Pyrrolidine compounds

Abstract

To provide compounds for use in the treatment of cardiovascular diseases.SOLUTION: A compound of formula (I'), or a pharmaceutically acceptable salt thereof, is provided.SELECTED DRAWING: None

Description

Technical Field

[0001] The present invention relates to pyrrolidine compounds, pharmaceutically acceptable salts thereof, pharmaceutical compositions, and therapeutic uses of these compounds. There have been dramatic improvements in the treatment of cardiovascular disease (CVD). Despite these treatment advances,

[0002] patients continue to experience cardiovascular events such as angina, myocardial infarction, stroke, etc., and will die if untreated. Dyslipidemia or lipid abnormalities remain major risk factors for CVD. Dyslipidemia can be divided into four common risk factors: elevated low-density lipoprotein cholesterol (LDL-c), decreased high-density lipoprotein cholesterol (HDL-c), elevated triglycerides (TG), and elevated lipoprotein(a) (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 may be used to filter the blood and remove LDL and Lp(a), but the effect is temporary and usually needs to be repeated every two weeks. There is no approved drug treatment to lower Lp(a) levels. Although the physiological function of Lp(a) is complex, an elevated Lp(a) plasma level has been reported to be an independent risk factor for CVD. There is a need for drug treatment for patients with elevated Lp(a).

[0003] Patients suffering from cardiovascular disease, especially those suffering from dyslipidemia or lipid abnormalities, desire additional treatment options. Dietary therapy, exercise, and / or statins, fibrates ​Patients who have been diagnosed with cardiovascular disease using the current standard of care, such as the use of one or more medications such as niacin or niacin. Additional treatment options are needed for patients whose cancer is not adequately controlled. Offers another treatment option for patients with VD. Reduces plasma Lp(a) levels Therefore, there is a need for pharmacologic acceptable compounds and treatment options.

[0004] A compound of formula I', [ka] During the ceremony, L is -CH2NHCH2-, -CH2NH-, -NH-, -S-, -S(O)-, -S (O)2-, -O-, -OCH2-, -OCH2CH2O-, -NHSO2NH-, [ka] is selected from the group consisting of R 1 , R 2 , R 3 , R 4 , R 5 , and R 6 are each independently converted from H and CH3 or a compound of formula I' selected from the group consisting of Pharmaceutically acceptable salts thereof are provided.

[0005] Compounds of Formula I'' [ka] ,or Pharmaceutically acceptable salts thereof are provided.

[0006] A compound of formula 1, [ka] or a pharma- ceutically acceptable salt thereof is provided.

[0007] One embodiment is a compound of formula 2, [Chemical formula] or a pharmaceutically acceptable salt thereof.

[0008] In one embodiment, it is a compound of formula I’, I’’, 1 or formula 2, and this compound is a pharmaceutically acceptable salt. In one embodiment, it is a compound of formula I’, formula 1, or formula 2, and this compound is a hydrochloride salt. In one embodiment, it is a compound of formula 1 or formula 2, and this compound is a tetrahydrochloride salt.

[0009] In one embodiment, it is a compound of formula I’, formula 1, or formula 2, and this compound is a hydrochloride salt selected from the group consisting of monohydrochloride, dihydrochloride, trihydrochloride, and tetrahydrochloride. In one embodiment, it is a compound of formula 1 or formula 2 as a zwitterion.

[0010] A compound of formula I’, wherein L is selected from the group consisting of -CH2NHCH2-, -CH2NH-, -N H-, -S-, -S(O)-, -S(O)2-, -O-, -OCH2-, -OCH2CH 2O-, -NHSO2NH-, or a pharmaceutically acceptable salt thereof is provided.

[0011] A compound of formula I’, wherein 1 and 3 are each H, and L is selected from the group consisting of -CH 2 NHCH 2 -, -CH2NH-, -NH-, -S-, -S(O)-, -S(O)2-, - O-, -OCH2-, -OCH2CH2O-, -NHSO2NH-, or a pharmaceutically acceptable salt thereof is provided.

[0012] ​​ A compound of formula I’, wherein R 1 and R 3 are each H, R 2 and R 4 are each CH3, and L is -CH2NHCH2- 、 -CH2NH-, -NH -、- S-、-S(O)-、-S(O)2 -, -O-, -OCH2-, -OCH2CH2O-, -NHSO2NH-, is selected from the group consisting of, a compound, or a pharmaceutically acceptable salt thereof is provided.

[0013] A compound of formula I’, wherein R 1 and R 3 are each H, R 2 and R 4 are each H, and L is -CH 2 NHCH 2 -, -CH 2 NH-, -NH-, -S- -, -S(O)-, -S(O) 2 -, -O-, -OCH2-, -OCH2CH2O-, -N HSO2NH-, is selected from the group consisting of, a compound, or a pharmaceutically acceptable salt thereof is provided.

[0014] A compound of formula I’, wherein L is

Chemical formula

[0015] A compound of formula I’, wherein R 1 , R 3 , and R 5 are each H, and L is

Chemical formula

[0016] A compound of formula I’, wherein R 1 , R 3 , and R 5 are each H, and R 2 , R 4 , and R 6 are each H, a compound, or a pharmaceutically acceptable salt thereof, is provided .

[0017] A compound of formula I’, wherein R 1 , R 3 , and R 5 are each H, and R 2 , R 4 , and R 6 are each CH3, a compound, or a pharmaceutically acceptable salt thereof, is provided.

[0018] A compound of formula I’, wherein L is

Chemical formula

[0019] In one embodiment, a compound of formula I’, formula I’’, formula 1, or formula 2, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier, diluent, or excipient , a pharmaceutical composition.

[0020] In one embodiment, a method of treating a patient in need of treatment for a cardiovascular disease, the method comprises administering an effective amount of a compound selected from the group consisting of formula I', formula 1, and 2, or a pharmaceutically acceptable salt thereof. In one embodiment, it is a method for treating a patient in need of treatment for cardiovascular disease, the method comprising administering an effective amount of a compound of formula I'', or a pharmaceutically acceptable salt thereof. In one embodiment, it is a method for treating a patient in need of treatment for elevated Lp(a) plasma levels, the method comprising administering an effective amount of a compound selected from the group consisting of formula I', formula 1, or formula 2, or a pharmaceutically acceptable salt thereof. In one embodiment, it is a method for treating a patient in need of treatment for elevated Lp(a) plasma levels, the method comprising administering an effective amount of a compound of formula I'', or a pharmaceutically acceptable salt thereof. In one embodiment, it is a compound selected from the group consisting of formula I', formula 1, and 2, or a pharmaceutically acceptable salt thereof for use in treatment. In one embodiment, it is a compound of formula I'', or a pharmaceutically acceptable salt thereof for use in treatment. In one embodiment, it is a compound selected from the group consisting of formula I', formula 1, and 2, or a pharmaceutically acceptable salt thereof for use in the treatment of cardiovascular disease. In one embodiment, it is a compound of formula I'', or a pharmaceutically acceptable salt thereof for use in the treatment of cardiovascular disease. In one embodiment, it is a compound selected from the group consisting of formula I', formula 1, and 2, or a pharmaceutically acceptable salt thereof for use in the treatment of elevated Lp(a) plasma levels. In one embodiment, it is a method for treating a patient in need of treatment for elevated Lp(a) plasma levels, the method comprising administering an effective amount of a compound of formula I'', or a pharmaceutically acceptable salt thereof. In one embodiment, it is a method for treating a patient in need of treatment for elevated Lp(a) plasma levels, the method comprising administering an effective amount of a compound of formula I'', or a pharmaceutically acceptable salt thereof. In one embodiment, it is a method for treating a patient in need of treatment for elevated Lp(a) plasma levels, the method comprising administering an effective amount of a compound of formula I'', or a pharmaceutically acceptable salt thereof. In one embodiment, it is a compound selected from the group consisting of formula I', formula 1, and 2, or a pharmaceutically acceptable salt thereof for use in treatment. In one embodiment, it is a compound of formula I'', or a pharmaceutically acceptable salt thereof for use in treatment.

[0021] In one embodiment, it is a compound selected from the group consisting of formula I', formula 1, and 2, or a pharmaceutically acceptable salt thereof for use in treatment. In one embodiment, it is a compound of formula I'', or a pharmaceutically acceptable salt thereof for use in treatment. In one embodiment, it is a compound selected from the group consisting of formula I', formula 1, and 2, or a pharmaceutically acceptable salt thereof for use in the treatment of cardiovascular disease. In one embodiment, it is a compound of formula I'', or a pharmaceutically acceptable salt thereof for use in the treatment of cardiovascular disease. In one embodiment, it is a compound selected from the group consisting of formula I', formula 1, and 2, or a pharmaceutically acceptable salt thereof for use in the treatment of elevated Lp(a) plasma levels.

[0022] In one embodiment, it is a compound selected from the group consisting of formula I', formula 1, and 2, or a pharmaceutically acceptable salt thereof for use in the treatment of cardiovascular disease. In one embodiment, it is a compound of formula I'', or a pharmaceutically acceptable salt thereof for use in the treatment of cardiovascular disease. In one embodiment, it is a compound selected from the group consisting of formula I', formula 1, and 2, or a pharmaceutically acceptable salt thereof for use in the treatment of elevated Lp(a) plasma levels. In one embodiment, it is a compound selected from the group consisting of formula I', formula 1, and 2, or a pharmaceutically acceptable salt thereof for use in the treatment of elevated Lp(a) plasma levels. In one embodiment, it is a compound selected from the group consisting of formula I', formula 1, and 2, or a pharmaceutically acceptable salt thereof for use in the treatment of elevated Lp(a) plasma levels.

[0023] In one embodiment, it is a compound selected from the group consisting of formula I', formula 1, and 2, or a pharmaceutically acceptable salt thereof for use in the treatment of elevated Lp(a) plasma levels. In one embodiment, it is a compound selected from the group consisting of formula I', formula 1, and 2, or a pharmaceutically acceptable salt thereof for use in the treatment of elevated Lp(a) plasma levels. In one embodiment, it is a compound selected from the group consisting of formula I', formula 1, and 2, or a pharmaceutically acceptable salt thereof for use in the treatment of elevated Lp(a) plasma levels.

[0024] In one embodiment, it is the use of a compound selected from the group consisting of Formula I', Formula 1, and 2, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for cardiovascular disease.

[0025] Lp(a) can exhibit both prothrombotic and antithrombotic properties, and atherothrombotic properties. Lp(a) inhibits fibrinolysis and accumulates in the vascular wall, which can induce thrombosis and atheromatous 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. Lp(a) plasma levels may be related to genetic predisposition.

[0026] 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 a unique apolipoprotein(a) (apo(a)) that is covalently bound to apoB via a disulfide bond. Apo(a) is synthesized in the liver. Aggregates of Lp(a) from apo(a) and LDL particles can occur within hepatocytes, on cell walls, or in plasma. Inhibition of the aggregation of LDL particles with apo(a) can reduce Lp(a) levels.

[0027] As used herein, the term "elevated Lp(a) plasma level" means a plasma level of about 50 mg / dL or higher. The compounds provided herein can be used in the treatment for reducing Lp(a) plasma levels.

[0028] As used herein, the term "pharmaceutically acceptable salt" refers to clinical and / or ​​​​​​​​​​Refers to salts of compounds that are acceptable for veterinary use. Examples of pharmaceutically acceptable salts and general methodologies for preparing them can be found in “Handbook of Pharmaceuti cal Salts: Properties, Selection and Use” P . Stahl, et al., 2nd Revised Edition, Wiley- VCH, 2011 and S.M. Berge, et al., “Pharmaceuti cal Salts”, Journal of Pharmaceutical Sci ences, 1977, 66(1), 1-19. The compounds of formula I’, I’’, 1 or also formula 2 can be zwitterions, monoacids, diacids, or triacid addition salts. The compounds of formula I’, I ’’, 1 or formula 2 can be monobase, dibase, or tribase addition salts.

