to 30% B within 15 min, hold at 30% B for 5 min; up to 95% B within 20 min, hold at 95% B for 10 min); Flow rate: 90 mL / min; Detector: UV 210 nm; RT = 40 min. Product-containing fractions were collected and evaporated in vacuo to give (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(6-chloropyridin-3-yl)propanoic acid. C 23 H 20 ClN2O4[M+H] + MS ESI calculated for 423.10, found 423.10.

[0209] Step 3: To a stirred solution of (S)-2-((((9H-fluoren-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) was added (4-(tert-butoxycarbonyl)phenyl)boronic acid (1.890 g, 8.51 mmol) and KPO (7.53 g, 35.5 mmol) under nitrogen at 25 °C. The resulting solution was stirred at 25 °C for 10 min. Pd(dtbpf)Cl (0.694 g, 1.064 mmol) was added to the solution, and then the mixture was stirred at 60 °C for 16 h. The reaction was cooled to room temperature, quenched with HO (200 mL), and extracted with EtOAc (2 × 500 mL). The combined organic layers were washed with brine (3 x 200 mL), dried over anhydrous NaSO and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by RP-flash using the following conditions: Column: Flash C 18(330 g); Mobile phase A: water (0.1% TFA), Mobile phase B: ACN; (Gradient: 5 min hold at 5% B, hold to 60% B within 15 min, hold at 60% B for 15 min; up to 95% B within 10 min, hold at 95% B for 10 min); Flow rate: 90 mL / min; Detector: UV 210 nm; RT = 55 min. Product-containing fractions were collected and evaporated in vacuo to give (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] + MS ESI calculated for 565.23, found 565.15; 1 H NMR (400MHz, methanol-d4) δ 8.66(d,J=1.9 Hz,1H),8.12-8.07(m,3H),7.99-7.92(m,3H),7.77(d,J=7.5 Hz,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). Synthetic Scheme 19 [ka]

[0210] Precursor to Bip4CO2H [ka]

[0211] (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4'-(tert-butoxycarbonyl)-[1,1'-biphenyl]-4-yl)propanoic acid Argon gas 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 KPO (8.19 g, 38.6 mmol) in THF (40 mL) for 10 minutes, followed by the addition of [1,1'-bis(di-tert-butylphosphino)ferrocene]dichloropalladium(II) (0.839 g, 1.287 mmol). The resulting mixture was stirred at 50 °C for 16 hours, after which it was diluted with EtOAc (300 mL), washed with saturated aqueous NaHCO (3 × 80 mL), brine (2 × 40 mL), dried over NaSO, and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with a gradient of 0% to 50% EtOAc in PE. Product-containing fractions were collected and rotary evaporated in vacuo. The residue was repurified by CombiFlash using the following conditions: Column: C18 gel column (330 g), 20-35 μm; Mobile phase A: 0.5% aqueous TFA; Mobile phase B: MeCN; (Gradient: 0% B, 10 min hold, to 62.3% B within 25 min, 62.3% B, 6.2 min hold; to 95% B within 2 min, 10 min hold 95% B); Flow rate: 90 mL / min; Detector: UV 254 and 210 nm; RT: 32.32 min. The product-containing fractions were collected and concentrated under reduced pressure to give (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] + MS ESI calculated for 564.23, found 564.15. 1 H NMR (300MHz, 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,2 H),4.51-4.11(m,4H),3.32-3.25(m,1H),3.03-2.95(m,1H),1.61(s,9H). Synthetic Scheme 20 [ka]

[0212] Precursor to Phe4AcPip [ka]

[0213] (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-(4-acetylpiperazin-1-yl)phenyl)propanoic acid 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) under nitrogen at 25 °C. The resulting solution was stirred at 100 °C for 10 min. 1-(piperazin-1-yl)ethan-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, quenched with HO (500 mL), and extracted with EtOAc (2 × 500 mL). The combined organic layers were washed with brine (3 × 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] + MS ESI calculated value of 392.21, found value of 392.25. 1H NMR (400MHz, 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). 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 hour 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 further purification. 15 H 22 N3O3[M+H] + MS ESI calculated for 292.16, found 292.20.

[0214] Step 3: To a stirred solution of (S)-3-(4-(4-acetylpiperazin-1-yl)phenyl)-2-(carboxyamino)propanoic acid (7 g, 20.87 mmol) in THF (25 mL) and water (25 mL) under nitrogen at 25 °C, Fmoc-OSu (6.34 g, 18.79 mmol) was added. NaHCO (8.77 g, 104 mmol) was then added. The resulting mixture was stirred at 25 °C for 2 h. The pH was adjusted to 5 with 1 N HCl and extracted with EtOAc (2 × 200 mL). The combined organic layers were washed with brine (3 × 100 mL), dried over anhydrous NaSO, 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: ACN; (Gradient: 5 min hold at 5% B, hold to 55% B within 15 min, hold at 55% B for 5 min; hold to 95% B within 20 min, hold at 95% B for 5 min); Flow rate: 90 mL / min; Detector: UV 210 nm; RT = 45 min. Product-containing fractions were collected and concentrated in vacuo to give (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-(4-acetylpiperazin-1-yl)phenyl)propanoic acid. 30 H 32 N3O5[M+H] + MS ESI calculated for 514.23, found 514.30. 1 H NMR (400MHz, 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,1) H),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). Synthetic Scheme 21 [ka]

[0215] Precursor to Phe4Pyrim5CO2H [ka]

[0216] (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-(5-(tert-butoxycarbonyl)pyrimidin-2-yl)phenyl)propanoic acid 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 BocO (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 in vacuo. The residue was purified by silica gel column chromatography, eluting with 0-60% EtOAc in PE, to give tert-butyl 2-chloropyrimidine-5-carboxylate. CH 12 ClN2O2[M+H] + MS ESI calculated for 215.05, found 214.95.

[0217] 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), (S)-3-(4-bromophenyl)-2-((tert-butoxycarbonyl)amino)propanoic acid (8.64 g, 28.0 mmol), PdCl(dppf) (1.364 g, 1.864 mmol), and KCO (7.73 g, 55.9 mmol) were added at room temperature under argon. The resulting mixture was stirred at 80 °C for 3 h, then the pH was adjusted to 4 with 1 N HCl and extracted with EtOAc (2 × 300 mL). The combined organic layers were washed with brine (3 × 50 mL), dried over anhydrous NaSO, 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] + MS ESI calculated for 444.21, found 444.35. 1 H NMR(400 MHz,CDCl3)δ 9.27(s,2H),8.42(d,J=7.8 Hz,2H),7.36(d,J=7.9 Hz,2H),4.69-4.68(m,1H),3.29-3.24(m,2H),1.63(s,9H),1.44(s,9H). 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 hour, the reaction was concentrated to give (S)-2-amino-3-(4-(5-(tert-butoxycarbonyl)pyrimidin-2-yl)phenyl)propanoic acid. C 18 H 22 N3O4[M+H] + MS ESI calculated for 344.15, found 344.20.

[0218] 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 HO (40 mL) was added NaHCO (4.89 g, 58.2 mmol) and Fmoc-OSu (3.54 g, 10.48 mmol) at room temperature under argon. After stirring at room temperature for 1 h, the pH was adjusted to 4 with 1 N HCl and the solution was extracted with EtOAc (2 × 300 mL). The combined organic layers were washed with brine (150 mL), dried over anhydrous NaSO, and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by RP flash using the following conditions: Column: C 18 Column (330 g); Mobile phase A: Water (0.05% TFA); Mobile phase B: MeCN; (Gradient: 5 min hold at 0% B, hold to 73% B within 30 min, hold at 73% B for 10 min; up to 95% B within 4 min, hold at 95% B for 10 min); Flow rate: 60 mL / min; Detector: UV 254 and 210 nm; RT: 35.32 min to give (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] + MS ESI calculated for 566.22, found 566.35. 1 H NMR(300 MHz,DMSO-d6)δ 12.85(s,1H),9.24(s,2H),8.37(d,J=8.0 Hz,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). Synthetic Scheme 22 [ka]

[0219] Precursor to PyrimAla4AcPip [ka]

[0220] (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(2-(4-acetylpiperazin-1-yl)pyrimidin-5-yl)propanoic acid Step 1: To a mixture of 5-bromo-2-fluoropyrimidine (7 g, 39.6 mmol) in DMF (70 ml), 1-(piperazin-1-yl)ethan-1-one (10.14 g, 79 mmol) and K2CO3 (10.93 g, 79 mmol) were added at room temperature. The reaction mixture was stirred at 100 °C for 2 hours and then cooled to room temperature. The reaction mixture was extracted with 500 mL of EtOAc, washed with HO (3 × 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% to 80% EtOAc in PE. Fractions containing the desired product were combined and concentrated under reduced pressure to give 4-(5-bromopyrimidin-2-yl)piperazine-1-carbaldehyde. C 10 H 14 BrNO [M+H] + MS ESI calculated values: 285.03, 287.03, found values: 284.95, 286.95. 1 H NMR(300 MHz,CDCl3)δ 8.32(s,2H),3.90-3.74(m,4H),3.74-3.64(m,2H),3.58-3.48(m,2H),2.15(s,3H). 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), tert-butyl (R)-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. Then, Zn (2.064 g, 31.6 mmol) was added. The resulting mixture was stirred at 30°C for 24 hours, after which the reaction mixture was filtered and washed with DCM. The organic layer was concentrated under reduced pressure. The residue was purified by column chromatography under the following conditions: 18 Purification by RP-flash using a column (330 g); mobile phase A: water (0.05% TFA); mobile phase B: MeCN; (gradient: 5 min hold at 0% B, hold to 82% B within 30 min, hold at 82% B for 6 min; hold to 95% B within 2 min, hold at 95% B for 3 min); flow rate: 60 mL / min; detector: UV 254 and 210 nm; RT: 35 min to give tert-butyl (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(2-(4-acetylpiperazin-1-yl)pyrimidin-5-yl)propanoate. 32 H 38 N5O5[M+H] + MS ESI calculated for 572.28, found 572.30. 1 H NMR(300 MHz,CDCl3)δ 8.19(s,2H),7.77(d,J=7.6 Hz,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),4.22-4.17(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). Step 3: To a mixture of tert-butyl (S)-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 hours and then concentrated under reduced pressure. The residue was purified by RP-flash using the following conditions: Column: C 18 Gel column (330 g); Mobile phase A: Water (0.05% TFA); Mobile phase B: MeCN; (Gradient: 5 min hold at 0% B, hold to 72% B within 30 min, hold at 72% B for 6 min; up to 95% B within 2 min, hold at 95% B for 10 min); Flow rate: 60 mL / min; Detector: UV 254 and 210 nm; RT: 35 min to give (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] + MS ESI calculated for 516.22, found 516.35. 1 H NMR(300 MHz,DMSO-d6)δ 8.31(s,2H),7.89(d,J=7.5 Hz,2H),7.81(d,J=8.6 Hz,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). Synthetic Scheme 23 [ka]

[0221] Precursor to sbMeW4F [ka]

[0222] (2S,3S)-2-amino-3-(4-fluoro-1H-indol-3-yl)butanoic acid A 1 L three-necked round-bottom flask purged with nitrogen and maintained under an inert nitrogen atmosphere was charged with 4-fluoro-1H-indole (10 g, 1.00 equiv.), l-threonine (10.6 g, 1.20 equiv.), DMSO (100 mL), and potassium phosphate buffer (0.2 M, 300 mL, pH = 7.4). 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.

[0223] To the above reaction mixture, THF (100 mL), sodium carbonate (23.56 g, 3.0 equiv.), and 2,5-dioxopyrrolidin-1-yl 9H-fluoren-9-ylmethyl carbonate (29.96 g, 1.20 equiv.) were added at 0° C. The resulting solution was stirred at room temperature overnight. The pH was adjusted to 4 with 3 M HCl, and the solid precipitate was removed by filtration. The resulting solution was extracted with EtOAc (3×500 mL). The organic fractions were combined, washed with brine (1 L), dried over anhydrous sodium sulfate, and concentrated under high 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(300 MHz,DMSO-d6)δ 12.60(s,1H),11.15(s,1H),7.87(d,J=7.6 Hz,2H),7.76-7.49(m,3H),7.47-7.34(m,2H),7.34-7.16(m,4H),7.03(td,J=7.9,5.0 Hz,1H),6.73(dd,J=11.8,7.7 Hz,1H),4.36(t,J=8.5 Hz,1H),4.31-4.02(m,3H),3.51(q,J=7.4 Hz,1H),1.31(d,J=7.0 Hz,4H),0.78(s,1H). Synthetic Scheme 24 [ka]

[0224] Precursor to sbMeW4Cl [ka]

[0225] (2S,3S)-2-amino-3-(4-chloro-1H-indol-3-yl)butanoic acid A 1 L three-necked round-bottom flask purged with nitrogen and maintained under an inert nitrogen atmosphere was charged with 4-chloro-1H-indole (10 g, 1.00 equiv.), l-threonine (14.09 g, 1.8 equiv.), DMSO (100 mL), and potassium phosphate buffer (0.2 M, 300 mL, pH = 7.4). 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 equiv.) 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.

[0226] To the above reaction mixture, THF (100 mL), sodium carbonate (20.9 g, 3.0 equiv.), and 2,5-dioxopyrrolidin-1-yl 9H-fluoren-9-ylmethyl carbonate (31.0 g, 1.40 equiv.) were added at 0° C. The resulting solution was stirred at room temperature overnight. The pH was adjusted to 4 with 3 M HCl, and the solid precipitate was removed by filtration. The resulting solution was extracted with EtOAc (3×500 mL). The organic fractions were combined, washed with brine (1 L), dried over anhydrous sodium sulfate, and concentrated under high vacuum. HPLC-MS: (ES, m / z): [M+1]: 475 Synthetic Scheme 25 [ka]

[0227] Precursor to sbMe1Nal [ka]

[0228] (2S,3S)-2-amino-3-(naphthalen-1-yl)butanoic acid 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. Cesium carbonate (477 g, 2.00 equiv.) was then added, and the solution was stirred at 20 °C for 3 h. The reaction was poured into ice HO (3 L) and extracted with EtOAc (500 mL × 3). The organic layer was washed with 3% LiCl solution (500 mL × 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. 1 H NMR(400 MHz, CDCl3): δ 7.77(d,J=7.6 Hz,1H),7.40(d,J=8.0 Hz,1H),7.31-7.36(m,10H),5.65-5.71 m,1H),5.13-5.31(m,3H),4.39-4.43(m,3H),4.22-4.25(m,1H),1.25(d,J=6.4 Hz,3H) Step 2: To a three-neck round-bottom flask, using an inert atmosphere of nitrogen, was added (2S,3R)-benzyl 2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-hydroxybutanoate (125 g, 1.00 equiv.) and DCE (750 mL). The reaction was cooled to 0 °C, followed by the addition of NIS (195 g, 3.00 equiv.) and PPh (228 g, 3.00 equiv.). The temperature was raised to 50 °C, and the reaction mixture was stirred for 3 h. The reaction was poured into ice HO (500 mL) and extracted with DCM (500 mL × 2). The organic layer 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). 1H NMR(400 MHz, CDCl3): δ 7.78(d,J=7.6 Hz,2H),7.68(d,J=7.2 Hz,2H),7.33-7.43(m,9H),5.27-5.68(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). Step 3: A three-necked round-bottom flask was charged with 2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-iodobutanoate, 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). A second three-necked round-bottom flask was charged with picolinimideamide.2HCl (42.2 g, 1.20 equiv.), NiCl.glyme (7.61 g, 0.25 equiv.), and DMA (750 mL) at 25 °C. 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 HO (3 L) and extracted with EtOAc (1 L × 2). The organic layer was dried over sodium sulfate and concentrated under vacuum at 40° C. The crude product was purified by reverse phase HPLC (MeCN:H2O). HPLC-MS: [M+23]: 564. 1 H NMR(400 MHz, 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). Step 4: 143 g of (2S)-benzyl 2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(naphthalen-1-yl)butanoate was separated by SFC. The organic layer was concentrated under vacuum at 35° C.

