Pyrrolidine compounds having Lp (a) reducing activity

Abstract

The present disclosure provides compounds of the formula: and pharmaceutically acceptable salts thereof, and compounds of the formula: and pharmaceutically acceptable salts thereof, as well as pharmaceutical compositions comprising these compounds, and their use in the treatment of cardiovascular disease and elevated Lp (a) plasma levels.

Description

Technical Field

[0001] This invention relates to pyrrolidine compounds, pharmaceutically acceptable salts thereof, pharmaceutical compositions and the therapeutic use of said compounds, particularly their use in reducing lipoprotein(a) (Lp(a)) plasma levels. Background Technology

[0002] Significant progress has been made in the treatment of cardiovascular disease (CVD). Despite these advancements, patients still experience cardiovascular events such as angina, myocardial infarction, and stroke, which can lead to death if left untreated. Dyslipidemia, or abnormal lipid levels, remains a major risk factor for CVD. Dyslipidemia can be categorized into four general risk factors: elevated low-density lipoprotein cholesterol (LDL-c), low high-density lipoprotein cholesterol (HDL-c), elevated triglycerides (TG), and elevated Lp(a). Various treatment options exist for elevated LDL-c, low HDL-c, and elevated triglycerides. For patients with elevated Lp(a) levels, approved treatment options are limited. In some cases, apheresis can be used to filter blood to remove LDL and Lp(a); however, the effects are temporary and usually need to be repeated every two weeks. Currently, there are no approved drug therapies specifically for lowering Lp(a) levels.

[0003] Lp(a) is a genetically determined, independent cardiovascular risk factor. Elevated serum Lp(a) levels greater than 50 mg / dL or 125 nmol / L, found in ~20% of the population, are associated with at least a 1.6-fold increased risk of first cardiovascular events (Kronenberg, F. Clin. Res. Cardiol. Suppl. 14, 5-12 (2019)) and a >1.42-fold increased risk of secondary events (Madsen, CM et al., Arterioscler. Thromb. Vasc. Biol. 40, 255-266 (2020)). Lp(a) may exhibit both prothrombotic and antithrombotic properties, as well as atherogenic and atherosclerotic thrombotic properties. Lp(a) may inhibit fibrinolysis and accumulate in the vessel wall to induce thrombosis and atherosclerotic lesions. Plasma Lp(a) levels vary considerably among individuals. Unlike other risk factors, Lp(a) plasma levels do not change significantly with diet and exercise.

[0004] Lp(a) is similar to LDL-c in that it includes an LDL lipid core and an attached apolipoprotein B (apoB), but unlike LDL-c, Lp(a) also contains a unique apolipoprotein (a) (apo(a)) that is bound to apoB via disulfide bonds. Apo(a) is synthesized in the liver. The assembly of Lp(a) from apo(a) and LDL particles can occur in hepatocytes, on the cell wall, or in plasma. Inhibition of the assembly of LDL particles with apo(a) reduces Lp(a) levels.

[0005] WO2020 / 247429 discloses a compound that inhibits Lp(a) formation by blocking the interaction between Apo(a) and ApoB.

[0006] Patients with cardiovascular disease, particularly those with dyslipidemia or abnormal lipid levels, require additional treatment options. Patients whose cardiovascular risk cannot be adequately managed using current standard of care therapies (such as diet, exercise, and / or the use of one or more medications, such as statins, fibrates, and niacin) require alternative treatment options. More specifically, there is a need for additional, pharmaceutically acceptable alternative compounds that inhibit Lp(a) formation and thus lower plasma Lp(a) levels. Summary of the Invention

[0007] Therefore, in the first aspect, compounds of formula I or pharmaceutically acceptable salts thereof are provided: in L is a C2-C6 alkylene group, a C2-C6 alkenyl group, or -(CH2). n NHCONH(CH2) n -; n is independently 2 to 4 each time it appears; Y is independently CH2, O, or S each time it appears; Z is independently H or C each time it appears. 1- C4 alkyl, OH, or cyclopropyl.

[0008] In the second aspect, compounds of formula II or pharmaceutically acceptable salts thereof are provided: in L is a C2-C3 alkylene or C2-C3 alkenylene; R 1 It is -NHC(O)R 2 -NHC(O)NHR 2 COOH, -C(O)NH-cyclopropyl, , , -O-phenyl, -NH-phenyl or phenyl, wherein the phenyl group is optionally substituted with one or two substituents selected from halogens and OCH3; R 2 It is a C1-C3 alkyl group; Y is CH2, O, or S; Z represents H and C. 1- C4 alkyl, OH, or cyclopropyl.

[0009] In a third aspect, a pharmaceutical composition is provided comprising a compound of formula I or II or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier, diluent, or excipient.

[0010] In a fourth aspect, a method for treating a patient’s cardiovascular disease is provided, comprising administering to the patient an effective amount of a compound of formula I or II or a pharmaceutically acceptable salt thereof.

[0011] In a fifth aspect, a method for treating elevated Lp(a) plasma levels in a patient is provided, comprising administering to the patient an effective amount of a compound of formula I or II or a pharmaceutically acceptable salt thereof.

[0012] In the sixth aspect, compounds of formula I or II or pharmaceutically acceptable salts thereof are provided for use in therapy.

[0013] In the seventh aspect, compounds of formula I or II or pharmaceutically acceptable salts thereof are provided for the treatment of cardiovascular diseases.

[0014] In the eighth aspect, compounds of formula I or II or pharmaceutically acceptable salts thereof are provided for the treatment of elevated Lp(a) plasma levels. Detailed Implementation

[0015] In one embodiment of the compound of formula I, Z is H each time it appears.

[0016] In one embodiment of the compound of formula I, Y is CH2 each time it appears.

[0017] In one embodiment, a compound of formula Ia is provided: Or a pharmaceutically acceptable salt thereof. Formula I encompasses Formula Ia, and references to Formula I below, such as in treatment methods and therapeutic uses, are also interpreted as references to Formula Ia.

[0018] In one embodiment of the compound of Formula I, L is a C2 alkylene group. In an alternative embodiment of the compound of Formula I, L is a C2 alkenyl group. In yet another alternative embodiment of the compound of Formula I, L is -(CH2)3NHCONH(CH2)3-.

[0019] In one embodiment, the compound of formula I is selected from: Or its pharmaceutically acceptable salt.

[0020] In one embodiment of the compound of formula II, Z is H.

[0021] In one embodiment of the compound of formula II, Y is CH2.

[0022] In one embodiment, a compound of formula IIa is provided: Or a pharmaceutically acceptable salt thereof. Formula II covers Formula IIa, and references to Formula II below, such as in treatment methods and therapeutic uses, are also interpreted as references to Formula IIa.

[0023] In one embodiment of the compound of formula II, R 1 It is -NHC(O)CH3, -NHC(O)NHCH2CH3, COOH, -C(O)NH-cyclopropyl, Or phenyl.

[0024] In one embodiment, the compound of formula II is selected from: Or its pharmaceutically acceptable salt.

