Pyrrolidine compounds having Lp(a)-reducing activity

Abstract

This disclosure is based on the formula: [Case 1] This invention provides compounds of TIFF2026524959000037.tif26128 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] The present invention relates to pyrrolidine compounds, pharmaceutically acceptable salts thereof, pharmaceutical compositions, and the therapeutic use of these compounds, particularly their use in reducing plasma levels of lipoprotein(a) (Lp(a)).

Background Art

[0002] There have been remarkable advancements in the treatment of cardiovascular disease (CVD). Despite these treatment advancements, patients continue to experience cardiovascular disease events such as angina, myocardial infarction, and stroke, which can be fatal if left untreated. Dyslipidemia or lipid abnormalities remain major risk factors for CVD. Dyslipidemia can be divided into four common risk factors: an increase in low-density lipoprotein cholesterol (LDL-c), a decrease in high-density lipoprotein cholesterol (HDL-c), an increase in triglyceride (TG), and an increase in Lp(a). There are various treatment regimens targeting high LDL-c, low HDL-c, and high triglycerides. There are few approved treatment options for patients with high Lp(a) concentrations. In some cases, apheresis can be used to filter the blood and remove LDL and Lp(a), however, the effect is temporary and typically needs to be repeated every two weeks. Currently, there is no drug treatment specifically approved for reducing Lp(a) levels.

[0003] Lp(a) is a genetically determined, independent cardiovascular risk factor. Elevated serum Lp(a) levels above 50 mg / dL or 125 nmol / L, found in approximately 20% of the population, are associated with at least a 1.6-fold increased risk of the first cardiovascular event (Kronenberg, F..Clin.Res.Cardiol.Suppl.14,5-12(2019)) and a more than 1.42-fold increased risk of the second event (Madsen, CM et al.Arterioscler.Thromb.Vasc.Biol.40,255-266(2020)). Lp(a) can exhibit both prothrombinogenic and antithrombotic properties, as well as both atherogenic and atherothrombotic properties. Lp(a) inhibits fibrinolysis and can accumulate in the vascular wall, inducing thrombus formation and atherosclerotic lesions. Plasma levels of Lp(a) vary substantially 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 contains an LDL lipid core with an associated apolipoprotein B (apoB), but unlike LDL-c, Lp(a) contains its own apolipoprotein (a) (apo(a)) bound to apoB via a disulfide bond. apo(a) is synthesized in the liver. Aggregations of apo(a) and Lp(a) from LDL particles can occur in hepatocytes, on the cell wall, or in plasma. Inhibition of LDL particle aggregation with apo(a) lowers Lp(a) levels.

[0005] International Publication No. 2020 / 247429 discloses compounds that inhibit Lp(a) formation by blocking the interaction between Apo(a) and ApoB.

[0006] Patients with cardiovascular disease, particularly those with dyslipidemia or dyslipidemia, require additional treatment options. Additional treatment options are needed for patients whose cardiovascular risk is not adequately managed with current standard treatments, including diet, exercise, and / or the use of one or more medications such as statins, fibrates, and niacin. More specifically, there is a need for further alternative, pharmaceutically acceptable compounds that inhibit Lp(a) formation and thus lower plasma Lp(a) levels. [Overview of the Initiative]

[0007] Therefore, in the first embodiment, Equation I:

[0008] [ka] (In the formula,

[0009] [ka] is phenyl,

[0010] [ka] or

[0011] [ka] And, In each occurrence, Z is independently H and C 1~4 It is alkyl, OH, or cyclopropyl, Y is, in each instance, independently CH2, O, or S. Compounds thereof, or pharmaceutically acceptable salts thereof, are provided.

[0012] In a second embodiment, a pharmaceutical composition is provided comprising a compound of formula I, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier, diluent, or excipient.

[0013] In a third aspect, there is provided a method of treating a cardiovascular disease in a patient, the method comprising administering to the patient an effective amount of a compound of formula I, or a pharmaceutically acceptable salt thereof.

[0014] In a fourth aspect, there is provided a method of treating an elevated Lp(a) plasma level in a patient, the method comprising administering an effective amount of a compound of formula I, or a pharmaceutically acceptable salt thereof.