[0229] Peak 1: (2S,3R)-benzyl 2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(naphthalen-1-yl)butanoate. 1 H NMR(400MHz,DMSO-d6):δ 8.11-8.12(m,2H),8.10-8.11(m,1H),7.88-7.90(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=8.0 Hz),4.17-4.25(m,4H),1.39(d,J=6.8 Hz, 3H). Peak 2: (2S,3S)-benzyl 2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(naphthalen-1-yl)butanoate. 1 H NMR(400MHz,DMSO-d6):δ 7.92-8.15(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=8.0 Hz),4.23-4.26(m,1H),4.05-4.18(m,3H),1.30(d,J=6.8 Hz,3H). Step 5: To a three-necked round-bottom flask was added (2S,3S)-benzyl 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 H three times and stirred under H (15 psi) at 25 °C for 12 h. The reaction was filtered through a pad of Celite and concentrated under vacuum at 35 °C. The crude product was triturated 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. 1H NMR(400 MHz,DMSO-d6)δ 12.78(s,1H),8.23(d,J=7.6 Hz,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=6.8 Hz,3H). Synthetic Scheme 26 [ka]

[0230] Precursors to TyrEtNAc [ka]

[0231] l-Tyrosine O-ethylacetamide or (S)-3-(4-(2-acetamidoethoxy)phenyl)-2-aminopropanoic acid Step 1: To a stirred solution of methyl (tert-butoxycarbonyl)-l-tyrosinate (10.0 g, 33.9 mmol), benzyl (2-bromoethyl)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 × 500 mL). The combined organic layers were washed with brine (3 × 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. 25 H 32 N2O7[M+Na] + MS ESI calculated for 495.22, found 495.10; 1H NMR(300 MHz,CDCl3)δ 7.38-7.32(m,5H),7.04(d,J=8.4 Hz,2H),6.81(d,J=8.4 Hz,2H),5.31(br,1H),5.21(s,2H),4.97(br,1H),4.56-4.53(m,1H),4.03(t,J=5.0 Hz,2H),3.72(s,3H),3.64-3.58(m,2H),3.06-3.01(m,2H),1.43(s,9H). Step 2: To a stirred solution of methyl (S)-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), Pd / C (3.60 g, 33.9 mmol, dry, 10% wt) was added at room temperature under a nitrogen atmosphere. The mixture was degassed with hydrogen three times and stirred at 20°C for 4 hours. DIPEA (17.74 mL, 102 mmol) was added to the mixture and stirred at 20°C for 1 hour. 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] + MS ESI calculated for 403.19, found 403.10; 1 H NMR(300 MHz,CDCl3)δ 7.05(d,J=8.4 Hz,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=5.0 Hz,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). Step 3: To a stirred solution of methyl (S)-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, 1N in water) at room temperature. The solution was stirred at 20° C. for 2 hours. The pH of the solution was adjusted to 3 with 1N HCl. The aqueous layer was extracted with EtOAc (2×250 mL). The combined organic layers were washed with brine (150 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure.

[0232] MS ESI [M+H] + :367.10. Step 4: To a stirred solution of (S)-3-(4-(2-acetamidoethoxy)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 hour. The solvent was concentrated under reduced pressure. MS ESI [M+H] + :267.05. Step 5: To a stirred mixture of (S)-3-(4-(2-acetamidoethoxy)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), Fmoc-OSu (7.62 g, 22.59 mmol) was added at room temperature. The mixture was stirred at 20 °C for 1 h. The pH value of the solution was adjusted to 3 with 1 N HCl. The aqueous layer was extracted with EtOAc (2 × 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) δ 7.79(d,J=7.6 Hz,2H),7.62-7.57(m,2H),7.42-7.26(m,4H),7.17-7.14(m,2H),6.83(d,J=8.4 Hz,2H),4.41-4.31(m,2H),4.29-4.10(m,2H),3.96(t,J=4.8 Hz,2H),3.51(t,J=5.4 Hz,2H),3.19-3.13(m,1H),2.92-2.84(m,1H),1.94(s,3H). Synthetic Scheme 27 [ka]

[0233] Precursor to Phe2Cl4COOH [ka]

[0234] (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-(tert-butoxycarbonyl)-2-chlorophenyl)propanoic acid Step 1: To a stirred mixture of 4-bromo-3-chlorobenzoic acid (25 g, 106 mmol) in THF (100 mL) was added BocO (27.8 g, 127 mmol) and DMAP (1.297 g, 10.62 mmol) at 25 °C under an argon-nitrogen atmosphere. The resulting mixture was stirred at 25 °C for 16 h. The reaction progress was monitored by LCMS. Upon completion, the reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluting with 0-10% EA in PE). The product-containing fractions were collected and rotary evaporated in vacuo to afford tert-butyl 4-bromo-3-chlorobenzoate (25 g, 86 mmol, 81% yield) as a colorless oil. 1 H NMR (400 MHz, chloroform-d) δ 8.04-8.03 (m, 1H), 7.73-7.65 (m, 2H), 1.59 (s, 9H). Step 2: To a stirred solution of nickel(II) chloride ethylene glycyl dimethyl ether complex (3.01 g, 13.72 mmol) in DMA (10 mL) was added 1,10-phenanthroline (2.97 g, 13.72 mmol) at room temperature under argon. The resulting solution was stirred at 50°C for 30 min. tert-Butyl 4-bromo-3-chlorobenzoate (20 g, 68.6 mmol), methyl (R)-2-((tert-butoxycarbonyl)amino)-3-iodopropanoate (22.58 g, 68.6 mmol), and Zn (8.97 g, 137 mmol) were added to the solution, and the resulting mixture was stirred at 30°C for 2 h. Upon completion, the reaction was quenched with HO (500 mL) and extracted with EA (2 x 500 mL). The combined organic layers were washed with brine (3 × 200 mL), dried over anhydrous NaSO, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography eluting with 0–60% EA in PE to give tert-butyl (S)-4-(2-((tert-butoxycarbonyl)amino)-3-methoxy-3-oxopropyl)-3-chlorobenzoate (10.1 g, 24.40 mmol, 35.6% yield) as an off-white solid. 20 H 27 ClNO6[MH] - MS ESI calculated for 412.16, found 412.10.

[0235] Step 3: To a stirred solution of tert-butyl (S)-4-(2-((tert-butoxycarbonyl)amino)-3-methoxy-3-oxopropyl)-3-chlorobenzoate (10.1 g, 24.40 mmol) in THF (70 mL) was added LiOH (1 M in water) (70 mL, 24.40 mmol) under argon at 25° C. The solution was stirred at 25° C. for 1 h. Upon completion, the solvent was concentrated under reduced pressure to give (S)-3-(4-(tert-butoxycarbonyl)-2-chlorophenyl)-2-((tert-butoxycarbonyl)amino)propanoic acid (8.5 g, 21.26 mmol, 87% yield) as a pale yellow solid. The crude product was used directly in the next step without further purification. 21 H 29 ClNO6Na[M+ACN+Na]+ MS ESI calculated for 463.14, found 463.20.

[0236] Step 4: To a stirred solution of (S)-3-(4-(tert-butoxycarbonyl)-2-chlorophenyl)-2-((tert-butoxycarbonyl)amino)propanoic acid (8.5 g, 21.26 mmol) in THF (70 mL) and HCl (4 M in dioxane) (70 mL, 852 mmol) at 25° C. The solution was stirred at 25° C. for 2 hours, and then the solvent was concentrated under reduced pressure to give (S)-2-amino-3-(4-(tert-butoxycarbonyl)-2-chlorophenyl)propanoic acid (5.5 g, 18.35 mmol, 86% yield) as a yellow solid. The crude product was used directly in the next step without further purification. 14 H 19 ClNO4[M+H] + MS ESI calculated for 300.09, found 300.15.

[0237] Step 5: To a stirred mixture of (S)-2-amino-3-(4-(tert-butoxycarbonyl)-2-chlorophenyl)propanoic acid (5.5 g, 18.35 mmol) and NaHCO (7.71 g, 92 mmol) in THF (40 mL) and water (40 mL), Fmoc-Su (5.57 g, 16.51 mmol) was added at room temperature. The mixture was stirred at 25 °C for 2 h. After completion, the pH value of the solution was adjusted to 3 with 1 N HCl. The aqueous layer was extracted with EA (2 × 500 mL). The combined organic layers were washed with brine (150 mL), dried over anhydrous NaSO, and filtered. The filtrate was concentrated under reduced pressure, 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: ACN; (Gradient: 5 min hold at 5% B, hold to 60% B within 25 min, hold at 60% B for 15 min; up to 95% B within 10 min, hold at 95% B for 10 min); Flow rate: 90 mL / min; Detector: UV 210 nm; RT = 30 min. Product-containing fractions were collected and rotary evaporated in vacuo to give (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-(tert-butoxycarbonyl)-2-chlorophenyl)propanoic acid (6.2303 g, 11.94 mmol, 65.0% yield) as an off-white solid. 29 H 28 ClNO6Na[M+Na] + MS ESI calculated for 544.16, found 544.20. 1 H NMR (300 MHz, methanol-d₄) δ 7.89 (s, 1H), 7.78-7.70 (m, 3H), 7.58-7.23 (m, 7H), 4.62-4.57 (m, 1H), 4.32-4.26 (m, 1H), 4.16-4.03 (m, 2H), 3.54-3.48 (m, 1H), 3.09-3.01 (m, 1H), 1.52 (s, 9H). Synthetic Scheme 28 [ka]

[0238] Precursor to OrnMe3 [ka]

[0239] (S)-4-((((9H-Fluoren-9-yl)methoxy)carbonyl)amino)-4-carboxy-N,N,N-trimethylbutan-1-aminium 2,2,2-trifluoroacetate Step 1: To a mixture of (S)-5-amino-2-((tert-butoxycarbonyl)amino)pentanoic acid (4.6 g, 19.80 mmol) in MeOH (50 mL) was added MeI (33.7 g, 238 mmol) and KHCO (1.983 g, 19.80 mmol) at ambient temperature. The resulting mixture was warmed to 50 °C and stirred for 12 hours. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo, and the residue was dissolved in DCM (80 mL) and then filtered again. The filtrate was concentrated in vacuo to give (S)-4-((tert-butoxycarbonyl)amino)-5-methoxy-N,N,N-trimethyl-5-oxopentan-1-aminium iodide (7.4 g, 16.00 mmol, 81% yield) as an off-pink solid. 14 H 29 N2O4[MI] + MS ESI calculated for 289.21, found 289.35.

[0240] Step 2: To a mixture of (S)-4-((tert-butoxycarbonyl)amino)-5-methoxy-N,N,N-trimethyl-5-oxopentan-1-aminium iodide (7.4 g, 16.00 mmol) in MeOH (48 mL) and THF (24 mL) was added LiOH (48.0 mL, 48.0 mmol, 1 M in water) at ambient temperature. The reaction was stirred at ambient temperature for 2 hours and then concentrated in vacuo. 48 mL of 1N HCl was then added. The solvent was concentrated in vacuo to give (S)-4-((tert-butoxycarbonyl)amino)-4-carboxy-N,N,N-trimethylbutan-1-aminium chloride (7.5 g, 14.48 mmol, 90% yield) as a white solid. C 13 H 27 N2O4[M-Cl]+ MS ESI calculated for 275.20, found 275.20.

[0241] Step 3: To a mixture of (S)-4-((tert-butoxycarbonyl)amino)-4-carboxy-N,N,N-trimethylbutan-1-aminium chloride (7.5 g, 14.48 mmol) in DCM (50 mL) was added TFA (25 mL, 324 mmol) at ambient temperature. The reaction was stirred at ambient temperature for 1 hour and then concentrated in vacuo to afford (S)-4-amino-4-carboxy-N,N,N-trimethylbutan-1-aminium 2,2,2-trifluoroacetate (10 g, 13.88 mmol, 96% yield) as an orange oil. CH 19 N2O2[M-CF3COO] + MS ESI calculated for 175.14, found 175.20.

[0242] Step 4: To a mixture of (S)-4-amino-4-carboxy-N,N,N-trimethylbutan-1-aminium 2,2,2-trifluoroacetate (10 g, 13.88 mmol) in THF (30 mL) and water (30 mL), NaHCO3 (9.33 g, 111 mmol) and Fmoc-OSu (4.21 g, 12.49 mmol) were added at ambient temperature. The reaction was stirred at ambient temperature for 2 h. The resulting solution was acidified to pH 3-4 with aqueous HCl and then filtered. The filtrate was purified by RP-flash chromatography under the following conditions: Column: Flash C 18 (330 g); Mobile phase A: water (0.05% TFA), Mobile phase B: ACN; (Gradient: 5 min hold at 0% B, hold to 33% B within 18 min, hold at 33% B for 7 min; hold to 95% B within 5 min, hold at 95% B for 5 min); Flow rate: 90 mL / min; Detector: UV 210 nm; RT = 40 min. Product-containing fractions were collected and lyophilized to give (S)-4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-carboxy-N,N,N-trimethylbutan-1-aminium 2,2,2-trifluoroacetate (6.2043 g, 12.15 mmol, 88% yield) as an off-white solid. C 23 H 29N2O4[M-CF3COO] + MS ESI calculated for 397.21, found 397.15. 1 H NMR(400 MHz,DMSO-d6)δ 12.85(br,1H),7.92-7.89(m,2H),7.74-7.63(m,3H),7.44-7.40(m,2H),7.33-7.31(m,2H) ,4.38-4.22(m,3H),4.02-3.98(m,1H),3.34-3.20(m,2H),3.09(s,9H),1.79-1.51(m,4H). 19 F-NMR (376 MHz, DMSO-d6) -73.64. Synthetic Scheme 29 [ka]

[0243] Precursor to DabMe3 [ka]

[0244] (S)-3-((((9H-Fluoren-9-yl)methoxy)carbonyl)amino)-3-carboxy-N,N,N-trimethylpropan-1-aminium 2,2,2-trifluoroacetate Step 1: To a mixture of (S)-4-amino-2-((tert-butoxycarbonyl)amino)butanoic acid (5.46 g, 25 mmol) in MeOH (20 mL) was added CHI (7.82 mL, 125 mmol) and KHCO (12.51 g, 125 mmol) at ambient temperature. The reaction was warmed to 35 °C for 12 h. The resulting mixture was filtered. The filtrate was concentrated in vacuo, and the residue was dissolved in ethanol and then filtered again. The filtrate was concentrated in vacuo to give (S)-3-((tert-butoxycarbonyl)amino)-3-carboxy-N,N,N-trimethylpropan-1-aminium iodide (13.3 g, 22.27 mmol, 89% yield) as an off-white solid. 12 H 25 IN2O4[MI] +MS ESI calculated for 261.18, found 261.25.

[0245] Step 2: To a mixture of (S)-3-((tert-butoxycarbonyl)amino)-3-carboxy-N,N,N-trimethylpropan-1-aminium iodide (13.3 g, 20.55 mmol) in DCM (60 mL) was added TFA (30 mL, 389 mmol) at ambient temperature. The resulting mixture was stirred at ambient temperature for 1 hour and then concentrated in vacuo to afford (S)-3-amino-3-carboxy-N,N,N-trimethylpropan-1-aminium 2,2,2-trifluoroacetate (10.5 g, 19.14 mmol, 93% yield) as an orange oil. CH 17 F3N2O4[M-CF3COO] + MS ESI calculated for 161.13, found 161.25.