[0025] In one embodiment, a method of treating a patient requiring treatment of cardiovascular disease is provided, comprising administering an effective amount of a compound of formula I or II or a pharmaceutically acceptable salt thereof. In another embodiment, a method of treating a patient requiring treatment of elevated Lp(a) plasma levels is provided, comprising administering an effective amount of a compound of formula I or II or a pharmaceutically acceptable salt thereof.

[0026] In one embodiment, a compound of formula I or II or a pharmaceutically acceptable salt thereof is provided for use in a therapeutic manner.

[0027] In one embodiment, a compound of formula I or II, or a pharmaceutically acceptable salt thereof, is provided for the treatment of cardiovascular disease. In another embodiment, a compound of formula I or II, or a pharmaceutically acceptable salt thereof, is provided for the treatment of elevated Lp(a) plasma levels.

[0028] In one embodiment, the use of a compound of formula I or II, or a pharmaceutically acceptable salt thereof, in the manufacture of a pharmaceutical agent for treating cardiovascular diseases is provided. In another embodiment, the use of a compound of formula I or II, or a pharmaceutically acceptable salt thereof, in the manufacture of a pharmaceutical agent for treating elevated Lp(a) plasma levels is provided.

[0029] Term "C" 1- C n "Alkyl" refers to a straight-chain or branched saturated hydrocarbon containing 1 to n carbon atoms. C 1- Examples of C4 alkyl groups include, but are not limited to, methyl, ethyl, propyl, butyl, and tert-butyl. 1- Examples of C3 alkyl groups include, but are not limited to, methyl, ethyl, and propyl.

[0030] Term "C" 2- "C6 alkylene" refers to a divalent straight-chain or branched saturated hydrocarbon containing 2 to 6 carbon atoms. The term "C6 alkylene"... 2- "C3 alkylene" refers to divalent straight-chain or branched saturated hydrocarbons containing 2 to 3 carbon atoms.

[0031] Term "C" 2- "C6-olefinic" refers to a divalent straight-chain or branched hydrocarbon containing 2 to 6 carbon atoms and at least one double bond. 2- "C3-olefinic" refers to a divalent straight-chain or branched hydrocarbon containing 2 to 3 carbon atoms and at least one double bond.

[0032] As used herein, the term "elevated Lp(a) plasma level" refers to an Lp(a) plasma level equal to or higher than approximately 50 mg / dL. The compounds described herein may be used for the treatment of lowering Lp(a) plasma levels.

[0033] As used in this article, the term "pharmaceuticalally acceptable salt" refers to a salt of a compound that is acceptable for clinical and / or veterinary use. Examples of pharmaceutically acceptable salts and common methods for their preparation can be found in "Handbook of Pharmaceutical Salts: Properties, Selection and Use," P. Stahl et al., 2nd revised edition, Wiley-VCH, 2011, and SM Berge et al., "Pharmaceutical Salts." Journal of Pharmaceutical Sciences, 1977, 66(1), 1-19. In particular, compounds of formula I or II can be zwitterions, monoacids, diacids or triacids addition salts.

[0034] The pharmaceutical compositions of the present invention can be prepared using pharmaceutically acceptable additives. The term "pharmaceutically acceptable" means one or more carriers, diluents, and / or excipients that are compatible with the other components of the composition and are pharmaceutically harmless to the patient. Examples of pharmaceutical compositions and methods of their preparation are well known to those skilled in the art and can be found, for example, in "Remington: The Science and Practice of Pharmacy," edited by Loyd, V. et al., 22nd edition, Mack Publishing Co., 2012.

[0035] As used herein, the term "effective dose" refers to a dose that is effective in treating the disorder. The effective dose for a particular patient can be determined by an experienced healthcare professional.

[0036] As used herein, the terms “treating,” “to treat,” or “treatment” include slowing, alleviating, preventing, or reversing the progression or severity of existing symptoms, disorders, conditions, or diseases. As used herein, “treating cardiovascular disease” refers to slowing, alleviating, preventing, or reversing the progression of heart or vascular disease.

[0037] As used herein, the term "patient" refers to a mammal. Preferably, the patient is a human.

[0038] The pharmaceutical composition may be formulated as tablets or capsules for oral administration, solutions for oral administration, or injectable solutions. In one embodiment, the composition is suitable for oral administration.

[0039] This invention considers all independent enantiomers, diastereomers, mixtures thereof, and racemates of compounds of formulas I and II, as well as pharmaceutically acceptable salts thereof.

[0040] Some abbreviations can refer to the following: "ACN" refers to acetonitrile; "Apo" refers to apolipoprotein; "BOC" refers to tert-butoxycarbonyl; "DCM" refers to dichloromethane; "DMA" refers to dimethylacetamide; "DMEM" refers to Dulbecco's modified Eagle's medium; "DMF" refers to dimethylformamide; "DMSO" refers to dimethyl sulfoxide; "EACA" refers to ε-aminocaproic acid or 6-aminocaproic acid; "ELISA" refers to enzyme-linked immunosorbent assay; "EtOAc" refers to ethyl acetate; "FBS" refers to fetal bovine serum; "HEC" refers to hydroxyethyl cellulose; "HEK" refers to human embryonic kidney; "HepG2" refers to human liver cancer cell line; "HEPES" refers to 4-(2-hydroxyethyl)-1-piperazine ethanesulfonic acid; "HLB" refers to hydrophilic-lipophilic balance; "hr / s" refers to hours per hour; "HRP" refers to horseradish peroxidase; "IC 50 "MeCN" refers to the reagent concentration that produces 50% of the maximum possible inhibitory response; "MeCN" refers to acetonitrile; "min" refers to minutes; "MTBE" refers to methyl tert-butyl ether; "RT" refers to room temperature; "TEA" refers to triethylamine; "TFA" refers to trifluoroacetic acid; "THF" refers to tetrahydrofuran; and "TMB" refers to 3,3',5,5'-tetramethylbenzidine.

[0041] Those skilled in the art can separate or resolve independent isomers, enantiomers and diastereomers at any convenient point in the synthesis of the following compounds by methods known to the art, such as selective crystallization or chiral chromatography.

[0042] Compounds of Formula I or II are readily converted into pharmaceutically acceptable salts and can be isolated as such. Salt formation can occur upon the addition of a pharmaceutically acceptable acid to form an acid addition salt or upon the addition of a pharmaceutically acceptable base to form a base addition salt. Salt formation can also occur simultaneously with the deprotection of nitrogen or oxygen, i.e., the removal of the protecting group. Examples, reactions, and conditions for salt formation are known to those skilled in the art.

[0043] Compounds of Formula I or II, or any of the described formulas, or salts thereof, can be prepared by various procedures, some of which are illustrated in the preparations and examples below. The specific synthetic steps of each route can be combined in different ways, or combined with steps from different routes, to prepare the compounds or salts of the present invention. The products of the steps in the following preparations can be recovered by conventional methods, including extraction, evaporation, precipitation, chromatography, filtration, preparation, and crystallization.

[0044] In the following figures, unless otherwise specified, all substituents are as defined above. Reagents and starting materials are readily available to those skilled in the art. The following figures, preparations, and examples are provided to further illustrate the invention without limiting its scope. Compounds of the above formulas or salts thereof can be prepared using starting materials or intermediates having the corresponding desired stereochemical configuration.

