Compounds for treating conditions associated with PCSK9 activity - Patents.com

Abstract

The present disclosure provides methods of treating hypercholesterolemia and other conditions associated with PCSK9 activity, e.g., atherosclerosis, atherosclerotic cardiovascular disease, coronary heart disease, metabolic syndrome, acute coronary syndrome, or related cardiovascular and cardiometabolic conditions, by orally administering to a subject an amount of a compound of Formula (I), where A" is selected from a pharma- ceutically acceptable anion, wherein the administered amount is from about 5 mg to about 300 mg of a compound of Formula (I). The present invention also relates to pharmaceutical compositions comprising a compound of Formula (I), including certain salts of the compound of Formula (I), and a permeation enhancer. [Formula 1] TIFF2024532129000025.tif55153

Description

[Technical field]

[0001] Related Applications This application claims priority to U.S. Provisional Patent Application No. 63 / 234,973, filed August 19, 2021, U.S. Provisional Patent Application No. 63 / 251,972, filed October 4, 2021, U.S. Provisional Patent Application No. 63 / 263,095, filed October 27, 2021, U.S. Provisional Patent Application No. 63 / 311,622, filed February 18, 2022, and U.S. Provisional Patent Application No. 63 / 371,685, filed August 17, 2022, the contents of each of which are incorporated herein by reference in their entirety.

[0002] The present disclosure relates to methods of treating hypercholesterolemia and other conditions associated with PCSK9 activity, such as, for example, atherosclerosis, atherosclerotic cardiovascular disease, coronary heart disease, metabolic syndrome, acute coronary syndrome, or related cardiovascular and cardiometabolic conditions. [Background technology]

[0003] Proprotein convertase subtilisin-kexin 9 (hereinafter referred to as "PCSK9"), also known as neuronal apoptosis-regulating convertase type I ("NARC-1"), is a proteinase K-like subtilase that has been identified as the ninth member of the secreted subtilase family. See Seidah et al., 2003 PNAS 100:928-933. PCSK9 belongs to the mammalian proprotein convertase family of serine proteases and contains an N-terminal signal sequence, a prodomain, a catalytic domain, and a C-terminal domain. See Seidah et al., 2012 Nat. Rev. Drug Discov. 11:367-383. Studies of PCSK9 transcriptional regulation have demonstrated that this regulation is regulated by sterol regulatory element binding proteins (Maxwell et al., 2003 J. Lipid Res. 44: 2109-2119), as seen with other genes involved in cholesterol metabolism, as is typical for other genes involved in lipoprotein metabolism (Dubuc et al., 2004 Arterioscler. Thromb. Vasc. Biol. 24: 1454-1459). Statins have been shown to upregulate PCSK9 expression in a manner that is in accordance with the cholesterol-lowering effects of the drug (supra). In addition, the PCSK9 promoter has been shown to have two conserved sites involved in cholesterol regulation, a sterol regulatory element and an Sp1 site (supra).

[0004] While in the endoplasmic reticulum, PCSK9 undergoes autocleavage between Gln-152 and Ser-153 residues as its sole catalytic activity. See Naureckiene et al., 2003 Arch.Biochem.Biophys.420:55-67; ​​Seidah et al., 2003 Proc.Natl.Acad.Sci.USA100:928-933. The prodomain remains tightly associated with the catalytic domain during subsequent transport through the trans-Golgi network. Autocleavage maturation has been demonstrated to be important for PCSK9 secretion and subsequent extracellular function (see Benjannet et al., 2012 J.Biol.Chem.287:33745-33755). Thus, several bodies of evidence demonstrate that PCSK9 specifically reduces the amount of hepatic LDLR protein, thereby impairing the liver's ability to clear low density lipoprotein ("LDL") cholesterol from the circulation.

[0005] Adenovirus-mediated overexpression of PCSK9 in mouse liver leads to accumulation of circulating low-density lipoprotein cholesterol ("LDL-C") due to dramatic loss of hepatic LDLR protein, without affecting LDLR mRNA abundance (Benjannet et al., 2004 J.Biol.Chem.279:48865-48875; Maxwell & Breslow, 2004 PNAS 101:7100-7105; Park et al., 2004 J.Biol.Chem.279:50630-50638; and Lalanne et al., 2005 J.Lipid Res.46:1312-1319). The effect of PCSK9 overexpression on circulating LDL-C levels in mice is entirely dependent on LDLR expression, further indicating that the regulation of LDL-9 by PCSK9 is mediated by downregulation of LDLR protein. In accordance with these findings, mice lacking PCPK9 or whose PCSK9 mRNA is reduced by antisense oligonucleotide inhibitors have higher levels of hepatic LDLR protein and a higher ability to clear circulating LDL-C (Rashid et al., 2005 PNAS 102:5374-5379; and Graham et al., 2007 J. Lipid Res. 48(4):763-767). In addition, reduction of PCSK9 levels by siRNA in cultured human hepatocytes also leads to higher LDLR protein levels and an increased ability to take up LDL-C (Benjannet et al., 2004 J. Biol. Chem. 279:48865-48875; and Lalanne et al., 2005 J. Lipid Res. 46:1312-1319). Taken together, these data indicate that PCSK9 action causes an increase in LDL-C by reducing LDLR protein levels.

[0006] It has been determined that some mutations in the gene PCSK9 are associated with familial hypercholesterolemia (autosomal dominant hypercholesterolemia, "ADH"), an inherited metabolic disorder characterized by a marked elevation of low-density lipoprotein ("LDL") particles in plasma, which may lead to premature circulatory failure. See Abifadel et al., 2003 Nature Genetics 34:154-156; Timms et al., 2004 Hum. Genet. 114:349-353; Leren, 2004 Clin. Genet. 65:419-422. A later published study on the S127R mutation by Abifadel et al. reported that patients with such mutations exhibit higher total cholesterol and apoB100 in plasma, which contribute to (1) the overproduction of apoB100-containing lipoproteins, such as low density lipoproteins ("LDL"), very low density lipoproteins ("VLDL"), and intermediate density lipoproteins ("IDL"), and (2) the associated reduction in clearance or conversion of such lipoproteins (Ouguerram et al., 2004 Arterioscler. Thromb. Vasc. Biol. 24:1448-1453).

[0007] Thus, there is no doubt that PCSK9 plays a role in regulating LDL. Expression or upregulation of PCSK9 is associated with increased plasma levels of LDL cholesterol, and corresponding inhibition or loss of expression of PCSK9 is associated with reduced LDL cholesterolemia levels. Reductions in LDL cholesterol levels associated with sequence mutations in PCSK9 have been found to confer a protective effect against coronary heart disease (Cohen, 2006 N. Engl. J. Med. 354:1264-1272).