[0246] Step 3: To a mixture of (S)-3-amino-3-carboxy-N,N,N-trimethylpropan-1-aminium 2,2,2-trifluoroacetate (10.5 g, 19.14 mmol) in THF (40 mL) and water (40 mL) was added NaHCO (8.04 g, 96 mmol) and Fmoc-OSu (5.81 g, 17.23 mmol). The resulting mixture was stirred at ambient temperature for 2 h, then acidified to pH 3-4 with aqueous HCl, and then filtered. The filtrate was purified by RP-flash chromatography under the following conditions: Column: Flash C 18 (330 g); Mobile phase A: water (0.05% TFA), Mobile phase B: ACN; (Gradient: 5 min hold at 0% B, hold to 33% B within 19 min, hold at 38% B for 7 min; hold to 95% B within 5 min, hold at 95% B for 5 min); Flow rate: 80 mL / min; Detector: UV 210 nm; RT = 41 min. Product-containing fractions were collected and lyophilized to give (S)-3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-carboxy-N,N,N-trimethylpropan-1-aminium 2,2,2-trifluoroacetate (5.2253 g, 10.52 mmol, 55% yield) as a pale yellow solid. C 24 H 27F3N2O6[M-CF3COO] + MS ESI calculated for 383.20, found 383.30. 1 H NMR(300 MHz,DMSO-d6)δ 7.91(d,J=7.5 Hz,2H),7.81-7.78(m,1H),7.74-7.71(m,2H),7.46-7.41(m,2H),7.37-7.32(m,2H),4.42-4.22( m,3H),4.07-4.00(m,1H),3.49-3.41(m,1H),3.34-3.28(m,1H),3.07(s,9H),2.22-2.04(m,2H). 19 F-NMR(282 MHz,DMSO-d6)δ 73.849. Synthetic Scheme 30 [ka]

[0247] Precursor to dProc4CH21Naph4F [ka]

[0248] (2R,4R)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-4-((4-fluoronaphthalen-1-yl)methyl)pyrrolidine-2-carboxylic acid Step 1: To a stirred solution of 1-(bromomethyl)-4-fluoronaphthalene (30 g, 125 mmol) in toluene (300 mL) was added PPh3 (36.2 g, 138 mmol) under a nitrogen atmosphere at 25 °C. The resulting solution was stirred at 110 °C for 16 h. The mixture was cooled to room temperature, and the solid was collected by filtration to give bromo((4-fluoronaphthalen-1-yl)methyl)triphenyl-15-phosphane (60 g, 120 mmol, 95% yield) as an off-white solid. 29 H 23 FP[M-Br+H] + MS ESI calculated for 421.14, found 421.20.

[0249] Step 2: To a stirred solution of bromo((4-fluoronaphthalen-1-yl)methyl)triphenyl-15-phosphane (46.4 g, 92 mmol) in DCM (1160 mL) was added K2CO3 (7.67 g, 55.5 mmol) under a nitrogen atmosphere at 25 °C. The resulting mixture was stirred at 25 °C for 50 min. 18-Crown-6 (0.978 g, 3.70 mmol) and 1-(tert-butyl) 2-methyl (R)-4-oxopyrrolidine-1,2-dicarboxylate (9 g, 37.0 mmol) were added to the mixture, and the resulting mixture was stirred at 45 °C for 16 h. The reaction was cooled to room temperature, quenched with HO (500 mL), and extracted with EA (2 × 250 mL). The combined organic layers were washed with brine (3 × 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% EA in PE to give 1-(tert-butyl) 2-methyl(R,Z)-4-((4-fluoronaphthalen-1-yl)methylene)pyrrolidine-1,2-dicarboxylate (8 g, 20.76 mmol, 56% yield) as an off-white solid. 22 H 25 FNO4[M-Boc+H] + MS ESI calculated for 286.17, found 286.10.

[0250] Step 3: To a stirred solution of 1-(tert-butyl) 2-methyl (R,Z)-4-((4-fluoronaphthalen-1-yl)methylene)pyrrolidine-1,2-dicarboxylate (8 g, 20.76 mmol) in MeOH (160 mL) was added Pd—C (2.21 g, 2.07 mmol, dry, 10% wt) at room temperature under a nitrogen atmosphere. The mixture was degassed with hydrogen three times and stirred at room temperature for 2 hours. The solid was filtered off, and the filtrate was concentrated under reduced pressure to give 1-(tert-butyl) 2-methyl (2R,4R)-4-((4-fluoronaphthalen-1-yl)methyl)pyrrolidine-1,2-dicarboxylate (7.9 g, 20.39 mmol, 98% yield) as a pale yellow oil. 22 H 27 FNO4[M-Boc+H] +MS ESI calculated for 288.18, found 288.20.

[0251] Step 4: To a stirred solution of 1-(tert-butyl) 2-methyl(2R)-4-((4-fluoronaphthalen-1-yl)methyl)pyrrolidine-1,2-dicarboxylate (8.8 g, 22.71 mmol) in DCM (80 mL) was added TFA (80 mL) at room temperature. The solution was stirred at 25° C. for 1 hour. The solvent was concentrated under reduced pressure to give the crude product. The crude product was separated by Prep-SFC using the following conditions: Column: Chiral Art Amylose-C NEO, 7 x 25 cm, 10 μm; Mobile phase A: CO2, Mobile phase B: MeOH (0.1% 2M NH3-MeOH); Flow rate: 220 mL / min; Gradient: Isocratic 30% B; Column temperature (°C): 35; Back pressure (bar): 100; Wavelength: 220 nm; RT1 (min): 8.23; RT2 (min): 9.4; Sample solvent: MeOH; Injection volume: 2 mL; Number of runs: 10. The fraction at 9.4 min was collected and concentrated under reduced pressure to give methyl (2R)-4-((4-fluoronaphthalen-1-yl)methyl)pyrrolidine-2-carboxylate (3.0 g, 10.44 mmol, 46% yield) as a pale yellow solid. 17 H 19 FNO2[M+H] + MS ESI calculated for 288.13, found 288.20.

[0252] Step 5: To a stirred solution of methyl (2R,4R)-4-((4-fluoronaphthalen-1-yl)methyl)pyrrolidine-2-carboxylate (3 g, 10.44 mmol) in THF (30 mL) was added LiOH (20.88 mL, 20.88 mmol, 1N in water) at room temperature. The solution was stirred at 25° C. for 2 hours. The pH of the solution was adjusted to 7 with 1N HCl. The solution was concentrated under reduced pressure to give (2R,4R)-4-((4-fluoronaphthalen-1-yl)methyl)pyrrolidine-2-carboxylic acid (2.85 g, 9.39 mmol, 90% yield) as a yellow oil. 16 H 17 FNO2[M+H] +MS ESI calculated for 274.12, found 274.10.

[0253] Step 6: To a stirred solution of (2R,4R)-4-((4-fluoronaphthalen-1-yl)methyl)pyrrolidine-2-carboxylic acid (2.85 g, 9.39 mmol) and NaHCO3 (2.365 g, 28.2 mmol) in THF (30 mL) and water (30 mL) was added Fmoc-OSu (3.17 g, 9.39 mmol) at room temperature. The mixture was stirred at 25 °C for 16 h. The pH of the mixture was adjusted to 3 with 1 N HCl. The solution was extracted with EA (3 × 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, and the residue was purified under the following conditions: 330 g C 18 Purification by column, 5% to 5% in 5 min, 5% to 77% in 30 min, 98% to 98% in 5 min, MeCN in water (0.05 TFA), RP-flash at RT = 35 min gave (2R,4R)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-4-((4-fluoronaphthalen-1-yl)methyl)pyrrolidine-2-carboxylic acid (2.9229 g, 5.84 mmol, 62.2% yield) as an off-white solid. 31 H 27 FNO4[M+H] + MS ESI calculated for 496.18, found 496.05. 1 H NMR(300 MHz,CD3OD)δ 8.11-8.09(m,2H),7.77-7.45(m,5H),7.37-7.10(m,7H),4.52-4.11(m,4H),3.31-3.00(m,4H),2.44-2.42(m,2H),1.88-1.85(m,1H). Synthetic Scheme 31 [ka]

[0254] Precursor to AlaPyrim4CONH2 [ka]

[0255] (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(2-carbamoylpyrimidin-5-yl)propanoic acid Step 1: To a stirred solution of 5-bromopyrimidine-2-carboxylic acid (4 g, 19.70 mmol) in DCM (80 mL) was added oxalyl chloride (5.00 g, 39.4 mmol) and DMF (0.153 mL, 1.970 mmol) at 0° C. The mixture was stirred at 25° C. for 3 h. Aqueous ammonia (3.45 g, 99 mmol, 28%) was then added to the solution and stirred at 25° C. for 1 h. The reaction was concentrated under reduced pressure and the residue was purified by RP-flash chromatography using the following conditions: Column: C 18 Gel column (330 g); Mobile phase A: Water (0.05% TFA); Mobile phase B: MeCN; (Gradient: 5 min hold at 0% B, 21% B within 15 min, 10 min hold at 21% B; up to 95% B within 20 min, 10 min hold at 95% B); Flow rate: 60 mL / min; Detector: UV 254 and 210 nm; Room temperature: 20 min afforded 5-bromopyrimidine-2-carboxamide (2.2 g, 10.89 mmol, 55.3% yield) as a white solid. CHBrNO [M+H] + MS ESI calculated values: 201.95, 203.95, found values: 202.00, 204.00. 1 H NMR(300 MHz,DMSO-d6)δ 9.13(s,2H),8.21(s,1H),7.86(s,1H). Step 2: A mixture of nickel(II) chloride ethylene glycol dimethyl ether complex (0.457 g, 2.079 mmol) and 1,10-phenanthroline (0.375 g, 2.079 mmol) in DMA (20 mL) was heated at 50° C. for 0.5 h. A solution of 5-bromopyrimidine-2-carboxamide (2.1 g, 10.40 mmol), tert-butyl (R)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-iodopropanoate (5.13 g, 10.40 mmol), and TBAI (3.84 g, 10.40 mmol) in DMA (20 mL) was added at 25° C. Zinc (1.359 g, 20.79 mmol) was then added and stirred at room temperature for 1 h. The reaction was quenched with water (300 mL) at room temperature. The mixture was extracted with EA (3 x 300 mL). The organic layer was washed with brine (200 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by column chromatography under the following conditions: 18 Gel column (330 g); Mobile phase A: water (0.05% TFA); Mobile phase B: MeCN; (Gradient: 5 min hold at 0% B, 41% B within 20 min, 10 min hold at 41% B; 95% B within 10 min, 5 min hold at 95% B); Flow rate: 60 mL / min; Detector: UV 254 and 210 nm; RT: 25 min. Purification by RP flash gave tert-butyl (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(2-carbamoylpyrimidin-5-yl)propanoate (1.6 g, 3.28 mmol, 31.5% yield) as a yellow solid. 27 H 29 N4O5[M+H] + MS ESI calculated for 489.21, found 489.15.

[0256] Step 3: To a stirred solution of tert-butyl (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(2-carbamoylpyrimidin-5-yl)propanoate (2.1 g, 4.30 mmol) in DCM (20 mL) was added TFA (20 mL, 260 mmol) at room temperature. The solution was stirred at 25° C. for 4 hours. The solvent was concentrated under reduced pressure and the residue was purified using the following conditions: Column: C 18 Purification by RP-flash using a gel column (330 g); mobile phase A: water (0.05% TFA); mobile phase B: MeCN; (gradient: 5 min hold at 0% B, hold to 23% B within 15 min, 10 min hold at 23% B; up to 95% B within 15 min, 10 min hold at 95% B); flow rate: 60 mL / min; detector: UV 254 and 210 nm; RT: 30 min afforded (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(2-carbamoylpyrimidin-5-yl)propanoic acid (1.13 g, 2.61 mmol, 60.8% yield) as an off-white solid. 23 H 21 N4O5[M+H] + MS ESI calculated for 433.14, found 433.00. 1 H NMR(400 MHz,CD3OD)δ 8.83(s,2H),7.80(d,J=7.6 Hz,2H),7.62-7.59(m,2H),7.41-7.25(m,4H),4.57-4.53(m,1H),4.40-4.08(m,3H),3.41-3.34(m,1H),3.13-3.07(m,1H). Synthetic Scheme 32 [ka]

[0257] Precursor to AlaPyrim4COOH [ka]

[0258] (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(2-(tert-butoxycarbonyl)pyrimidin-5-yl)propanoic acid Step 1: To a solution of (((9H-fluoren-9-yl)methoxy)carbonyl)-L-serine (10 g, 30.5 mmol) in DMF (150 mL) was added NaHCO (12.83 g, 153 mmol), 3-bromoprop-1-ene (11.09 g, 92 mmol) at 0 °C under Ar atmosphere. The resulting mixture was stirred at room temperature for 12 h. The mixture was diluted with water (500 mL) and extracted with EA (1000 mL). The combined organic layers were washed with brine (3 × 500 mL), dried over anhydrous NaSO and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography, eluting with 0-50% EA in PE to give allyl (((9H-fluoren-9-yl)methoxy)carbonyl)-L-serinate (9.6 g, 26.1 mmol, 85% yield) as a colorless oil. 21 H 21 NO5Na[M+Na] + MS ESI calculated for 390.14, found 390.20. 1 H NMR(300 MHz,CDCl3)δ 7.80-7.21(m,8H),5.99-5.67(m,2H),5.40-5.19(m,2H),4.69-4.67(m,2 H),4.43-4.41(m,3H),4.22-4.20(m,1H),4.04-3.79(m,2H),2.17(s,1H). Step 2: To a mixture of allyl (((9H-fluoren-9-yl)methoxy)carbonyl)-L-serinate (9.6 g, 26.1 mmol) in DCM (960 mL), imidazole (3.56 g, 52.3 mmol), PhP (11.65 g, 44.4 mmol), and iodine (9.95 g, 39.2 mmol) were added successively at room temperature. The reaction was stirred at room temperature for 4 hours. The resulting solution was quenched with saturated NaSO (300 mL) and extracted with DCM (500 mL). The organic layers were combined, washed with brine (2 × 400 mL), dried over anhydrous NaSO, and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography eluting with a gradient of 0% to 25% EA in PE to give allyl (R)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-iodopropanoate (6.3 g, 13.15 mmol, 50.3% yield) as a white solid. 21 H 20 INO4Na[M+Na] + MS ESI calculated for 500.04, found 500.15. 1 H NMR(400 MHz,CDCl3)δ 7.78-7.76(m,2H),7.63-7.61(m,2H),7.41-7.39(m,2H),7.33-7.31(m,2H),5.94-5.92(m,1H),5.68-5.66(m,1H), 5.49-5.22(m,2H),4.74-4.73(m,2H),4.62-4.60(m,1H),4.41-4.40(m,2H),4.25-4.24(m,1H),3.72-3.52(m,2H). Step 3: To a mixture of 5-bromopyrimidine-2-carboxylic acid (5 g, 24.63 mmol) in t-BuOH (75 mL) was added DMAP (0.301 g, 2.463 mmol) and BocO (6.45 g, 29.6 mmol) at room temperature under argon. The reaction was stirred at 50 °C for 16 h. The reaction was cooled to room temperature and concentrated in vacuo. The residue was purified by silica gel column chromatography eluting with 0-20% EA in PE to give tert-butyl 5-bromopyrimidine-2-carboxylate (3.55 g, 13.58 mmol, 55.1% yield) as a white solid. CH 11 BrN2O2[M-tBu+H] + MS ESI calculated values ​​for 203.00, 205.00, found values ​​202.95, 204.95.