[0045] Diagram 1

[0046] In Figure 1, step A depicts the conversion of compound (1) to compound (2). Compound (1) is reacted with (E)-1,4-dibromobut-2-ene in the presence of bis(trimethylsilyl)aminolithium as a base and a solvent such as THF to give compound (2).

[0047] Step B describes the formation of compound (3) by reacting compound (2) with potassium acetate and a solvent such as MeCN.

[0048] Step C shows the reaction in which the oxazolidinone in compound (3) is hydrolyzed with LiOH / H2O2 in the presence of a solvent such as THF to obtain compound (4).

[0049] Step D describes the formation of tert-butyl ester, compound (5), by reacting compound (4) with 2-tert-butyl-1,3-diisopropylisourea in a solvent such as 2-methyltetrahydrofuran.

[0050] Step E shows the conversion of compound (5) into compound (6) in the presence of an iridium catalyst and a solvent such as acetone.

[0051] Step F describes the oxidation of the aldehyde in compound (6) with selenium oxide (IV) in the presence of H2O2 to obtain compound (7). In step 2F, compound (7) is converted into compound (7a) (the compound of formula II) by acid deprotection of compound (7) with excess HCl / diethyl ether to obtain the HCl salt compound.

[0052] Step G shows a method for reacting compound (7) with cyclopropylamine in the presence of ethyl chloroformate and a solvent such as DCM to give compound (8). In step 2G, compound (8) is converted into compound (8a) (the compound of formula II) by acid deprotection of compound (8) with excess HCl / diethyl ether to give the HCl salt compound.

[0053] Diagram 2

[0054] In step A of Formula 2, the aldehyde compound (6) is reduced to the alcohol compound (9) in an alcohol solvent using a reducing agent such as sodium borohydride. In step B, the alcohol compound (9) is converted to a methanesulfonyl chloride in the presence of a base such as TEA in a solvent such as DCM. In step C, compound (10) is reacted with aniline (optionally substituted with one or two substituents selected from halogens and OCH3 (described by "R")) using a base (which may include potassium carbonate and sodium iodide) and a solvent such as DCM to give compound (11). BOC deprotection of (11) in step D using HCl gives compound (11a) (the compound of Formula II).

[0055] Methanesulfonate (10) can alternatively be reacted with potassium cyanide in a solvent such as DMF in step E to give cyano compound (13). The reaction with HCl in step F converts the cyano group to a carboxylic acid and removes the BOC group to give compound (13a) (a compound of formula II).

[0056] Methanesulfonate (10) can also be reacted with pyrazole in step G in the presence of a base such as potassium carbonate and a solvent such as MeCN to give compound (12), and then deprotected by BOC in step H to give compound (12a) (the compound of formula II).

[0057] Diagram 3

[0058] In Figure 3, step A describes a method for converting compound (14) into compound (15) by reacting compound (14) with compound (1) in the presence of bis(trimethylsilyl)aminolithium and a solvent such as THF to obtain compound (15).

[0059] Step B shows the reaction in which the oxazolidinone in compound (15) is cleaved in a solvent such as THF using LiOH / H2O2 to obtain compound (16).

[0060] Step C shows the acid deprotection by using excess HCl and diethyl ether to obtain the HCl salt, compound (17) (the compound of formula II).

[0061] Step D shows an alternative route in which the double bond in compound (16) is reduced by hydrogenation in an alcohol solvent such as ethanol with a palladium catalyst and hydrogen to obtain compound (31).

[0062] Step E shows the acid deprotection of compound (31) by using excess HCl and diethyl ether to give the HCl salt, compound (32) (the compound of formula II).

[0063] Diagram 4

[0064] In Figure 4, step A depicts a reductive amination reaction in which compound (6) is converted into compound (18) by reacting with dibenzylamine in the presence of sodium triacetoxyborohydride and a solvent such as DCM, to obtain compound (18).

[0065] Step B shows the conversion of compound (18) into compound (19) in a hydrogenation reaction in the presence of a palladium on carbon catalyst and a solvent such as ethanol.

[0066] Step C shows the effect of reacting with acyl chloride [R] 2 The amine in compound (19) is converted into the amide in compound (21) by reacting with a base such as TEA and a solvent such as DCM.

[0067] Step D shows the formation of compound (20) by reacting 2 equivalents of compound (19) with 1 equivalent of 1,1'-carbonyldiimidazole and a solvent such as THF.

[0068] Step E shows isocyanate (R) 2 The reaction of -N=C=O) with the amine in compound (19) in the presence of a base such as TEA and a solvent such as DCM yields compound (22).

[0069] Steps 2C, 2D, and 2E describe the removal of the BOC of compounds (21), (20), and (22) respectively by using excess HCl to obtain HCl salt compounds (24) (compound of formula II), (23) (compound of formula I), and (25) (compound of formula II), respectively. 2 As defined in Formula II.

[0070] Diagram 5

[0071] In Figure 5, step A depicts the alkylation of compound (1) to give compound (26). Compound (1) is reacted with allyl bromide in the presence of bis(trimethylsilyl)aminolithium as a base and a solvent such as THF to give compound (26).

[0072] Step B shows a hydrolysis reaction in which the oxazolidinone in compound (26) is cleaved in a solvent such as THF using LiOH / H2O2 to give compound (27).

[0073] Step C shows an esterification reaction in which compound (27) is reacted with N,N-dimethylformamide di-tert-butyl acetal in the presence of a solvent such as toluene to give compound (28).

[0074] Step D shows the metathesis reaction that forms compound (29) by contacting compound (28) with a second-generation Grubbs catalyst and a solvent such as DCM.

[0075] Step E describes the acid deprotection of compound (29) by using excess HCl and diethyl ether to give the HCl salt, compound (30) (the compound of formula II).

[0076] Preparation 1 ( 3R )-3-[2-[( 4S )-4-benzyl-2-oxo-oxazolidine-3-yl]-2-oxo-ethyl]pyrrolidine-1-carboxylic acid Uncle Ding ester TEA (56.5 g, 77.9 mL, 559 mmol) was added to (R)-N-boc-3-pyrrolidineacetic acid (CAS Registry No.) maintained at 10°C. 204688-60-8 53.8 g (235 mmol) was added to a solution of tervapotranilyl chloride (33.7 g, 34.2 mL, 279 mmol) in THF (540 mL). After 5 minutes, lithium chloride (11.8 g, 279 mmol) and (4S)-4-benzyloxazolidin-2-one (40.0 g, 223 mmol) in THF (540 mL) were added. The mixture was allowed to warm to room temperature and stirred for 24 hours. After 24 hours, an aqueous HCl solution (500 mL) was added to separate the organic phase from the aqueous phase. The organic phase was washed with an aqueous NaOH solution (500 mL) and a saturated NaCl solution (500 mL), dried over MgSO4, filtered, and concentrated under vacuum. The residue was suspended in a mixture of MeOH and H2O (1:2, 575 mL) and stirred at room temperature for 16–18 hours. The solid was filtered off, washed with hexane (2 × 150 mL), and dried to give the title compound (65.7 g, 76%) as a white solid. ES / MS (m / z): 333 [M+H]- tert-butyl .