[0008] In clinical trials, reductions in LDL cholesterol levels are directly related to the rate of coronary events (Law et al., 2003 BMJ 326:1423-1427). Modest reductions in plasma LDL cholesterol levels over a lifetime have been found to correlate with substantial reductions in the incidence of coronary events (Cohen et al., 2006 N.Engl.J.Med.354:1264-1272). This is also true for populations with a high prevalence of non-lipid-related cardiovascular risk factors (see above). Thus, there is a great deal of benefit to be gained from managing LDL cholesterol level control.

[0009] Thus, the identification of compounds and / or drugs effective in treating cardiovascular disease, including antagonizing the role of PCSK9 in LDL regulation, is highly desirable. However, because PCSK9 generally circulates in the blood and has the most modest binding affinity to cell surface LDL receptors, attempts to exploit this mechanism for the treatment of diseases related to high serum LDL levels have thus far focused on the use of large biomolecules, such as antibodies. Although either PCSK9-specific siRNA or monoclonal antibody (mAb) therapy can lower LDL-C in patients with hypercholesterolemia, both of these types of therapy are administered by injection. The therapeutic potential of small peptides or molecules as drugs targeting PCSK9 is only just beginning to be explored. See, e.g., Tombling et al., Atherosclerosis 330(2021)52-60. In addition, few compounds are suitable for formulation into dosage forms to utilize the oral route of administration of such compounds, a route that is highly desirable for addressing therapies of conditions in which modulation of PCSK9 activity can play a role.

[0010] WO 2019 / 246349 discloses cyclic peptide compounds useful for the treatment of cardiovascular disease and conditions associated with PCSK9 activity. The present disclosure advances the state of the art by providing a method for treating hypercholesterolemia and other conditions associated with PCSK9 activity, preferably comprising oral administration of an identified PCSK9 inhibitor. Novel salt forms of the PCSK9 inhibitors are also provided herein. [Prior art documents] [Patent documents]

[0011] [Patent Document 1] International Publication No. 2019 / 246349 [Non-patent literature]

[0012] [Non-Patent Document 1] Seidah et al.,2003 PNAS 100:928-933 [Non-Patent Document 2] Seidah et al.,2012 Nat. Rev. Drug Discov.11:367-383 [Non-Patent Document 3] Dubuc et al.,2004 Arterioscler.Thromb.Vasc.Biol.24:1454-1459 [Non-Patent Document 4] Maxwell et al.,2003 J.Lipid Res.44:2109-2119 [Non-Patent Document 5] Naureckiene et al.,2003 Arch.Biochem.Biophys.420:55-67 [Non-Patent Document 6] Seidah et al.,2003 Proc.Natl.Acad.Sci.USA100:928-933 [Non-Patent Document 7] Benjannet et al.,2012 J.Biol.Chem.287:33745-33755 [Non-licensed document 8] Benjannet et al., 2004 J.Biol.Chem.279:48865-48875

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[0013] The present disclosure provides a method of treating hypercholesterolemia and other conditions associated with PCSK9 activity, such as, for example, atherosclerosis, atherosclerotic cardiovascular disease, coronary heart disease, metabolic syndrome, acute coronary syndrome, or related cardiovascular and cardiometabolic conditions, comprising the step of: [ka] [In the formula, A - is a pharma- ceutically acceptable anion. to a subject in need thereof, wherein the administered amount is from about 5 mg to about 300 mg of a compound of formula (I).

[0014] The present disclosure also provides a method of lowering LDL-C in a subject in need thereof, comprising administering to said subject a compound represented by formula (I), - is a pharma- ceutically acceptable anion] to a subject, wherein the amount is from about 5 mg to about 300 mg of a compound of formula (I).

[0015] The present disclosure provides a method of treating atherosclerotic cardiovascular disease in a subject in need of such treatment, comprising administering to a subject a compound of formula (I), - is a pharma- ceutically acceptable anion] to a subject, wherein the dosage is an amount of a compound of formula (I) in an amount of from about 5 mg to about 300 mg.

[0016] The present disclosure also provides a method of inhibiting PCSK9 activity in a subject in need thereof, comprising administering to said subject a compound of formula (I), -is a pharma- ceutically acceptable anion] to a subject, wherein the amount is from about 5 mg to about 300 mg of a compound of formula (I).

[0017] The present invention also relates to certain salts of the compounds of formula (I).

[0018] The present invention also relates to pharmaceutical compositions comprising a compound of formula (I), including certain salts of the compound of formula (I), and a permeation enhancer. [Brief description of the drawings]

[0019] [Figure 1] 1 shows the pharmacokinetics of a single dose of Compound 1, a compound of formula (I), at various doses ranging from about 10 to about 300 mg. [Diagram 2] Summary of plasma pharmacokinetic statistics following administration of single oral doses of 10-300 mg of Compound 1 to healthy male participants. [Diagram 3] 1 shows the % change from baseline LDL-C for formulations of the compound of formula (I), as well as known anti-PCSK9 monoclonal antibodies, and anti-PCSK9 siRNA activity, and placebo. [Figure 4] 1 shows the reduction in plasma levels of free PCSK9 compared to baseline following a single dose of Compound 1. [Diagram 5] The difference in stability between Compound 1, Compound 2, and Compound 3 is shown. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0020] The present disclosure relates to methods of treating hypercholesterolemia and other conditions associated with PCSK9 activity, such as atherosclerosis, atherosclerotic cardiovascular disease, coronary heart disease, metabolic syndrome, acute coronary syndrome, or related cardiovascular and cardiometabolic conditions. [ka] [In the formula, A - is a pharma- ceutically acceptable anion. The compound is orally administered to a subject in need of treatment.

[0021] In one embodiment, the present disclosure provides a method of treating hypercholesterolemia in a subject in need thereof, comprising administering to a subject a compound represented by formula (I), - is a pharma- ceutically acceptable anion] to a subject, wherein the amount is about 5 mg to about 300 mg of a compound of formula (I).

[0022] In one embodiment, the disclosure provides a method of lowering LDL-C in a subject in need thereof, comprising administering to said subject a compound of formula (I), - is a pharma- ceutically acceptable anion] to a subject, wherein the amount is about 5 mg to about 300 mg of a compound of formula (I).

[0023] In one embodiment, the present disclosure provides a method of treating atherosclerotic cardiovascular disease in a subject in need of such treatment, comprising administering to a subject a compound of formula (I), - is a pharma- ceutically acceptable anion] to a subject, wherein the amount is from about 5 mg to about 300 mg of a compound of formula (I).

[0024] In one embodiment, the disclosure generally relates to a method of inhibiting PCSK9 activity in a subject in need thereof, comprising administering to said subject a compound represented by formula (I), -is a pharmaceutically acceptable anion] to a subject in an amount, the amount being about 5 mg to about 300 mg of a compound of formula (I). As used herein, "inhibit" or "antagonize" refers to providing a compound of formula (I) to one or more diseased tissues that opposes the effect of, inhibits, counteracts, neutralizes, or reduces one or more activities or functions of PCSK9 in the diseased tissue or tissues. In some embodiments, the method for inhibiting PCSK9 activity is for the treatment of a condition associated with PCSK9 activity, as described above, or alternatively, for providing therapy for a disease, disorder, or condition that would benefit from the effects of a PCSK9 antagonist.