[0029] The pharmaceutical compositions of the present invention can be prepared using pharmaceutically acceptable additives. The term “pharmaceutically acceptable” refers to 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 “Remingto n: The Science and Practice of Pharmacy”, Loyd, V., et al. Eds., 22nd Ed., Mack Publish ing Co., 2012.

[0030] As used herein, the term “effective amount” refers to a dosage that is effective in treating a disorder. The effective amount for a particular patient can be determined by a skilled medical practitioner . ​​

[0031] As used herein, the terms "treating," "treat," or "treatment" include delaying, reducing, preventing, or reversing the progression or severity of an existing symptom, disorder, condition, or disease. As used herein, "treating a cardiovascular disease" means delaying, reducing, preventing, or reversing the progression of a disease of the heart or blood vessels. There is provided a method of treating

[0032] myocardial infarction, which includes administering to a patient in need thereof a compound of Formula I', Formula 1, or Formula 2.

[0033] As used herein, the term "patient" refers to a mammal. Preferably, the patient is a human.

[0033] The pharmaceutical composition can be formulated as tablets or capsules for oral administration, solutions for oral administration, or injectable solutions. In one embodiment, the composition is

[0034] suitable for oral administration.

[0034] Certain abbreviations are defined as follows: "Apo" refers to apolipoprotein, "BOC" refers to tert-butoxycarbonyl, "BSA" refers to bovine serum albumin, "DAD" refers to diode array detector, "DCM" refers to dichloromethane or methylene chloride, "de" refers to diastereomeric excess, "DMEA" refers to dimethylethylamine, "DMEM" refers to Dulbecco's modified Eagle's medium, "DMF" refers to N,N-dimethylformamide, "DMSO" refers to dimethylsulfoxide, "ee" refers to enantiomeric excess, "EACA" Refers to caproic acid or 6-aminocaproic acid, "ELISA" refers to enzyme-linked immunosorbent assay, "equiv" refers to equivalent, "Et2O" refers to diethyl ether "EtOAc" refers to ethyl acetate, "EtOH" refers to ethanol or ethyl alcohol, "Ex" refers to Example, "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-hydroxy ethyl)-1-piperazineethanesulfonic acid, "HPLC" refers to high performance liquid chromatography "HRP" refers to horseradish peroxidase, "IC 50 " refers to the concentration of the agent that produces 50% of the maximum inhibitory response possible with that agent, "min" refers to minute (singular or plural), "MeOH" refers to methanol or methyl alcohol "MTBE" refers to methyl tert-butyl ether, "RP-HPLC / MS" refers to reverse phase high performance liquid chromatography with mass spectrometry, "RT" refers to room temperature "SFC" refers to supercritical fluid chromatography, "SPA" refers to scintillation proximity assay, "t (R) " refers to retention time, "THF" refers to tetrahydro furan, "TMB" refers to 3,3’,5,5’-tetramethylbenzidine, "T ris" refers to tris(hydroxymethyl)aminomethane.

[0035] Individual isomers, enantiomers, and diastereomers can be selectively crystallized or chiral chromatographed by those skilled in the art at any convenient point in the synthesis of the compounds listed below or chiral chromatography techniques at any convenient point in the synthesis of the compounds listed below. It may be separated or divided by methods such as chromatography.

[0036] Compounds of formula I’, formula I’’, formula 1, or formula 2 can be readily converted and 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. The salts can also be formed simultaneously upon deprotection of nitrogen or oxygen, i.e., removal of the protecting group. Examples, reactions, and conditions for salt formation are known to those skilled in the art. Compounds selected from the group consisting of formula I’, formula I’’, formula 1, and formula 2, or salts thereof, can be prepared by various procedures, some of which are illustrated in the following preparations and examples. For each of the described routes for preparing the compounds or salts of the present invention, the specific synthetic steps can be combined in different ways or combined with steps from different routes. The products of each step in the following preparations can be recovered by conventional methods including extraction, evaporation, precipitation, chromatography, filtration, trituration, and crystallization. In the following schemes, all substituents are as 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 present invention. Compounds of formula I’, I’’, formula 1, and formula 2, or salts thereof, can be prepared by using starting materials or intermediates having the corresponding desired stereochemical configuration. In the following schemes, all substituents are as 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 present invention. Compounds of formula I’, I’’, formula 1, and formula 2, or salts thereof, can be prepared by using starting materials or intermediates having the corresponding desired stereochemical configuration. In the following schemes, all substituents are as 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 present invention. Compounds of formula I’, I’’, formula 1, and formula 2, or salts thereof, can be prepared by using starting materials or intermediates having the corresponding desired stereochemical configuration. In the following schemes, all substituents are as 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 present invention. Compounds of formula I’, I’’, formula 1, and formula 2, or salts thereof, can be prepared by using starting materials or intermediates having the corresponding desired stereochemical configuration.

[0037] Compounds selected from the group consisting of formula I’, formula I’’, formula 1, and formula 2, or salts thereof, can be prepared by various procedures, some of which are illustrated in the following preparations and examples. For each of the described routes for preparing the compounds or salts of the present invention, the specific synthetic steps can be combined in different ways or combined with steps from different routes. The products of each step in the following preparations can be recovered by conventional methods including extraction, evaporation, precipitation, chromatography, filtration, trituration, and crystallization. Compounds selected from the group consisting of formula I’, formula I’’, formula 1, and formula 2, or salts thereof, can be prepared by various procedures, some of which are illustrated in the following preparations and examples. For each of the described routes for preparing the compounds or salts of the present invention, the specific synthetic steps can be combined in different ways or combined with steps from different routes. The products of each step in the following preparations can be recovered by conventional methods including extraction, evaporation, precipitation, chromatography, filtration, trituration, and crystallization. Compounds selected from the group consisting of formula I’, formula I’’, formula 1, and formula 2, or salts thereof, can be prepared by various procedures, some of which are illustrated in the following preparations and examples. For each of the described routes for preparing the compounds or salts of the present invention, the specific synthetic steps can be combined in different ways or combined with steps from different routes. The products of each step in the following preparations can be recovered by conventional methods including extraction, evaporation, precipitation, chromatography, filtration, trituration, and crystallization. Compounds selected from the group consisting of formula I’, formula I’’, formula 1, and formula 2, or salts thereof, can be prepared by various procedures, some of which are illustrated in the following preparations and examples. For each of the described routes for preparing the compounds or salts of the present invention, the specific synthetic steps can be combined in different ways or combined with steps from different routes. The products of each step in the following preparations can be recovered by conventional methods including extraction, evaporation, precipitation, chromatography, filtration, trituration, and crystallization. Compounds selected from the group consisting of formula I’, formula I’’, formula 1, and formula 2, or salts thereof, can be prepared by various procedures, some of which are illustrated in the following preparations and examples. For each of the described routes for preparing the compounds or salts of the present invention, the specific synthetic steps can be combined in different ways or combined with steps from different routes. The products of each step in the following preparations can be recovered by conventional methods including extraction, evaporation, precipitation, chromatography, filtration, trituration, and crystallization. Compounds selected from the group consisting of formula I’, formula I’’, formula 1, and formula 2, or salts thereof, can be prepared by various procedures, some of which are illustrated in the following preparations and examples. For each of the described routes for preparing the compounds or salts of the present invention, the specific synthetic steps can be combined in different ways or combined with steps from different routes. The products of each step in the following preparations can be recovered by conventional methods including extraction, evaporation, precipitation, chromatography, filtration, trituration, and crystallization.

[0038] In the following schemes, all substituents are as 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 present invention. Compounds of formula I’, I’’, formula 1, and formula 2, or salts thereof, can be prepared by using starting materials or intermediates having the corresponding desired stereochemical configuration. In the following schemes, all substituents are as 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 present invention. Compounds of formula I’, I’’, formula 1, and formula 2, or salts thereof, can be prepared by using starting materials or intermediates having the corresponding desired stereochemical configuration. In the following schemes, all substituents are as 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 present invention. Compounds of formula I’, I’’, formula 1, and formula 2, or salts thereof, can be prepared by using starting materials or intermediates having the corresponding desired stereochemical configuration. In the following schemes, all substituents are as 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 present invention. Compounds of formula I’, I’’, formula 1, and formula 2, or salts thereof, can be prepared by using starting materials or intermediates having the corresponding desired stereochemical configuration. In the following schemes, all substituents are as 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 present invention. Compounds of formula I’, I’’, formula 1, and formula 2, or salts thereof, can be prepared by using starting materials or intermediates having the corresponding desired stereochemical configuration. In the following schemes, all substituents are as 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 present invention. Compounds of formula I’, I’’, formula 1, and formula 2, or salts thereof, can be prepared by using starting materials or intermediates having the corresponding desired stereochemical configuration.

[0039] Scheme 1 shows the preparation of intermediates that can provide access to the compounds of the present invention. Protected pyrrolidin-3-ylacetic acid derivative A is first converted to acyloxazolidinone B . This is achieved by first converting A to an acid chloride and reacting it with (4S)- 4-benzyloxazolidin-2-one at 10 °C in the presence of lithium chloride. Benzyl bromide derivatives and bases such as lithium bis(trimethylsilyl)amide alkylate acyloxazolidinone B at 0 °C to give intermediate C diastereoselectively. One skilled in the art will recognize that the stereochemistry of the oxazolidinone substitution affects the diastereoselectivity of the alkylation, and that the use of oxazolidinone derivative B with the opposite stereochemical configuration or a racemic stereochemical configuration in this synthesis will give either the opposite or diastereoselectivity, respectively. The conversion of acyloxazolidinone C to acid intermediate D is achieved at 5 - 15 °C using an aqueous solution of LiOH in THF and H2O2. Acid intermediate D is optionally isolated as an ammonium salt. Acid intermediate D is protected as a tert-butyl ester, for example, by reacting it with tert-butyl -1,3-diisopropylisourea at elevated temperature to give intermediate E. Scheme 1 also shows the conversion of acid A to methyl ester F, which is achieved by reacting A with iodomethane in the presence of a carbonate base. Intermediate F is then alkylated with a benzyl bromide derivative using a base such as lithium bis(trimethylsilyl)amide at -78 °C to give intermediate G. Intermediate G is then treated with lithium bis(trimethyl

[0040] Using a base such as (tert-butyldimethylsilyl)amide, alkylate again with iodomethane and then hydrolyze the ester with sodium hydroxide at high temperature to obtain acidic intermediate H. Acid intermediate H is, for example, obtained by reaction with tert-butyl-1,3-diisopropylisourea at high temperature to give the tert-butyl ester and intermediate I. In particular, intermediates D, E, H, and I where R is bromine or -NO2 are particularly useful for further conversions in the preparation of compounds of formula I’. Intermediate D where R

[0041] is -H can be prepared either by alkylation of B with benzyl bromide followed by hydrolysis of the acyloxazolidinone, or a by stirring intermediate D where R is bromide with palladium on carbon under a hydrogen atmosphere. Deprotection of the pyrrolidine nitrogen on intermediate D when R a is -H gives the compound of formula I. Scheme 2 shows the conversion of an important intermediate (J, prepared as described in Scheme 1) to the penultimate compound of the present invention. The bromide is converted to aldehyde K using syngas (1:1 CO / H2), palladium(II) acetate, butyldi-1-adamantylphosphine, and a N,N,N’,N’-tetramethylethylenediamine at high temperature. Aldehyde K is then subjected to reductive amination of K with a reducing agent such as ammonia and sodium triacetoxyborohydride or sodium cyanoborohydride. a When R is -H, deprotection of the pyrrolidine nitrogen on intermediate D gives the compound of formula I. [Chemical formula]

[0042] Scheme 2 shows the conversion of an important intermediate (J, prepared as described in Scheme 1) to the penultimate compound of the present invention. The bromide is converted to aldehyde K using syngas (1:1 CO / H2), palladium(II) acetate, butyldi-1-adamantylphosphine, and N,N,N’,N’-tetramethylethylenediamine at high temperature. Aldehyde K is then subjected to reductive amination of K with a reducing agent such as ammonia and sodium triacetoxyborohydride or sodium cyanoborohydride. It is converted to a mixture of N and O with 2 or 3 equivalents respectively, and then N and O are separated by chromatography. Alternatively, the dimer compound O is prepared by converting the aldehyde K to the aldoxime L and reducing L to the amine M by flow hydrogenation using a sponge nickel catalyst at high temperature, and then preparing it by reductive amination of the amine M with the aldehyde K. The intermediate P is prepared by reductive amination of the dimer intermediate O with 3-fluoro-5-methoxybenzaldehyde.