[0077] Preparation 2 ( 3R )-3-[( E,1S )-5-acetoxy-1-[( 4S[4-Benzyl-2-oxo-oxazolidine-3-carbonyl]pent-3-enyl]pyrrolidine-1-carboxylic acid Uncle Ding ester Will( 3R )-3-[( E,1S )-1-[( 4S [4-Benzyl-2-oxo-oxazolidine-3-carbonyl]-5-bromo-pent-3-enyl]pyrrolidine-1-carboxylic acid Uncle Ding The ester (5.59 g, 10.7 mmol) was dissolved in MeCN (75 mL) and potassium acetate (1.6 g, 16 mmol) was added. The mixture was stirred at 70 °C for 16–18 hours. The solvent was evaporated, and the residue was dissolved in diethyl ether and washed with H₂O. The organic layer was dried over Na₂SO₄, filtered, and concentrated under vacuum to give the title compound (5.36 g, 99+%) as a pale yellow foamy material. ES / MS (m / z): 401 (M+H-Boc).

[0078] Preparation 3 ( 3R )-3-[( E,1S )-1-tert-butoxycarbonyl-5-hydroxy-pent-3-enyl]pyrrolidine-1-carboxylic acid Uncle Ding ester ( E,2S )-2-[( 3R 1-Tert-Butoxycarbonylpyrrolidine-3-yl]-6-hydroxy-hex-4-enoic acid (0.28 g, 0.935 mmol) was purged under N2, followed by the addition of 2-methyltetrahydrofuran (5.5 mL) and 2- tert-butyl -1,3-Diisopropylisourea (830 µL), and the reaction mixture was stirred at room temperature for 16-18 hours. Add an additional 2- tert-butyl -1,3-Diisopropylisourea (150 mL) was reacted and the reaction was continued with stirring at room temperature for approximately 4 hours. The reaction was then diluted with EtOAc and filtered through diatomaceous earth. The solid was further washed with diethyl ether (x3), and the filtrate was evaporated. The residue was subjected to three more cycles of dissolving in diethyl ether, filtering, and then drying under vacuum. The material was then purified by silica gel chromatography, eluting with DCM / EtOAc (3:1) to give the title compound (0.26 g, 70%) as a yellow viscous oil. ES / MS m / z: 298 (MH, negative ionization mode).

[0079] Preparation 4 (3R )-3-[( 1S [1-tert-butoxycarbonyl-5-oxo-pentyl]pyrrolidine-1-carboxylic acid Uncle Ding ester Will( 3R )-3-[( E,1S )-1-tert-butoxycarbonyl-5-hydroxy-pent-3-enyl]pyrrolidine-1-carboxylic acid Uncle Ding The ester (0.195 g, 0.56 mmol) was dissolved in acetone and H₂O. A dichloro(pentamethylcyclopentadienyl)iridium(III) dimer catalyst (0.006 g, 0.008 mmol) was added, and the reaction mixture was stirred at room temperature for 16–18 hours. The acetone was evaporated, and the residue was separated between diethyl ether and H₂O. After separation, the organic matter was dried over Na₂SO₄, evaporated, and dried under vacuum for approximately 1 hour to give a pale yellow viscous oil (0.186 g). The crude product was purified by silica gel chromatography eluting with DCM / EtOAc (9:1), evaporated, and dried under vacuum to give the title compound (0.14 g, 75%) as a very viscous pale yellow oil. ES / MS m / z: 244 (M+H- 2x tert-butyl ).

[0080] Preparation 5 (3R)-3-[(1S)-1-tert-butoxycarbonyl-5-hydroxypentyl]pyrrolidine-1-carboxylic acid Uncle Ding ester Will( 3R )-3-[( 1S [1-tert-butoxycarbonyl-5-oxo-pentyl]pyrrolidine-1-carboxylic acid Uncle Ding The ester (0.58 g, 1.631 mmol) was dissolved in EtOH (7 mL). Sodium borohydride (0.049 g, 1.29 mmol) was added, and the reaction mixture was stirred at room temperature for 0.5 h. The reaction mixture was diluted with EtOAc and washed with aqueous solutions of H2O and NH4Cl. The organic matter was dried over Na2SO4, evaporated, and dried under vacuum to give a pale yellow viscous oil (0.55 g). The crude product was purified by silica gel chromatography, eluted with DCM / EtOAc (2.7:1), evaporated, and dried under vacuum to give the title compound (0.48 g, 80%) as the pale yellow viscous oil. ES / MS m / z: 246 (M+H- 2x tert-butyl ).

[0081] Preparation of 6 ( 3R )-3-[( 1S )-1-tert-butoxycarbonyl-5-methylsulfonyloxy-pentyl]pyrrolidine-1-carboxylic acid Uncle Ding ester Under N2, ( 3R )-3-[( 1S [1-tert-butoxycarbonyl-5-hydroxy-pentyl]pyrrolidine-1-carboxylic acid Uncle Ding The ester (0.24 g, 0.671 mmol) and TEA (160 µL) were dissolved in anhydrous DCM. The mixture was cooled to 0 °C and methanesulfonyl chloride (70 µL) was added dropwise. The reaction mixture was stirred at 0 °C for 1 hour. The reaction mixture was diluted with DCM and washed with H2O. After separation, the organic matter was dried over Na2SO4, evaporated, and dried under vacuum to give the title compound (0.29 g, 90%) as a very viscous, pale yellow oil. ES / MS m / z: 324 (M+H- 2x tert-butyl ).

[0082] Preparation 7 ( 3R )-3-[( 1S )-1-tert-butoxycarbonyl-5-(3-fluoro-5-methoxy-aniline)pentyl]pyrrolidine-1-carboxylic acid Uncle Ding ester 3-Fluoro-5-methoxy-aniline and ( 3R )-3-[( 1S )-1-tert-butoxycarbonyl-5-methylsulfonyloxy-pentyl]pyrrolidine-1-carboxylic acid Uncle Ding The ester (0.162 g, 0.371 mmol) was purged under N2 with potassium carbonate (0.075 g, 0.542 mmol) and sodium iodide (0.008 g, 0.053 mmol). DMF was added and the reaction mixture was heated at 85 °C for 16–18 h. The reaction mixture was poured into H2O and extracted with diethyl ether. The organic matter was washed with H2O (x2), dried over Na2SO4, evaporated, and dried under vacuum. The crude product was purified by silica gel chromatography eluted with DCM / diethyl ether (95:5) to give a very viscous, pale yellow oil (0.07 g). This product was further purified by reversed-phase chromatography (12 g column; A: water, B: MeCN; 70% 2CVs, 70–80% 4CVs, 80% 4CVs) to give the title compound (0.038 g, 22.91%) as the pale yellow viscous oil. ES / MSm / z: 481 (M+H).