[0259] Step 4: A mixture of nickel(II) chloride ethylene glycol dimethyl ether complex (63.6 mg, 0.289 mmol) and pyridine-2-carboximidamide hydrochloride (91 mg, 0.579 mmol) in DMA (20 mL) was heated at 50° C. for 1 hour. A solution of allyl (R)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-iodopropanoate (1.38 g, 2.89 mmol), tert-butyl 5-bromopyrimidine-2-carboxylate (750 mg, 2.89 mmol), and TBAI (2.14 g, 5.79 mmol) in DMA (25 mL) was added at 25° C. Then, zinc (378 mg, 5.79 mmol) was added and stirred at 25° C. for 2 hours. The reaction was quenched with HO (200 mL) and extracted with EA (2 × 400 mL). The combined organic layers were washed with brine (3 × 200 mL), dried over anhydrous NaSO, and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with 0% to 45% EA in PE to afford tert-butyl (S)-5-(2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(allyloxy)-3-oxopropyl)pyrimidine-2-carboxylate (570 mg, 1.022 mmol, 35.3% yield) as a colorless semi-solid.30 H 32 N3O6[M+H] + MS ESI calculated for 530.22, found 530.25.

[0260] Step 5: To a stirred solution of tert-butyl (S)-5-(2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(allyloxy)-3-oxopropyl)pyrimidine-2-carboxylate (1.44 g, 2.72 mmol) in THF (25 mL) was added Pd(PPh3)4 (0.157 g, 0.136 mmol) and phenylsilane (0.588 g, 5.44 mmol) at room temperature. The resulting mixture was stirred at 25 °C for 1 h. The solvent was concentrated under reduced pressure, and the residue was purified under the following conditions: 330 g of C 18 Purification by column, 2% to 2% in 5 min, 2% to 50% in 30 min, and RP-flash with MeCN (0.05% TFA) in water gave (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(2-(tert-butoxycarbonyl)pyrimidin-5-yl)propanoic acid (1.01 g, 2.051 mmol, 75% yield) as a white solid. 27 H 28 N3O6[M+H] + MS ESI calculated for 490.19, found 490.15. 1 H NMR(300 MHz,DMSO-d6)δ 8.86(s,2H),7.86-7.85(m,3H),7.69-7.52(m,2H),7.46-7.24(m,4H),4.33-4.3 2(m,1H),4.20-4.18(m,3H),3.23-3.21(m,1H),2.98-2.97(m,1H),1.55(s,9H). Synthetic Scheme 33 [ka]

[0261] Precursor to dProc3Bn4F [ka]

[0262] (2R,3S)-1-(((9H-Fluoren-9-yl)methoxy)carbonyl)-3-(4-fluorobenzyl)pyrrolidine-2-carboxylic acid Step 1: To a solution of ethyl (R)-N-(but-3-en-1-yl)-N-(1-phenylethyl)glycinate (5 g, 19.13 mmol) in THF (50 mL) was added LDA (9.57 mL, 19.13 mmol, 2N in THF) at −78° C. The resulting solution was stirred at −78° C. for 1 hour. A solution of zinc(II) bromide (12.92 g, 57.4 mmol) in THF (50 mL) was added at −78° C. The resulting mixture was stirred at room temperature for 1 hour. Tris(dibenzylideneacetone)dipalladium(0) (0.526 g, 0.574 mmol), tri-o-tolylphosphane (0.757 g, 2.487 mmol), and 1-fluoro-4-iodobenzene (5.52 g, 24.87 mmol) were added at room temperature. The resulting mixture was stirred at room temperature for 16 hours. The reaction was quenched with aqueous NH4Cl (2M, 50 mL) and extracted with EA (3 x 200 mL). The combined organic layers were washed with brine (3 x 60 mL), dried over Na2SO4, and filtered. The filtrate was concentrated in vacuo to give a yellow oil. The residue was purified by silica gel column chromatography eluting with EA / PE (1 / 10) to give (2R,3S)-3-(4-fluorobenzyl)-1-((R)-1-phenylethyl)pyrrolidine-2-carboxylate (2.5 g, 4.57 mmol, 23.9% yield) as a yellow oil. C 22 H 27 FNO2[M+H] + MS ESI calculated for 356.19, found 356.25.

[0263] Step 2: To a solution of ethyl (2R,3S)-3-(4-fluorobenzyl)-1-((R)-1-phenylethyl)pyrrolidine-2-carboxylate (2 g, 5.63 mmol) in t-BuOH (40 mL) was added Pd / C (0.599 g, 5.63 mmol, dry, 10% wt) at room temperature. The reaction mixture was degassed with nitrogen three times and stirred under hydrogen at room temperature for 3 hours. The mixture was filtered. The filter cake was washed with MeOH (3 x 100 mL). The filtrate was concentrated under reduced pressure. The residue was purified by the following conditions: 330 g C 18 Purification by RP flash using a column, 2% to 2% in 5 min, 2% to 30% in 25 min, 98% to 98% in 5 min, aqueous MeCN (0.05% TFA), RT = 30 min, gave ethyl (2R,3S)-3-(4-fluorobenzyl)pyrrolidine-2-carboxylate (0.6 g, 2.388 mmol, 42.4% yield) as a colorless oil. 14 H 19 FNO2[M+H] + MS ESI calculated for 252.13, found 252.25. 1 H NMR(400 MHz,DMSO-d6)7.28-7.25(m,2H),7.17-7.12(m,2H),4.45(d,J=7.6 Hz,1H),4.25-4.15(m,2H),3.46-3.39(m,1H),3.21-3.18(m,1H),2.84-2.72(m, 2H),2.44-2.38(m,1H),1.94-1.93(m,1H),1.68-1.64(m,1H),1.26-1.22(m,3H). Step 3: To a mixture of ethyl (2R,3S)-3-(4-fluorobenzyl)pyrrolidine-2-carboxylate (2 g, 7.96 mmol) in THF (20 mL), LiOH (23.88 mL, 23.88 mmol, 1N in water) was added at room temperature. The resulting mixture was stirred at room temperature for 18 hours. The pH value of the reaction solution was adjusted to 7 with HCl (1 M). The mixture was used directly in the next step. 12 H 15 FNO2[M+H] + MS ESI calculated for 224.10, found 224.05.

[0264] Step 4: To a mixture of (2R,3S)-3-(4-fluorobenzyl)pyrrolidine-2-carboxylic acid (5 g, 22.40 mmol), NaHCO3 (5.64 g, 67.2 mmol) in THF (40 mL) and water (40 mL), Fmoc-OSu (7.56 g, 22.40 mmol) was added at room temperature. The resulting mixture was stirred at room temperature for 12 hours. The pH value of the reaction solution was adjusted to 5 with HCl (0.1 M). The reaction was extracted with EA (3 x 100 mL). The combined organic layers were washed with brine (3 x 150 mL), dried over Na2SO4, and filtered. The filtrate was concentrated under vacuum to give a pink oil. The residue was purified by the following conditions: 330 g C 18 Purification by RP flash using a column, 2% to 2% in 5 min, 2% to 500% in 30 min, 98% to 98% in 5 min, MeCN (0.05% TFA) in water, RT=35 min gave 10.7 g of crude product. The crude product was separated by SFC using the following conditions: Column: Chiralpak IH, 3 × 25 cm, 5 μm; Mobile phase A: CO , Mobile phase B: MeOH; Flow rate: 120 mL / min; Gradient: Isocratic 30% B; Column temperature (°C): 35; Back pressure (bar): 100; Wavelength: 220 nm; RT1 (min): 3.13; RT2 (min): 3.72; Sample solvent: MeOH:ACN:DCM = 1:1:1 (0.1% DEA); Injection volume: 3 mL; Number of runs: 82. The fraction at 3.13 min was collected and concentrated under reduced pressure to give (2R,3S)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-3-(4-fluorobenzyl)pyrrolidine-2-carboxylic acid (5.1912 g, 11.65 mmol, 52% yield) as an off-white solid. 27 H 25 FNO4[M+H] + MS ESI calculated for 446.17, found 446.10. 1H NMR(400 MHz,DMSO-d6)δ 7.93-7.88(m,2H),7.70-7.64(m,2H),7.44-7.40(m,2H),7.35-7.26(m,4H),7.13-7.08(m,2H),4.42-4.15(m,4H), 3.56-3.53(m,1H),3.29-3.24(m,1H),3.02-2.91(m,1H),2.51-2.50(m,1H),2.34-2.29(m,1H),1.74-1.70(m,2H). Synthetic Scheme 34 [ka]

[0265] Precursor to dProc3Bn4Cl [ka]

[0266] (2R,3S)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-3-(4-chlorobenzyl)pyrrolidine-2-carboxylic acid Step 1: To a solution of (R)-1-(4-methoxyphenyl)ethan-1-amine (10 g, 66.1 mmol) in DMF (170 mL) was added K2CO3 (18.28 g, 132 mmol) at room temperature, and the resulting mixture was stirred at room temperature for 10 minutes. Then, 4-bromobut-1-ene (8.04 g, 59.5 mmol) was added. After stirring at room temperature for 18 hours, the resulting mixture was diluted with water (300 mL) and extracted with EA (3 x 300 mL). The combined organic layers were washed with brine (3 x 150 mL), dried over Na2SO4, and filtered. The filtrate was concentrated in vacuo. The residue was purified by silica gel column chromatography eluting with EA / PE (2 / 3) to give (R)-N-(1-(4-methoxyphenyl)ethyl)but-3-en-1-amine (8.00 g, 39.0 mmol, 58.9% yield) as a colorless oil. 13 H 20 NO[M+H] +MS ESI calculated for 206.15, found 206.20.

[0267] Step 2: To a mixture of (R)-N-(1-(4-methoxyphenyl)ethyl)but-3-en-1-amine (10 g, 48.7 mmol) in THF (40 mL) and DMPU (40 mL) was added ethyl 2-bromoacetate (8.13 g, 48.7 mmol) and K2CO3 (7.40 g, 53.6 mmol) at room temperature. The resulting mixture was stirred at room temperature for 18 h. The reaction was diluted with water (100 mL) and extracted with EA (3 x 100 mL). The combined organic layers were washed with brine (3 x 150 mL), dried over Na2SO4, and filtered. The filtrate was concentrated in vacuo. The residue was purified by silica gel column chromatography eluting with EA / PE (1 / 5) to give ethyl (R)-N-(but-3-en-1-yl)-N-(1-(4-methoxyphenyl)ethyl)glycinate (10 g, 34.3 mmol, 70.5% yield) as a colorless oil. 17 H 26 NO3[M+H] + MS ESI calculated for 292.18, found 292.25. 1 H NMR(400 MHz,CDCl3)δ 7.28(d,J=8.4 Hz,2H),6.84(d,J=8.4 Hz,2H),5.81-5.70(m,1H),5.04-5.03(m,1H),4.99-4.94(m,1H),4.16-4.10(m,2H),4.00-3.97(m,1H),3.80(s,3H),3.40(d,J=17.2 Hz,1H),3.25(d,J=17.2 Hz,1H),2.68-2.64(m,2H),2.22-2.16(m,2H),1.33-1.32(m,3H),1.27-1.23(m,3H). Step 3: To a solution of ethyl (R)-N-(but-3-en-1-yl)-N-(1-(4-methoxyphenyl)ethyl)glycinate (2.6 g, 8.92 mmol) in THF (20 mL) was added LDA (6.69 mL, 13.38 mmol, 2N in THF) at −78° C. The resulting mixture was stirred at −78° C. for 1 hour. A solution of zinc(II) bromide (6.03 g, 26.8 mmol) in THF (20 mL) was added at −78° C. The resulting mixture was stirred at room temperature for 1 hour, and then Pd(dba) (0.245 g, 0.268 mmol), P(o-tolyl) (0.353 g, 1.160 mmol), and 1-chloro-4-iodobenzene (2.77 g, 11.60 mmol) were added at room temperature. The resulting mixture was stirred at room temperature for 16 hours. The reaction was quenched with aqueous NH4Cl (2 M, 50 mL) and extracted with EA (3 x 200 mL). The combined organic layers were washed with brine (3 x 160 mL), dried over Na2SO4, and filtered. The filtrate was concentrated in vacuo. The residue was purified by silica gel column chromatography eluting with EA / PE (1 / 10) to give ethyl (2R,3S)-3-(4-chlorobenzyl)-1-((R)-1-(4-methoxyphenyl)ethyl)pyrrolidine-2-carboxylate (1.20 g, 2.99 mmol, 33.5% yield) as a yellow oil. 23 H 29 ClNO3[M+H] + MS ESI calculated for 402.18, found 402.25.

[0268] Step 4: A solution of ethyl (2R,3S)-3-(4-chlorobenzyl)-1-((R)-1-(4-methoxyphenyl)ethyl)pyrrolidine-2-carboxylate (1 g, 2.488 mmol) in TFA (30 mL) was stirred at 80° C. for 18 hours. The reaction was cooled to room temperature and concentrated in vacuo. The residue was purified using the following conditions: 18Purification by RP flash using MeCN (0.05% TFA) in water, column, 330 g, 2%-2% in 5 min, 2%-30% in 20 min, 98%-98% in 5 min, gave ethyl (2R,3S)-3-(4-chlorobenzyl)pyrrolidine-2-carboxylate (0.3 g, 1.120 mmol, 45% yield) as a yellow oil. 14 H 19 ClNO2[M+H] + MS ESI calculated for 268.10, found 268.15.

[0269] Step 5: To a solution of ethyl (2R,3S)-3-(4-chlorobenzyl)pyrrolidine-2-carboxylate (4.3 g, 16.06 mmol) in THF (50 mL) was added a solution of LiOH (1.154 g, 48.2 mmol) in water (50 mL) at room temperature. The resulting mixture was stirred at room temperature for 18 hours. The pH value of the reaction solution was adjusted to 7.0 with HCl (1 M). The resulting mixture was used directly in the next step. 12 H 15 ClNO2[M+H] + MS ESI calculated for 240.07, found 240.10.