Chemical Structure

[0043] Scheme 3 shows the further use of the bromide intermediate J for preparing the penultimate compound of the present invention. The bromide J is converted to the boronic acid Q using tetrahydroxydiboron, chloro(2-dicyclo hexylphosphino-2’,4’,6’-tri-i-propyl-1,1’-biphenyl )(2’-amino-1,1’-biphenyl-2-yl)palladium(II), X-PHOS, and potassium acetate. The boronic acid Q is then converted to the phenol T using H2O2 at 5 ℃. The phenol T is combined with the bromide J using copper(I) iodide, N,N-dimethylglycine hydrochloride, and cesium carbonate at high temperature to obtain the biphenyl ether U. The phenol T also reacts with 1,2-dibromoethane and a carbonate base at high temperature to obtain V. The aldehyde intermediate K (prepared from the bromide J described in Scheme 2) is reduced to the alcohol R with sodium borohydride at 0 °C and then subjected to the Mitsunobu reaction to obtain the intermediate S. (I), N,N-dimethylglycine hydrochloride, and cesium carbonate at high temperature to obtain the biphenyl ether U. The phenol T also reacts with 1,2-dibromoethane and a carbonate base at high temperature to obtain V. The aldehyde intermediate K (prepared from the bromide J described in Scheme 2) is reduced to the alcohol R with sodium borohydride at 0 °C and then subjected to the Mitsunobu reaction to obtain the intermediate S. (prepared from the bromide J described in Scheme 2) is reduced to the alcohol R with sodium borohydride at 0 °C and then subjected to the Mitsunobu reaction to obtain the intermediate S.

Chemical Structure

[0044] Scheme 4 shows the further use of bromide intermediate J for preparing the penultimate compound of the present invention. 2 equivalents of bromide J are coupled with potassium thioacetate using bis(dibenzylideneacetone)palladium, tripotassium phosphate, 1,1'-bis(diphenylphosphino)ferrocene to obtain sulfide W. Sulfide W is then converted to either sulfone X or sulfoxide Y using 1 equivalent or 2 equivalents of m-chloroperoxybenzoic acid respectively.

[0045]

Chemical formula

Chemical formula

[0046] Intermediates N, O, P, S, U, V, W, X, Y, CC, DD, and E in Schemes 2 - 5 By overall deprotection of E, a compound of formula I’ is obtained. The pyrrolidine protecting group (Pg in Schemes 1 - 5 ) is -BOC, and the ester (Pg in Schemes 1 - 5 1 ) is tert-butyl 2 ester, the overall deprotection is achieved in one step using a solution of HCl in an organic solvent such as diethyl ether, dioxane, or isopropanol. Upon deprotection the pyrrolidine nitrogen in the compound of formula I’ can be methylated by reductive amination with paraformaldehyde and sodium triacetoxyborohydride.

[0047] Preparation 1 tert-Butyl (3R)-3-[2-[(4S)-4-benzyl-2-oxo-oxazolidin-3-yl]-2-oxo-ethyl]pyrrolidine-1-carboxylate

Chemical formula

[0048] Preparation 2 tert-Butyl (3S)-3-[2-[(4R)-4-benzyl-2-oxo-oxazolidin-3-yl]-2-oxo-ethyl]pyrrolidine-1-carboxylate [Chemical formula] ​​​​​​​​​2-[(3S)-1-tert-Butoxycarbonylpyrrolidin-3-yl]acetic acid and using (4R)-4-benzyloxazolidin-2-one, prepare the title compound essentially as described in Preparation 1. Purify the product by silica gel chromatography using a gradient of 10 - 50% EtOAc in hexane. ES / MS (m / z ): 333 (M + H - tert-butyl)

[0049] Preparation 3 tert-Butyl (3R)-3-(2-methoxy-2-oxo-ethyl)pyrrolidine-1 -carboxylate

Chemical formula

[0050] Formulation 4 tert-Butyl (3R)-3-[1-[(3-bromophenyl)methyl]-2-methoxy -2-oxo-ethyl]pyrrolidine-1-carboxylate

Chemical Structure

[0051] Formulation 5 3-(3-Bromophenyl)-2-[(3R)-1-tert-butoxycarbonylpyrro lidin-3-yl]-2-methyl-propanoic acid

Chemical Structure

[0052] Preparation 6 tert - butyl (3R)-3-[(1S)-2-[(4S)-4-benzyl-2-oxo -ethyl]pyrrolidine-1-carboxylate A solution of lithium bis(trimethylsilyl)amide (1 M in THF, 818 mL, 818 mmol, 1.2 equiv) was placed in THF (1325 mL) in a 3 L three-necked round-bottom flask in an ice bath under nitrogen. tert-Butyl (3R)-3-[2-[(4S)-4-benzyl -2-oxo-oxazolidin-3-yl]-2-oxo-ethyl]pyrrolidine-1-carboxylate (265 g, 682 mmol) in 0 °C was added mechanically with stirring over 29 minutes. The mixture was stirred at 0.6 °C for 47 minutes. Then, a solution of 1-bromo-3-(bromomethyl)benzene (190 g, 760 mmol, 1.12 equiv) in THF (450 mL) was added over 28 minutes, and the reaction temperature was raised to 5.1 °C. The mixture was warmed to room temperature with stirring overnight. The reaction mixture was cooled using an ice / water bath, and then a saturated aqueous solution of NH4Cl (1 L) was added in 4 portions at a rate such that the reaction temperature was maintained below 21 °C. Water (1 L) was added to the mixture, and it was extracted with MTBE (3.5 L). The organic layer was washed with a mixture of water (1 L) and saturated NaCl aqueous solution (500 mL), and then with saturated NaCl aqueous solution (500 mL). The organic matter was dried over Na2SO4, filtered, and then concentrated in vacuo. Hexane (1 L) was added to the residue, and it was concentrated in vacuo and then dried under high vacuum overnight to obtain the title compound as an orange oil (416 g, >100%), and the purity was estimated to be 90 wt% based on the theoretical yield. ES / MS (m / z): 501 / -2-oxo-oxazolidin-3-yl]-2-oxo-ethyl]pyrrolidine-1-carboxylate at 0 °C was added mechanically with stirring over 29 minutes. The mixture was stirred at 0.6 °C for 47 minutes. Then, a solution of 1-bromo-3-(bromomethyl)benzene (190 g, 760 mmol, 1.12 equiv) in THF (450 mL) was added over 28 minutes, and the reaction temperature was raised to 5.1 °C. The mixture was warmed to room temperature with stirring overnight. The reaction mixture was cooled using an ice / water bath, and then a saturated aqueous solution of NH4Cl (1 L) was added in 4 portions at a rate such that the reaction temperature was maintained below 21 °C. Water (1 L) was added to the mixture, and it was extracted with MTBE (3.5 L). The organic layer was washed with a mixture of water (1 L) and saturated NaCl aqueous solution (500 mL), and then with saturated NaCl aqueous solution (500 mL). The organic matter was dried over Na2SO4, filtered, and then concentrated in vacuo. Hexane (1 L) was added to the residue, and it was concentrated in vacuo and then dried under high vacuum overnight to obtain the title compound as an orange oil (416 g, >100%), and the purity was estimated to be 90 wt% based on the theoretical yield. ES / MS (m / z): 501 / over 28 minutes, and the reaction temperature was raised to 5.1 °C. The mixture was warmed to room temperature with stirring overnight. The reaction mixture was cooled using an ice / water bath, and then a saturated aqueous solution of NH4Cl (1 L) was added in 4 portions at a rate such that the reaction temperature was maintained below 21 °C. Water (1 L) was added to the mixture, and it was extracted with MTBE (3.5 L). The organic layer was washed with a mixture of water (1 L) and saturated NaCl aqueous solution (500 mL), and then with saturated NaCl aqueous solution (500 mL). The organic matter was dried over Na2SO4, filtered, and then concentrated in vacuo. Hexane (1 L) was added to the residue, and it was concentrated in vacuo and then dried under high vacuum overnight to obtain the title compound as an orange oil (416 g, >100%), and the purity was estimated to be 90 wt% based on the theoretical yield. ES / MS (m / z): 501 / L) of 1-bromo-3-(bromomethyl)benzene (190 g, 760 mmol, 1.12 equiv) was added over 28 minutes, and the reaction temperature was raised to 5.1 °C. The mixture was warmed to room temperature with stirring overnight. The reaction mixture was cooled using an ice / water bath, and then a saturated aqueous solution of NH4Cl (1 L) was added in 4 portions at a rate such that the reaction temperature was maintained below 21 °C. Water (1 L) was added to the mixture, and it was extracted with MTBE (3.5 L). The organic layer was washed with a mixture of water (1 L) and saturated NaCl aqueous solution (500 mL), and then with saturated NaCl aqueous solution (500 mL). The organic matter was dried over Na2SO4, filtered, and then concentrated in vacuo. Hexane (1 L) was added to the residue, and it was concentrated in vacuo and then dried under high vacuum overnight to obtain the title compound as an orange oil (416 g, >100%), and the purity was estimated to be 90 wt% based on the theoretical yield. ES / MS (m / z): 501 / 12 equiv) was added over 28 minutes, and the reaction temperature was raised to 5.1 °C. The mixture was warmed to room temperature with stirring overnight. The reaction mixture was cooled using an ice / water bath, and then a saturated aqueous solution of NH4Cl (1 L) was added in 4 portions at a rate such that the reaction temperature was maintained below 21 °C. Water (1 L) was added to the mixture, and it was extracted with MTBE (3.5 L). The organic layer was washed with a mixture of water (1 L) and saturated NaCl aqueous solution (500 mL), and then with saturated NaCl aqueous solution (500 mL). The organic matter was dried over Na2SO4, filtered, and then concentrated in vacuo. Hexane (1 L) was added to the residue, and it was concentrated in vacuo and then dried under high vacuum overnight to obtain the title compound as an orange oil (416 g, >100%), and the purity was estimated to be 90 wt% based on the theoretical yield. ES / MS (m / z): 501 / overnight. The reaction mixture was cooled using an ice / water bath, and then a saturated aqueous solution of NH4Cl (1 L) was added in 4 portions at a rate such that the reaction temperature was maintained below 21 °C. Water (1 L) was added to the mixture, and it was extracted with MTBE (3.5 L). The organic layer was washed with a mixture of water (1 L) and saturated NaCl aqueous solution (500 mL), and then with saturated NaCl aqueous solution (500 mL). The organic matter was dried over Na2SO4, filtered, and then concentrated in vacuo. Hexane (1 L) was added to the residue, and it was concentrated in vacuo and then dried under high vacuum overnight to obtain the title compound as an orange oil (416 g, >100%), and the purity was estimated to be 90 wt% based on the theoretical yield. ES / MS (m / z): 501 / using an ice / water bath, and then a saturated aqueous solution of NH4Cl (1 L) was added in 4 portions at a rate such that the reaction temperature was maintained below 21 °C. Water (1 L) was added to the mixture, and it was extracted with MTBE (3.5 L). The organic layer was washed with a mixture of water (1 L) and saturated NaCl aqueous solution (500 mL), and then with saturated NaCl aqueous solution (500 mL). The organic matter was dried over Na2SO4, filtered, and then concentrated in vacuo. Hexane (1 L) was added to the residue, and it was concentrated in vacuo and then dried under high vacuum overnight to obtain the title compound as an orange oil (416 g, >100%), and the purity was estimated to be 90 wt% based on the theoretical yield. ES / MS (m / z): 501 / using an ice / water bath, and then a saturated aqueous solution of NH4Cl (1 L) was added in 4 portions at a rate such that the reaction temperature was maintained below 21 °C. Water (1 L) was added to the mixture, and it was extracted with MTBE (3.5 L). The organic layer was washed with a mixture of water (1 L) and saturated NaCl aqueous solution (500 mL), and then with saturated NaCl aqueous solution (500 mL). The organic matter was dried over Na2SO4, filtered, and then concentrated in vacuo. Hexane (1 L) was added to the residue, and it was concentrated in vacuo and then dried under high vacuum overnight to obtain the title compound as an orange oil (416 g, >100%), and the purity was estimated to be 90 wt% based on the theoretical yield. ES / MS (m / z): 501 / The mixture was extracted with MTBE (3.5 L). The organic layer was washed with a mixture of water (1 L) and saturated NaCl aqueous solution (500 mL), and then with saturated NaCl aqueous solution (500 mL). The organic matter was dried over Na2SO4, filtered, and then concentrated in vacuo. Hexane (1 L) was added to the residue, and it was concentrated in vacuo and then dried under high vacuum overnight to obtain the title compound as an orange oil (416 g, >100%), and the purity was estimated to be 90 wt% based on the theoretical yield. ES / MS (m / z): 501 / The organic layer was washed with a mixture of water (1 L) and saturated NaCl aqueous solution (500 mL), and then with saturated NaCl aqueous solution (500 mL). The organic matter was dried over Na2SO4, filtered, and then concentrated in vacuo. Hexane (1 L) was added to the residue, and it was concentrated in vacuo and then dried under high vacuum overnight to obtain the title compound as an orange oil (416 g, >100%), and the purity was estimated to be 90 wt% based on the theoretical yield. ES / MS (m / z): 501 / The organic matter was dried over Na2SO4, filtered, and then concentrated in vacuo. Hexane (1 L) was added to the residue, and it was concentrated in vacuo and then dried under high vacuum overnight to obtain the title compound as an orange oil (416 g, >100%), and the purity was estimated to be 90 wt% based on the theoretical yield. ES / MS (m / z): 501 / The residue was concentrated in vacuo and then dried under high vacuum overnight to obtain the title compound as an orange oil (416 g, >100%), and the purity was estimated to be 90 wt% based on the theoretical yield. ES / MS (m / z): 501 / to obtain the title compound as an orange oil (416 g, >100%), and the purity was estimated to be 90 wt% based on the theoretical yield. ES / MS (m / z): 501 / ​​503 (M + H - tert - butyl).