[0083] Preparation of 8 (5S)-6-tert-butoxy-5-[( 3R )-1-tert-butoxycarbonylpyrrolidine-3-yl]-6-oxo-hexanoic acid Towards( 3R )-3-[( 1S [1-tert-butoxycarbonyl-5-oxo-pentyl]pyrrolidine-1-carboxylic acid Uncle Ding The ester (0.15 g, 0.422 mmol) was added to a solution of selenium dioxide (IV) (0.003 g, 0.270 mmol) in THF (2 mL), followed by the addition of H2O2 (34% aqueous solution, 100 µL), and the reaction mixture was heated at 70 °C for approximately 2.5 h. The reaction mixture was diluted with EtOAc and washed with H2O. After separation, the organic matter was washed with H2O, dried over Na2SO4, evaporated, and dried under vacuum to give a clear viscous oil (160 mg). The crude product was purified by silica gel chromatography eluting with DCM / diethyl ether (9:1), evaporated, and dried under vacuum to give the title compound (0.125 g, 80%) as the clear viscous oil. ES / MS m / z: 260 (M+H- 2x Uncle Butyl ).

[0084] Preparation 9 ( 3R )-3-[( 1S )-1-tert-butoxycarbonyl-5-(cyclopropylamino)-5-oxo-pentyl]pyrrolidine-1-carboxylic acid Uncle Ding ester ( 5S )-6-tert-butoxy-5-[( 3RA solution of 1-tert-butoxycarbonylpyrrolidine-3-yl]-6-oxohexanoic acid (0.215 g, 0.579 mmol) and TEA (170 µL) in anhydrous DCM (3.5 mL) was purged under N2. Ethyl chloroformate (80 µL) was added dropwise and the mixture was stirred for 0.75 h. Next, cyclopropylamine (60 µL) was added dropwise and the reaction mixture was stirred at room temperature for about 2.5 h. The reaction mixture was diluted with DCM and washed with H2O + 10% Na2CO3. After phase separation, the organic phase was washed with H2O, dried over Na2SO4, evaporated and dried under vacuum to give a pale yellow viscous oil (250 mg). The crude product was purified by silica gel chromatography eluting with DCM / diethyl ether (4:1), evaporated and dried under vacuum to give the title compound (0.13 g, 55%) as a very viscous pale yellow oil. ES / MS m / z: 411 (M+H).

[0085] Preparation 10 ( 3R )-3-[( 1S )-1-tert-butoxycarbonyl-5-cyano-pentyl]pyrrolidine-1-carboxylic acid Uncle Ding ester ( 3R )-3-[( 1S )-1-tert-butoxycarbonyl-5-methylsulfonyloxy-pentyl]pyrrolidine-1-carboxylic acid Uncle Ding The ester (0.146 g) and potassium cyanide (0.028 g, 0.335 mmol) were purged under N2. Anhydrous DMF (1.2 mL) was added, and the reaction mixture was heated at 90 °C for 16–18 h. The reaction mixture was diluted with diethyl ether and washed with H2O. After separation, the organic matter was washed with H2O (x2), dried over Na2SO4, evaporated, and dried under vacuum to give a pale yellow viscous oil (80 mg). The crude product was purified by silica gel chromatography eluting with DCM / EtOAc (95:5); and dried under vacuum to give the title compound (0.058 g, 47%) as a very viscous, transparent oil. ES / MS m / z: 255 (M+H- 2x tert-butyl ).

[0086] Preparation 11 ( 3R )-3-[( 1S )-1-tert-butoxycarbonyl-5-pyrazol-1-yl-pentyl]pyrrolidine-1-carboxylic acid Uncle Ding ester ( 3R )-3-[( 1S )-1-tert-butoxycarbonyl-5-methylsulfonyloxy-pentyl]pyrrolidine-1-carboxylic acid Uncle Ding A mixture of ester (0.155 g, 0.356 mmol) and pyrazole (0.04 g, 0.588 mmol) was purged under N2 in a refrigerator with potassium carbonate (0.095 g, 0.687 mmol). MeCN (2.5 mL) was added, and the reaction mixture was heated at 75 °C for 16–18 h, followed by heating under N2 atmosphere for approximately 16 h. The reaction was then heated at 80 °C under N2 atmosphere for another 16–18 h. The reaction mixture was then diluted with EtOAc and washed with H2O. After separation, the organic matter was dried over Na2SO4, evaporated, and dried under vacuum to give a very pale yellow viscous oil (150 mg). The crude product was purified by silica gel chromatography eluting with DCM / EtOAc (9:1); the solvent was evaporated, and the residue was dried under vacuum to give the title compound (0.071 g, 50%) as a very viscous, transparent oil. ES / MS m / z: 408 (M+H).

[0087] Preparation 12 ( 3R )-3-[( 1S 1-Terbutoxycarbonyl-5-(ethylcarbamoylamino)pentyl]pyrrolidine-1-carboxylic acid Uncle Ding ester In an N2 atmosphere, ( 3R )-3-[( 1S [5-amino-1-tert-butoxycarbonyl-pentyl]pyrrolidine-1-carboxylic acid Uncle Ding A mixture of ester (0.145 g, 0.407 mmol) and TEA (120 µL) was dissolved in anhydrous DCM. Isocyanoethane (60 µL) was then added dropwise, and the reaction mixture was stirred at room temperature for approximately 1.5 hours. The reaction mixture was diluted with more DCM and washed with H₂O. The organic matter was dried over Na₂SO₄, evaporated, and dried under vacuum to give a pale yellow viscous oil (190 mg). The crude product was purified by silica gel chromatography eluting with DCM / MeOH (65:35). The solvent was evaporated, and the residue was dried under vacuum to give the title compound (0.124 g, 80%) as a very viscous, transparent oil. ES / MS m / z: 428 (M+H).

[0088] Preparation 13 1-[( E)-4-bromobut-2-enoxy]-3-fluoro-5-methoxy-benzene (E)-1,4-dibromobut-2-ene (CAS Registry No.) 821-06-7 ; 0.00192 g, 1.92 mmol) and 3-fluoro-5-methoxy-phenol (CAS registry number) 850793-25-8 A mixture of 0.833 g and 0.833 g was dissolved in MeCN (50 mL) in a refrigerator under a nitrogen atmosphere. Potassium carbonate (0.00135 g, 0.0097 mmol) was added in one step, and the final reaction mixture was stirred at 60 °C for approximately 4.5 hours. The solvent was evaporated, and the residue was separated into phases between diethyl ether and H₂O. After phase separation, the organic matter was washed with water, dried over Na₂SO₄, filtered, and evaporated to give a pale yellow viscous oil (2 g). The crude product was purified by silica gel chromatography eluting with DCM / hexane (1:9) to give the title compound (0.702 g, 45%) as a very pale yellow oil. 1H NMR(400.13 MHz, CDCl3) d 6.29-6.25 (m, 3H), 6.14-6.09 (m, 1H), 6.02-5.96 (m,1H), 4.53 (dd, J= 1.1, 5.1 Hz, 2H), 4.01 (dd, J= 0.8, 7.3 Hz, 2H), 3.79 (s, 3H).

[0089] Preparation 14 2-[( E [4-Bromobut-2-enyl]isoindoline-1-one (E)-1,4-dibromobut-2-ene (CAS Registry No.) was subjected to a nitrogen atmosphere. 821-06-7 1.12 g) was dissolved in anhydrous THF (20 mL). Then, sodium hydride (60% mineral oil, 0.175 g) was added at room temperature, and the mixture was stirred for 0.25 hours. Solid isoindolin-1-one (CAS registry number) was added in a single batch. 480-91-1(0.551 g, 4.13 mmol) and the reaction mixture was stirred at room temperature for approximately 4.5 hours. The reaction mixture was diluted with diethyl ether, washed with H₂O (x2), dried over Na₂SO₄, evaporated, and stirred under vacuum to give a crude residue as a yellow solid (1 g). The crude product was purified by silica gel chromatography eluting with EtOAc (100 mL). The EtOAc was evaporated, and the residue was dried under vacuum to give the title compound (0.22 g, 19%) as a very viscous, pale yellow oil. ES / MS m / z: 266, 268 (M+H).