[0015] In a fifth aspect, there is provided a compound of formula I, or a pharmaceutically acceptable salt thereof, for use in therapy.

[0016] In a sixth aspect, there is provided a compound of formula I, or a pharmaceutically acceptable salt thereof, for use in the treatment of a cardiovascular disease.

[0017] In a seventh aspect, there is provided a compound of formula I, or a pharmaceutically acceptable salt thereof, for use in the treatment of an elevated Lp(a) plasma level.

MODE FOR CARRYING OUT THE INVENTION

[0018] In one embodiment of the compound of formula I, Z is H at each occurrence.

[0019] In one embodiment of the compound of formula I, Y is CH2 at each occurrence.

[0020] In one embodiment, formula II:

[0021]

CHEMICAL FORMULA

[0022] In some embodiments, the compound of formula I is

[0023] [ka] Select from, or a pharmaceutically acceptable salt thereof.

[0024] In one embodiment, a method is provided for treating a patient requiring treatment for cardiovascular disease, comprising administering an effective amount of a compound of formula I or a pharmaceutically acceptable salt thereof. In another embodiment, a method is provided for treating a patient requiring treatment for elevated Lp(a) plasma levels, comprising administering an effective amount of a compound of formula I or a pharmaceutically acceptable salt thereof.

[0025] In one embodiment, a compound of formula I, or a pharmaceutically acceptable salt thereof, is provided for use in therapy.

[0026] In one embodiment, a compound of formula I, or a pharmaceutically acceptable salt thereof, is provided for use in the treatment of cardiovascular disease. In another embodiment, a compound of formula I, or a pharmaceutically acceptable salt thereof, is provided for use in the treatment of elevated Lp(a) plasma levels.

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

[0028] "C 1~4 The term "alkyl" refers to a straight-chain or branched-chain saturated hydrocarbon containing 1 to 4 carbon atoms. 1~4 Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, butyl, and tert-butyl.

[0029] As used herein, the term “elevated Lp(a) plasma levels” means Lp(a) plasma levels of approximately 50 mg / dL or higher. The compounds provided herein can be used for therapeutic purposes to lower Lp(a) plasma levels.

[0030] As used herein, the term “pharmaceutically acceptable salt” refers to a salt of a compound that is acceptable for clinical and / or veterinary use. Examples of pharmaceutically acceptable salts and general methodologies for preparing them can be found in “Handbook of Pharmaceutical Salts: Properties, Selection and Use” P. Stahl, et al., 2nd Revised Edition, Wiley-VCH, 2011 and SMBerge, et al., “Pharmaceutical Salts,” Journal of Pharmaceutical Sciences, 1977, 66(1), 1-19. In particular, the compound of formula I may be a zwitterionic, monoacid, diacid, or triacid addition salt.

[0031] The pharmaceutical compositions of the present invention may 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 not pharmaceutically harmful to the patient. Examples of pharmaceutical compositions and processes for 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," Loyd, V., et al. Eds., 22nd Ed., Mack Publishing Co., 2012.

[0032] As used herein, the term “effective dose” refers to the dosage that is effective in treating a disorder. The effective dose for a particular patient can be determined by a skilled healthcare professional.

[0033] As used herein, the terms “to treat,” “to cure,” or “to treat” include slowing, reducing, preventing, or reversing the progression or severity of an existing symptom, disorder, condition, or disease. As used herein, “to treat a cardiovascular disease” means slowing, reducing, preventing, or reversing the progression of a disease of the heart or blood vessels.

[0034] As used herein, the term “patient” refers to a mammal; preferably, the patient is a human.

[0035] The pharmaceutical composition can be formulated as tablets or capsules for oral administration, as a solution for oral administration, or as an injectable solution. In one embodiment, the composition is suitable for oral administration.

[0036] The present invention aims to provide all individual enantiomers, diastereomers, mixtures thereof, and racemates of the compound of formula I, as well as pharmaceutically acceptable salts thereof.