[0053] Preparation 7 tert - butyl (3S)-3 - [(1R)-2 - [(4R)-4 - benzyl - 2 - oxo -oxazolidin - 3 - yl]-1 - [(3 - bromophenyl)methyl]-2 - oxo -ethyl]pyrrolidine - 1 - carboxylate

Chemical formula

[0054] Preparation 8 tert - butyl (3R)-3 - [(1S)-2 - [(4S)-4 - benzyl - 2 - oxo -oxazolidin - 3 - yl]-1 - [(3 - nitrophenyl)methyl]-2 - oxo -ethyl]pyrrolidine - 1 - carboxylate

Chemical formula

[0055] Preparation 9 tert-Butyl (3R)-3-[(1S)-1-benzyl-2-[(4S)-4-ben dyl-2-oxo-oxazolidin-3-yl]-2-oxo-ethyl]pyrrolidine-1 -carboxylate

Chem.

[0056] Preparation 10 (2S)-3-(3-Bromophenyl)-2-[(3R)-1-tert-butoxycar bonylpyrrolidin-3-yl]propanoic acid

Chem.

[0057] Preparation 11 Ammonium; (2S)-3-(3-bromophenyl)-2-[(3R)-1-tert -butoxycarbonylpyrrolidin-3-yl]propanoate

Chemical formula

[0058] Preparation 12 Ammonium; (2R)-3-(3-bromophenyl)-2-[(3S)-1-tert -butoxycarbonylpyrrolidin-3-yl]propanoate

Chemical formula

[0059] Preparation 13 (2S)-2-[(3R)-1-tert-butoxycarbonylpyrrolidin-3-yl] -3-(3-nitrophenyl)propanoic acid

Chemical formula

[0060] Preparation 14 (2S)-2-[(3R)-1-tert-butoxycarbonylpyrrolidin-3-yl] -3-phenyl-propanoic acid

Chem.

[0061] Preparation 15 (2R)-2-[(3S)-1-tert-butoxycarbonylpyrrolidin-3-yl] -3-phenyl-propanoic acid

Chem.

[0062] Preparation 16 tert-butyl (3R)-3-[(1S)-1-[(3-bromophenyl)methyl]- 2-tert-butoxy-2-oxo-ethyl]pyrrolidine-1-carboxylate [Chemical Formula] Add ammonium; (2S)-3-(3-bromophenyl)-2-[(3R)-1 -tert-butoxycarbonylpyrrolidin-3-yl]propanoate (500 g, 1 210 mmol), 2-methyltetrahydrofuran (4000 mL), and then KHSO4 solution (1 M in water, 3000 mL) to the reactor. Stir the mixture for 30 minutes, during which the pH is measured to be 2 - 3. Separate the phases of the reaction mixture and extract the aqueous layer with 2-methyltetrahydrofuran ( 1000 mL). Combine the organic phases and wash them with saturated aqueous NaCl solution. Dry the organic phase over MgSO4 and filter it. Transfer the solution to the reactor and add 2-tert -butyl-1,3-diisopropylisourea (618.2 g, 3024 mmol, 2.5 equivalents). Stir the mixture at 65 °C for 3 hours and then add an additional 2-tert-butyl-1, 3-diisopropylisourea (247.3 g, 1210 mmol, 1 equivalent). Stir the mixture at 65 °C overnight. Cool the mixture to room temperature. Filter off the solid and wash the organic layer with saturated aqueous NaHCO3 solution (1000 mL). Dry the organic layer over MgSO4 and filter it, then concentrate it in vacuo. Add MTBE (2000 mL) to the residue and filter off the solid matter. Concentrate the filtrate to obtain the title compound (483 g, 88%) as a white solid. E S / MS (m / z): 342 / 344 (M + H - 2×tert-butyl)

[0063] Preparation 17 tert-butyl (3R)-3-[1-[(3-bromophenyl)methyl]-2-ter t-butoxy-1-methyl-2-oxo-ethyl]pyrrolidine-1-carboxylate

Chemical Structure

[0064] Preparation 18 tert-butyl (3R)-3-[(1S)-2-tert-butoxy-1-[(3-nitro (2S)-2-[(3R)-1-tert-Butoxycarbonylpyrrolidin-3-yl]-3-(3-nitrophenyl)propanoic acid

Chemical formula

[0065] Preparation 19 tert-Butyl (3R)-3-[(1S)-2-tert-butoxy-1-[(3-h ydroxyphenyl)methyl]-2-oxo-ethyl]pyrrolidine-1-carboxylate

Chemical formula

[0066] Preparation 20 tert-Butyl (3R)-3-[2-tert-butoxy-1-[(3-formylphenyl)methyl]-1-methyl-2-oxo-ethyl]pyrrolidine-1-carboxylate

Chemical formula

[0067] Preparation 21 tert-Butyl (3R)-3-[(1S)-2-tert-butoxy-1-[[3- hydroxyiminomethyl]phenyl]methyl]-2-oxo-ethyl]pyrrolidine-1- carboxylate

Chemical formula

[0068] Preparation 22 tert-Butyl (3R)-3-[(1S)-1-[[3-(aminomethyl)phenyl] methyl]-2-tert-butoxy-2-oxo-ethyl]pyrrolidine-1-carboxylate rate

Chemical Structure

[0069] tert-Butyl (3R)-3-[(1S)-2-tert-butoxy-1-[[3- [Hydroxyiminomethyl]phenyl]methyl]-2-oxo-ethyl]pyrrolidine-1 -Carboxylate (49 g, 103 mmol, 88% w / w purity) was dissolved in ammonia (Me OH, 7M solution, 606 mL) and equilibrated with hydrogen gas using column equilibration parameters. and recovering the effluent from the reactor column. The column was flushed again with ammonia (7 M solution in MeOH) for 20 min, and the reactor column was The reactor column eluate fractions are combined and the mixture is concentrated in vacuo to give The title compound is obtained as an oil (46.9 g, 88% yield, purity 78% w / w). MS(m / z): 405(M+H).

[0070] Preparation 23 tert-Butyl(3R)-3-[(1S)-1-[(3-aminophenyl)methyl]- 2-tert-Butoxy-2-oxo-ethyl]pyrrolidine-1-carboxylate [ka] Palladium on carbon (10% w / w, 380 mg, 0.36 mmol, 0.05 equiv.) of tert-butyl (3R)-3-[(1S)-2-tetramethylphenyl]phenylacetamide in EtOAc (71 mL) rt-Butoxy-1-[(3-nitrophenyl)methyl]-2-oxo-ethyl]pyrrol To a stirred solution of 1,2-diphenyl-1-carboxylate (3 g, 7.1 mmol) was added under a nitrogen atmosphere. The mixture is purged with hydrogen and stirred under a balloon of hydrogen at room temperature overnight. The reaction mixture is filtered through a pad of algal earth. The filtrate is concentrated in vacuo to give the title compound (2.62 g, 94%) is obtained as a white solid. ES / MS (m / z): 291 (M+H - BO C).

[0071] Preparation 24 tert - butyl (3R)-3 - [(1S)-2 - tert - butoxy - 1 - [[3 - ( hydroxymethyl)phenyl]methyl]-2 - oxo - ethyl]pyrrolidine - 1 - carbo xylate [Chemical formula] Sodium borohydride (0.208 g, 5.50 mmol, 1.2 eq) is added at 0 °C to a solution of tert - butyl (3R)-3 - [(1S)-2 - tert - butoxy - 1 - [[3 - ( formylphenyl)methyl]-2 - oxo - ethyl]pyrrolidine - 1 - carboxylate (1.85 g, 4.58 mmol) in MeOH (25 mL). After 10 minutes , the solvent is evaporated. Saturated aqueous NaHCO3 is added and the aqueous layer is extracted with EtOAc. The organic matter is washed with water and saturated aqueous NaCl. The organic matter is dried over MgSO4 and concentrated in vacuo to give the title compound (2.94 g, 97.5%) as a colorless oil. ES / MS (m / z): 428 (M + Na).