[0025] The following details regarding compounds of formula (I), their pharma- ceutically acceptable anions, amounts thereof, subjects to be treated, oral administration, oral dosage forms, formulations, pharma- ceutically acceptable excipients, LDL-C lowering, PCSK9 inhibition, and the like, are relevant to all of the methods of the disclosure described above or below.

[0026] Compound (I), also referred to as "Compound A", [ka] [In the formula, A - is a pharma- ceutically acceptable anion. The compounds of formula (I) are used in all of the methods of the present disclosure. As used herein, "pharmacologically acceptable anion" refers to an anion suitable for forming pharma- ceutically acceptable salts.

[0027] The term "salt(s)" and the use of the phrase "pharmaceutically acceptable salt" as used herein include either acid salts formed with inorganic and / or organic acids, basic salts formed with inorganic and / or organic bases, or zwitterionic quaternary ammonium complexes. Salts of the compounds of the invention may be formed by methods known to those skilled in the art, for example, by reacting the compounds of the invention with an equivalent or similar amount of acid or base in a medium in which the salt precipitates or in an aqueous medium, followed by lyophilization.

[0028] The compounds of the invention contain a positively charged tetracoordinate nitrogen atom and can be stabilized by addition of anions to form salts or through the formation of anions in different parts of the molecule to produce zwitterions, sometimes referred to as inner salts. Thus, the compounds of the invention can be prepared in the form of quaternary ammonium salts or quaternary ammonium zwitterions.

[0029] Thus, the presentation of the structure of the compounds of the present invention also includes all other forms of such compounds as described above, whether in salt or zwitterion form. Therefore, one aspect of the present invention is the provision of the compounds of the present invention in the form of pharma- ceutically acceptable salts or zwitterions. Those skilled in the art will recognize examples where the compounds of the present invention may form such salts, including when the tetracoordinate nitrogen is quaternized and the charged nitrogen form may be stabilized by an associated anion. The term "pharma-ceutically acceptable salt" refers to any salt (including salts and internal salts as zwitterions) that is biologically or otherwise highly desirable (e.g., non-toxic to the recipient and not otherwise toxic).

[0030] The formation of pharma- ceutically acceptable salts from basic (or acidic) pharmaceutical compounds is generally described, for example, by S. Berge et al., Journal of Pharmaceutical Sciences (1977) 66(1) 1-19; P. Gould, International J. of Pharmaceutics (1986) 33 201-217; Anderson et al., The Practice of Medicinal Chemistry (1996), Academic Press, New York; in The Orange Book (Food & Drug Administration, Washington, DC on their website); and P. Heinrich Stahl, Camille G. Wermuth (Eds.), Handbook of Pharmaceutical Salts: Properties, Selection, and Use, (2002) Int'l. Union of Pure and Applied Chemistry, pp. 330-331, the disclosures of which are incorporated herein in their entirety.

[0031] The present disclosure contemplates all available salts of compounds of formula (I), including salts that are generally recognized as safe for use in the preparation of pharmaceutical formulations and those that may be formed presently within the purview of those skilled in the art and that are later classified as being "generally recognized as safe" for use in the preparation of pharmaceutical formulations, and are referred to herein as "pharmaceutical acceptable salts."

[0032] Examples of pharma- ceutically acceptable acid salts include acetate, including trifluoroacetate, adipate, alginate, ascorbate, aspartate, benzoate, benzenesulfonate, bisulfate, borate, butyrate, citrate, camphorate, camphorsulfonate, caprate (also known as decanoate), cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, fumarate, glucoheptanoate, glycerophosphate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, and the like. Examples of suitable salts include, but are not limited to, phosphate, 2-hydroxyethanesulfonate, lactate, maleate, methanesulfonate, methylsulfate, 2-naphthalenesulfonate, nicotinate, nitrate, oxalate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, salicylate, succinate, sulfate, sulfonate (such as those mentioned herein), tartrate, thiocyanate, toluenesulfonate (also known as tosylate), undecanoate, and the like. Thus, the pharma- ceutically acceptable anion corresponding to each of these salts, according to the present disclosure, is A - It could be.

[0033] Further examples of pharma- ceutically acceptable salts and corresponding pharma- ceutically acceptable anions that can be used in the present disclosure include, but are not limited to, fluoride, chloride, bromide, iodide, acetate and caprate anions.In one embodiment of the present invention, the pharma- ceutically acceptable anion is selected from chloride, acetate or caprate anions.As used herein, caprate and decanoate are used interchangeably.

[0034] In one embodiment of the invention, the pharma- ceutically acceptable anion is chloride and the compound of formula (I) has the following structure: [ka] It is.

[0035] In one embodiment, Compound 1 is an amorphous form of the chloride salt of the compound of Formula (I). In Figure 5, "API Amorphous Chloride" refers to Compound 1.

[0036] In one embodiment of the invention, the pharma- ceutically acceptable anion is caprate anion and the compound of formula (I) has the following structure: [ka] and is referred to herein as "Compound 2." In one embodiment, Compound 2 is an amorphous form of the caprate salt of the compound of Formula (I).

[0037] In one embodiment of the invention, the pharma- ceutically acceptable anion is acetate and the compound of formula (I) has the following structure: [ka] and is referred to herein as "Compound 3." In one embodiment, Compound 3 is an amorphous form of the acetate salt of the compound of Formula (I).

[0038] In one embodiment, the method of the present invention comprises administering a compound of formula (I), which is [ka] [ka] or [ka] is selected from.

[0039] The preparation of the amorphous chloride salt (Compound 1) utilizes an acidification step whereby chloride is introduced by adding hydrochloric acid to the product after supercritical fluid chromatography and evaporation. Figure 5 demonstrates the risk associated with adding excess HCl to a given batch of Compound 1 (referred to as "API amorphous chloride"), as such excess hydrochloric acid, being a strong acid, can result in increased chemical degradation of the molecule, as observed by the higher impurity levels shown for the amorphous chloride salts other than either the amorphous acetate or amorphous caprate salts in Figure 5. Although the stability of the amorphous chloride salt varies depending on the process used to synthesize the salt, the amorphous acetate and amorphous caprate salts are not subject to such risks and demonstrate stability regardless of the synthetic process used.

[0040] The present disclosure also provides a method of inhibiting PCSK9 activity in a subject in need thereof, comprising administering to said subject a compound of formula (I), - is a pharma- ceutically acceptable anion] to a subject, wherein the amount is from about 5 mg to about 300 mg of a compound of formula (I).