[0037] Certain abbreviations may 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, "DMSO" refers to dimethyl sulfoxide, "EACA" refers to epsilon-aminocaproic acid or 6-aminocaproic acid, "ELISA" refers to enzyme-linked immunosorbent assay, "Ã" refers to ethyl acetate, and "FBS" refers to fetal bovine serum. "HEC" refers to hydroxyethyl cellulose, "HEK" refers to human embryonic kidney, "HepG2" refers to human hepatocellular carcinoma cell line, "HEPES" refers to 4-(2-hydroxyethyl)-1-piperazine ethanesulfonic acid, "HLB" refers to hydrophilic lipophilic balance, "hr / s" refers to hour / hours, "HRP" refers to horseradish peroxidase, and "IC 50" refers to the concentration of the drug that produces 50% of the maximum possible inhibitory response for that drug, "MeCN" refers to acetonitrile, "min" refers to minutes (minutes / s), "MTBE" refers to methyl tert-butyl ether, "RT" refers to room temperature, "TFA" refers to trifluoroacetic acid, "THF" refers to tetrahydrofuran, and "TMB" refers to 3,3',5,5'-teramethylbenzidin.

[0038] Individual isomers, enantiomers, and diastereomers may be separated or divided by methods known to those skilled in the art, such as selective crystallization techniques or chiral chromatography, at any convenient point in the synthesis of the compounds listed below.

[0039] The compounds of formula I are readily convertible to pharmaceutically acceptable salts and can be isolated as pharmaceutically acceptable salts. Salt formation can be carried out by adding a pharmaceutically acceptable acid to form an acid addition salt, or by adding a pharmaceutically acceptable base to form a base addition salt. Salts can also be formed 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.

[0040] Compounds of formula I, or any of the illustrated formulas, or salts thereof, may be prepared by various procedures, some of which are illustrated in the following preparations and examples. The compounds or salts of the present invention may be prepared by combining specific synthesis steps of each described pathway in different ways, or by combining steps of different pathways. The products of each step in the following preparations can be recovered by conventional methods, including extraction, evaporation, precipitation, chromatography, filtration, grinding, and crystallization.

[0041] In the following schemes, all substituents are as previously defined unless otherwise indicated. Reagents and starting materials are readily available to those skilled in the art. Without limiting the scope of the present invention, the following schemes, preparations, and examples are provided to further illustrate the invention. Compounds of the formulas shown above, or salts thereof, can be prepared by using starting materials or intermediates having the corresponding desired stereochemical configuration.

[0042] Scheme 1

[0043] [ka]

[0044] In Scheme 1, step A represents the conversion of compound (1) to compound (3). Compound (1) and a bromobenzene compound (2), such as 1,3,5-tris(bromomethyl)benzene, 1,3-bis(bromomethyl)benzene, or 1,4-bis(bromomethyl)benzene, are reacted in the presence of lithium bis(trimethylsilyl)amide as a base and a solvent such as THF to obtain compound (3).

[0045] Step B describes a hydrolysis reaction in which oxazolidinone of compound (3) is cleaved using LiOH / H2O2 in the presence of a solvent such as THF to obtain compound (4).

[0046] Step C shows the deoxidation of compound (4) in which the BOC protecting group is removed in the presence of HCl, and the amine is obtained as an HCl salt [compound (5) - compound of formula I].

[0047] Scheme 2

[0048] [ka]

[0049] In Scheme 2, step A represents the conversion of compound (6) to compound (7). Compound (6) is reacted with 4-fluoro-3-nitrobenzyl bromide in the presence of lithium bis(trimethylsilyl)amide as a base and a solvent such as THF to obtain compound (7).

[0050] Step B describes a hydrolysis reaction in which oxazolidinone of compound (7) is cleaved using LiOH / H2O2 in the presence of a solvent such as THF to obtain compound (8).

[0051] Step C involves the reaction of compound (8) with 2-tert-butyl-1,3-diisopropylisourea in the presence of a solvent such as 2-methyltetrahydrofuran, resulting in the formation of compound (9), which is a tert-butyl ester.

[0052] Step D shows the reaction of 2-phenylethaneamine with compound (9) in the presence of DMA as a solvent, followed by the addition of tert-butyl(3R)-3-[(1S)-1-tert-butoxycarbonyl-3-oxopropyl]pyrrolidine-1-carboxylate to produce compound (10).

[0053] Step E is a deprotection process in which the BOC group in compound (10) is removed under acidic conditions (e.g., HCl) to obtain compound (11) (compound of formula I).