[0072] Preparation 25 [3 - [(2S)-3 - tert - butoxy - 2 - [(3R)-1 - tert - butoxy carbonylpyrrolidin - 3 - yl]-3 - oxo - propyl]phenyl]boronic acid [Chemical formula] tert - butyl (3R)-3 - [(1S)-1 - [(3 - bromophenyl)methyl] -2 - tert - butoxy - 2 - oxo - ethyl]pyrrolidine - 1 - carboxylate ( ​16.4 g, 36.1 mmol), tetrahydroxydiboron (5.00 g, 54.1 m mol, 1.5 equiv), chloro(2-dicyclohexylphosphino-2’,4’,6’- triisopropyl-1,1’-biphenyl)[2-(2’-amino-1,1’-bipheny ryl)]palladium(II) (XPhos Pd G2, 0.145 g, 0.180 mm ol, 0.005 equiv), 2-dicyclohexylphosphino-2’,4’,6’-trii sopropylbiphenyl (XPhos, 0.176 g, 0.361 mmol, 0.01 equiv ), potassium acetate (10.6 g, 108 mmol, 3 equiv), EtOH (246 mL) and ethylene glycol (6.10 mL, 108 mmol, 3 equiv) are mixed. The mixture is purged with nitrogen for 5 minutes. The mixture is stirred at 90 °C overnight. The reaction is cooled to room temperature, 2-methyl tetrahydrofuran (158 mL) is added, and the mixture is filtered. It is concentrated and used without further purification to give the residue of Preparation 26. ES / MS (m / z): 308 (M+H - 2x t ert-butyl).

[0073] Preparation 26 tert-butyl (3R)-3-[(1S)-2-tert-butoxy-1-[(3-h ydroxyphenyl)methyl]-2-oxo-ethyl]pyrrolidine-1-carboxylate

Chemical formula

[0074] Preparation 27 tert-Butyl (3R)-3-[(1S)-2-tert-butoxy-1-[[3- [[[3-[(2S)-3-tert-butoxy-2-[(3R)-1-tert-but oxycarbonylpyrrolidin-3-yl]-3-oxo-propyl]phenyl]methyl]a mino]methyl]phenyl]methyl]-2-oxo-ethyl]pyrrolidine-1-carboxy late

Chemical Structure

[0075] Preparation 28 tert-Butyl (3R)-3-[(1S)-1-[[3-[[bis[[3-[(2S)-3-tert-butoxy-2-[(3R)-1-tert-butoxycarbonylpyrrolidin-3-yl]-3-oxopropyl]phenyl]methyl]amino]methyl]phenyl]methyl]-2-tert-butoxy-2-oxoethyl]pyrrolidine-1-carboxylate and tert-Butyl (3R)-3-[(1S)-2-tert-butoxy-1-[[3-[[[[3-[(2S)-3-tert-butoxy-2-[(3R)-1-tert-butoxycarbonylpyrrolidin-3-yl]-3-oxopropyl]phenyl]methyl]amino]methyl]phenyl]methyl]-2-oxoethyl]pyrrolidine-1-carboxylate tert-Butyl (3R)-3-[(1S)-1-[[3-[[bis[[3-[(2S)-3-tert-butoxy-2-[(3R)-1-tert-butoxycarbonylpyrrolidin-3-yl]-3-oxopropyl]phenyl]methyl]amino]methyl]phenyl]methyl]-2-tert-butoxy-2-oxoethyl]pyrrolidine-1-carboxylate and tert-Butyl (3R)-3-[(1S)-2-tert-butoxy-1-[[3-[[[[3-[(2S)-3-tert-butoxy-2-[(3R)-1-tert-butoxycarbonylpyrrolidin-3-yl]-3-oxopropyl]phenyl]methyl]amino]methyl]phenyl]methyl]-2-oxoethyl]pyrrolidine-1-carboxylate tert-Butyl (3R)-3-[(1S)-1-[[3-[[bis[[3-[(2S)-3-tert-butoxy-2-[(3R)-1-tert-butoxycarbonylpyrrolidin-3-yl]-3-oxopropyl]phenyl]methyl]amino]methyl]phenyl]methyl]-2-tert-butoxy-2-oxoethyl]pyrrolidine-1-carboxylate and tert-Butyl (3R)-3-[(1S)-2-tert-butoxy-1-[[3-[[[[3-[(2S)-3-tert-butoxy-2-[(3R)-1-tert-butoxycarbonylpyrrolidin-3-yl]-3-oxopropyl]phenyl]methyl]amino]methyl]phenyl]methyl]-2-oxoethyl]pyrrolidine-1-carboxylate tert-Butyl (3R)-3-[(1S)-1-[[3-[[bis[[3-[(2S)-3-tert-butoxy-2-[(3R)-1-tert-butoxycarbonylpyrrolidin-3-yl]-3-oxopropyl]phenyl]methyl]amino]methyl]phenyl]methyl]-2-tert-butoxy-2-oxoethyl]pyrrolidine-1-carboxylate and tert-Butyl (3R)-3-[(1S)-2-tert-butoxy-1-[[3-[[[[3-[(2S)-3-tert-butoxy-2-[(3R)-1-tert-butoxycarbonylpyrrolidin-3-yl]-3-oxopropyl]phenyl]methyl]amino]methyl]phenyl]methyl]-2-oxoethyl]pyrrolidine-1-carboxylate tert-Butyl (3R)-3-[(1S)-1-[[3-[[bis[[3-[(2S)-3-tert-butoxy-2-[(3R)-1-tert-butoxycarbonylpyrrolidin-3-yl]-3-oxopropyl]phenyl]methyl]amino]methyl]phenyl]methyl]-2-tert-butoxy-2-oxoethyl]pyrrolidine-1-carboxylate and tert-Butyl (3R)-3-[(1S)-2-tert-butoxy-1-[[3-[[[[3-[(2S)-3-tert-butoxy-2-[(3R)-1-tert-butoxycarbonylpyrrolidin-3-yl]-3-oxopropyl]phenyl]methyl]amino]methyl]phenyl]methyl]-2-oxoethyl]pyrrolidine-1-carboxylate tert-Butyl (3R)-3-[(1S)-1-[[3-[[bis[[3-[(2S)-3-tert-butoxy-2-[(3R)-1-tert-butoxycarbonylpyrrolidin-3-yl]-3-oxopropyl]phenyl]methyl]amino]methyl]phenyl]methyl]-2-tert-butoxy-2-oxoethyl]pyrrolidine-1-carboxylate and tert-Butyl (3R)-3-[(1S)-2-tert-butoxy-1-[[3-[[[[3-[(2S)-3-tert-butoxy-2-[(3R)-1-tert-butoxycarbonylpyrrolidin-3-yl]-3-oxopropyl]phenyl]methyl]amino]methyl]phenyl]methyl]-2-oxoethyl]pyrrolidine-1-carboxylate tert-Butyl (3R)-3-[(1S)-1-[[3-[[bis[[3-[(2S)-3-tert-butoxy-2-[(3R)-1-tert-butoxycarbonylpyrrolidin-3-yl]-3-oxopropyl]phenyl]methyl]amino]methyl]phenyl]methyl]-2-tert-butoxy-2-oxoethyl]pyrrolidine-1-carboxylate and tert-Butyl (3R)-3-[(1S)-2-tert-butoxy-1-[[3-[[[[3-[(2S)-3-tert-butoxy-2-[(3R)-1-tert-butoxycarbonylpyrrolidin-3-yl]-3-oxopropyl]phenyl]methyl]amino]methyl]phenyl]methyl]-2-oxoethyl]pyrrolidine-1-carboxylate tert-Butyl (3R)-3-[(1S)-1-[[3-[[bis[[3-[(2S)-3-tert-butoxy-2-[(3R)-1-tert-butoxycarbonylpyrrolidin-3-yl]-3-oxopropyl]phenyl]methyl]amino]methyl]phenyl]methyl]-2-tert-butoxy-2-oxoethyl]pyrrolidine-1-carboxylate and tert-Butyl (3R)-3-[(1S)-2-tert-butoxy-1-[[3-[[[[3-[(2S)-3-tert-butoxy-2-[(3R)-1-tert-butoxycarbonylpyrrolidin-3-yl]-3-oxopropyl]phenyl]methyl]amino]methyl]phenyl]methyl]-2-oxoethyl]pyrrolidine-1-carboxylate tert-Butyl (3R)-3-[(1S)-1-[[3-[[bis[[3-[(2S)-3-tert-butoxy-2-[(3R)-1-tert-butoxycarbonylpyrrolidin-3-yl]-3-oxopropyl]phenyl]methyl]amino]methyl]phenyl]methyl]-2-tert-butoxy-2-oxoethyl]pyrrolidine-1-carboxylate and tert-Butyl (3R)-3-[(1S)-2-tert-butoxy-1-[[3-[[[[3-[(2S)-3-tert-butoxy-2-[(3R)-1-tert-butoxycarbonylpyrrolidin-3-yl]-3-oxopropyl]phenyl]methyl]amino]methyl]phenyl]methyl]-2-oxoethyl]pyrrolidine-1-carboxylate

Chemical Structure

[0076] From the above silica gel chromatography, tert-butyl (3R)-3-[(1S )-2-tert-butoxy-1-[[3-[[[[3-[(2S)-3-tert-b utoxy-2-[(3R)-1-tert-butoxycarbonylpyrrolidin-3-yl]- 3-oxo-propyl]phenyl]methyl]amino]methyl]phenyl]methyl]-2- oxo-ethyl]pyrrolidine-1-carboxylate (82.50 g, 34%) was also obtained as a colorless oil. ES / MS (m / z): 792 (M + H), HPLC showed 90 wt % purity.

[0077] tert-butyl (3R)-3-[(1S)-1-[[3-[[bis[[3-[(2S) -3-tert-butoxy-2-[(3R)-1-tert-butoxycarbonylpyrrol idine-3-yl]-3-oxo-propyl]phenyl]methyl]amino]methyl]phenyl yl]methyl]-2-tert-butoxy-2-oxo-ethyl]pyrrolidine-1-carbo Method 2 for the preparation of xylate In a round-bottom flask, tert-butyl (3R)-3-[(1S)-2-tert-butoxy -1-[[3-[[[[3-[(2S)-3-tert-butoxy-2-[(3R)-1 -tert-butoxycarbonylpyrrolidin-3-yl]-3-oxo-propyl]phenyl yl]methyl]amino]methyl]phenyl]methyl]-2-oxo-ethyl]pyrrolidine -1-carboxylate (90% purity, 81.5 g, 92.6 mmol), ter t-butyl (3R)-3-[(1S)-2-tert-butoxy-1-[(3-formyl phenyl)methyl]-2-oxo-ethyl]pyrrolidine-1-carboxylate (8 5% purity, 50.6 g, 106 mmol, 1.15 eq), 2-propanol (652 m L) and acetic acid (5.31 mL, 92.6 mmol, 1 eq) were placed in a mixture and stirred for 30 minutes . Sodium triacetoxyborohydride (2 eq, 185 mmol, 39.3 g) was added to the mixture, stirred at room temperature for 2 hours, and then the reaction mixture was concentrated in vacuo. The residue was added to water (200 mL) and MTBE (300 mL), and then concentrated aqueous ammonium hydroxide solution was added to adjust the pH to 9 - 10. The organic phase was separated, dried over MgSO4 , filtered, and concentrated to dryness. The residue was purified by silica gel chromatography using a gradient of 20 - 40% EtOAc in hexane to give tert-butyl (3R)-3 -[(1S)-1-[[3-[[biS[[3-[(2S)-3-tert-butoxy- 2-[(3R)-1-tert-butoxycarbonylpyrrolidine-3-yl]-3-oxo -propyl]phenyl]methyl]amino]methyl]phenyl]methyl]-2-tert -butoxy-2-oxo-ethyl]pyrrolidine-1-carboxylate (89 g, 82% ) as a white solid.