[0090] Preparation 15 3-[( E )-4-bromobut-2-enyl]-1,3-benzoxazol-2-one In the refrigerator, under N2, ( E 1,4-Dibromobut-2-ene (CAS Registry Number) 821-06-7 ; 1.5 g, 7.02 mmol) and 3 H -1,3-Benzoxazol-2-one (CAS Registry Number) 59-49-4 0.69 g) was dissolved in MeCN (40 mL). Potassium carbonate (0.99 g, 7.16 mmol) was then added in a single batch, and the reaction mixture was stirred at 50 °C for approximately 4.25 h. The solvent was evaporated, and the residue was separated between diethyl ether and water. After phase separation, the organic matter was washed with H₂O, dried over Na₂SO₄, filtered, and evaporated to give a crude product (1.5 g) as a brown solid. The crude product was purified by silica gel chromatography eluting with DCM / hexane (3:1) to give the title compound (0.72 g, 50%) as a very pale yellow solid. ES / MS m / z: 268, 270 (M+H).

[0091] Preparation of 16 ( 3R )-3-[( 1S 1-tert-butoxycarbonyl-5-(dibenzylamino)pentyl]pyrrolidine-1-carboxylic acid Uncle Ding ester Will( 3R )-3-[( 1S [1-tert-butoxycarbonyl-5-oxo-pentyl]pyrrolidine-1-carboxylic acid Uncle DingA mixture of ester (0.15 g, 0.52 mmol) and dibenzylamine (0.12 mL, 0.61 mmol) was dissolved in DCM (3 mL) and stirred at room temperature for 30 min. Sodium triacetoxyborohydride (0.175 g, 0.80 mmol) was added in a single addition and stirred at room temperature for 16–18 h. The mixture was diluted with DCM, and an aqueous solution of NaHCO3 was added to separate the phase. The organic phase was dried over Na2SO4, filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography eluting with 5% diethyl ether / DCM to give the title compound (0.17 g, 54% yield) as a pale yellow viscous oil. ES / MS (m / z): 537 (M+H).

[0092] Preparation of 17 ( 3R )-3-[( 1S [5-amino-1-tert-butoxycarbonyl-pentyl]pyrrolidine-1-carboxylic acid Uncle Ding ester (3R)-3-[(1S)-1-tert-butoxycarbonyl-5-(dibenzylamino)pentyl]pyrrolidine-1-carboxylic acid Uncle Ding The ester (0.170 g, 0.30 mmol) was dissolved in EtOH (3 mL) and purged with N2. Palladium / carbon (5% w / w, 140 mg, 0.06 mmol) was added and purged with hydrogen. The mixture was pressurized with 70 psi hydrogen and stirred at room temperature for 4.5 h, filtered through a diatomaceous earth pad and washed with DCM. The filtrate was evaporated to give the title compound (0.12 g, 99+% yield) as a dark brown viscous oil. ES / MS (m / z): 357 (M+H).

[0093] Preparation of 18 (3R)-3-[(1S)-5-[[(5S)-6-tert-butoxy-5-[(3R)-1-tert-butoxycarbonylpyrrolidine-3-yl]-6-oxo-hexyl]carbamoylamino]-1-tert-butoxycarbonyl-pentyl]pyrrolidine-1-carboxylic acid Uncle Ding ester (3R)-3-[(1S)-5-amino-1-tert-butoxycarbonyl-pentyl]pyrrolidine-1-carboxylic acid under N2 Uncle DingThe ester (0.12 g, 0.29 mmol) was dissolved in anhydrous THF (3 mL). 1,1'-carbonyldiimidazole (0.025 g, 0.15 mmol) was added and the mixture was stirred at 70 °C for 5 hours. The reaction mixture was poured into water and extracted with diethyl ether (x2). The combined organic matter was washed with H₂O, dried over Na₂SO₄, filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography with sequential elution of DCM / diethyl ether (3:1) and EtOAc to give the title compound (0.050 g, 49%). 1H NMR (400.21 MHz, CDCl3): 3.67-3.37 (m, 4H), 3.32-3.07 (m, 6H), 2.99-2.81 (m, 2H), 2.35-2.21(m, 2H), 2.18-2.08 (m, 2H), 1.99-1.86 (m, 2H), 1.69-1.23 (m, 50H).

[0094] Preparation of 19 (E,2S,7S)-2,7- pair[ ( 3R )-1-tert-butoxycarbonylpyrrolidine-3-yl]oct-4-enic acid Second Uncle Ding ester (3R)-3-[(1S)-1-tert-butoxycarbonylbut-3-enyl]pyrrolidine-1-carboxylic acid Uncle Ding The ester (0.204 g, 0.627 mmol) was purged in a refrigerator under a nitrogen atmosphere. Then, anhydrous DCM was added, followed by Grubbs catalyst® M2a (C848, CAS Registry No.). # 246047-72-3 (0.055 g, 0.065 mmol). Next, the reaction mixture was heated under reflux for 16–18 hours. The solvent was diluted with DCM and washed with H2O. The organic matter was dried over Na2SO4 and evaporated to dryness to give a dark brown viscous oil (0.3 g). The residue was purified by silica gel chromatography eluting with DCM / EtOAc (95:5). The solvent was evaporated and the residue was dried under vacuum to give the title compound (0.11 g, 55%) as a brown foamy substance. ES / MS m / z: 623 (M+H).

[0095] Preparation 20 ( 2S, 7S )-2,7-double[( 3R )-1-tert-butoxycarbonylpyrrolidine-3-yl]octanediic acid Second Uncle Ding ester (E,2S,7S)-2,7- pair[ ( 3R )-1-tert-butoxycarbonylpyrrolidine-3-yl]oct-4-enic acid Second Uncle Ding A mixture of ester (400 mg, 0.64 mmol) and palladium catalyst (5% by mass on carbon, 700 mg, 0.33 mmol) was purged under a nitrogen atmosphere. EtOH (7 mL) was added and the mixture was purged with hydrogen under vigorous stirring. The mixture was stirred under a hydrogen balloon at room temperature for 2 hours and then filtered through a diatomaceous earth pad. The pad was washed with DCM and the filtrate was evaporated. The residue was dissolved in DCM and dried over Na2SO4, filtered, and concentrated. The residue was purified by silica gel chromatography using 10% EtOAc / DCM to give the title compound (365 mg, 91%) as a white foamy solid. ES-MS m / z 525 (M+H-Boc).