[0054] Preparation 1 Di-tert-butyl 3,3'-((2S,2'S)-1,3-phenylenebis(3-((S)-4-benzyl-2-oxoxazolidine-3-yl)-3-oxopropane-1,2-diyl))(3R,3'R)-bis(pyrrolidine-1-carboxylate)

[0055] [ka]

[0056] Lithium bis(trimethylsilyl)amide (1 M, 4.6 mL in THF) was added at 0°C to a solution of tert-butyl(3R)-3-[2-[(4S)-4-benzyl-2-oxo-oxazolidine-3-yl]-2-oxo-ethyl]pyrrolidine-1-carboxylate (essentially prepared as described in International Publication No. 2020 / 247429, 1.7 g, 4.4 mmol) in THF (12.9 mL). The mixture was stirred at 0°C for 30 minutes. A solution of 1,3-bis(bromomethyl)benzene (0.5 g, 1.8 mmol) in THF (5.5 mL) was added dropwise, and the resulting mixture was allowed to reach room temperature and stirred overnight. Saturated NH4Cl was added, and the mixture was extracted with ELISA. The organic layer was washed with H2O and diatomaceous earth. The organic layer was dried over MgSO4, and the solution was concentrated under vacuum to obtain the title compound (1.5 g, 93%) as a dark yellow oil. ES / MS (m / z): 780 [M+H-BOC].

[0057] Preparation 2 (2S,2'S)-3,3'-(1,3-phenylene)bis(2-((R)-1-tert-butoxycarbonyl)pyrrolidine-3-yl)propanoic acid)

[0058] [ka]

[0059] A mixture of hydrogen peroxide (6.9 mL, 68 mmol) and lithium hydroxide (0.8 g, 34 mmol) was added at 0°C to a solution of di-tert-butyl 3,3'-((2S,2'S)-1,3-phenylenebis(3-((S)-4-benzyl-2-oxoxazolidine-3-yl)-3-oxopropane-1,2-diyl))(3R,3'R)-bis(pyrrolidine-1-carboxylate) (1.5 g, 1.7 mmol) in THF (17 mL). The mixture was stirred at 0°C for 5 minutes, then stirred overnight at room temperature. The reaction was quenched by adding 40% NaHSO3 in water at 0°C and allowed to reach room temperature. The pH was adjusted to approximately 3 with 1N HCl in water. The mixture was extracted with ELISA. The organic matter was washed with saturated aqueous NaC1, dried over MgSO4, filtered, and evaporated to dryness. The resulting residue was purified by reverse-phase chromatography: Column: XBridge C 18 (Trademark) (19 × 100 mm, 5 μm); Mobile phase: 20 mM ammonium bicarbonate in water - pH 9 (A) / MeCN (B); Elution conditions: ammonium bicarbonate with a 25% gradient over 6 minutes; Flow rate: 25 mL / min; Room temperature; The recovered material was eluted by MS (ESI+ mode) to obtain the title compound (90 mg, 9%). ES / MS (m / z): 562 [M + H].

[0060] Preparation 3 Tri-tert-butyl 3,3',3”-((2S,2'S,2”'S)-benzene-1,3,5-triyltris(3-((S)-4-benzyl-2-oxoxazolidine-3-yl)-3-oxopropane-1,2-diyl))(3R,3'R,3”R)-tris(pyrrolidine-1-carboxylate)

[0061] [ka]

[0062] Lithium bis(trimethylsilyl)amide (1 M in THF, 5.7 mL, 5.7 mmol) was added to a solution of tert-butyl(3R)-3-[2-[(4S)-4-benzyl-2-oxo-oxazolidine-3-yl]-2-oxo-ethyl]pyrrolidine-1-carboxylate (essentially prepared as described in International Publication No. 2020 / 247429, 2 g, 5.22 mmol) in THF (12 mL) and cooled to 0°C. The mixture was stirred for 30 minutes, then a solution of 1,3,5-tris(bromomethyl)benzene (600 mg, 1.63 mmol) in THF (5 mL) was added. The mixture was warmed to room temperature and stirred overnight. Saturated NH4Cl was added, and the mixture was diluted with ethyl acetate. The phases were separated, the organic phase was dried with MgSO4, filtered, and the filtrate was concentrated under vacuum to obtain the title compound. This compound was then used in preparation 4 without further characterization or purification.