[0078] Preparation 29 tert-butyl (3R)-3-[(1S)-2-tert-butoxy-1-[[3- [[3-[(2S)-3-tert-butoxy-2-[(3R)-1-tert-butoxy [[1-[(3R)-3-tert-Butoxycarbonylpyrrolidin-3-yl]-3-oxopropyl]phenyl]methyl- (3-Fluoro-5-methoxyphenyl)methyl]amino]methyl]phenyl]methyl] -2-oxoethyl]pyrrolidine-1-carboxylate

Chemical Structure

[0079] Preparation 30 tert-Butyl (3R)-3-[1-[[3-[[Bis[[3-[3-tert-but oxycarbonylpyrrolidin-3-yl]-3-oxopropyl]phenyl]methyl-Xylyl-2-[(3R)-1-tert-butoxycarbonylpyrrolidin-3-yl]-2 -methyl-3-oxo-propyl]phenyl]methyl]amino]methyl]phenyl]meth yl]-2-tert-butoxy-1-methyl-2-oxo-ethyl]pyrrolidine-1-ca rboxylate

Chemical formula

[0080] Preparation 31 tert-butyl (3R)-3-[(1S)-2-tert-butoxy-1-[[3- 3-[(2S)-3-tert-butoxy-2-[(3R)-1-tert-butoxyca rbonylpyrrolidin-3-yl]-3-oxo-propyl]phenoxy]phenyl]meth yl]-2-oxo-ethyl]pyrrolidine-1-carboxylate

Chemical formula

[0081] Preparation 32 tert-Butyl (3R)-3-[(1S)-2-tert-butoxy-1-[[3- 2-[3-[(2S)-3-tert-butoxy-2-[(3R)-1-tert-but oxycarbonylpyrrolidin-3-yl]-3-oxo-propyl]phenoxy]ethoxy ​(Phenyl)methyl]-2-oxo-ethyl]pyrrolidine-1-carboxylate [Chemical formula] tert-Butyl (3R)-3-[(1S)-2-tert-butoxy-1-[(3- (Hydroxyphenyl)methyl]-2-oxo-ethyl]pyrrolidine-1-carboxylate tert-Butyl (275 mg, 0.70 mmol) is dissolved in DMF (1.2 mL). Cesium carbonate (285 mg, 0.86 mmol) and 1,2-dibromoethane (0.030 mL, 0.34 mmol) are added, and the mixture is stirred at room temperature for 3 days. Over the next 9 days, cesium carbonate or potassium carbonate and 1,2-dibromoethane are added 3 more times, and the temperature is gradually raised to 70 °C and 110 °C. The mixture is cooled to room temperature, and a saturated aqueous solution of NH4Cl is added . The aqueous layer is extracted with EtOAc. The organic phase is washed with saturated NaCl aqueous solution and water . The organic phase is dried over MgSO4, filtered, and the solution is concentrated in vacuo. The residue is purified by silica gel chromatography using a gradient of 0–3 0% EtOAc in hexane to give the title compound (33.4 mg, 11%) as a colorless oil. ES / MS (m / z): 709 (M+H-BOC).

[0082] Preparation 33 tert-Butyl (3R)-3-[(1S)-2-tert-butoxy-1-[[3- [3-[(2S)-3-tert-butoxy-2-[(3R)-1-tert-butoxy carbonylpyrrolidin-3-yl]-3-oxo-propyl]phenoxy]methyl]phen yl]methyl]-2-oxo-ethyl]pyrrolidine-1-carboxylate [Chemical formula] ​ Triphenylphosphine (0.5733 g, 2.164 mmol, 1.5 eq) was added to a solution of tert-butyl (3R)-3-[(1S)-2-tert-butoxy-1-[[3- tert-butyl (3R)-3-[(1S)-2-tert-butoxy-1-[(3-hydroxyphenyl)methyl]-2-oxo-ethyl]pyrrolidine-1-carboxylate (585 mg, 1.443 mmol) and tert-butyl (3R)-3-[(1S)-2-tert-butoxy-1-[(3-hydroxyphenyl)methyl]-2-oxo-ethyl]pyrrolidine-1-carboxylate (0.8472 g, 2.164 mmol, 1.5 eq) in THF (14 mL). tert-butyl (3R)-3-[(1S)-2-tert-butoxy-1-[(3-hydroxyphenyl)methyl]-2-oxo-ethyl]pyrrolidine-1-carboxylate (585 mg, 1.443 mmol) and tert-butyl (3R)-3-[(1S)-2-tert-butoxy-1-[(3-hydroxyphenyl)methyl]-2-oxo-ethyl]pyrrolidine-1-carboxylate (0.8472 g, 2.164 mmol, 1.5 eq) in THF (14 mL). tert-butyl (3R)-3-[(1S)-2-tert-butoxy-1-[(3-hydroxyphenyl)methyl]-2-oxo-ethyl]pyrrolidine-1-carboxylate (585 mg, 1.443 mmol) and tert-butyl (3R)-3-[(1S)-2-tert-butoxy-1-[(3-hydroxyphenyl)methyl]-2-oxo-ethyl]pyrrolidine-1-carboxylate (0.8472 g, 2.164 mmol, 1.5 eq) in THF (14 mL). tert-butyl (3R)-3-[(1S)-2-tert-butoxy-1-[(3-hydroxyphenyl)methyl]-2-oxo-ethyl]pyrrolidine-1-carboxylate (585 mg, 1.443 mmol) and tert-butyl (3R)-3-[(1S)-2-tert-butoxy-1-[(3-hydroxyphenyl)methyl]-2-oxo-ethyl]pyrrolidine-1-carboxylate (0.8472 g, 2.164 mmol, 1.5 eq) in THF (14 mL). tert-butyl (3R)-3-[(1S)-2-tert-butoxy-1-[(3-hydroxyphenyl)methyl]-2-oxo-ethyl]pyrrolidine-1-carboxylate (585 mg, 1.443 mmol) and tert-butyl (3R)-3-[(1S)-2-tert-butoxy-1-[(3-hydroxyphenyl)methyl]-2-oxo-ethyl]pyrrolidine-1-carboxylate (0.8472 g, 2.164 mmol, 1.5 eq) in THF (14 mL). tert-butyl (3R)-3-[(1S)-2-tert-butoxy-1-[(3-hydroxyphenyl)methyl]-2-oxo-ethyl]pyrrolidine-1-carboxylate (585 mg, 1.443 mmol) and tert-butyl (3R)-3-[(1S)-2-tert-butoxy-1-[(3-hydroxyphenyl)methyl]-2-oxo-ethyl]pyrrolidine-1-carboxylate (0.8472 g, 2.164 mmol, 1.5 eq) in THF (14 mL). After purging with nitrogen, diethyl azodicarboxylate (0.34 mL, 2.2 mmol, 1.5 eq) was added dropwise. The mixture was stirred overnight. The reaction was filtered through a pad of diatomaceous earth and washed with DCM and MeOH. The filtrate was concentrated in vacuo. The residue obtained by SFC using the following parameters was purified: column - Chiracel® OD (5 μm, 2 x 25 cm), mobile phase - solvent A = CO2, solvent B = MeOH + DMEA (1.0% v / v), gradient - isocratic 80:20 A:B, flow rate - 80 mL / min, pressure - 120 bar, column temperature - 40 °C. Title compound (315 mg, 28%). ES / MS (m / z): 679 (M + H - BOC). After purging with nitrogen, diethyl azodicarboxylate (0.34 mL, 2.2 mmol, 1.5 eq) was added dropwise. The mixture was stirred overnight. The reaction was filtered through a pad of diatomaceous earth and washed with DCM and MeOH. The filtrate was concentrated in vacuo. The residue obtained by SFC using the following parameters was purified: column - Chiracel® OD (5 μm, 2 x 25 cm), mobile phase - solvent A = CO2, solvent B = MeOH + DMEA (1.0% v / v), gradient - isocratic 80:20 A:B, flow rate - 80 mL / min, pressure - 120 bar, column temperature - 40 °C. Title compound (315 mg, 28%). ES / MS (m / z): 679 (M + H - BOC). After purging with nitrogen, diethyl azodicarboxylate (0.34 mL, 2.2 mmol, 1.5 eq) was added dropwise. The mixture was stirred overnight. The reaction was filtered through a pad of diatomaceous earth and washed with DCM and MeOH. The filtrate was concentrated in vacuo. The residue obtained by SFC using the following parameters was purified: column - Chiracel® OD (5 μm, 2 x 25 cm), mobile phase - solvent A = CO2, solvent B = MeOH + DMEA (1.0% v / v), gradient - isocratic 80:20 A:B, flow rate - 80 mL / min, pressure - 120 bar, column temperature - 40 °C. Title compound (315 mg, 28%). ES / MS (m / z): 679 (M + H - BOC). After purging with nitrogen, diethyl azodicarboxylate (0.34 mL, 2.2 mmol, 1.5 eq) was added dropwise. The mixture was stirred overnight. The reaction was filtered through a pad of diatomaceous earth and washed with DCM and MeOH. The filtrate was concentrated in vacuo. The residue obtained by SFC using the following parameters was purified: column - Chiracel® OD (5 μm, 2 x 25 cm), mobile phase - solvent A = CO2, solvent B = MeOH + DMEA (1.0% v / v), gradient - isocratic 80:20 A:B, flow rate - 80 mL / min, pressure - 120 bar, column temperature - 40 °C. Title compound (315 mg, 28%). ES / MS (m / z): 679 (M + H - BOC). After purging with nitrogen, diethyl azodicarboxylate (0.34 mL, 2.2 mmol, 1.5 eq) was added dropwise. The mixture was stirred overnight. The reaction was filtered through a pad of diatomaceous earth and washed with DCM and MeOH. The filtrate was concentrated in vacuo. The residue obtained by SFC using the following parameters was purified: column - Chiracel® OD (5 μm, 2 x 25 cm), mobile phase - solvent A = CO2, solvent B = MeOH + DMEA (1.0% v / v), gradient - isocratic 80:20 A:B, flow rate - 80 mL / min, pressure - 120 bar, column temperature - 40 °C. Title compound (315 mg, 28%). ES / MS (m / z): 679 (M + H - BOC). After purging with nitrogen, diethyl azodicarboxylate (0.34 mL, 2.2 mmol, 1.5 eq) was added dropwise. The mixture was stirred overnight. The reaction was filtered through a pad of diatomaceous earth and washed with DCM and MeOH. The filtrate was concentrated in vacuo. The residue obtained by SFC using the following parameters was purified: column - Chiracel® OD (5 μm, 2 x 25 cm), mobile phase - solvent A = CO2, solvent B = MeOH + DMEA (1.0% v / v), gradient - isocratic 80:20 A:B, flow rate - 80 mL / min, pressure - 120 bar, column temperature - 40 °C. Title compound (315 mg, 28%). ES / MS (m / z): 679 (M + H - BOC). After purging with nitrogen, diethyl azodicarboxylate (0.34 mL, 2.2 mmol, 1.5 eq) was added dropwise. The mixture was stirred overnight. The reaction was filtered through a pad of diatomaceous earth and washed with DCM and MeOH. The filtrate was concentrated in vacuo. The residue obtained by SFC using the following parameters was purified: column - Chiracel® OD (5 μm, 2 x 25 cm), mobile phase - solvent A = CO2, solvent B = MeOH + DMEA (1.0% v / v), gradient - isocratic 80:20 A:B, flow rate - 80 mL / min, pressure - 120 bar, column temperature - 40 °C. Title compound (315 mg, 28%). ES / MS (m / z): 679 (M + H - BOC). After purging with nitrogen, diethyl azodicarboxylate (0.34 mL, 2.2 mmol, 1.5 eq) was added dropwise. The mixture was stirred overnight. The reaction was filtered through a pad of diatomaceous earth and washed with DCM and MeOH. The filtrate was concentrated in vacuo. The residue obtained by SFC using the following parameters was purified: column - Chiracel® OD (5 μm, 2 x 25 cm), mobile phase - solvent A = CO2, solvent B = MeOH + DMEA (1.0% v / v), gradient - isocratic 80:20 A:B, flow rate - 80 mL / min, pressure - 120 bar, column temperature - 40 °C. Title compound (315 mg, 28%). ES / MS (m / z): 679 (M + H - BOC).