[0096] Preparation 21 ( 3R )-3-[( E,1S )-1-[( 4S [4-Benzyl-2-oxo-oxazolidine-3-carbonyl]-5-(3-fluoro-5-methoxy-phenoxy)pent-3-enyl]pyrrolidine-1-carboxylic acid Uncle Ding ester Under N2, ( 3R )-3-[2-[( 4S )-4-benzyl-2-oxo-oxazolidine-3-yl]-2-oxo-ethyl]pyrrolidine-1-carboxylic acid Uncle Ding The ester (0.815 g; 2.1 mmol) was dissolved in anhydrous THF (9 mL) and cooled to approximately 0 °C. Bis(trimethylsilyl)aminolithium (2.3 mL, 1 M) was added dropwise, and the mixture was stirred for approximately 1 hour. Solid 1-[( E[-4-bromobut-2-enoxy]-3-fluoro-5-methoxybenzene (0.607 g, 2.21 mmol) was reacted and stirred at 0 °C for approximately 1 hour, then allowed to stand at room temperature for approximately 16–18 hours. The reaction mixture was poured into saturated NaHCO3 and extracted with diethyl ether. The organic matter was further washed with H2O, dried over Na2SO4, and the solvent was evaporated to give a yellow viscous oil (1.25 g). The crude material was purified by silica gel chromatography eluting with DCM / EtOAc (96:4). The solvent was evaporated, and the residue was dried under vacuum to give the title compound (0.53 g, 40%) as a white foamy substance. ES / MS m / z: 483 (M+H-Boc).

[0097] Prepare preparations 22-26 in Table 1 using appropriate reagents and reaction times adjusted to achieve completion, similar to the method used in preparation 21.

[0098] Table 1

[0099] Preparation 27 (E,2S)-2-[(3R)-1-tert-butoxycarbonylpyrrolidine-3-yl]-6-hydroxy-hex-4-enoic acid (3R)-3-[(E,1S)-5-acetoxy-1-[(4S)-4-benzyl-2-oxo-oxazolidine-3-carbonyl]pent-3-enyl]pyrrolidine-1-carboxylic acid Uncle Ding The ester (5.36 g, 10.7 mmol) solution in THF (80 mL) was cooled to 0 °C. H₂O₂ (34 wt%, 9.5 mL, 110 mmol) was added, followed by a solution of lithium hydroxide (750 mg, 30.7 mmol) in H₂O (14 mL). The reaction mixture was stirred for 16–18 hours while reaching room temperature. The mixture was then poured into H₂O, and EtOAc and 1 M HCl aqueous solution were added and shaken vigorously. The organic phase was separated and washed with H₂O (x2). The organic phase was dried over Na₂SO₄, filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography eluting with 3% MeOH / DCM containing 0.25% acetic acid to give a heavy oil containing a small amount of acetic acid. The residue was then dissolved in DCM and washed with H₂O (x2). The organic phase was dried over Na₂SO₄, filtered, and concentrated under vacuum to give the title compound (2.1 g) as the heavy oil. ES / MS (m / z): 298 (MH).

[0100] Prepare preparations 28-33 in Table 2 using appropriate reagents and reaction times adjusted to achieve completion, similar to the method used in preparation 27.

[0101] Table 2

[0102] Preparation of 32 ( 2S )-2-[(3R)-1-tert-butoxycarbonylpyrrolidine-3-yl]-5-phenyl-pentanoic acid A solution of (E,2S)-2-[(3R)-1-tert-butoxycarbonylpyrrolidine-3-yl]-5-phenyl-pent-4-enoic acid (155 mg, 0.449 mmol) in ethanol (7 mL) was purged with nitrogen, and then palladium (5% by mass on carbon, 205 mg, 0.096 mmol) was added. The reaction was purged with hydrogen and then stirred under a hydrogen balloon for 3.5 h. The mixture was filtered through a diatomaceous earth pad, washed with DCM, and the filtrate was concentrated under reduced pressure. The residue was placed in DCM, dried over Na2SO4, filtered, and concentrated under reduced pressure to give the title compound (149 mg, 96%) as a white foam. ES-MS m / z 292 (M+H- tert-butyl ).

[0103] Preparation of 34 (3R)-3-[(1S)-1-tert-butoxycarbonylbut-3-enyl]pyrrolidine-1-carboxylic acid Uncle Ding ester N,N-dimethylformamide di-tert-butyl acetal (42.0 mmol, 90% by mass, 11.2 mL, 9.49 g) was added to a solution of (2S)-2-[(3R)-1-tert-butoxycarbonylpyrrolidine-3-yl]pent-4-enoic acid (D, 10.5 mmol, 10.5 mmol, 2.83 g) in toluene (105 mL) at 80 °C, and the mixture was heated at 80 °C for 1 hour. More N,N-dimethylformamide di-tert-butyl acetal (42.0 mmol, 90% by mass, 11.2 mL, 9.49 g) was added, and the mixture was heated at 80 °C for 3 hours. The reaction mixture was concentrated, and the residue was purified by silica gel chromatography (5-20% acetone / hexane gradient) to give the title compound (1.8 g, 53%) as a colorless oil. ES-MS m / z 348 (M + Na+ ).

[0104] Preparation of 35 (3R)-3-[(1S)-5-acetamido-1-tert-butoxycarbonyl-pentyl]pyrrolidine-1-carboxylic acid Uncle Ding ester (3R)-3-[(1S)-5-amino-1-tert-butoxycarbonyl-pentyl]pyrrolidine-1-carboxylic acid Uncle Ding Acetyl chloride (25 µL, 27.5 mg, 0.35 mmol) was added dropwise to a mixture of ester (110 mg, 0.308 mmol) and TEA (100 µL, 72.5 mg, 0.72 mmol) in DCM (2 mL), and the mixture was stirred at room temperature for 2 hours. The mixture was diluted with DCM and washed with water. The organic matter was dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography using a 33-100% diethyl ether / DCM gradient to give the title compound (75 mg, 68%) as a pale yellow oil. ES-MS m / z 299 (M-BOC+H).

[0105] Preparation of 36 (3R)-3-[(1S)-1-tert-butoxycarbonyl-5-(3-fluoro-5-methoxy-phenoxy)pentyl]pyrrolidine-1-carboxylic acid Uncle Ding ester (3R)-3-[(1S)-1-tert-butoxycarbonyl-5-hydroxy-pentyl]pyrrolidine-1-carboxylic acid Uncle Ding A mixture of ester (265 mg, 0.74 mmol), 3-fluoro-5-methoxy-phenol (129 mg, 0.91 mmol), and triphenylphosphine (241 mg, 0.92 mmol) was purged under nitrogen atmosphere, followed by the addition of anhydrous THF (4 mL) and cooling in an ice / water bath. Diisopropyl azodicarbonate (190 µL, 196 mg, 0.97 mmol) was added dropwise, and the reaction was stirred under cooling for 0.5 h. The reaction was stirred overnight at room temperature, then poured into a 10% aqueous solution of Na₂CO₃ and extracted with diethyl ether. The organic matter was dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography with DCM as eluent, followed by two further purifications by reversed-phase HPLC to give the title compound (96 mg, 27%) as a brown oil. ES-MS m / z 382 (M-BOC+H).