[0063] Preparation 4 (2S,2'S,2''S)-3,3',3''-(benzene-1,3,5-triyl)tris(2-((R)-1-(tert-butoxycarbonyl)pyrrolidine-3-yl)propanoic acid)

[0064] [ka]

[0065] The title compound was prepared using tri-tert-butyl 3,3',3”-((2S,2'S,2”'S)-benzene-1,3,5-triyltris(3-((S)-4-benzyl-2-oxoxazolidine-3-yl)-3-oxopropane-1,2-diyl))(3R,3'R,3”R)-tris(pyrrolidine-1-carboxylate) essentially as described in Preparation 2. ES / MS m / z: 702 [M+H-Boc].

[0066] Preparation 5 tert-butyl(3R)-3-[(1S)-2-[(4S)-4-benzyl-2-oxo-oxazolidine-3-yl]-1-[[4-[(2S)-3-[(4S)-4-benzyl-2-oxo-oxazolidine-3-yl]-2-[(3R)-1-tert-butoxycarbonylpyrrolidine-3-yl]-3-oxo-propyl]phenyl]methyl]2-oxo-ethyl]pyrrolidine-1-carboxylate

[0067] [ka]

[0068] The title compound was prepared using 1,4-bis(bromomethyl)benzene essentially as described in Preparation 1, and purified by silica gel chromatography using a gradient of 0–60% SiO2 in hexane. ES / MS 779[M+H-Boc].

[0069] Preparation 6 tert-butyl(3R)-3-[(1S)-2-[(4S)-4-benzyl-2-oxo-oxazolidine-3-yl]-1-[(4-fluoro-3-nitrophenyl)methyl]-2-oxo-ethyl]pyrrolidine-1-carboxylate

[0070] [ka]

[0071] A solution of lithium bis(trimethylsilyl)amide (1M) in THF (14 mL, 14 mmol) was added at 0°C to a solution of tert-butyl(3R)-3-[2-[(4S)-4-benzyl-2-oxo-oxazolidine-3-yl]-2-oxo-ethyl]pyrrolidine-1-carboxylate (essentially prepared as described in International Publication No. 2020 / 247429, 4.5 g, 12 mmol) in THF (81 mL). The mixture was stirred for 30 minutes. A solution of 4-fluoro-3-nitrobenzyl bromide (3.0 g, 13 mmol) in THF (14 mL) was added dropwise, and the resulting mixture was allowed to reach room temperature. The mixture was quenched with saturated NH4Cl and extracted with ELISA. The combined organic phase was washed with saturated aqueous NaCl, dried over anhydrous MgSO4, filtered, and evaporated to dryness. The residue was purified using silica gel flash chromatography with a gradient of 8–40% acetone in hexane to obtain the title compound (4.3 g, 48%) as a brown oily substance. ES / MS m / z: 486 [M-tBu+H].

[0072] Preparation 7 (2S)-2-[(3R)-1-tert-butoxycarbonylpyrrolidine-3-yl]-3-(4-fluoro-3-nitrophenyl)propanoic acid

[0073] [ka]

[0074] The title compound was prepared using tert-butyl(3R)-3-[(1S)-2-[(4S)-4-benzyl-2-oxo-oxazolidine-3-yl]-1-[(4-fluoro-3-nitrophenyl)methyl]-2-oxo-ethyl]pyrrolidine-1-carboxylate, essentially as described in Preparation 2, and purified by silica gel chromatography using a gradient of 20–80% in hexane (1% acetic acid in toluene). ES / MS m / z 283(M-BOC+H); 327(M-tBu+H).