[0083] Preparation 34 tert-butyl (3R)-3-[(1S)-2-tert-butoxy-1-[[3- 3-[(2S)-3-tert-butoxy-2-[(3R)-1-tert-butoxycarbonylpyrrolidin-3-yl]-3-oxo-propyl]phenyl]sulfanylphenyl 3-[(2S)-3-tert-butoxy-2-[(3R)-1-tert-butoxycarbonylpyrrolidin-3-yl]-3-oxo-propyl]phenyl]sulfanylphenyl Nyl]methyl]-2-oxo-ethyl]pyrrolidine-1-carboxylate [ka] Dry toluene (0.9 mL) and acetone (1.8 mL) were dissolved in tert-butyl(3 R)-3-[(1S)-1-[(3-bromophenyl)methyl]-2-tert-butoxy 1-(2-oxo-ethyl)pyrrolidine-1-carboxylate (796 mg, 1.75 m mol), potassium phosphate tribasic (229 mg, 1.06 mmol), bis(dibenzylidene (diphenylacetone)palladium (50 mg, 0.09 mmol), 1,1'-bis(diphenyl phosphino)ferrocene (70 mg, 0.12 mmol) and potassium thioacetate (10 The resulting mixture was stirred for 5 min under nitrogen atmosphere. Sonicate for 20 min and then stir at 110 °C for 6 h. Add saturated aqueous NH4Cl and EtO Ac is added. The organic layer is washed with water. The organic phase is dried over MgSO4, filtered and Concentrate in vacuo. Silica gel chromatography using a gradient of 0-40% EtOAc in hexanes. The residue is purified by chromatography to give the title compound as a yellow oil (370 mg, 52%). ES / MS(m / z):781(M+H).

[0084] Preparation 35 tert-Butyl (3R)-3-[(1S)-2-tert-butoxy-1-[[3-[ 3-[(2S)-3-tert-butoxy-2-[(3R)-1-tert-butoxy] [Carbonylpyrrolidin-3-yl]-3-oxo-propyl]phenyl]sulfinylphenyl Nyl]methyl]-2-oxo-ethyl]pyrrolidine-1-carboxylate [ka] tert-Butyl (3R)-3-[(1S)-2-tert-butoxy-1-[[3- [3-[(2S)-3-tert-butoxy-2-[(3R)-1-DCM (2 mL) of tert-butoxycarbonylpyrrolidin-3-yl]-3-oxo-propyl]phenyl sulfanylphenyl]methyl]-2-oxo-ethyl]pyrrolidine-1-carbo xylate (111 mg, 0.14 mmol) is dissolved in DCM (2 mL). 3-Chloro peroxybenzoic acid (34 mg, 0.14 mmol, 1 equivalent) is added. The mixture is stirred at room temperature for 3 hours. Saturated aqueous NaHCO3 is added, followed by DCM, and the layers are separated . The organic phase is washed with NaOH solution (3% w / v in water) and water. The organic phase is dried over MgSO4 and then filtered, and the solution is concentrated in vacuo. The residue is purified by silica gel chromatography using a gradient of 0 - 60% EtOAc in hexane to give the title compound (97.2 mg, 87%) as a colorless oil. ES / MS (m / z): 697( M + H - BOC).

[0085] Preparation 36 tert-Butyl (3R)-3-[(1S)-2-tert-butoxy-1-[[3- 3-[(2S)-3-tert-butoxy-2-[(3R)-1-tert-butoxycarbonyl pyrrolidin-3-yl]-3-oxo-propyl]phenyl]sulfonylphenyl methyl]-2-oxo-ethyl]pyrrolidine-1-carboxylate

Chemical formula

[0086] Preparation 37 tert-Butyl (3R)-3-[(1S)-2-tert-butoxy-1-[[3- [3-[(2S)-3-tert-butoxy-2-[(3R)-1-tert-butoxy carbonylpyrrolidin-3-yl]-3-oxo-propyl]anilino]methyl]phenyl yl]methyl]-2-oxo-ethyl]pyrrolidine-1-carboxylate

Chemical formula

[0087] Preparation 38 (2S)-3-[3-[3-[(2S)-3-tert-Butoxy-2-[(3R)-1 -tert-Butoxycarbonylpyrrolidin-3-yl]-3-oxo-propyl]ani lino]phenyl]-2-[(3R)-1-tert-Butoxycarbonylpyrrolidin-3 -yl]propanoic acid

Chemical Structure

[0088] Preparation 39 Di - tert - butyl 3,3’ - ((2S,2’S) - ((sulfonylbis(azanediyl ))bis(3,1 - phenylene))bis(3 - (tert - butoxy) - 3 - oxoprop an - 1,2 - diyl))(3R,3’R) - bis(pyrrolidine - 1 - carboxylate)

Chemical Structure

[0089] Example 1 (2S)-3-[3-[[Bis[[3-[(2S)-2-carboxy-2-[(3R)- pyrrolidin-3-yl]ethyl]phenyl]methyl]amino]methyl]phenyl]-2- [(3R)-pyrrolidin-3-yl]propanoic acid, tetrahydrochloride

Chemical formula

[0090] Example 2 (2S)-3-[3-[[Bis[[3-[(2S)-2-carboxy-2-[(3R)- pyrrolidin-3-yl]ethyl]phenyl]methyl]amino]methyl]phenyl]-2- [(3R)-pyrrolidin-3-yl]propanoic acid [Chemical formula] In a round-bottom flask, tert-butyl (3R)-3-[(1S)-1-[[3-[[bi S[[3-[(2S)-3-tert-butOxy-2-[(3R)-1-tert- butoxycarbonylpyrrolidin-3-yl]-3-oxo-propyl]phenyl]methyl amino]methyl]phenyl]methyl]-2-tert-butoxy-2-oxo-ethyl pyrrolidine-1-carboxylate (499.3 g, 423.3 mmol), 1,4- dioxane (1997 mL), and hydrochloric acid solution (12 M in water, 529.1 mL, 15 equivalents ) are added. The mixture is stirred at 40 °C for 1 hour, then the mixture is concentrated in vacuo to remove 1,4- dioxane, resulting in an aqueous slurry. The mixture is filtered through a propylene filter to remove insoluble particles. The pH of the filtrate is adjusted to 9 - 10 using a solution of NaOH (2 M in water). The mixture is stirred at room temperature overnight. The resulting solid is slowly filtered off using filter paper (slow filtration, using low vacuum). The solid is washed with water and dried in vacuo at 45 °C to obtain the title compound (281 g, 88%) as a white crystalline solid. ES / MS (m / z): 711 (M+H); 1 1H-NMR (500 MHz, D2O) δ 7.3 3 (t, J = 7.6 Hz, 3H), 7.27 (d, J = 7.8 Hz, 3H), 7.13( d, J = 7.8 Hz, 3H), 7.09 (s, 3H), 4.20 (s, 6H), 3.54 (dd, J = 7.9, 11.6 Hz, 3H), 3.39 - 3.34 (m, 3H), 3.2 3 - 3.17 (m, 3H), 3.02 - 2.98 (m, 3H), 2.84 (dd, J = 4 .6, 13.7 Hz, 3H), 2.76 (dd, J = 10.6, 13.3 Hz, 3H), 2.60 (td, J = 9.9, 4.8 Hz, 3H), 2.48 (td, J = 17.3, 9 .6 Hz, 3H), 2.12 - 2.07 (m, 3H), 1.73 - 1.65 (m, 3H) 。

[0091] Example 3 (2S)-3-[3-[[[3-[(2S)-2-Carboxy-2-[(3R)-pyrrol idin-3-yl]ethyl]phenyl]methylamino]methyl]phenyl]-2-[(3R )-pyrrolidin-3-yl]propanoic acid, trihydrochloride

Chem.

[0092] Example 4 (2S)-3-[3-[[[3-[(2S)-2-carboxy-2-[(3R)-pyrrol idin-3-yl]ethyl]phenyl]methyl-[(3-fluoro-5-methoxy-phen yl)methyl]amino]methyl]phenyl]-2-[(3R)-pyrrolidin-3-yl]prop anoic acid

Chemical formula

[0093] Example 5 (2S)-3-[3-[[[3-[(2S)-2-carboxy-2-[(3R)-pyrrol idin-3-yl]ethyl]phenyl]methyl-[(3-fluoro-5-methoxy-phen yl)methyl]amino]methyl]phenyl]-2-[(3R)-pyrrolidin-3-yl]prop anoic acid, trihydrochloride

Chemical formula

[0094] Example 6 3-[3-[[Bis[[3-[2-carboxy-2-[(3R)-pyrrolidin-3-yl propyl]phenyl]methyl]amino]methyl]phenyl]-2-methyl-2-[(3 R)-pyrrolidin-3-yl]propanoic acid, tetrahydrochloride

Chemical Structure

[0095] Example 7 (2S)-3-[3-[3-[(2S)-2-Carboxy-2-[(3R)-pyrrolidin -3-yl]ethyl]phenoxy]phenyl]-2-[(3R)-pyrrolidin-3-yl propanoic acid, dihydrochloride

Chemical formula

[0096] Example 8 (2S)-3-[3-[2-[3-[(2S)-2-Carboxy-2-[(3R)-pyrro lidine-3-yl]ethyl]phenoxy]ethoxy]phenyl]-2-[(3R)-pyrro lidine-3-yl]propanoic acid, dihydrochloride

Chemical formula

[0097] Example 9 (2S)-3-[3-[[3-[(2S)-2-carboxy-2-[(3R)-pyrrolidin -3-yl]ethyl]phenoxy]methyl]phenyl]-2-[(3R)-pyrrolidin -3-yl]propanoic acid, dihydrochloride

Chemical Structure

[0098] Example 10 (2S)-3-[3-[3-[(2S)-2-carboxy-2-[(3R)-pyrrolidin -3-yl]ethyl]phenyl]sulfanylphenyl]-2-[(3R)-pyrrolidin -3-yl]propanoic acid, dihydrochloride

Chemical Structure

[0099] Example 11 (2S)-3-[3-[3-[(2S)-2-carboxy-2-[(3R)-pyrrolidin -3-yl]ethyl]phenyl]sulfinylphenyl]-2-[(3R)-pyrrolidin -3-yl]propanoic acid, dihydrochloride

Chemical Structure

[0100] Example 12 (2S)-3-[3-[3-[(2S)-2-Carboxy-2-[(3R)-pyrrolidin -3-yl]ethyl]phenyl]sulfonylphenyl]-2-[(3R)-pyrrolidine- 3-yl]propanoic acid, dihydrochloride

Chemical Structure

[0101] Example 13 (2S)-3-[3-[[3-[(2S)-2-Carboxy-2-[(3R)-pyrrolidi ne-3-yl]ethyl]anilino]methyl]phenyl]-2-[(3R)-pyrrolidine- 3-yl]propanoic acid, trihydrochloride

Chemical Structure

[0102] Example 14 (2S)-3-[3-[[3-[(2S)-2-carboxy-2-[(3R)-pyrrolidin yl]ethyl]anilino]phenyl]-2-[(3R)-pyrrolidin-3-yl propanoic acid, dihydrochloride [Chemical formula] (2S)-3-[3-[3-[(2S)-3-tert-butoxy-2-[(3R)- 1-tert-butoxycarbonylpyrrolidin-3-yl]-3-oxo-propyl]a nilino]phenyl]-2-[(3R)-1-tert-butoxycarbonylpyrrolidin- 3-yl]propanoic acid (9 g, 12.7 mmol), isopropanol (27 mL), and HCl (5.5 M solution in isopropanol) are mixed and stirred at room temperature for 2.5 hours . Heat the reaction mixture to 60 °C for 2.5 hours, then cool to room temperature and stir for 3 days. Heat the reaction mixture again to 60 °C for 2 hours, cool to room temperature, and dry the reaction mixture to dryness in vacuo. Sonicate Using MTBE, the solid residue is kneaded, filtered, washed with MTBE, and then the solid is dried in vacuo. The solid is mixed with aqueous hydrochloric acid, heated at 80 °C overnight, and then at room temperature, cooled, and concentrated to dryness in vacuo. The residue is dissolved in a minimum amount of water, and the pH is adjusted to 7.5 by adding aqueous NaOH. The mixture is stirred at room temperature for 3 h, and then the precipitated solid is filtered off. The solid is dissolved in aqueous HCl (1 N), stirred at room temperature for 15 min, and then the water is removed in vacuo. The residue is dried in vacuo at 45 °C overnight to give the title compound (5.4 g, 81%). ES / MS (m / z): 452 (M + H).