[0041] The compounds of formula (I) are compounds that have properties that antagonize PCSK9 function and are therefore PCSK9-specific antagonists or inhibitors. The compounds of formula (I) are disclosed in WO 2019 / 246349, the entire disclosure of which is incorporated herein by reference, as are methods for making the compounds.

[0042] The compounds of formula (I) are represented using conventional stereochemical notation for some asymmetric carbon centers. Thus, a solid black "wedge" bond represents a bond that protrudes out of the plane of the representation, while a "dashed wedge" bond represents a bond that recesses into the plane of the representation. As is conventional, a solid flat line represents all spatial configurations for the bond depicted. Thus, unless a specific stereochemical notation is provided, such representation contemplates all stereochemical and spatial orientations of structural features.

[0043] The compound of formula (I) is stable. As used herein, "stable" refers to a compound that can be prepared and isolated and maintains its structure and properties or remains essentially unchanged for a sufficient period of time to allow the compound to be used for the purposes described herein (e.g., therapeutic administration to a subject).

[0044] The compound of formula (I) is bioavailable, particularly orally bioavailable.As used herein, "bioavailable" refers to the ability of the compound of formula (I) to be absorbed and utilized by the body.As used herein, "orally bioavailable" means that the compound of formula (I) can be absorbed and utilized by the body when taken by mouth.

[0045] As used herein, the term "treat" or "treatment" refers to the inhibition or alleviation of a disease, condition, or disorder in a subject experiencing or exhibiting the pathology or symptoms of the disease, condition, or disorder. For example, inhibition of a disease, condition, or disorder refers to arresting further development of the pathology and / or symptoms of the disease, condition, or disorder. Furthermore, alleviation of a disease, condition, or disorder refers to the reversal of the pathology and / or symptoms, e.g., a reduction in the severity of the disease.

[0046] "Prevent," "preventing," or "prevention," as used herein, includes the prevention of at least one symptom associated with or caused by the disease, condition, or disorder being prevented.

[0047] As used herein, "subject" refers to an animal, preferably a mammal, particularly a human or non-human, including domestic animals, including but not limited to cows, horses, sheep, pigs, goats, rabbits, cats, dogs, and other mammals in need of treatment. In some embodiments, the subject is a human.

[0048] As used herein, the terms "administration" and variations thereof (e.g., "administering") with respect to a compound of formula (I) refer to providing the compound to a subject in need of treatment. As used herein, "orally" and variations thereof (e.g., "oral") refer to administration via the mouth, i.e., administration of a compound of formula (I) via the mouth.

[0049] Administering a compound of formula (I) to a subject includes both self-administration and administration to the subject by another person. A subject may be in need of or desire treatment for an existing disease or medical condition, or in need of or desire prophylactic treatment to prevent or reduce the risk of developing a disease or medical condition. As used herein, a subject "in need" of treatment for an existing condition or prophylactic treatment includes both a medical professional's determination of need and a patient's desire for such treatment.

[0050] Unless otherwise specified or clear from the context, as used herein, the term "about" is understood as within 10%, within 9%, within 8%, within 7%, within 6%, within 5%, within 4%, within 3%, within 2%, within 1%, within 0.5%, within 0.1%, within 0.05% or within 0.01% of the specified value.

[0051] In one embodiment, the amount administered to the subject is about 5 mg to about 300 mg of the compound of formula (I). Natural numbers and half integers between 5 and 300 mg are included in the present invention. In one embodiment, the amount administered is about 10 mg to about 300 mg of the compound of formula (I). In one embodiment, the amount administered is about 10 mg or about 20 mg of the compound of formula (I). In one embodiment, the amount administered is about 5 mg, about 6 mg, about 10 mg, about 12 mg, about 15 mg, about 18 mg, about 20 mg, about 24 mg, about 25 mg, about 30 mg, about 35 mg, or about 100 mg of the compound of formula (I). In one embodiment, the amount administered is about 10 mg, about 12 mg, about 15 mg, about 18 mg, about 20 mg, about 24 mg, about 25 mg, or about 30 mg of the compound of formula (I). In one embodiment, the amount administered is about 10 mg, about 10.5 mg, about 11 mg, about 11.5 mg, about 12 mg, about 12.5 mg, about 13 mg, about 13.5 mg, about 14 mg, about 14.5 mg, about 15 mg, about 15.5 mg, about 16 mg, about 16.5 mg, about 17 mg, about 17.5 mg, about 18 mg, about 18.5 mg, about 19 mg, about 19.5 mg, about 20 mg, about 20.5 mg, about 21 mg, about 21.5 mg, about 22 mg, about 22.5 mg, about 23 mg, about 23.5 mg, about 24 mg, about 24.5 mg, about 25 mg, about 25.5 mg, about 26 mg, about 26.5 mg, about 27 mg, about 27.5 mg, about 28 mg, about 28.5 mg, about 29 mg, about 29.5 mg, or about 30 mg of a compound of formula (I). In one embodiment, the amount administered is a daily dose of about 5 mg to about 300 mg.In one embodiment, the amount administered is about 10 mg, about 10.5 mg, about 11 mg, about 11.5 mg, about 12 mg, about 12.5 mg, about 13 mg, about 13.5 mg, about 14 mg, about 14.5 mg, about 15 mg, about 15.5 mg, about 16 mg, about 16.5 mg, about 17 mg, about 17.5 mg, about 18 mg, about 18.5 mg, about 19 mg, about 19.5 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 36 mg, about 37 mg, about 38 mg, about 39 mg, about 40 mg, about 41 mg, about 42 mg, about 43 mg, about 44 mg, about 45 mg, about 46 mg, about 47 mg, about 48 mg, about 49 mg, about 50 mg, about 51 mg, about 52 mg, about 53 mg, about 54 mg, about 55 mg, about 56 mg, about 57 mg, about 58 mg, about 59 mg, about 60 mg, about 61 mg, about 62 mg, about 63 mg, about 64 mg, about 65 mg, about 66 mg, about 67 mg, about 68 mg, about 69 mg, about 70 mg, about 71 mg, about 72 mg, about 75 mg, about 76 mg, about 77 mg, about 78 mg, about 79 mg, about 80 mg, about 81 mg, about 82 mg, about 83 mg, about 84 mg, about 85 mg, about 86 mg, about 87 mg, about 88 mg, about 89 mg, about 90 mg, about 91 mg, about 92 mg, about 93 mg, about 9 g, about 20.5 mg, about 21 mg, about 21.5 mg, about 22 mg, about 22.5 mg, about 23 mg, about 23.5 mg, about 24 mg, about 24.5 mg, about 25 mg, about 25.5 mg, about 26 mg, about 26.5 mg, about 27 mg, about 27.5 mg, about 28 mg, about 28.5 mg, about 29 mg, about 29.5 mg, or about 30 mg of the compound of formula (I). In one embodiment, the amount administered is about 5 mg, about 6 mg, about 10 mg, about 12 mg, about 15 mg, about 18 mg, about 20 mg, about 24 mg, about 25 mg, or about 30 mg of the compound of formula (I). In one embodiment, the amount administered is a daily dose of about 10 mg, about 12 mg, about 15 mg, about 18 mg, about 20 mg, about 24 mg, about 25 mg, or about 30 mg of a compound of formula (I).