[0075] Preparation 8 tert-butyl(3R)-3-[(1S)-2-tert-butoxy-1-[(4-fluoro-3-nitrophenyl)methyl]-2-oxo-ethyl]pyrrolidine-1-carboxylate

[0076] [ka]

[0077] To a suspension of (2S)-2-[(3R)-1-tert-butoxycarbonylpyrrolidine-3-yl]-3-(4-fluoro-3-nitrophenyl)propanoic acid (1.078 g, 2.819 mmol) in 2-methyltetrahydrofuran (17 mL), 2-tert-butyl-1,3-diisopropylisourea (1.37 g, 6.48 mmol) was added, and the mixture was heated at 55°C for 30 minutes. Additional 2-tert-butyl-1,3-diisopropylisourea (1.37 g, 6.48 mmol) was added, and the mixture was heated at 55°C for 40 minutes. After 40 minutes, additional 2-tert-butyl-1,3-diisopropylisourea (0.8917 g, 4.229 mmol) was added, and the mixture was heated at 55°C for 1 hour. The reaction mixture was allowed to reach room temperature, MTBE was added, the white solid was filtered, washed with MTBE, and discarded. The filtrate was evaporated to dryness. The residue was purified by silica gel flash column chromatography with elution using hexane in acetone (7-30% gradient) to obtain the title compound (1 g, 81%) as a pale yellow oil. ES / MS 461[M+23].

[0078] Preparation 9 tert-butyl(3R)-3-[(1S)-1-[(4S)-4-benzyl-2-oxo-oxazolidine-3-carbonyl]buta-3-enyl]pyrrolidine-1-carboxylate

[0079] [ka]

[0080] The title compound was prepared using allyl bromide essentially as described in Preparation 6 and purified by silica gel chromatography using a gradient of 0–100% acetone in hexane. ES-MS m / z 373 (M-tBu+H).

[0081] Preparation 10 (2S)-2-[(3R)-1-tert-butoxycarbonylpyrrolidine-3-yl]penta-4-enoic acid

[0082] [ka]

[0083] The title compound was prepared essentially as described in Preparation 7 using tert-butyl(3R)-3-[(1S)-1-[(4S)-4-benzyl-2-oxo-oxazolidine-3-carbonyl]buta-3-enyl]pyrrolidine-1-carboxylate and purified by silica gel chromatography using a gradient of 20-80% ELISA in (hexane + 1% acetic acid). ES-MS m / z 214(M-tBu+H).

[0084] Preparation 11 tert-butyl(3R)-3-[(1S)-1-tert-butoxycarbonylbuta-3-enyl]pyrrolidine-1-carboxylate

[0085] [ka]

[0086] The title compound was prepared using (2S)-2-[(3R)-1-tert-butoxycarbonylpyrrolidine-3-yl]penta-4-enoic acid essentially as described in Preparation 8, and purified by silica gel chromatography using a gradient of 0–30% acetone in hexane. ES-MS m / z 214 (M-2xtBu+H).

[0087] Preparation 12 tert-butyl(3R)-3-[(1S)-1-tert-butoxycarbonyl-3-oxopropyl]pyrrolidine-1-carboxylate

[0088] [ka]

[0089] To a mixture of tert-butyl(3R)-3-[(1S)-1-tert-butoxycarbonylbuta-3-enyl]pyrrolidine-1-carboxylate (260 mg, 0.7990 mmol) in 0°C THF (7.7 mL) and water (3.2 mL), sodium periodate (0.68 g, 3.20 mmol) was added. Then, osmium tetroxide (4% by mass in water, 711 mg, 0.11 mmol) was added, and the resulting mixture was stirred at 0°C for 20 minutes, then at room temperature overnight. The mixture was quenched with water and extracted with MTBE. The combined organic phase was washed with brine, dried over MgSO4, filtered, and evaporated to dryness. The residue was purified by silica gel chromatography (0-100% ethyl hexane) to obtain 112 mg (43%) of the title compound as a colorless oil. ES-MS m / z 216(M-2xtBu+H).