[0270] Step 6: To a stirred solution of (2R,3S)-3-(4-chlorobenzyl)pyrrolidine-2-carboxylic acid (5.5 g, 22.95 mmol) in THF (100 mL) and water (100 mL) was added NaHCO (5.78 g, 68.8 mmol) and Fmoc-OSu (7.74 g, 22.95 mmol) at 25° C. The resulting mixture was stirred at 25° C. for 2 h. The pH of the mixture was adjusted to 3 with 1 N HCl and then extracted with EA (3×200 mL). The organic fraction was washed with brine (2×100 mL), dried over NaSO, and filtered. The filtrate was concentrated under vacuum. The residue was purified by RP flash using the following conditions: 18Column, 330 g, 2% to 2% in 5 min, 2% to 60% in 40 min, 98% to 98% in 5 min, MeCN in water (0.05% TFA). The crude product was separated by SFC using the following conditions: Column: Chiralpak IG 3 x 25 cm, 5 μm; Mobile phase A: CO2, Mobile phase B: MeOH; Flow rate: 110 mL / min; Gradient: Isocratic 50% B; Back pressure (bar): 100; Wavelength: 254 nm; RT1 (min): 10.1; RT2 (min): 16.8; Sample solvent: DCM / MeOH = 1:1 (0.1% AcOH); Injection volume: 5 mL; Number of runs: 30. The 10.1 min fraction was collected and concentrated under reduced pressure to give (2R,3S)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-3-(4-chlorobenzyl)pyrrolidine-2-carboxylic acid (7.8731 g, 16.87 mmol, 73.5% yield) as an off-white solid. 27 H 23 ClNO4[MH] - MS ESI calculated for 460.14, found 460.05. 1 H NMR(400 MHz,DMSO-d6)δ 12.86(s,1H),7.90(d,J=7.5 Hz,2H),7.68-7.65(m,2H),7.44-7.27(m,8H),4.42-4.17(m,4H),3.56-3.30(m,1H),3. 28-3.26(m,1H),2.95-2.88(m,1H),2.51(s,1H),2.35-2.31(m,1H),1.75-1.69(m,2H). Synthetic Scheme 35 [ka]

[0271] Precursor to dProc4Bn4Cl [ka]

[0272] (2R,4R)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-4-(4-chlorobenzyl)pyrrolidine-2-carboxylic acid Step 1: To a mixture of 1-(bromomethyl)-4-chlorobenzene (10 g, 48.7 mmol) in toluene (100 mL) was added triphenylphosphane (14.04 g, 53.5 mmol) at ambient temperature. The resulting mixture was stirred at 110° C. for 6 hours and then cooled to ambient temperature. The solid was collected by filtration to give (4-chlorobenzyl)triphenylphosphonium bromide (18 g, 38.5 mmol, 79% yield) as a white solid. 25 H 21 ClP[M-Br] + MS ESI calculated for 387.11, found 387.15. 1 H NMR(300 MHz,CDCl3)δ 7.82-7.73(m,9H),7.64-7.58(m,6H),7.16-7.12(m,2H),7.04(d,J=8.1 Hz,2H),5.61(d,J=14.7 Hz,2H). Step 2: To a mixture of (4-chlorobenzyl)triphenylphosphonium bromide (40.9 g, 87 mmol) and K2CO3 (12.07 g, 87 mmol) in DCM (400 mL) was added 18-crown-6 (0.462 g, 1.747 mmol) at room temperature. The resulting mixture was stirred at room temperature for 1 hour. 1-(tert-butyl) 2-methyl (R)-4-oxopyrrolidine-1,2-dicarboxylate (8.5 g, 34.9 mmol) was added, and the resulting mixture was stirred at 48 °C for 48 hours. The reaction was cooled to ambient temperature, then diluted with water (200 mL), and extracted with EA (3 × 200 mL). The combined organic layers were washed with brine (3 × 150 mL), dried over Na2SO4, and filtered. The filtrate was concentrated in vacuo. The residue was purified by silica gel column chromatography eluting with EA / PE (1 / 6) to give 1-(tert-butyl) 2-methyl(R,Z)-4-(4-chlorobenzylidene)pyrrolidine-1,2-dicarboxylate (7.0 g, 19.90 mmol, 56.9% yield) as a yellow solid. 18 H 23 ClNO4[M+H] + MS ESI calculated for 352.12, found 352.15. 1H NMR(400 MHz,CDCl3)δ 7.34-7.26(m,2H),7.16-7.09(m,2H),6.42-6.33(m,1H),4.63-4.26(m,3 H),3.71(s,3H),3.19-3.12(m,1H),2.91-2.77(m,1H),1.47-1.43(m,9H). Step 3: To a mixture of 1-(tert-butyl) 2-methyl (R,Z)-4-(4-chlorobenzylidene)pyrrolidine-1,2-dicarboxylate (1 g, 2.84 mmol) in EA (10 mL) and toluene (10 mL) was added PtO (0.065 g, 0.284 mmol) at room temperature. The reaction mixture was degassed with hydrogen three times and stirred under hydrogen at ambient temperature for 16 hours. The reaction was filtered, and the filtrate was concentrated in vacuo. The residue was purified by silica gel column chromatography eluting with EA / PE (1 / 5) to give 1-(tert-butyl) 2-methyl (2R,4R)-4-(4-chlorobenzyl)pyrrolidine-1,2-dicarboxylate (0.7 g, 1.978 mmol, 69.6% yield) as a colorless oil. 18 H 25 ClNO4[M+H] + MS ESI calculated for 354.14, found 354.15. 1 H NMR(400 MHz,CDCl3)δ 7.30-7.26(m,2H),7.11-7.08(m,2H),4.28-4.15(m,1H),3.76-3.60(m,4H),3.17-3.1 2(m,1H),2.75-2.60(m,2H),2.47-2.33(m,2H),2.08-2.05(m,1H),1.48-1.41(m,9H). Step 4: To a solution of 1-(tert-butyl) 2-methyl (2R,4R)-4-(4-chlorobenzyl)pyrrolidine-1,2-dicarboxylate (1 g, 2.83 mmol) in DCM (10 mL) was added TFA (2 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 hours and then concentrated in vacuo to give a colorless oil, which was used directly in the next step. 13 H 17 ClNO2[M+H] +MS ESI calculated for 254.09, found 254.10.

[0273] Step 5: To a mixture of methyl (2R,4R)-4-(4-chlorobenzyl)pyrrolidine-2-carboxylate (1 g, 3.94 mmol) in THF (10 mL) was added a mixture of LiOH (0.283 g, 11.82 mmol) in water (10 mL) at room temperature. The resulting mixture was stirred at room temperature for 18 hours. The pH value of the reaction solution was adjusted to 7.0 with HCl (1 M) and then used directly in the next step. 12 H 15 ClNO2[M+H] + MS ESI calculated for 240.07, found 240.05.

[0274] Step 6: To a mixture of (2R,4R)-4-(4-chlorobenzyl)pyrrolidine-2-carboxylic acid (9 g, 37.5 mmol) in water (140 mL) and THF (140 mL), Fmoc-OSu (12.67 g, 37.5 mmol) and NaHCO (15.77 g, 188 mmol) were added at room temperature. The resulting mixture was stirred at room temperature for 2 hours. The pH value of the reaction solution was adjusted to 3 with HCl (1 M) and extracted with EA (3 × 300 mL). The combined organic layers were washed with brine (3 × 100 mL), dried over NaSO, and filtered. The filtrate was concentrated in vacuo to give a yellow oil. The residue was purified by RP-flash using the following conditions: C18 column, 330 g, 2 to 2% in 5 min, 2% to 50% in 30 min, 98% to 98% in 5 min, MeCN (0.05% TFA) in water to give 14 g of crude product. The crude product was separated by SFC using the following conditions: Column: Chiralpak IG 3 x 25 cm, 5 μm; Mobile phase A: CO2, Mobile phase B: MeOH:MeCN = 1:1; Flow rate: 100 mL / min; Gradient: Isocratic 45% B; Column temperature (°C): 35; Back pressure (bar): 100; RT1 (min): 4; RT2 (min): 5.43; Sample solvent: MeOH; Injection volume: 2 mL; Number of runs: 15. The fraction at 5.43 min was collected and concentrated under reduced pressure to give (2R,4R)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-4-(4-chlorobenzyl)pyrrolidine-2-carboxylic acid (7.6433 g, 16.55 mmol, 44.1% yield) as an off-white solid. 27 H 23 ClNO4[MH] - MS ESI calculated for 460.14, found 460.05. 1 H NMR(300 MHz,DMSO-d6)δ 7.88-7.70(m,2H),7.68-7.57(m,2H),7.43-7.19(m,8H),4.27-4.24(m,2H),4.15-4.01(m,2H), 3.52-3.36(m,1H),3.05-3.00(m,1H),2.68-2.50(m,2H),2.37-2.25(m,2H),1.72-1.42(m,1H). Synthetic Scheme 36 [ka]

[0275] Precursor to dProc3CH24Pal [ka]

[0276] (2R,3S)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-3-(pyridin-4-ylmethyl)pyrrolidine-2-carboxylic acid Step 1: To a solution of ethyl (R)-N-(but-3-en-1-yl)-N-(1-(4-methoxyphenyl)ethyl)glycinate (5 g, 17.16 mmol) in THF (50 mL) was added LDA (8.58 mL, 17.16 mmol, 2N in THF) at −78° C. The resulting mixture was stirred at −78° C. for 1 hour. A solution of zinc(II) bromide (11.59 g, 51.5 mmol) in THF (50 mL) was added at −78° C. The resulting mixture was stirred at room temperature for 1 hour, and then Pd(dba) (0.471 g, 0.515 mmol), P(o-tolyl) (0.679 g, 2.231 mmol), and 4-iodopyridine (4.57 g, 22.31 mmol) were added at room temperature. The resulting mixture was stirred at room temperature for 18 hours, after which it was quenched with aqueous NH4Cl (2M, 150 mL) and extracted with EA (3 x 200 mL). The combined organic layers were washed with brine (3 x 150 mL), dried over Na2SO4, and filtered. The filtrate was concentrated in vacuo. The residue was purified by silica gel column chromatography eluting with MeOH / DCM (1 / 4) to give ethyl (2R,3S)-1-((R)-1-(4-methoxyphenyl)ethyl)-3-(pyridin-4-ylmethyl)pyrrolidine-2-carboxylate (2.0 g, 5.43 mmol, 31.6% yield) as a yellow oil. C 22 H 29 N2O3[M+H] + MS ESI calculated for 369.21, found 369.15.

[0277] Step 2: A mixture of ethyl (2R,3S)-1-((R)-1-(4-methoxyphenyl)ethyl)-3-(pyridin-4-ylmethyl)pyrrolidine-2-carboxylate (4 g, 10.86 mmol) in TFA (80 mL) was stirred at 80° C. for 12 hours. The reaction was cooled to room temperature and concentrated in vacuo. The residue was purified using the following conditions: 18 Purification by RP flash using MeCN (0.05% TFA) in water (column, 330 g, 2% to 2% in 5 min, 2% to 30% in 20 min, 98% to 98% in 5 min) gave ethyl (2R,3S)-3-(pyridin-4-ylmethyl)pyrrolidine-2-carboxylate (1.2 g, 5.12 mmol, 47.2% yield) as a yellow oil. 13 H 19 N2O2[M+H] + MS ESI calculated for 235.14, found 235.10.

[0278] Step 3: To a solution of ethyl (2R,3S)-3-(pyridin-4-ylmethyl)pyrrolidine-2-carboxylate (2 g, 8.54 mmol) in THF (30 mL) was added a solution of LiOH (0.613 g, 25.6 mmol) in water (30 mL) at room temperature. The resulting mixture was stirred at room temperature for 18 hours. The pH of the reaction solution was adjusted to 7.0 with HCl (1 M) and used directly in the next step. 11 H 15 N2O2[M+H] + MS ESI calculated for 207.11, found 207.00.

[0279] Step 4: To a mixture of (2R,3S)-3-(pyridin-4-ylmethyl)pyrrolidine-2-carboxylic acid (6 g, 29.1 mmol) in THF (60 mL) and water (60 mL), Fmoc-OSu (9.81 g, 29.1 mmol) and NaHCO3 (12.22 g, 145 mmol) were added at room temperature. The resulting mixture was stirred at room temperature for 2 hours. The pH value of the reaction solution was adjusted to 5 with HCl (1 M) and then extracted with EA (3 x 200 mL). The combined organic layers were washed with brine (3 x 50 mL), dried over Na2SO4, and filtered. The filtrate was concentrated under vacuum. The residue was purified by RP flash using the following conditions: C 18 Column: 330 g, 2% to 2% in 5 min, 2% to 50% in 40 min, 98% to 98% in 5 min, MeCN (0.05% TFA) in water. The crude product was separated by SFC using the following conditions: Column: Chiral art Cellulose-SZ 3 × 25 cm, 5 μm; Mobile phase A: CO2, Mobile phase B: EtOH; Flow rate: 100 mL / min, Gradient: Isocratic 10% B; Column temperature (°C): 35; Back pressure (bar): 100; Wavelength: 220 nm; RT1 (min): 4.88; RT2 (min): 6.55; Sample solvent: MeOH; Injection volume: 1.8 mL. The 6.55 min fraction was collected and concentrated under reduced pressure to give (2R,3S)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-3-(pyridin-4-ylmethyl)pyrrolidine-2-carboxylic acid (7.5851 g, 17.52 mmol, 60.2% yield) as a yellow solid. 26 H 25 N2O4[M+H] + MS ESI calculated for 429.17, found 429.10. 1 H NMR(400 MHz,CD3OD)δ 8.76-8.73(m,2H),8.00-7.98(m,2H),7.82-7.78(m,2H),7.66-7.61(m,2H),7.43-7.39(m,2H),7.34-7.30(m,2H), 4.44-4.19(m,4H),3.70-3.68(m,1H),3.42-3.34(m,1H),3.33-3.32(m,1H),3.32-3.21(m,2H),2.93-1.94(m,2H). Synthetic Scheme 37 [ka]

[0280] Precursor to orbOHorPhe4COOH [ka]

[0281] (2S,3R)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-(tert-butoxycarbonyl)phenyl)-3-hydroxycypropanoic acid Step 1: Ethyl 2-((diphenylmethylene)amino)acetate (19.44 g, 72.7 mmol) was dissolved in THF (150 mL). The mixture was cooled to −70° C., and LDA (36.4 mL, 72.7 mmol, 2 M in THF) was added under argon. The reaction was stirred at −70° C. for 1 h. To the mixture was then added tert-butyl 4-formylbenzoate (10 g, 48.5 mmol) under argon at −70° C. The reaction was stirred at −70° C. for 2 h. The resulting solution was quenched with aqueous NH4Cl (100 mL) and extracted with EA (3×500 mL). The organic layers were combined, washed with brine (2×300 mL), dried over anhydrous Na2SO4, and filtered. The residue was concentrated in vacuo to give tert-butyl 4-(2-((diphenylmethylene)amino)-3-ethoxy-1-hydroxy-3-oxopropyl)benzoate (30 g, 41.2 mmol, 85% yield) as an orange semi-solid. 29 H 32 NO5[M+H] + MS ESI calculated for 474.22, found 474.30.

[0282] Step 2: To a mixture of tert-butyl 4-(2-((diphenylmethylene)amino)-3-ethoxy-1-hydroxy-3-oxopropyl)benzoate (30 g, 41.2 mmol) in THF (200 mL) and water (200 mL) was added AcOH (47.1 mL, 824 mmol) at ambient temperature. The reaction was stirred at ambient temperature for 16 hours. The resulting solution was extracted with EtOAc (3 x 300 mL). The aqueous layers were combined and concentrated in vacuo to afford tert-butyl 4-(2-amino-3-ethoxy-1-hydroxy-3-oxopropyl)benzoate (16.8 g, 38.0 mmol, 92% yield) as a white solid. 16 H 24 NO5[M+H] + MS ESI calculated for 310.16, found 310.10.

[0283] Step 3: To a mixture of tert-butyl 4-(2-amino-3-ethoxy-1-hydroxycyano-3-oxopropyl)benzoate (16.8 g, 38.0 mmol) in THF (152 mL) and water (76 mL) was added LiOH (1 M, 76 mL, 76 mmol) at ambient temperature. The reaction was stirred at ambient temperature for 2 hours. The resulting solution was acidified to pH 5 with dilute HCl (1 M) and concentrated in vacuo to afford 2-amino-3-(4-(tert-butoxycarbonyl)phenyl)-3-hydroxycyanopropanoic acid (21.5 g, 37.5 mmol, 99% yield) as a white semi-solid. 14 H 20 NO5[M+H] + MS ESI calculated for 282.13, found 282.10.