[0052] In one embodiment, the amount of formula (I) administered to the subject is about 10 mg to about 30 mg of the compound of formula (I). In another embodiment, the amount administered to the subject is about 12 mg to about 27 mg of the compound of formula (I). In yet another embodiment, the amount administered to the subject is about 15 mg to about 25 mg of the compound of formula (I). In one embodiment, the amount administered to the subject is about 10 mg to about 20 mg of the compound of formula (I). In yet another embodiment, the amount administered to the subject is about 15 mg to about 20 mg of the compound of formula (I).

[0053] In one embodiment, the amount administered to the subject is about 10 mg to about 30 mg of compound 1. In another embodiment, the amount administered to the subject is about 12 mg to about 27 mg of compound 1. In yet another embodiment, the amount administered to the subject is about 15 mg to about 25 mg of compound 1. In one embodiment, the amount administered to the subject is about 10 mg to about 20 mg of compound 1. In yet another embodiment, the amount administered to the subject is about 15 mg to about 20 mg of compound 1. In one embodiment, the amount is about 10 mg, about 10.5 mg, about 11 mg, about 11.5 mg, about 12 mg, about 12.5 mg, about 13 mg, about 13.5 mg, about 14 mg, about 14.5 mg, about 15 mg, about 15.5 mg, about 16 mg, about 16.5 mg, about 17 mg, about 17.5 mg, about 18 mg, about 18.5 mg, about 19 mg, about 19.5 mg of compound 1. , about 20 mg, about 20.5 mg, about 21 mg, about 21.5 mg, about 22 mg, about 22.5 mg, about 23 mg, about 23.5 mg, about 24 mg, about 24.5 mg, about 25 mg, about 25.5 mg, about 26 mg, about 26.5 mg, about 27 mg, about 27.5 mg, about 28 mg, about 28.5 mg, about 29 mg, about 29.5 mg, or about 30 mg daily dose of Compound 1. In one embodiment, the amount is about 15 mg, about 15.5 mg, about 16 mg, about 16.5 mg, about 17 mg, about 17.5 mg, about 18 mg, about 18.5 mg, about 19 mg, about 19.5 mg, about 20 mg, about 20.5 mg, about 21 mg, about 21.5 mg, about 22 mg daily dose of Compound 1. In yet another embodiment, the dosage administered to a subject in need thereof is about 15 mg, about 17.5 mg, 18 mg, about 20 mg, or about 22 mg of Compound 1.

[0054] In one embodiment, the amount administered to the subject is about 10 mg to about 30 mg of compound 2. In another embodiment, the amount administered to the subject is about 12 mg to about 27 mg of compound 2. In yet another embodiment, the amount administered to the subject is about 15 mg to about 25 mg of compound 2. In one embodiment, the amount administered to the subject is about 10 mg to about 20 mg of compound 2. In yet another embodiment, the amount administered to the subject is about 15 mg to about 20 mg of compound 2. In one embodiment, the amount is about 10 mg, about 10.5 mg, about 11 mg, about 11.5 mg, about 12 mg, about 12.5 mg, about 13 mg, about 13.5 mg, about 14 mg, about 14.5 mg, about 15 mg, about 15.5 mg, about 16 mg, about 16.5 mg, about 17 mg, about 17.5 mg, about 18 mg, about 18.5 mg, about 19 mg, about 19.5 mg of compound 2. , about 20 mg, about 20.5 mg, about 21 mg, about 21.5 mg, about 22 mg, about 22.5 mg, about 23 mg, about 23.5 mg, about 24 mg, about 24.5 mg, about 25 mg, about 25.5 mg, about 26 mg, about 26.5 mg, about 27 mg, about 27.5 mg, about 28 mg, about 28.5 mg, about 29 mg, about 29.5 mg, or about 30 mg daily dose of Compound 2. In one embodiment, the amount is about 15 mg, about 15.5 mg, about 16 mg, about 16.5 mg, about 17 mg, about 17.5 mg, about 18 mg, about 18.5 mg, about 19 mg, about 19.5 mg, about 20 mg, about 20.5 mg, about 21 mg, about 21.5 mg, about 22 mg daily dose of Compound 2. In yet another embodiment, the dosage administered to a subject in need thereof is about 15 mg, about 17.5 mg, 18 mg, about 20 mg, or about 22 mg of Compound 2.

[0055] In one embodiment, the amount administered to the subject is about 10 mg to about 30 mg of compound 3. In another embodiment, the amount administered to the subject is about 12 mg to about 27 mg of compound 3. In yet another embodiment, the amount administered to the subject is about 15 mg to about 25 mg of compound 3. In one embodiment, the amount administered to the subject is about 10 mg to about 20 mg of compound 3. In yet another embodiment, the amount administered to the subject is about 15 mg to about 20 mg of compound 3. In one embodiment, the amount is about 10 mg, about 10.5 mg, about 11 mg, about 11.5 mg, about 12 mg, about 12.5 mg, about 13 mg, about 13.5 mg, about 14 mg, about 14.5 mg, about 15 mg, about 15.5 mg, about 16 mg, about 16.5 mg, about 17 mg, about 17.5 mg, about 18 mg, about 18.5 mg, about 19 mg, about 19.5 mg of compound 3. , about 20 mg, about 20.5 mg, about 21 mg, about 21.5 mg, about 22 mg, about 22.5 mg, about 23 mg, about 23.5 mg, about 24 mg, about 24.5 mg, about 25 mg, about 25.5 mg, about 26 mg, about 26.5 mg, about 27 mg, about 27.5 mg, about 28 mg, about 28.5 mg, about 29 mg, about 29.5 mg, or about 30 mg daily dose of Compound 3. In one embodiment, the amount is about 15 mg, about 15.5 mg, about 16 mg, about 16.5 mg, about 17 mg, about 17.5 mg, about 18 mg, about 18.5 mg, about 19 mg, about 19.5 mg, about 20 mg, about 20.5 mg, about 21 mg, about 21.5 mg, about 22 mg daily dose of Compound 3. In yet another embodiment, the dosage administered to a subject in need thereof is about 15 mg, about 17.5 mg, 18 mg, about 20 mg, or about 22 mg of Compound 3.

[0056] In one embodiment, the method of the invention comprises administering an oral dosage form comprising an amount of a compound of formula (I). In a further embodiment, the method comprises administering a single oral dosage form comprising an amount of a compound of formula (I). In a further embodiment, the method comprises administering a single oral dosage form comprising an amount of a compound of formula (I) once daily.