[0090] Preparation 13 Di-tert-butyl 3,3'-((2S,2'S)-(1-phenethyl-1H-benzo[d]imidazole-2,5-diyl)bis(3-(tert-butoxy)-3-oxopropane-1,2-diyl))(3R,3'R)-bis(pyrrolidine-1-carboxylate)

[0091] [ka]

[0092] 2-phenylethanamine (18 μmol, 0.118 mmol, 100% by mass) was added to a mixture of tert-butyl(3R)-3-[(1S)-2-tert-butoxy-1-[(4-fluoro-3-nitrophenyl)methyl]-2-oxo-ethyl]pyrrolidine-1-carboxylate (50 mg, 0.114 mmol) and DMA (980 μL). The reaction mixture was incubated at 90°C for 2 hours. Na2S2O4 (2M aqueous solution, 295 μL, 0.59 mmol) was added, followed by tert-butyl(3R)-3-[(1S)-1-tert-butoxycarbonyl-3-oxo-propyl]pyrrolidine-1-carboxylate (118 μmol, 0.118 mmol). The reaction mixture was stirred at room temperature for approximately 2 hours, then at 90°C, and subsequently purified using an HLB cartridge (6 g, 2 CV of water, 2 CV of ACN). The organic layer was separated, and the solvent was removed under N2 to obtain the title compound (28.5 mg, 31%). This compound was used without characterization to prepare Example 4. [Examples]

[0093] (2S,2'S)-3,3'-(1,3-phenylene)bis(2-((R)-pyrrolidine-3-yl)propanoic acid) dihydrochloride

[0094] [ka]

[0095] HCl (2M, 2mL, 4 mmol in diethyl ether) was added to a solution of (2S,2'S)-3,3'-(1,3-phenylene)bis(2-((R)-1-tert-butoxycarbonyl)pyrrolidine-3-yl)propanoic acid (90 mg, 0.16 mmol) in DCM (2.4 mL), and the mixture was stirred overnight at room temperature. The mixture was then concentrated to dryness under nitrogen flow at 40°C, the solid was ground with MTBE, sonicated, filtered, washed with MTBE, and dried under high vacuum. The solid was dissolved in the smallest possible amount of water and concentrated to dryness under nitrogen flow at 40°C. The solid was dried in an oven at 45°C under high vacuum to obtain the title compound as a pale yellow solid (69 mg, 99%). ES / MS m / z: 361 [M+H]. [Examples]

[0096] (2S,2'S,2''S)-3,3',3''-(benzene-1,3,5-triyl)tris(2-((R)-pyrrolidine-3-yl)propanoic acid) trihydrochloride

[0097] [ka]

[0098] The title compound was prepared using (2S,2'S,2''S)-3,3',3''-(benzene-1,3,5-triyl)tris(2-((R)-1-tert-butoxycarbonyl)pyrrolidine-3-yl)propanoic acid, essentially as described in Example 1. ES / MS (m / z): 502 [M+H]. [Examples]

[0099] (2S)-3-[4-[(2S)-2-carboxy-2-[(3R)-pyrrolidine-3-yl]ethyl]phenyl]-2-[(3R)-pyrrolidine-3-yl]propanoic acid; dihydrochloride

[0100] [ka]

[0101] A mixture of hydrogen peroxide (30% by mass in water, 2.04 mL, 18 mmol) and lithium hydroxide (218 mg, 9.1 mmol) was added at 0°C to a solution of tert-butyl(3R)-3-[(1S)-2-[(4S)-4-benzyl-2-oxo-oxazolidine-3-yl]-1-[[4-[(2S)-3-[(4S)-4-benzyl-2-oxo-oxazolidine-3-yl]-2-[(3R)-1-tert-butoxycarbonylpyrrolidine-3-yl]-3-oxo-propyl]phenyl]methyl]-2-oxo-ethyl]pyrrolidine-1-carboxylate (2.0 g, 2.3 mmol) in THF (23 mL). The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was quenched with NaOH (1N), and the crude product was washed with ELISA. The aqueous phase was acidified with HCl (1N) and extracted with HCl, followed by DCM. The combined organic extract phase was dried over MgSO4 and evaporated under reduced pressure. The residue was purified by silica gel chromatography using elution with hexane (10-80%) in acetone to obtain a clear oily substance, which was foamed under vacuum. The foamy substance was dissolved in DCM and HCl (4M in 1,4-dioxane, 1 mL, 4 mmol) was added. The reaction mixture was sealed and stirred overnight at room temperature, then concentrated to obtain a white solid. The white solid was purified by reversed-phase chromatography [column: 30 × 100 mm, 5 μm; flow rate: 85 mL / min; mobile phase: 20% MeCN: 0.4% (v / v) TFA isocratic in water] to obtain the title compound as a TFA salt. The TFA salt was diluted with a 5N aqueous HCl solution and concentrated under reduced pressure. The resulting oily substance was then suspended in HCl (4M in 1,4-dioxane). This white suspension was filtered, and the solid was dried overnight in a vacuum oven to obtain the title compound (192 mg, 19%) as an HCl salt. ES / MS m / z: 361 [M+H]. [Examples]