[0284] Step 4: To a mixture of 2-amino-3-(4-(tert-butoxycarbonyl)phenyl)-3-hydroxycypropanoic acid (21.5 g, 37.5 mmol) in THF (200 mL) and water (200 mL) was added NaHCO (15.73 g, 187 mmol) and Fmoc-OSu (11.37 g, 33.7 mmol) at ambient temperature. The reaction was stirred at ambient temperature for 4 h. The resulting solution was acidified to pH 5 with dilute HCl (1 M) and extracted with EtOAc (3 × 500 mL). The organic layers were combined, washed with brine (2 × 200 mL), dried over anhydrous NaSO, and filtered. The residue was purified by RP Flash with the following conditions: Column: Flash C 18 (300 g); Mobile phase A: water (0.03% TFA), Mobile phase B: ACN; (Gradient: 5 min hold at 2% B, hold to 52% B within 25 min, hold at 52% B for 5 min; hold to 95% B within 5 min, hold at 95% B for 5 min); Flow rate: 50 mL / min; Detector: UV 210 nm; RT = 45 min. Product-containing fractions were collected and concentrated in vacuo to give 2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-(tert-butoxycarbonyl)phenyl)-3-hydroxycypropanoic acid (17.3 g, 32.6 mmol, 87% yield) as an orange solid. C 29 H 30 NO7[M+H-tBu] + MS ESI calculated for 448.19, found 448.05.

[0285] Step 5: To a mixture of 2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-(tert-butoxycarbonyl)phenyl)-3-hydroxycypropanoic acid (17 g, 32.0 mmol) in DMF (150 mL) was added NaHCO (8.07 g, 96 mmol) and (bromomethyl)benzene (8.21 g, 48.0 mmol) at ambient temperature. The reaction was stirred at ambient temperature for 16 h. The resulting solution was diluted with water (100 mL) and extracted with EtOAc (3 × 200 mL). The organic layers were combined, washed with brine (2 × 150 mL), dried over anhydrous NaSO, and filtered. The filtrate was concentrated in vacuo to give the crude product. The residue was purified by silica gel chromatography eluting with a gradient of 0% to 28% EtOAc in PE to give tert-butyl 4-(2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(benzyloxy)-1-hydroxy-3-oxopropyl)benzoate (14.8 g, 24.93 mmol, 78% yield) as a white solid. 36 H 36 NO7[M+H-tBu] + MS ESI calculated for 538.24, found 538.10.

[0286] Step 6: tert-Butyl 4-(2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(benzyloxy)-1-hydroxy-3-oxopropyl)benzoate (14.8 g, 24.93 mmol) was purified using a Prep-SFC-HPLC column: Chiralpak IH, 7 x 25 cm, 10 μm; mobile phase A: CO, mobile phase B: MeOH; flow rate: 250 mL / min; gradient: isocratic 30% B; column temperature (°C): 35; back pressure (bar): 100; wavelength: 220 nm; RT1 (min): 8.95. The collected fractions were combined and concentrated under vacuum. The residue was lyophilized to give tert-butyl 4-((1R,2S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(benzyloxy)-1-hydroxy-3-oxopropyl)benzoate (1.4 g, 2.358 mmol, 9.46% yield) as a white solid. 36H 36 NO7[M+H] + MS ESI calculated for 594.24, found 594.15.

[0287] Step 7: To a solution of tert-butyl 4-((1R,2S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(benzyloxy)-1-hydroxy-3-oxopropyl)benzoate (1.4 g, 2.358 mmol) in THF (15 mL) and EtOAc (15 mL) was added Pd / C (300 mg, 0.282 mmol, 10% wt, dry) under a nitrogen atmosphere. The suspension was degassed under vacuum and purged several times with H. The reaction mixture was stirred under 2 atmospheres of H at room temperature for 6 hours. The resulting suspension was filtered through diatomaceous earth and washed with THF / EtOH (1:1, 2 x 40 mL). The filtrate was concentrated in vacuo. The residue was purified by RP Flash with the following conditions: Column: Flash C 18 (120 g); Mobile phase A: water (0.05% TFA), Mobile phase B: ACN; (Gradient: 3 min hold at 2% B, hold to 60% B within 27 min, hold at 60% B for 5 min; hold to 95% B within 5 min, hold at 95% B for 5 min); Flow rate: 60 mL / min; Detector: UV 210 nm; RT = 45 min. Product-containing fractions were collected and concentrated in vacuo to give (2S,3R)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-(tert-butoxycarbonyl)phenyl)-3-hydroxycypropanoic acid (1.0498 g, 2.085 mmol, 88% yield) as a brown solid. C 29 H 29 NO7Na[M+Na] + MS ESI calculated for 526.19, found 526.25. 1 H NMR(300 MHz,CD3OD)δ 7.87(d,J=8.2 Hz,2H),7.77(d,J=7.5 Hz,2H),7.57-7.19(m,8H),5.42(d,J=2.8 Hz,1H),4.55(d,J=2.8 Hz,1H),4.35-4.20(m,1H),4.08-3.97(m,2H),1.50(s,9H). Synthetic Scheme 38 [ka]

[0288] Precursor to Phe44PyNH3 [ka]

[0289] (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-(2-((tert-butoxycarbonyl)amino)pyridin-4-yl)phenyl)propanoic acid To a stirred solution of (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-iodophenyl)propanoic acid (15 g, 29.2 mmol) in THF (176 mL) and water (88 mL) was added tert-butyl (4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-yl)carbamate (14.03 g, 43.8 mmol), potassium phosphate (18.61 g, 88 mmol), and Pd(dppf)Cl (1.904 g, 2.92 mmol) at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at 50 °C for 2 h. The pH of the mixture was adjusted to 3 with 1 N HCl. The mixture was extracted with EA (3 × 100 mL). The combined organic layers were washed with brine (100 mL) and dried over anhydrous Na2SO4. The residue was purified by silica gel column chromatography eluting with DCM / MeOH=10:1 to give (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-(2-((tert-butoxycarbonyl)amino)pyridin-4-yl)phenyl)propanoic acid (4.8015 g, 7.70 mmol, 26.4% yield) as a brown solid. 34 H 34 N3O6[M+H] + MS ESI calculated for 580.24, found 580.45. 1H NMR(400 MHz,DMSO-d6)δ 9.95(s,1H),8.28(d,J=5.3 Hz,1H),8.05(s,1H),7.88-7.75(m,3H),7.68-7.60(m,4H),7.46-7.25(m, 7H),4.29-4.10(m,4H),3.17-3.15(m,1H),3.00-2.89(m,1H),1.50(s,9H). Synthetic Scheme 39 [ka]

[0290] Precursor to Bip4CONH2 [ka]

[0291] (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4'-carbamoyl-[1,1'-biphenyl]-4-yl)propanoic acid To a stirred solution of (S)-2-((((9H-Fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-iodophenyl)propanoic acid (15 g, 29.2 mmol) in THF (246 mL) was added PdCl2(dtbpf) (2.86 g, 4.38 mmol) and tribasic potassium phosphate (88 mL, 88 mmol, 1 N in water) at room temperature. The solution was stirred at 50 °C for 2 h. The resulting solution was cooled to room temperature. The pH was adjusted to 3 with 1H HCl and extracted with EA (3 × 250 mL). The organic layers were combined, washed with brine (4 × 200 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was recrystallized from EtOH (100 mL) to give (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4'-carbamoyl-[1,1'-biphenyl]-4-yl)propanoic acid (7.5221 g, 14.12 mmol, 48.3% yield) as a brown solid. 31 H 27 N2O5[M+H] +MS ESI calculated for 507.18, found 507.45. 1 H NMR(400 MHz,DMSO-d6)δ 12.87(s,1H),8.02(s,1H),8.07-7.96(m,2H),7.94-7.82(m,2H),7.78-7.60(m,7H),7.39 -7.29(m,5H),7.29-7.19(m,2H),4.20-4.13(m,4H),3.17-3.13(m,1H),2.93-2.90(m,1H). Synthetic Scheme 40 [ka]

[0292] Precursor to SbOHPhe4CONH2 [ka]

[0293] (2S,3S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-carbamoylphenyl)-3-hydroxycypropanoic acid 4-Formylbenzoic acid (200 g, 1.33 mol), pyridine (322 mL, 4.00 mol), and NH4HCO3 (368 g, 4.66 mol, 3.50 equiv.) were added to THF (1.20 L). Boc2O (377 g, 1.73 mol, 397 mL) was added to the reaction mixture at 0 °C. The reaction mixture was stirred at 25 °C for 4 h. The pH of the reaction mixture was adjusted to 5 with HCl (12 M). After filtration and washing the filter cake with HO, the crude product was triturated with isopropanol and HO at 25 °C for 1 h to give the carboxamide intermediate.

[0294] To a solution of buffer (NaHPO (0.10 M, 6.70 L), NaHPO (0.10 M, 6.70 L) was added glycine (187 g, 2.50 mol), pyridoxal-5'-phosphate (PLP) (662 mg, 2.68 mmol), T2G aldolase (3.0 g), and carboxamide intermediate (100 g, 670 mmol). The mixture was stirred at 25 °C for 2 h. The amino acid intermediate was collected by filtration and washed with water.

[0295] The amino acid intermediate (120 g, 535 mmol) was added to a mixture of THF (480 mL) and HO (960 mL). NaCO (113 g, 1.07 mol) was added to the mixture. FmocOSu (180 g, 535 mmol) was added to the mixture. The mixture was stirred at 25 °C for 12 h. The pH of the mixture was adjusted to 1 with HCl (12 M). After filtration and washing the filter cake with HO, the crude product was triturated with acetone:ethyl acetate (1:1) at 25 °C for 2 h to give the title compound.

[0296] Synthetic Scheme 41 [ka]

[0297] Precursors for RbOH4Pal and sbOH4Pal [ka]

[0298] (2S,3R)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-hydroxy-3-(pyridin-4-yl)propanoic acid and (2S,3S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-hydroxy-3-(pyridin-4-yl)propanoic acid To a solution of buffer (KH2PO4, 1.0 M, 15%) at 25°C was added glycine (140 g, 1.87 mol), pyridoxal-5'-phosphate (PLP) (922 mg, 3.73 mmol), T2G aldolase (15.0 g), and isonicotinaldehyde (100 g, 933 mmol). The mixture was stirred at 25°C for 2 hours. The product was collected after filtration and washing with water.

[0299] To a mixture of the hydroxy-substituted amino acid intermediate in THF (1.5 L) was added NaCO (174 g, 1.65 mol) and FmocOSu (277 g, 823 mmol) at 25 °C. The mixture was stirred at 25 °C for 12 h. The pH of the mixture was adjusted to 4 by the addition of HCl (12 M). The mixture was filtered, the filter cake was washed with HO, and the crude product was triturated with acetonitrile. The Fmoc-protected product was purified by SFC to give the two title compounds.

[0300] Synthetic Scheme 42 [ka]

[0301] Precursor to Ala3Isoxa5Me [ka]

[0302] (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(5-methylisoxazol-3-yl)propanoic acid To a mixture of the aldehyde (30.0 g, 270 mmol) in pyridine (300 mL) at 25° C., malonic acid (56.2 g, 540 mmol) and piperidine (4.60 g, 54.0 mmol) were added. The reaction mixture was stirred at 100° C. for 12 hours. The reaction mixture was concentrated under reduced pressure to give the crude product. The crude product was triturated with MTBE at 25° C. for 60 minutes to give the carboxylic acid intermediate.

[0303] To the mixture of carboxylic acid intermediates in 7 M ammonium carbonate buffer, Anabaena variabilis phenylalanine ammonia-lyase (AvPAL, 1.45 g) was added at 25 °C. The reaction mixture was stirred at 37 °C for 12 h. The reaction mixture was concentrated under reduced pressure to give the amino acid intermediate.

[0304] To a mixture of the amino acid intermediate (32.0 g) in HO (160 mL) and THF (160 mL) at 25° C. was added NaCO (39.8 g, 376 mmol) and FmocOSu (63.4 g, 188 mmol). The reaction mixture was stirred at 25° C. for 12 h and concentrated under reduced pressure. The crude residue was acidified to pH=3 with 2 N HCl (aq) (180 mL). The mixture was filtered and the filter cake was washed with water. The filtrate was concentrated under reduced pressure to give the crude product. The crude product was triturated with acetonitrile to give the title compound.

[0305] Precursor to Ala2Oxa (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(oxazol-2-yl)propanoic acid The Fmoc-protected precursor to Ala2Oxa, (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(oxazol-2-yl)propanoic acid, was prepared using a similar method as above, starting from oxazole-2-carbaldehyde.

[0306] Preparation of final compounds: A. Generalized Procedure for Synthesizing Linear Peptide Precursors The peptides in Table 1 were synthesized using standard solid-phase synthesis with Fmoc / tBu chemistry, as exemplified in Chan, W.C.; White, P.D. "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, N.L. "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.

[0307] During peptide chain elongation, the α-amino group of each amino acid was protected with a 9H-fluoren-9-ylmethoxycarbonyl (Fmoc) group. To avoid side reactions during the chain elongation step, any reactive amino acid side chains also carried acid-labile protecting groups, which effectively mask the reactive group until removed upon treatment with strong acid. After each coupling step, the Fmoc group on the N-terminal amino acid was removed with piperidine or 4-methylpiperidine, and the resin was thoroughly washed, preparing it for the coupling of the next Fmoc-protected amino acid derivative.

[0308] The side chain protecting groups used were tert-butyl (tBu) for 3PalCOH, 3PalPhCOH, aIT, aMeD, aMeS, BipCOH, daMeD, daMeS, FbcpA, FpcCCA, FptCCA, hS, PheCOOH, PhePyrimCOH, ProcOH, PyrimAlaPhCOH, S, and T; tert-butoxycarbonyl (Boc) for AccN, AlaPip, AlaPiperaz, aMeDab, aMeK, Dab, Dap, daMeDab, daMeK, K, PipH, and Orn; trityl (Trt) for Q; 2,2,4,6,7-pentamethyldihydrobenzofuran-5-sulfonyl (Pbf) for R; and β-methylpentyl ester (OMpe) for D.

[0309] Fmoc-protected amino acids were typically obtained from sources such as Sigma-Aldrich, Novabiochem, Chem-Impex and Combi-Block.

[0310] B. Synthetic Procedures Used to Prepare Cyclic Peptides Synthetic Scheme 43 [ka]

[0311] Solid-phase synthesis of peptides Protocol A The peptide was synthesized on a Liberty Blue™ synthesizer from CEM Corporation using standard solid phase synthesis with Fmoc / tBu chemistry as outlined in Scheme 44 above.

[0312] An amide bond was formed between the free amino terminus of the resin-bound protected peptide and the carboxylic acid of the Fmoc-protected amino acid using N,N'-diisopropylcarbodiimide (DIC) and ethyl cyano(hydroxyimino)acetate (Oxyma) as coupling agents.

[0313] H-Gly-loaded 2-chlorotrityl resin (200-400 mesh, 0.79 mmol / g loading, 1% cross-linked polystyrene, Novabiochem) was used for the synthesis. All amino acids were dissolved in DMF (N,N-dimethylformamide) at a concentration of 0.2 M. Amino acids were activated with an equimolar amount of Oxyma solution (0.5 M in DMF) and a two-fold molar excess of DIC solution (1.0 M in DMF). Alternatively, amino acids were dissolved in DMF (N,N-dimethylformamide) at a concentration of 0.125 M. Amino acids were activated with an equimolar amount of Oxyma Pure solution (0.125 M in DMF; with 0.05 M DIEA) and a two-fold molar excess of DIC solution (0.25 M in DMF). Reactions were typically performed on a 25 μmol scale.

[0314] All synthesis cycles included deprotection of Fmoc-amino acids with 20% piperidine in DMF (microwave-assisted heating at 90 °C for 2 min) and coupling with Fmoc-protected amino acid / DIC / Oxyma (5, 5, and 10 equivalents, respectively; microwave-assisted heating at 90 °C for 2 or 4 min), which could be repeated twice for difficult couplings. Cycles of Fmoc-deprotection and Fmoc-protected amino acid coupling were repeated with the desired monomer until the complete linear peptide was formed.