[0057] In one embodiment, an amount is a therapeutically or prophylactically effective amount of a compound of formula (I). As used herein, "therapeutically effective" or "prophylactically effective" with respect to an amount refers to the amount required at the intended dosage to achieve a desired therapeutic and / or prophylactic effect for a desired period of time. The desired effect can be, for example, relief, amelioration, reduction or cessation of at least one symptom associated with the treatment condition. For example, when treating hypercholesterolemia, lowering LDL-C is a desired effect. As one skilled in the art will appreciate, the amount can vary according to a variety of factors, including, but not limited to, the disease state, the age, sex and weight of the individual, and the ability of the PCSK9 antagonist to induce a desired effect in the individual. The response can be demonstrated by in vitro assays, in vivo non-human animal studies, and / or further supported by clinical trials.

[0058] In one embodiment, oral administration comprises administering a single oral dosage form comprising an amount of the compound of formula (I). In one embodiment, oral administration comprises administering one or more oral dosage forms each comprising an amount of the compound of formula (I) or a portion thereof. In one embodiment, oral administration comprises administering a single oral dosage form comprising an amount of the compound of formula (I) once per day. In one embodiment, oral administration comprises administering one or more oral dosage forms each comprising an amount of the compound of formula (I) or a portion thereof once per day. In one embodiment, oral administration comprises administering a single oral dosage form comprising an amount of the compound of formula (I) two or more times per day, for example, two, three or four times per day. In one embodiment, oral administration comprises administering one or more oral dosage forms each comprising an amount of the compound of formula (I) two or more times per day, for example, two, three or four times per day. The oral dosage forms may be administered with or without fasting, i.e., with or without food. In one embodiment of the invention, a subject in need of treatment fasts about 30 minutes prior to administration of a compound of formula (I).

[0059] In one embodiment, the single oral dosage form is administered once a day for at least 14 days. In one embodiment, the single oral dosage form is administered once a day for 14 days. In one embodiment, the single oral dosage form is administered once a day for as long as the subject is in need of treatment.

[0060] As used herein, "oral dosage form" refers to a pharmaceutical formulation comprising a compound of formula (I) and at least one pharma- ceutically acceptable excipient suitable for administration through the mouth of a subject. As used herein, the terms "oral dosage form" and "pharmaceutical composition" are intended to encompass both the combination of the specified components in the specified amounts and any product resulting directly or indirectly from the combination of the specified components in the specified amounts. An oral dosage form may contain a total amount of a compound of formula (I), for example, from about 5 mg to about 300 mg, which may or may not be a daily dose. An oral dosage form may contain a portion of the daily dose of a compound of formula (I).

[0061] The oral dosage forms according to the present disclosure may be solid, semi-solid or liquid. Such oral dosage forms include, but are not limited to, powders, dispersible granules, minitablets and beads (e.g., may be used for tableting, encapsulation or direct administration), pills, tablets, coated tablets, dragees, hard and soft capsules including gelatin capsules, lozenges, fast-dissolving tablets, aqueous, alcoholic or oily solutions, gels, syrups, emulsions or suspensions. The oral dosage forms according to the present disclosure may further include one or more coatings that modify the release characteristics, for example, coatings that provide delayed release or formulations with sustained release characteristics. Formulations that are intended to be converted to suspensions or solutions immediately prior to use are also included in the present disclosure, including, but not limited to, lyophilized formulations and liquid formulations absorbed into a solid absorbing medium. In one embodiment, the oral dosage form is a liquid-filled capsule, for example, a hard gelatin capsule filled with a compound of formula (I) in a combination of Labrasol® and propylene glycol, for example, in a ratio of 2:1. In one embodiment, the oral dosage form is a hard gelatin capsule filled with a compound of formula (I), e.g., a combination of Labrasol® and propylene glycol, e.g., in a 2:1 ratio, and encapsulated in an enteric capsule, e.g., HPMC Vcaps® enteric capsule (Capsugel®, Lonza). In one embodiment, the oral dosage form is a suspension, e.g., a compound of formula (I) suspended in a combination of OraBlend SF and propylene glycol, e.g., in a 2:1 ratio. In one embodiment, the oral dosage form is a dry-filled enteric coated capsule, e.g., dry-filled HPMC Vcaps® enteric capsule. In one embodiment, the oral dosage form is a tablet. In one embodiment, the oral dosage form is a tablet. In a further embodiment, the oral dosage form is a film-coated tablet.

[0062] As understood by those skilled in the art, a pharma- ceutically acceptable excipient is any component that makes a composition suitable for a specific route of administration or aids in the processing of a drug into a dosage form without exerting a beneficial pharmaceutical effect itself. In general, a composition comprises two or more pharma-ceutically acceptable excipients, and one or more pharma-ceutically acceptable excipients are selected based on the form of the oral dosage form. Examples of pharma-ceutically acceptable excipients and methods of manufacturing such oral dosage forms as described above can be found in A. Gennaro (ed.), Remington: The Science and Practice of Pharmacy, 20th Edition, (2000), Lippincott Williams & Wilkins, Baltimore, MD.

[0063] Pharmaceutically acceptable excipients suitable for use in the present disclosure include carriers (e.g., lactose, starch, starch derivatives, talc, stearic acid or its salts, etc. for pills, tablets, dragees and hard gelatin capsules; fats, waxes, semi-solid and liquid polyols, natural or hardened oils, etc. for soft gelatin capsules; solutions (emulsions or syrups), fillers, disintegrants, binders, lubricants, pressurization aids, wetting agents, stabilizing agents, emulsifiers, absorption enhancers, penetration enhancers, permeation enhancers, dispersing agents, preservatives, sweeteners, colorants, flavors, fragrances, thickening agents, diluents, buffer substances, solvents, solubilizers, agents to achieve a depot effect, agents to modify osmotic pressure, Examples of suitable pharmacopoeias include, but are not limited to, water, physiologically acceptable sodium chloride solution, alcohols, glycerol, polyols, sucrose, invert sugar, glucose, mannitol, vegetable oils, etc. for salts, coating materials and / or antioxidants. A particular pharmacopoeiasable excipient(s) and amount(s) thereof are selected for use in an oral dosage form to provide a desired amount of a compound of formula (I) in an acceptable amount of oral dosage form such that the oral dosage form can provide effective therapeutic serum levels for an acceptable period of time in a subject to which it is administered, and such that the oral dosage form maintains biological activity during storage within an acceptable temperature range for an acceptable period of time.