[0106] Preparation of 37 (2S)-2-[(3R)-1-tert-butoxycarbonylpyrrolidine-3-yl]-6-(2-oxo-1,3-benzoxazol-3-yl)hexanoic acid A mixture of (E,2S)-2-[(3R)-1-tert-butoxycarbonylpyrrolidine-3-yl]-6-(2-oxo-1,3-benzoxazol-3-yl)hex-4-enoic acid (52 mg, 0.12 mmol) in ethanol (2 mL) was purged with nitrogen. Then, palladium (5% by mass on carbon, 75 mg, 0.035 mmol) was added, the reaction was purged with hydrogen, and maintained under a hydrogen balloon for 2 hours. The mixture was filtered through a diatomaceous earth pad, which was washed with DCM. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel chromatography using 3% MeOH / DCM to give the title compound (42 mg, 82%) as a white foam. ES-MS m / z 319 (M-BOC+H).

[0107] Example 1 (2S) -6-(3-Fluoro-5-methoxy-aniline)-2-[( 3R )-pyrrolidine-3-yl]hexanoic acid; dihydrochloride Add concentrated HCl (35% aqueous solution, 0.3 mL) to (3R)-3-[(1S)-1-tert-butoxycarbonyl-5-(3-fluoro-5-methoxy-aniline)pentyl]pyrrolidine-1-carboxylic acid Uncle Ding The ester (0.038 g, 0.079 mmol) was in a solution of diethyl ether (3 mL). The reaction mixture was stirred at room temperature for 16–18 hours. The ether phase was removed by pipette, and the aqueous phase was washed with diethyl ether (x2). H2O was then evaporated with a stream of N2 and dried under vacuum to give the title compound (0.029 g, 95.17%) as a light brown solid. ES / MS m / z: 325 (M+H).

[0108] Examples 2-17 in Table 3 were prepared in a manner substantially as described in Example 1, using suitable starting materials protected with BOC and tert-butyl ester while deprotecting amine and acid functional groups, or using suitable starting materials protected with unprotected carboxylic acids and BOC.

[0109] Table 3 a. Starting material used: (3R)-3-[(1S)-1-tert-butoxycarbonyl-5-cyano-pentyl]pyrrolidine-1-carboxylic acid Uncle Ding ester.

[0110] In vitro Lp(a) assembly detection The ability of the compounds to inhibit Lp(a) particle formation in vitro was assessed using a cell-free assembly assay. Conditioned medium (supplemented with 10% FBS, 20 mM HEPES, and 1x penicillin / streptomycin in DMEM) was collected from confluent wild-type HepG2 cells (the source of endogenously expressed ApoB) and from a stable HEK293 cell line expressing human Apo(a) protein with 17 Kringle repeats (selected on 1 mg / ml genistein) after 24 hours of culture at 37°C and 5% CO2. In vitro assembly assays were performed by combining equal portions of HepG2 and HEK293 conditioned medium with serially added test compounds (final concentrations 0.01–100 nM). The reaction was incubated at 37°C for 2 hours and then terminated by adding 6-aminocaproic acid (EACA) to a final concentration of 150 mM. Lp(a) was detected using a sandwich ELISA with anti-Lp(a) capture antibody and HRP-labeled anti-ApoB detection antibody. ELISA was performed using TMB colorimetry, terminated with 1 N sulfuric acid, and the signal was read at 450 nm using a Molecular Devices reader. The percentage inhibition of Lp(a) formed under each experimental condition was determined by setting the assembly reaction without inhibitors (with a matched DMSO concentration of 1%) as 0% inhibition and the assembly reaction with minimal HepG2 conditioned medium (50-fold dilution) as 100% inhibition. Data were fitted to a 4-parameter curve to determine the IC50 values ​​summarized in Table 4. 50 Values. Adding the test compounds from the examples to conditioned medium containing ApoB and Apo(a) resulted in concentration-dependent inhibition of in vitro Lp(a) formation as summarized in Table 4. The results indicate that these compounds inhibit the assembly of Lp(a) by Apo(a) and LDL particles. Example <![CDATA[IC 50 nM (SEM, n)]]> 1 12.8 (5.02, n = 4) 2 161 (28.1, n = 4) 3 110 (9.76, n = 4) 4 61.5 (18.6, n = 4) 5 27.6 (2.39, n = 2) 6 66.6 (8.33, n = 2) 7 137 (10.9, n = 4) 8 215 (10.5, n = 2) 9 26.9 (8.26, n = 4) 10 61.5 (4.57, n = 4) 11 194 (3.89, n = 2) 12 96.4 (43.8, n = 2) 13 685 (n = 1) 14 618 (n = 1) 15 0.749 (0.315, n = 5) 16 1.33 (0.892, n = 3) 17 0.638 (0.134, n = 4)

Claims

1. A compound of the formula: ###0001### or a pharmaceutically acceptable salt thereof. wherein L is C2-C6alkylene, C2-C6alkenylene or -(CH2) n NHCONH(CH2) n -; n is independently at each occurrence 2 to 4; Y is independently at each occurrence CH2, O, or S; Z is independently at each occurrence H, C 1- C4alkyl, OH, or cyclopropyl.

2. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein Z is H at each occurrence.

3. The compound according to claim 1 or claim 2, or a pharmaceutically acceptable salt thereof, wherein Y is CH2 at each occurrence.

4. The compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, wherein L is C2 alkylene.

5. The compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, wherein L is C2 alkenylene.

6. The compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, wherein L is -(CH2)3NHCONH(CH2)3-.

7. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of: ###0002### ###0003### 。 8. A compound of the formula: ###0004### or a pharmaceutically acceptable salt thereof. wherein L 1 is C2-C3alkylene or C2-C3alkenylene; R 1 is -NHC(O)R 2 , -NHC(O)NHR 2 , COOH, -C(O)NH-cyclopropyl, , , -O-phenyl, -NH-phenyl or phenyl, wherein said phenyl is optionally substituted with one or two substituents selected from halogen and OCH3; R 2 is C1-C3 alkyl; Y is CH2, O, or S; Z is H, C 1- C4alkyl, OH or cyclopropyl.

9. The compound according to claim 8, or a pharmaceutically acceptable salt thereof, wherein Z is H.

10. The compound according to claim 8 or claim 9, or a pharmaceutically acceptable salt thereof, wherein Y is CH2.

11. The compound or pharmaceutically acceptable salt thereof of any one of claims 8 to 10, wherein R 1 is -NHC(O)CH3, -NHC(O)NHCH2CH3, COOH, -C(O)NH-cyclopropyl, or phenyl.

12. A pharmaceutical composition comprising a compound according to any one of claims 1 to 11, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier, diluent, or excipient.

13. A method of treating cardiovascular disease in a patient comprising administering to the patient an effective amount of a compound according to any one of claims 1 to 11, or a pharmaceutically acceptable salt thereof.

14. A method of treating elevated plasma levels of Lp(a) in a patient comprising administering to the patient an effective amount of a compound according to any one of claims 1 to 11, or a pharmaceutically acceptable salt thereof.

15. A compound according to any one of claims 1 to 11, or a pharmaceutically acceptable salt thereof, for use in therapy.

16. A compound according to any one of claims 1 to 11, or a pharmaceutically acceptable salt thereof, for use in the treatment of cardiovascular disease.

17. A compound according to any one of claims 1 to 11, or a pharmaceutically acceptable salt thereof, for use in the treatment of elevated plasma levels of Lp(a).

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