[0315] Solid Phase Synthesis of Peptides Protocol B Alternatively, peptides were synthesized manually on a Biotage® Syro II peptide synthesizer using standard solid-phase synthesis with Fmoc / tBu chemistry, as summarized in Scheme 43 above. HATU and DIPEA were used as coupling agents to create an amide bond between the free amino terminus of the resin-bound protected peptide and the carboxylic acid of the Fmoc-protected amino acid. H-Gly-loaded 2-chlorotrityl resin (200-400 mesh, 0.79 mmol / g loading, 1% cross-linked polystyrene, Novabiochem) was used for the synthesis. All amino acids were dissolved in 1:1 DMF:NMP at 0.2 M concentration. Reactions were typically performed on a 12 μmol scale.

[0316] All synthesis cycles included the following: (1) Coupling (repeated twice) with Fmoc-protected amino acid / HATU / DIPEA (4, 4, and 8 equivalents, respectively; room temperature; 15 min). The mixture was filtered, and the peptidyl resin was washed with DMF (2 × 1 mL). (2) Fmoc deprotection (repeated three 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 × 1 mL). Cycles of Fmoc deprotection and Fmoc-protected amino acid coupling were repeated with the desired monomer until the complete linear peptide was formed.

[0317] Selective cleavage and macrolactamization of protected peptides To cleave the protected linear peptide from the solid support, the peptidyl resin (approximately 16 mg) was treated with 25% hexafluoroisopropanol (HFIP) in DCM for 20 minutes at room temperature, filtered, and the solvent was removed under reduced pressure. The resulting residue was dissolved in DMF (5 mL). HATU (0.44 equiv.) and DIPEA (2.5 equiv.) were added. The mixture was stirred at room temperature for 5 minutes. An additional 0.66 equiv. of HATU was then added. Completion of the macrolactamization was monitored by UPLC-MS, and the solvent was removed under reduced pressure.

[0318] Final side chain deprotection 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 minutes. Cold diethyl ether (15 mL) was added to the solution. The peptide was precipitated by centrifugation (3200 rpm, -10 °C). The precipitate was washed with diethyl ether (2 × 10 mL) and dried under vacuum overnight to give the crude deprotected cyclic peptide as a solid.

[0319] HPLC purification Purification was performed by preparative reverse-phase high-performance liquid chromatography (RP-HPLC) on a Waters X-Bridge Prep C18 OBD Prep column (130 Å, 500 Hz, column size 19 x 100 mm) using a Waters MS-Directed AutoPurification HPLC / MS system. Mobile phase: (A) HPLC 0.16% TFA in water and (B) HPLC 0.16% TFA in acetonitrile; flow rate: 25 mL / min; UV wavelength λ = 215 nm; gradient: 25 to 50% B over 5 min. Alternatively, purification was performed on a Waters CSH-C18 column (19 x 250 mm, 5 μM) using an Agilent 1290 Infinity II 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: 25 mL / min; UV wavelength λ = 215 nm; gradient: 20% B over 2.5 min, 55% B over 2.5–20 min. UV-absorbing fractions containing the target m / z ions were collected, and fractions containing the product were confirmed by LC / MS.

[0320] The purity of the fractions was confirmed by UPLC, which was measured using a reverse-phase Waters Acquity UPLC-MS system. Column: Waters XSelect CSH C18 column (130 Å, 2.5 μm, 2.1 × 50 mm column size). Mobile phase: (A) HPLC 0.05% TFA in water and (B) HPLC 0.05% TFA in acetonitrile; injection volume: 1 μL; flow rate: 1 mL / min; UV wavelength λ = 215 nm; gradient: 5 to 100% B in 5 min. Lyophilization of the combined fractions containing the pure peptide afforded the final cyclized product as a powder.

[0321] Synthetic Scheme 44 (SEQ ID NOs: 51 and 214, respectively, in order of appearance) [ka]

[0322] R 11 -CH2-C 11 -Ca and C 11 For certain compounds where is phenyl, their synthesis was completed using a final aryl / heteroaryl coupling. The aryl / heteroaryl bromide precursors were synthesized in the solid phase synthesis of peptides protocol using the general procedure described in Synthetic Scheme 6.

[0323] In a N2-filled glovebox, a solution of aryl bromide (0.007 mmol) in 150 μL of DMF was added to arylboronic acid or pinacol ester (0.07 mmol) at room temperature, followed by an aqueous solution of K2HPO4 (1.25 M, 48 μL) and a solution of QPhos Pd G3 (1.15 mg) in DMF (15 μL). The resulting mixture was heated to 60 °C for 21 h. The reaction mixture was then filtered and purified by the HPLC purification procedure described above.

[0324] Synthetic Scheme 45 [ka]

[0325] R 1 R 1e For certain compounds that are -C(O)NH-CHCH-O-, further transformations were carried out to give the final compounds. These transformations are exemplified in the schemes above and the procedures below.

[0326] tert-Butyl 4-(((2R,9S,12S,15S,21S,24S,30S,32aS,34S,41S,44S,47S,49aR)-21-((1H-indol-3-yl)methyl)-9-(2-((tert-butoxycarbonyl)amino)ethyl)-24-(tert-butoxymethyl)-34-fluoro-3 in CHCl (10 mL) 0-((S)-1-(4-fluoro-1H-indol-3-yl)ethyl)-18,18,27,27-tetramethyl-5,8,11,14,17,20,23,26,29,32,37,40,43,46,49-pentadecaoxo-12-(3-oxo-3-(tritylamino)propyl)-2,44-bis(pyridin-4-ylmethyl)-15-(pyridin-4-ylmethyl) Imidin-5-ylmethyl)-47-(4-(2-(((vinyloxy)carbonyl)amino)ethoxy)benzyl)octatetraoctahydro-1H-dipyrrolo[1,2-a:1',2'-e1][1,4,7,10,13,16,19,22,25,28,31,34,37,40,43]pentadecaazacyclopentatetracontin-41-yl)methyl)benzoate (I-45A, 0.606 g, 0.24 mmol) was bubbled with N, and Pd(PPh) (0.021 g, 0.018 mmol) and phenylsilane (0.118 mL, 0.960 mmol) were added. N was bubbled again, followed by stirring at room temperature for 2 h. The solvent was removed, and the crude mixture of I-45B was used in the next step without purification.

[0327] To the acid monomer R1eCOOH (0.055 mmol) at room temperature, I-45B (0.011 mmol, 0.82 mL in DMF), HATU (0.4 M in DMF, 121 μL), and DIEA (2 M in NMP, 48 μL) were added. The resulting mixture was stirred at room temperature for 2 h and then evaporated to dryness to give I-45C. I-45C was treated with 1.00 mL of TFA cleavage cocktail (90% TFA, 2% TIPS, 8% HO) at room temperature for 0.5 h. The reaction mixture was then transferred to 10 mL of cold Et2O, centrifuged, decanted, and dried under vacuum. DMSO (1 mL) was added, and the crude mixture was purified by reverse-phase HPLC to give the final compound.

[0328] Synthetic Scheme 46 [ka]

[0329] R 1 C 1 and C 1 For certain compounds where is aryl or heteroaryl, further transformations were carried out to give the final compounds. These transformations are exemplified in the schemes above and the procedures below.

[0330] In a glovebox, to ArB(OH) or boronic ester (0.095 mmol, 10.7 equiv.) at room temperature was added I-46A (0.009 mmol, 1 equiv.) in 190 μL DMF, followed by KHPO (1.25 M aqueous solution, 58 μL, 8 equiv.) and QPhos Pd G (0.19 mg in 19 μL DMF). The resulting mixture was heated at 60 °C for 21 h. The reaction mixture was then filtered, dissolved in DMSO (1 mL), and purified by reverse-phase HPLC to give the final compound.

[0331] Biological Assays: Procedure for IL-6 assay in MRC5 cells Inhibition of IL-1β-induced IL-6 secretion in MRC5 cells was assessed. 2X EC 80Recombinant human IL-1β (BioLegend 579404) was prepared in EMEM (ATCC 30-2003) containing seeding medium, 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). 20 μL of IL-1β was added to a 384-well collagen-coated plate (Corning 354664) and preincubated for 1 h at ambient temperature with 200 nL of compound dispensed using an ECHO 555 liquid handler. Human lung fibroblast MRC5 cells (ATCC CCL-171) were seeded at a density of 3,000 cells / 20 μL / well. Cells were prepared by passage three times in collagen-coated T175 flasks (Greiner 661950) in EMEM (ATCC 30-2003) growth medium 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). Cells were then harvested in the seeding medium after 5 min of digestion with 0.25% trypsin-EDTA (Gibco 25200-056). A 384-well collagen-coated plate containing a final volume of 40 μL was incubated overnight at 37°C, 5% CO2. Five μL of conditioned medium was transferred to a 384-well AlphaLISA plate (PerkinElmer 6005350) for IL-6 detection using the 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 the 384-well AlphaLISA plate and incubated at ambient temperature for 1 hour. Protecting the donor bead mix from light, 25 μL was added to the plate and incubated at ambient temperature for 30 minutes.AlphaLISA plates were read on an EnVision multimode plate reader (Perkin Elmer model 2104) using the AlphaScreen setting (laser excitation at 680 nm and emission at 570 nm). Dose-response curves and IC. 50 Values ​​were analyzed using a four-parameter logistic equation in Spotfire software (Tibco, Palo Alto, CA).

[0332] The amino acid sequences of Example Nos. 1 to 213 and 215 to 285 (SEQ ID NOs. 1 to 213 and 215 to 285), biological activity (MRC IC 50 ), calculated monoisotopic mass, molecular formula, calculated molecular weight and mass spectral data (M+H), (M+2H / 2) or (M+3H / 3) are provided in Table 1 below. [Table 2]

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Claims

1. A compound of formula (I) 【Chemical 95】 R １ is R １ｅ -C(O)NH-CH ２ CH ２ -O-, C １ ~C ４ Alkyl, halo, or C １ and R １ｅ－ teeth, (a) C １ ~C ４ alkyl; or (b) C Ｙ１ (where C Ｙ１ teeth, (i) C ３ ~C ６ cycloalkyl; (ii) phenyl; or (iii) a 5- to 6-membered monocyclic heteroaryl, wherein the heteroaryl contains 1 to 2 heteroatoms independently selected from the group consisting of N, O, and S; Here, C Ｙ１ is unsubstituted or C １ ~C ３ 1 to 3 R selected from the group consisting of alkyl, halo, and piperazinyl Ｙ１ is substituted with a substituent, C １ teeth, (i) a 5-6 membered monocyclic aryl or heteroaryl, wherein said heteroaryl contains 1-2 heteroatoms independently selected from the group consisting of N, O and S; (ii) a 3- to 6-membered monocyclic or 5- to 8-membered bicyclic cycloalkyl; or (iii) a 5- to 6-membered monocyclic saturated heterocycloalkyl containing 1 to 2 heteroatoms independently selected from the group consisting of N, O, and S; Here, C １ is unsubstituted or is selected from halo, C １ ~C ３ Alkyl, C １ ~C ３ Fluoroalkyl, carboxy, C １ ~C ３ Alkoxy, C ２ ~C ３ Acyl, —C(O)NH ２ and -C(O)N(CH ３ ) ２ 1 to 3 R independently selected from the group consisting of Ｃ１ is substituted with a substituent, R ２ teeth, (i) a 5- to 6-membered monocyclic aryl or heteroaryl, wherein the heteroaryl contains 1-2 heteroatoms independently selected from the group consisting of N, O, and S; and (ii) 3- to 8-membered monocyclic or bicyclic cycloalkyl; Here, R ２ is unsubstituted or is selected from halo, amino, hydroxy, C １ ~C ３ Alkyl, C １ ~C ３ Fluoroalkyl and C １ ~C ３ 1 to 3 R independently selected from the group consisting of alkoxy ２ａ is substituted with a substituent, R ２ｂ is H or hydroxy, R ３ is F or hydroxy, R ４ teeth, (i) naphthyl; or (ii) a 9- to 10-membered heteroaryl, wherein the heteroaryl contains 1 to 2 heteroatoms independently selected from the group consisting of N, O, and S; Here, R ４ is unsubstituted or is 1 to 2 R independently selected from the group consisting of halo ４ａ is substituted with a substituent, R ５ａ is H, C １ ~C ３ Alkyl, H ２ N (CH ２ ) ｍ - or HOCH ２ - and R ５ｂ is H, C １ ~C ３ Alkyl, H ２ N (CH ２ ) ｍ - or HOCH ２ - and Or, R ５ａ and R ５ｂ together with the carbon atoms to which they are attached, form C ３ ~C ６ forming a cycloalkyl or a 4- to 6-membered saturated heterocycloalkyl containing one N atom, R ６ａ and R ６ｂ Each of the groups is independently H, —(CH ２ ) ｎ１ CH ３ , -(CH ２ ) ｎ２ -OH, or -(CH ２ ) ｎ２ CO ２ H, R ７ａ is H, C １ ~C ３ Alkyl, HOCH ２ -, H ２ N (CH ２ ) ｐ -, HO ２ CCH ２ -, H ２ NC(O)CH ２ -, CH ３ OCH ２ - or PhCH ２ - and R ７ｂ is H, C １ ~C ３ Alkyl, HOCH ２ -, H ２ N (CH ２ ) ｐ -, HO ２ CCH ２ -, H ２ NC(O)CH ２ -, CH ３ OCH ２ - or PhCH ２ - and Or, R ７ａ and R ７ｂ together with the carbon atoms to which they are attached form a 4- to 6-membered saturated heterocycloalkyl containing one N atom; R ８ａ is HO-(CH ２ ) ｑ -, CH ３ -O-(CH ２ ) ｑ -, CH ３ CH ２ -O-(CH ２ ) ｑ -, PhCH ２ -O-(CH ２ ) ｑ -, C １～ C ３ Alkyl, C １ ~C ３ Fluoroalkyl, H ２ N-(CH ２ ) ｒ -, (CH ３ ) ３ N-(CH ２ ) ｒ -, H ２ NC(NH)N(H)-(CH ２ ) ｒ -, H ２ NC(O)N(H)-(CH ２ ) ｒ -, HO ２ C-(CH ２ ) ｒ -, (CH ３ ) SO ２ - (CH ２ ) ｒ -, C ８ａ , or C ８ａ－ CH ２ - and Here, C ８ａ teeth, (i) a 5- to 6-membered monocyclic aryl or heteroaryl, wherein said heteroaryl contains 1 to 2 heteroatoms independently selected from the group consisting of N, O, and S; (ii) a 5- to 6-membered monocyclic saturated heterocycloalkyl, wherein the heterocycloalkyl contains 1 to 2 heteroatoms independently selected from the group consisting of N, O, and S; or (iii) C ３ ~C ６ is cycloalkyl, Here, C ８ａ is unsubstituted or is selected from halo, amino, hydroxy, C １ ~C ３ Alkyl, C １ ~C ３ Fluoroalkyl, C １ ~C ３ Alkoxy, H ２ N-(CH ２ ) ｓ -, H ２ NC(O)-(CH ２ ) ｓ -, H ２ C=CH-CH ２ 1 to 3 R independently selected from the group consisting of O- and phenyl Ｃ８ａ is substituted with a substituent, R ８ｂ is H, methyl or hydroxy, R ９ is HO-(CH ２ ) ｔ -, H ２ N-(CH ２ ) ｕ -, H ２ NC(NH)N(H)-(CH ２ ) ｕ -, H ２ NC(O)N(H)-(CH ２ ) ｕ - or C ９ and Here, C ９ is a 5-6 membered saturated heterocycloalkyl containing 1-2 heteroatoms independently selected from the group consisting of N, O and S; Here, C ９ is unsubstituted or is halo, C １ ~C ３ Alkyl, C １ ~C ３ Fluoroalkyl and C １ ~C ３ 1 to 2 R independently selected from the group consisting of alkoxy Ｃ９ is partially substituted with R １０ is H or methyl, R １１ is H, -CH ２ -C １１ , or -CH ２ -C １１ -C ａ and C １１ teeth, (i) phenyl; or (ii) a 5- to 6-membered monocyclic aryl or heteroaryl, wherein the heteroaryl contains 1 to 2 heteroatoms independently selected from the group consisting of N, O, and S; Here, C １１ is unsubstituted or is selected from halo, hydroxy, amino, C １ ~C ３ Alkyl, C １ ~C ３ Fluoroalkyl, carboxy, C １ ~C ３ Alkoxy and C ２ ~C ３ 1 to 3 R independently selected from the group consisting of acyl Ｃ１１ is substituted with a substituent, C ａ is a 5- to 6-membered monocyclic heteroaryl, said heteroaryl containing 1-2 heteroatoms independently selected from the group consisting of N, O and S; Here, C ａ is unsubstituted or is selected from halo, hydroxy, amino, C １ ~C ３ Alkyl, C １ ~C ３ Fluoroalkyl, carboxy, C １ ~C ３ Alkoxy and C ２ ~C ３ 1 to 3 R independently selected from the group consisting of acyl Ｃａ is substituted with a substituent, R １２ is H or -CH ２ C １２ and Here, C １２ teeth, (i) phenyl, or (ii) a 5- to 6-membered monocyclic heteroaryl, wherein the heteroaryl contains 1 to 2 heteroatoms independently selected from the group consisting of N, O, and S; Here, C １２ is unsubstituted or is selected from halo, hydroxy, amino, C １ ~C ３ Alkyl, C １ ~C ３ Fluoroalkyl, carboxy, C １ ~C ３ Alkoxy and C ２ ~C ３ 1 to 3 R independently selected from the group consisting of acyl Ｃ１２ is substituted with a substituent, R １３ is H or methyl, R １４ is a halo, R １５ is —OH or —NH ２ and R １６ is halo, hydroxy, C １ ~C ３ Alkyl, C １ ~C ３ Fluoroalkyl, or C １ ~C ３ is an alkoxy, R １７ is H, hydroxy or methyl, each occurrence of subscript m is independently 1, 2, 3, or 4; the subscript n1 is 0, 1, 2, or 3; the subscript n2 is 0, 1, or 2; each occurrence of subscript p is independently 2, 3, or 4; the subscript q is 0, 1 or 2; the subscript r is 0, 1, 2, or 3; each occurrence of subscript s is independently 1 or 2; the subscript t is 0, 1, or 2; the subscript u is 0, 1, 2, or 3; the subscript v is 0, 1 or 2; the subscript w is 0, 1, or 2; X １ and X ２ are independently C(H) or N; X ３ and X ４ are independently C(H), C(Cl), C(F) or N; The compound, or a pharmaceutically acceptable salt thereof.