[0064] In one embodiment, the pharmaceutical composition of the present invention contains a diluent selected from polyethylene glycol (of various molecular weights above 3000), microcrystalline cellulose, mannitol, starch, dicalcium phosphate, calcium carbonate, sodium carbonate, lactose, or a combination thereof. In one embodiment, the pharmaceutical composition of the present invention contains a diluent selected from macrogol (PEG4000), microcrystalline cellulose, mannitol, lactose, or a combination thereof. In a further embodiment, the diluent is selected from macrogol (PEG4000), microcrystalline cellulose, or lactose. In one embodiment, the pharmaceutical composition of the present invention contains a disintegrant selected from croscarmellose sodium, crospovidone, or sodium starch glycolate. In a further embodiment, the disintegrant is croscarmellose sodium. In one embodiment, the pharmaceutical composition of the present invention contains a lubricant selected from silicon dioxide, starch, talc, magnesium stearate, or tricalcium phosphate. In a further embodiment, the lubricant is selected from silicon dioxide or tricalcium phosphate. In one embodiment, the pharmaceutical composition of the present invention contains a lubricant selected from magnesium stearate, sodium stearyl fumarate, or both. In one embodiment, the pharmaceutical composition of the present invention contains a solubilizer selected from propylene glycol, polysorbate 80, sorbitol, Cremophor EL, castor oil, corn oil, cottonseed oil, sunflower oil, sesame oil, soybean oil, peppermint oil, olive oil, miglyol, glycerin, or combinations thereof, hi a further embodiment, the solubilizer is propylene glycol.

[0065] In one embodiment, the oral dosage form further comprises a permeation enhancer. As used herein, "permeation enhancer" refers to a pharma- ceutically acceptable excipient that improves the absorption of an active agent, such as a compound of formula (I), from the gastrointestinal tract. Permeation enhancers facilitate size-limited passage through tight junctions between intestinal epithelial cells, resulting in the absorption of cell-impermeable compounds. (DJ Drucker, Advances in oral peptide therapeutics, Nat Rev Drug Discov, 19, pp 277-289 (2020)). Suitable permeation enhancers include, but are not limited to, sodium caprate, Labrasol®, sodium salcaprozate (SNAC), and combinations thereof. Labrasol® is also known as caprylcaproyl macrogol-8 glycerides and is manufactured by Gattefosse, Saint Priest, Lyon, France. In one embodiment, the oral dosage form comprises Labrasol®. In one embodiment, the oral dosage form comprises sodium caprate. When present in an oral dosage form, a permeation enhancer is used in an amount of up to about 1800 mg, in an amount of up to about 720 mg, in an amount of up to about 540 mg, in an amount of up to about 360 mg, in an amount ranging from about 90 mg to about 360 mg, in an amount ranging from about 180 to 360 mg, or in an amount of 90 mg, 180 mg, or 360 mg. In one embodiment of the invention, the oral dosage form comprises a permeation enhancer in an amount of up to about 360 mg, in an amount ranging from about 90 mg to about 360 mg, in an amount ranging from about 180 to about 360 mg, or in an amount of 90 mg, 180 mg, or 360 mg. In one embodiment, the oral dosage form of the invention comprises a permeation enhancer in an amount of 90 mg, 180 mg, or 360 mg. In one embodiment, the oral dosage form of the invention comprises a permeation enhancer in an amount of 180 mg or 360 mg.

[0066] When present in an oral dosage form, sodium caprate is used in an amount up to about 360 mg, in an amount ranging from about 90 mg to about 360 mg, in an amount ranging from about 180 to about 360 mg, or in an amount of 90 mg, 180 mg, or 360 mg. In one embodiment, an oral dosage form of the present invention comprises the permeation enhancer sodium caprate in an amount of 90 mg, 180 mg, or 360 mg. In one embodiment, 180 mg of sodium caprate is used in the oral dosage form. In one embodiment, 360 mg of sodium caprate is used in the oral dosage form.

[0067] In one embodiment of the present invention, dry-filled capsules or tablets may be used to administer the compound of formula (I) to a subject in need thereof. In the pharmaceutical composition of the present invention, a permeation enhancer may be included. In one embodiment, the amount of the permeation enhancer, such as sodium caprate, may range from 1% to 75% by weight. As used herein, weight % refers to the weight percentage of the component relative to the total weight of the pharmaceutical composition. In another embodiment, the amount of the permeation enhancer in the pharmaceutical composition is about 18% to about 65% by weight. For tablets, the amount of the permeation enhancer, such as sodium caprate, may range from about 22% to about 65% by weight. Oral dosage forms may be manufactured by standard methods, including wet granulation and dry granulation.

[0068] [Table 1]

[0069] In one embodiment, the present invention provides a method for the preparation of a compound of formula (I): [ka] [In the formula, A - is a pharma- ceutically acceptable anion. and a permeation enhancer. In a further embodiment, the permeation enhancer is sodium caprate. In another embodiment, the pharmaceutical composition further comprises a diluent. In a further embodiment, the composition comprises two or more diluents, and the two or more diluents comprise a combination of microcrystalline cellulose, macrogol (PEG4000), and lactose.

[0070] In one embodiment of the present invention, the pharmaceutical composition comprises: a) 1% to 7% by weight of the compound of formula (I) based on the total weight of the pharmaceutical composition; b) about 1% to 75% by weight of the permeation enhancer based on the total weight of the pharmaceutical composition; c) at least one diluent; and d) optionally including a glidant and / or lubricant. In one embodiment, about 18% to 74% by weight of the permeation enhancer based on the total weight of the pharmaceutical composition is present in the pharmaceutical composition. In another embodiment of the present invention, the pharmaceutical composition comprises: a) about 1% to about 7% by weight of the compound of formula (I) based on the total weight of the pharmaceutical composition; b) about 22% to about 67% by weight of the permeation enhancer selected from sodium caprate or Labrasol® based on the total weight of the pharmaceutical composition; c) at least one diluent or solubilizer selected from PEG4000, microcrystalline cellulose, propylene glycol, and lactose; d) optionally including a glidant; and e) optionally including a lubricant.

[0071] In one embodiment of the present invention, the pharmaceutical composition comprises: a) about 2% to 6% by weight of a compound of formula (I) based on the total weight of the pharmaceutical composition; b) about 18% to 74% by weight of a permeation enhancer which is sodium caprate based on the total weight of the pharmaceutical composition; c) at least one diluent selected from PEG4000, microcrystalline cellulose, or lactose; d) 0% to about 3% by weight of a lubricant which is silicon dioxide based on the total weight of the pharmaceutical composition; e) 0% to about 2% by weight of a lubricant which is magnesium stearate based on the total weight of the pharmaceutical composition; and, optionally, f) at least one disintegrant.

[0072] In one embodiment, the subject has a history of hypercholesterolemia treatment with one or more statin drugs, which may or may not have been discontinued. In other words, the subject treated with a compound of formula (I) is currently being treated or has previously been treated with statin therapy. In one embodiment, the subject is statin naive. In other words, the subject has never been treated with statin therapy. In one embodiment, the subject is concurrently treated with statin therapy, which may or may not have achieved therapeutic goals.