[0103] Example 15 (2S)-3-[3-[[3-[(2S)-2-Carboxy-2-[(3R)-1-methyl pyrrolidin-3-yl]ethyl]anilino]phenyl]-2-[(3R)-1-methyl pyrrolidin-3-yl]propanoic acid, dihydrochloride

Chemical Structure

[0104] Example 16 (2S,2’S)-3,3’-[Sulfonylbis(azanediyl-3,1-phenylene) bis{2-[(3R)-pyrrolidin-3-yl]propanoic acid}, dihydrochloride

Chemical Structure

[0105] Example 17 (2S)-3-Phenyl-2-[(3R)-pyrrolidin-3-yl]propanoic acid, hydrochloride

Chemical Structure

[0106] Example 18 (2R)-3-Phenyl-2-[(3S)-pyrrolidin-3-yl]propanoic acid, hydrochloride [ka] (2R)-2-[(3S)-1-tert-butoxycarbonylpyrrolidin-3-yl ]-3-phenyl-propanoic acid to prepare the title compound essentially as described in Example 6. The isolated product is stirred with HCl (2M in ether) and water for 3 h to prepare the pure product. Concentrate in air, then triturate with MTBE and dry in vacuum at 40°C. ES / MS (m / z):220(M+H).

[0107] Radiolabeling and low temperature (i.e. non-radioactive) for in vitro Apo(a) binding assays Synthesis of labeled) standards Preparation 40 1-(3-benzyloxyphenyl)imidazolidin-2-one [ka] Argon-sparged DMF (20 mL) was dissolved in ethylene urea (1.4 g, 16 mmol) l), 1-benzyloxy-3-iodobenzene (4.9 g, 16 mmol, 1.0 equiv. ), cuprous iodide (0.61 g, 3.1 mmol, 0.20 equiv.), potassium dihydrogen phosphate The resulting suspension was added to a mixture of N ,N'-dimethylethylenediamine (0.33 mL, 0.28 g, 3.1 mmol, 0. 20 equiv) is added. The blue suspension is heated at 120 °C for 3 h in a microwave reactor. The reaction mixture is cooled and filtered through a pad of silica gel, and the pad is flushed with EtOAc. The filtrate is concentrated. The residue is purified by silica gel chromatography using a 0 - 100% gradient of 5:20:75 MeOH:acetone:EtOAc in hexane to give the title compound (1.10 g, 26%) as a yellow solid. ES / MS (m / z): 2 69 (M + H)

[0108] Preparation 41 (2S)-3-[3-[3-(3-benzyloxyphenyl)-2-oxo-imidazolidin-1-yl]phenyl]-2-[(3R)-1-tert-butoxycarbonylpyrrolidin-3-yl]propanoic acid A mixture of 1-(3-benzyloxyphenyl)imidazolidin-2-one (3.33 g, 12. 4 mmol, 1.50 equiv) and ammonium; (2S)-3-(3-bromophenyl)-

Chemical Structure

[0109] Preparation 42 tert-Butyl (3R)-3-[(1S)-1-[[3-[3-(3-benzyloxy phenyl)-2-oxo-imidazolidin-1-yl]phenyl]methyl]-2-tert- butoxy-2-oxo-ethyl]pyrrolidine-1-carboxylate

Chemical Structure

[0110] Preparation 43 tert-Butyl (3R)-3-[(1S)-2-tert-butoxy-1-[[3- 3-(3-hydroxyphenyl)-2-oxo-imidazolidin-1-yl]phenyl] methyl]-2-oxo-ethyl]pyrrolidine-1-carboxylate

Chem.

[0111] Preparation 44 tert-Butyl (3R)-3-[(1S)-2-tert-butoxy-2-oxo-1 -[[3-[2-oxo-3-[3-(tritylthiomethoxy)phenyl]imidazolid in-1-yl]phenyl]methyl]ethyl]pyrrolidine-1-carboxylate

Chem.

[0112] Preparation 45 (2S)-3-[3-[2-oxo-3-[3-(tritritiomethoxy)phenyl]i midazolidin-1-yl]phenyl]-2-[(3R)-pyrrolidin-3-yl]prop enoic acid, hydrochloride

Chemical Structure

[0113] Preparation 46 1-(3 - methoxyphenyl)imidazolidin - 2 - one

Chemical Structure

[0114] Preparation 47 (2S)-2 - [(3R)-1 - tert - butoxycarbonylpyrrolidin - 3 - yl] -3 - [3 - [3 - (3 - methoxyphenyl)-2 - oxo - imidazolidin - 1 - yl phenyl]propanoic acid

Chemical Structure

[0115] Preparation 48 (2S)-3-[3-[2-oxo-3-[3-(methoxy)phenyl]imidazolidine -1-yl]phenyl]-2-[(3R)-pyrrolidin-3-yl]propanoic acid, hydrochloride [ka] (2S)-2-[(3R)-1-tert-butoxycarbonylpyrrolidin-3-yl ]-3-[3-[3-(3-methoxyphenyl)-2-oxo-imidazolidine-1-yl Preparation 45 was prepared using 1,2-diphenylpropanoic acid, essentially as described in Preparation 45. Compound prepared: ES / MS(m / z): 410(M+H).

[0116] Biological assays In vitro Apo(a) binding assay The in vitro binding affinity of compounds to the intended target human Apo(a) protein was determined by competitive assay. The human Apo(a) tag, which contains 17 kringle repeats, is tested in a binding assay. Proteins were avidly isolated from conditioned medium of transiently transfected HEK-293F cells. All reagents were prepared in 50 mM Tris-HCl pH 7.4, 0.1% B Binding assays are performed in each well of a clear bottom plate. In the tube, (1) a dilution series of test compounds (final concentrations 0.32 to 10,000 nM), (2) Ap (a) Protein (6 ng / well), (3) Resuspended wheat germ agglutinin polyvinyl chloride Ruen SPA beads (20 mg / mL), and (4) radioactive ligand, tritium-labeled. (2S)-3-[3-[2-oxo-3-[3-(trithiomethoxy)phenyl]i [(3R)-pyrrolidin-3-yl]propane Performed by adding 50 μl each of phosphoric acid and its hydrochloride (final concentration 0.52 nM). The plate is incubated at room temperature for 60 minutes and counted with a TRILUX LSC. Non-specific binding and binding in the presence of 10 μM cold (i.e., non-radiolabeled) ligand are defined as and the specific binding is determined by subtracting the binding of (2S)-3-[3-[2-oxo-3-[3-(methoxy)phenyl imidazolidin-1-yl]phenyl]-2-[(3R)-pyrrolidin-3-yl]propanoic acid, hydrochloride. The data is analyzed by fitting to a standard single-site binding model to determine the IC of the exemplary test compound. These results are summarized in Table 1, showing that examples of test compounds bind to human Apo(a) protein. Inhibition 50 of the aggregation of LDL particles by apo(a) via binding to the Apo(a) protein supports a decrease in Lp(a) levels.

Table 1

[0117] In Vitro Lp(a) Aggregation Assay ​​​​​​​​Equal amounts of HepG2 and HEK293 conditioned media are combined with the test compound added in a dilution series (final concentration 0.01 - 10 0 nM) to perform an in vitro assembly assay. The reaction is incubated at 37 °C for 2 hours and then stopped by adding 6-aminocaproic acid (EAC A) to a final concentration of 150 mM. Lp(a) is detected using a sandwich ELISA with an anti-Lp(a) capture antibody and an HRP-conjugated anti-ApoB detection antibody . The ELISA is developed using TMB and stopped using 1 N sulfuric acid, and the signal is read at 450 nm on a Molecular Devices plate reader. The percent inhibition of Lp(a) formed under each test condition is set such that the assembly reaction without inhibitor (DMSO concentration matching 1 %) is set to 0% inhibition and the minimum amount of HepG2 conditioned media present ( 50-fold dilution) is set to 100% inhibition. To determine the IC values summarized in Table 2, the data is fit to a 4-parameter curve. As summarized in Table 2, the addition of exemplary test compounds to conditioned media containing ApoB and Apo(a) results in concentration-dependent inhibition of Lp(a) formation in vitro. The results indicate that these compounds inhibit the assembly of Lp(a) from Apo(a) 50 and LDL particles. In Vivo Lp(a) Inhibition in Mice The ability of a compound to lower steady-state Lp(a) levels in vivo is evaluated in a transgenic mouse model capable of producing humanized Lp(a ) particles. Lp(a)

Table 2

[0118] ​​​​The in vivo effects of the disruptor compound are tested in female double transgenic mice, 7 - 17 months old, expressing human apoB - 100 and human apo(a) containing 17 kringle repeats: B6.SJL - Tg(APOB)1102Sgy Tg(A lb - LPA)32Arte. The mice are housed under standard light cycles (12 - hour light / 12 - hour dark), at room temperature 72 ± 8°F, relative humidity 30 - 70%, and allowed free access to water and normal solid diet (Harlan Tecklad diet 2014). The mice are randomized into treatment groups (n = 5 / group) three to five days prior to the study, using BRAT (Block Randomization Allocation Tool), based on body weight and baseline plasma Lp(a) concentration. The mice are administered vehicle (oral vehicle: 10 mL / kg, 1% H EC, 0.25% Tween 80, 0.01% antifoaming agent, subcutaneous vehicle: 5 mL / kg saline) or the test compound at various doses, twice daily (6:30 am and 3 :30 pm) for five days (or subcutaneously if described). Blood is collected into heparin - coated capillary tubes via tail bleeding. Twenty microliters of each blood sample is applied to a DBS card (Whatman catalog number : WB12 9243) for drug exposure analysis. The remaining blood samples are centrifuged to separate plasma. The Lp(a) concentration in plasma is measured using a sandwich ELISA as described in the in vitro Lp(a) aggregation assay. The % inhibition for each dose group is determined by setting the average Lp(a) level of the vehicle control group to 0% inhibition. Table 3 shows the results of a dose - response study in which tail bleed samples were taken 8 hours after oral administration on day 3 in the morning. Ed 50 ​​​(50 (the calculated value of an effective amount) to inhibit Lp(a) is determined by analysis of the threshold minimum effective amount. Table 4 shows the results of a single-dose study in which tail bleed samples were taken 4 hours after oral administration (or subcutaneous administration if stated) on day 3 in the morning (or 8 hours if stated). These results shown in Tables 3 and 4 indicate that the compound is effective in lowering plasma Lp(a) levels in vivo, supporting the proposition that the compound can be used to lower Lp(a) plasma concentrations. [Table 3] [Table 4]

Claims

[Claim 1] The invention described in the present specification.

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