2. The group 【Chemistry 96】 but, 【Chemistry 97】 2. The compound of claim 1, wherein:

3. The compound of formula (I) is of formula (IA) 【Chem.98】 2. The compound of claim 1, wherein:

4. C １ is phenyl, pyrimidinyl or piperazinyl, wherein C １ is unsubstituted or has 1 to 2 R Ｃ１ is substituted with a substituent, R ２ is pyridyl or bicyclo[1.1.1]pentanyl, where R ２ is unsubstituted or has 1 to 2 R ２ａ is substituted with a substituent, R ４ is indolyl or naphthyl, where R ４ is unsubstituted or has one R ４ａ is substituted with a substituent, C ８ａ is phenyl, pyridyl, pyrimidinyl, pyridazinyl, tetrahydropyranyl, piperidinyl, morpholinyl or piperazinyl, wherein C ８ａ is unsubstituted or has 1 to 2 R Ｃ８ａ is replaced by C ９ is morpholinyl, where C ９ is unsubstituted or contains one R Ｃ９ is replaced by R １１ but, -CH ２ -C １１ (where C １１ is phenyl, pyridyl, pyrimidinyl or pyrazinyl, where C １１ is unsubstituted or has one R Ｃ８ is replaced by -CH ２ -C １１ -C ａ (where C １１ is phenyl, and C １１ is unsubstituted or one R Ｃ１１ and C ａ is pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, isoxazolyl or imidazolyl, where C ａ is unsubstituted or has one R Ｃａ substituted with R １２ But -CH ２ C １２ (where C １２ is phenyl or pyridyl, where C １２ is unsubstituted or has one R Ｃ１２ substituted with 2. The compound of claim 1 or a pharmaceutically acceptable salt thereof.

5. X １ and X ２ is C(H), and R １ but, C １ where C １ is phenyl or bicyclo[1.1.1]pentanyl, Here, C １ is substituted with carboxy, or R １ｅ -C(O)NH-CH ２ CH ２ -O- (where R １ｅ is C １ ~C ４ 2. The compound of claim 1, wherein R is 1 or 2; and R is 2 or 3; or a pharmaceutically acceptable salt thereof.

6. X １ and X ２ is C(H), and R １ is phenyl substituted with carboxy; 2. The compound of claim 1 or a pharmaceutically acceptable salt thereof.

7. R ２ｂ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein:

8. R ２ is unsubstituted or is substituted with 1 to 3 R ２ａ is a 5- to 6-membered monocyclic aryl or heteroaryl substituted with a substituent, and X ３ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein:

9. R ２ 9. The compound of claim 8, or a pharmaceutically acceptable salt thereof, wherein is unsubstituted or substituted pyridyl.

10. R ２ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein: is unsubstituted bicyclo[1.1.1]pentanyl.

11. R ３ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein is fluoro.

12. R ４ The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein is 4-fluoroindolyl.

13. R ５ａ and R ５ｂ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein: is methyl.

14. R ６ａ is H, -(CH ２ ) ｎ１ CH ３ , -(CH ２ ) ｎ２ -OH or -(CH ２ ) ｎ２ CO ２ H, R ６ｂ is H or methyl, the subscript n1 is 1, 2, or 3, and the subscript n2 is 0, 1, or 2; 2. The compound of claim 1 or a pharmaceutically acceptable salt thereof.

15. R ６ａ is —OH or —CH ２ CO ２ H, and R ６ｂ is H, 15. The compound of claim 14 or a pharmaceutically acceptable salt thereof.

16. X ４ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein: is C(H).

17. R ７ａ and R ７ｂ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein: is methyl.

18. R ８ａ But 1 to 3 R Ｃ８ａ phenyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, piperidinyl, tetrahydropyranyl, or morpholinyl, which may or may not be substituted by a substituent; and R ８ｂ is H, 2. The compound of claim 1 or a pharmaceutically acceptable salt thereof.

19. R ８ａ 19. The compound of claim 18, or a pharmaceutically acceptable salt thereof, wherein is unsubstituted pyridyl, pyrimidinyl, or pyrazinyl.

20. R ９ But, H ２ N-(CH ２ ) ｕ - and the subscript u is 1 or 2, or a pharmaceutically acceptable salt thereof.

21. R １０ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein:

22. R １１ is H, R １２ But -CH ２ C １２ (where C １２ is phenyl or pyridyl, where C １２ is unsubstituted or contains one R Ｃ１２ substituted with 2. The compound of claim 1 or a pharmaceutically acceptable salt thereof.

23. R １１ but, -CH ２ -C １１ (where C １１ is phenyl, pyridyl, pyrimidinyl or pyrazinyl, where C １１ is unsubstituted or contains one R Ｃ１１ replaced by ); or -CH ２ -C １１ -C ａ (where C １１ is phenyl, and C １１ is unsubstituted or contains one R Ｃ１１ and C ａ is pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, isoxazolyl or imidazolyl, where C ａ is unsubstituted or contains one R Ｃａ substituted with R １２ is H, 2. The compound of claim 1 or a pharmaceutically acceptable salt thereof.

24. X １ X ２ , X ３ and X ４ is C(H), R １ is phenyl substituted with carboxy; R ２ is a 5-6 membered monocyclic aryl or heteroaryl that is unsubstituted or ... unsubstituted or is unsubstituted ２ａ a 5- to 6-membered monocyclic aryl or heteroaryl substituted with a substituent; R ３ But it is fluoro, R ４ is 4-fluoroindolyl, R ５ａ and R ５ｂ is methyl, R ６ａ is —OH or —CH ２ CO ２ H, R ６ｂ is H, R ７ａ and R ７ｂ is methyl, R ８ａ is unsubstituted or is substituted with 1 to 3 R Ｃ８ａ phenyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, piperidinyl, tetrahydropyranyl, or morpholinyl, each of which is substituted by a substituent; R ８ｂ is H, R ９ But, H ２ N-(CH ２ ) ｕ - and R １０ is H, and the subscript u is 1 or 2; 2. The compound of claim 1 or a pharmaceutically acceptable salt thereof.

25. R １１ is H, and R １２ But -CH ２ C １２ (where C １２ is phenyl or pyridyl, where C １２ is unsubstituted or has one R Ｃ１２ substituted with 25. The compound of claim 24 or a pharmaceutically acceptable salt thereof.

26. R １１ but, -CH ２ -C １１ (where C １１ is phenyl, pyridyl, pyrimidinyl or pyrazinyl, and C １１ is unsubstituted or contains one R Ｃ１１ replaced by ); or -CH ２ -C １１ -C ａ (where C １１ is phenyl, and C １１ is unsubstituted or contains one R Ｃ１１ and C ａ is pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, isoxazolyl or imidazolyl, where C ａ is unsubstituted or contains one R Ｃａ substituted with R １２ is H, 25. The compound of claim 24 or a pharmaceutically acceptable salt thereof.

27. R １ But CH ３ C(O)NH-CH ２ CH ２ —O—, 5-CO ２ H-pyrimidin-2-yl, 4-CH ３ C(O)-piperazin-1-yl, 4-COH-cyclohex-4-yl, 4-CO ２ H-phenyl, or bicyclo[1.1.1]pentane-1-carboxylic acid; R ２ is pyridin-4-yl, pyridazin-4-yl, bicyclo[1.1.1]pentan-1-yl, or cyclobutyl; R ４ is 4-fluoroindol-3-yl, 4-chloroindol-3-yl, or naphth-1-yl; R ５ａ But CH ３ , HOCH ２ -, H ２ NCH ２ CH ２ CH ２ CH ２ - or H ２ NCH ２ CH ２ - and R ５ｂ But CH ３ , HOCH ２ - or H ２ NCH ２ CH ２ - and Or, R ５ａ and R ５ｂ together with the carbon atoms to which they are attached form an azetidinyl ring, R ６ａ But -CH ２ CO ２ H, -OH, -H, -CO ２ H, —CH ２ OH, CH ３ , or -CH ２ CH ３ and R ６ｂ is H or CH ３ - and R ７ａ But CH ３ , HOCH ２ -, H ２ NCH ２ CH ２ -, H ２ NCH ２ CH ２ CH ２ CH ２ - or -CH ２ CO ２ H, R ７ｂ But CH ３ , HOCH ２ -, H ２ NCH ２ CH ２ -, H ２ NCH ２ CH ２ CH ２ CH ２ - or -CH ２ CO ２ H, Or, R ７ａ and R ７ｂ together with the carbon atoms to which they are attached form an azetidinyl or piperidinyl ring, R ８ａ Amino, hydroxy, methyl, H ２ NC(NH)N(H)CH ２ CH ２ -, H ２ NC(O)-N(H)CH ２ CH ２ -, H ２ N CH ２ CH ２ -, phenyl, pyridin-4-yl, pyridin-3-yl, pyridin-2-yl, pyrimidin-5-yl, pyrimidin-2-yl, pyrazin-2-yl, pyridazin-3-yl, pyridazin-4-yl, piperidin-4-yl, tetrahydropyran-4-yl, or morpholin-4-yl; R ８ｂ is H or CH ３ and R ９ But, HO-, H ２ N-, H ２ NCH ２ -, H ２ NCH ２ CH ２ -, H ２ NCH ２ CH ２ CH ２ -, H ２ NC(NH)N(H)CH ２ CH ２ - or morpholin-4-yl, R １０ is H or methyl, R １１ But -CH ２ Ph, -CH ２ -(4-bromophenyl), -CH ２ -(pyrimidin-5-yl), -CH ２ -4-(pyrimidin-5-yl)phenyl, -CH ２ -4-(2-aminopyrimidin-5-yl)phenyl, -CH ２ -4-(pyridin-4-yl)phenyl, CH ２ -4-(pyridin-3-yl)phenyl, -CH ２ -4-(5-aminopyrazin-2-yl)phenyl, -CH ２ -4-(2-aminopyrimidin-5-yl)phenyl, -CH ２ -4-(2-methoxypyrimidin-5-yl)phenyl, -CH ２ -4-(pyridin-2-yl)phenyl, -CH ２ -4-[(3-methyl)-isoxazol-4-yl]phenyl, -CH ２ -4-[(1-methyl)imidazol-2-yl]phenyl, or -CH ２ -4-[(1-methyl)imidazol-4-yl]phenyl, and R １２ is H, -CH ２ Ph, -CH ２ -(4-Fphenyl), or -CH ２ -(4-pyridin-4-yl), 2. The compound of claim 1 or a pharmaceutically acceptable salt thereof.

28. The compound of formula (I) is of formula (IB) 【Chem.99】 2. The compound of claim 1, wherein:

29. R ５ａ is methyl or HOCH ２ - and R ６ａ is H, —OH or —CH ２ CO ２ H, R ８ａ but, (i) C ８ (where C ８ is unsubstituted pyridyl, pyrimidinyl, or pyrazinyl; or (ii)CH ３ CH ２ -O- and R ８ｂ is H, methyl, R １１ is H or -CH ２ Ph, R １２ but, (i) H, or (ii) —CH ２ C １２ (where C １２ is phenyl or pyridyl, where C １２ is unsubstituted or substituted with one halo, and the subscript u is 1 or 2; 29. The compound of claim 28.

30. 2. The compound of claim 1, selected from the group consisting of SEQ ID NOs: 1-213 and 215-385, or a pharmaceutically acceptable salt thereof.

31. (SEQ ID NOs: 22, 29, 41, 42, 44, 48, 51, 67, 72, 99, 101, 218 and 381 (in order of appearance): 【Chemistry 100】 【change】 【change】 【change】 【change】 2. The compound of claim 1, selected from the group consisting of: or a pharmaceutically acceptable salt thereof.

32. 32. A pharmaceutical composition comprising the compound of any one of claims 1 to 31 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

33. 32. A method for treating atherosclerosis, comprising administering a therapeutically effective amount of the compound of any one of claims 1 to 31 or a pharmaceutically acceptable salt thereof to a subject in need of such treatment.

34. 32. A method for treating vascular inflammation, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of any one of claims 1 to 31 or a pharmaceutically acceptable salt thereof.

35. 32. A method for treating an inflammatory disorder, comprising administering a therapeutically effective amount of a compound of any one of claims 1 to 31 or a pharmaceutically acceptable salt thereof to a subject in need of such treatment.

36. 36. The method of claim 33, 34, or 35, wherein an effective amount of the compound or a pharmaceutically acceptable salt thereof is orally administered to the subject.

37. 32. Use of a compound according to any one of claims 1 to 31, or a pharmaceutically acceptable salt thereof, in therapy.

38. 32. Use of a compound according to any one of claims 1 to 31 or a pharmaceutically acceptable salt thereof for treating atherosclerosis.

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

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