[0102] (2S,2'S)-3,3'-(1-phenethyl-1H-benzo[d]imidazole-2,5-diyl)bis(2-((R)-pyrrolidine-3-yl)propanoate) trihydrochloride

[0103] [ka]

[0104] The title compound was prepared using di-tert-butyl 3,3'-((2S,2'S)-(1-phenethyl-1H-benzo[d]imidazole-2,5-diyl)bis(3-tert-butoxy)-3-oxopropane-1,2-diyl))(3R,3'R)-bis(pyrrolidine-1-carboxylate) essentially as described in Example 1, and purified using an HLB cartridge (eluted with water and ACN). ES / MS m / z 505[M+H].

[0105] In vitro Lp(a) aggregation assay The ability of the compounds to inhibit Lp(a) particle formation in vitro was evaluated by a cell-free aggregation assay. Conditioning media (DMEM supplemented with 10% FBS, 20 mM HEPES, and 1x penicillin / streptomycin) were collected from confluent wild-type HepG2 cells (a source of endogenously expressed ApoB) and from HEK293 stable cell lines expressing human Apo(a) containing 17 kringle repeats (selected with 1 mg / mL Geneticin) after 24 hours of incubation at 37°C and 5% CO2. In vitro aggregation assays were performed by combining the test compounds with equal volumes of HepG2 and HEK293 conditioning media in a dilution series (final concentrations 0.01–100 nM). The reaction mixtures were incubated at 37°C for 2 hours, followed by the addition of EACA to a final concentration of 150 mM to stop the reaction. Lp(a) was detected using a sandwich ELISA with an anti-Lp(a) capture antibody and an HRP-conjugated anti-ApoB detection antibody. The ELISA was colorimetrically developed using TMB and stopped with 1N sulfuric acid, and the signal was read at 450 nm with a Molecular Devices plate reader. The inhibition percentage of Lp(a) formed under each test condition was set to 0% inhibition for the aggregate reaction in the absence of the inhibitor (corresponding to a 1% DMSO concentration) and to 100% inhibition for the minimum amount of HepG2 conditioned medium present (50-fold dilution). To determine the IC50 values ​​summarized in Table 1, the data were fitted to a four-parameter curve. As summarized in Table 1, the addition of exemplary test compounds to conditioned media containing ApoB and Apo(a) resulted in concentration-dependent inhibition of Lp(a) formation in vitro. The results indicate that these compounds inhibit the aggregation of Lp(a) from Apo(a) and LDL particles.

[0106] [Table 1]

Claims

1. formula: 【Chemistry 1】 (In the formula, 【Chemistry 2】 Phenyl, 【Transformation 3】 or 【Chemistry 4】 And, In each occurrence, Z is independently H and C １～４ It is alkyl, OH, or cyclopropyl, Y, in each appearance, independently, CH ２ (It is either O or S) Compounds thereof, or pharmaceutically acceptable salts thereof.

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

3. Y is CH in each appearance ２ The compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof.

4. formula: 【Transformation 5】 (In the formula, 【Transformation 6】 is phenyl, 【Transformation 7】 or 【Transformation 8】 (is) The compound according to claim 1, or a pharmaceutically acceptable salt thereof.

5. The aforementioned compound, 【Chemistry 9】 Selected from, The compound according to claim 1, or a pharmaceutically acceptable salt thereof.

6. A pharmaceutical composition comprising a compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier, diluent, or excipient.

7. A method for 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 5, or a pharmaceutically acceptable salt thereof.

8. A method for treating an elevation of Lp(a) plasma levels in a patient, comprising administering to the patient an effective amount of a compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof.

9. A compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, for use in therapy.

10. A compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, for use in the treatment of cardiovascular disease.

11. A compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, for use in the treatment of elevated Lp(a) plasma levels.

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