[0073] In one embodiment, one or more additional pharmacologically effective agents may be administered in combination with the compound of formula I. As used herein, "one or more additional pharmacologically effective agents" is intended to mean one or more pharmacologically effective agents that are effective in the body, including prodrugs that are different from the compound of formula I and are converted into a pharmacologically effective form after administration, and also including free acids, free bases, and pharma- ceutically acceptable salts of the additional pharmacologically effective agents. In general, one or more suitable additional pharmacologically effective agents, including but not limited to antihypertensive agents, lipid antiatherosclerotic agents such as lipid-modifying compounds, antidiabetic agents, and / or antiobesity agents, may be used in some form of combination with the compound of formula I in a single oral dosage form (fixed-dose combination drug), or administered to the subject in one or more separate dosage formulations. This may allow for simultaneous or sequential administration (co-administration of separate effective agents) of the compound of formula (I) and one or more additional pharmacologically effective agents.

[0074] Examples of additional pharmacologically active agents that may be used include angiotensin converting enzyme inhibitors (e.g., alacepril, benazepril, captopril, ceronapril, cilazapril, delapril, enalapril, enalaprilat, fosinopril, imidapril, lisinopril, moveltipril, perindopril, quinapril, ramipril, spirapril, temocapril, or trandolapril), angiotensin II receptor antagonists (e.g., losartan, i.e., COZAAR®, valsartan, candesartan, olmesartan, tel ... any of these drugs used in combination with hydrochlorothiazide, such as rutan and HYZAAR®, neutral endopeptidase inhibitors (e.g., thiorphan and phosphoramidon), aldosterone antagonists, aldosterone synthase inhibitors, renin inhibitors (e.g., urea derivatives of dipeptides and tripeptides (U.S. Pat. No. 5,116,835), amino acids and derivatives (U.S. Pat. Nos. 5,095,119 and 5,104,869), amino acid chains linked by non-peptide bonds (U.S. Pat. No. 5,114,937), dipeptide derivatives and tripeptide derivatives, peptidyl aminodiols and peptidyl beta-aminoacyl aminodiol carbamates, and small molecule renin inhibitors (diol sulfonamides and sulfinyls), N-morpholino derivatives, N-heterocyclic alcohols and pyrrole imidazolones, pepstatin derivatives and fluoro- and chloro-derivatives of statone-containing peptides, enalklein, RO42-5892, A65317, CP80794, ES1005, ES8891, SQ34017, aliskiren (2(S),4(S),5(S),7(S)-N-(2 -carbamoyl-2-methylpropyl)-5-amino-4-hydroxy-2,7-diisopropyl-8-[4-methoxy-3-(3-methoxypropoxy)-phenyl]-octanamide hemifumarate) SPP600, SPP630 and SPP635), endothelin receptor antagonists, phosphodiesterase-5 inhibitors (e.g., sildenafil, tadalfil and vardenafil), vasodilators, calcium channel blockers (e.g., amlodipine, nifedipine, verapamil, diltiazem, gallopamil, niludipine, nimodipine, nicardipine),Potassium channel activators (e.g., nicorandil, pinacidil, cromakalim, minoxidil, aprilkarim, loprazolam), diuretics (e.g., hydrochlorothiazide), sympatholytics, beta adrenergic blockers (e.g., propranolol, atenolol, bisoprolol, carvedilol, metoprolol or metoprolol tartrate), alpha adrenergic blockers (e.g., doxazosin, prazotin or alpha methyldopa), central alpha adrenergic agonists, peripheral vasodilators (e.g., hydrazine), lactone prodrug forms lipid-lowering agents, for example HMG-CoA reductase inhibitors, such as simvastatin and lovastatin, which are marketed in the U.S. as ZOCOR® and MEVACOR® and function as inhibitors after administration; atorvastatin (particularly the calcium salt sold under the trademark LIPITOR®), rosuvastatin (particularly the calcium salt sold under the trademark CRESTOR®), pravastatin (particularly the sodium salt sold under the trademark PRAVACHOL®), fluvastatin (particularly the sodium salt sold under the trademark LESCOL®), pharmacokinetic and pharmacokinetic parameters may vary, for example, with respect to the sodium salts thereof, pharmacokinetic and pharmacokinetic parameters, such as acetaminophen (sodium salts thereof, for example acetaminophen), ...pharmacokinetic and pharmacokinetic parameters, such as pharmacokinetic and pharmacokinetic parameters, such as pharmacokinetic and pharmacokinetic parameters, such as pharmacokinetic and pharmacokinetic parameters, such as pharmacokinetic and pharmacokinetic parameters, such as pharmacokinetic and pharmacokinetic parameters, such as pharmacokinetic and pharmacokinetic parameters, such as pharmacokinetic and pharmacokinetic parameters, such as pharmacokinetic and pharmacokinetic parameters, such as pharmacokinetic and pharmacokinetic parameters, such as pharmacokinetic and pharmacokinetic parameters, such as pharmacokinetic and niacin receptor agonists such as sifran, and niacin receptor partial agonists, metabolic modifiers including insulin and insulin mimetics (e.g., insulin degludec, insulin glargine, insulin lispro), dipeptidyl peptidase-IV (DPP-4) inhibitors (e.g., sitagliptin, alogliptin, linagliptin, vildagliptin), (i) PPARγ agonists such as the glitazones (e.g., pioglitazone, AMG131, MBX2044, mitoglitazone, lobeglitazone, IDR-105, rosiglitazone and balaglitazone),and (1) PPAR alpha / gamma dual agonists (e.g., ZYH2, ZYH1, GFT505, tiglitazar, muraglitazar, aleglitazar, soderglitazar, and naveglitazar), (2) PPAR alpha agonists such as fenofibric acid derivatives (e.g., gemfibrozil, clofibrate, ciprofibrate, fenofibrate, and bezafibrate), (3) selective PPAR gamma modulators (SPPAR gamma M's), (e.g., International Publication Nos. WO 02 / 060388, WO 02 / 08188, WO 2004 / 019869, WO 2004 / 023661, WO 2004 / 023666, WO 2004 / 023667, WO 2004 / 023668, WO 2004 / 023669, WO 2004 / 023666, WO 2004 / 023667 ... and (4) other PPAR ligands, including PPARγ partial agonists; (ii) metformin and its pharma- ceutically acceptable salts, particularly biguanides such as metformin hydrochloride and its extended release formulations such as Glumetza™, Fortamet™ and GlucophageXR™; and (iii) protein tyrosine phosphatase-1B (PTP-1B) inhibitors (e.g., ISIS-113715 and TTP814), insulin, insulin-like peptides ... or insulin analogues (e.g., insulin detemir, insulin glulisine, insulin degludec, insulin glargine, insulin lispro and their respective inhaled formulations), leptin and leptin derivatives and leptin agonists, amylin and amylin analogues (e.g., pramlintide), sulfonylurea and non-sulfonylurea insulin secretagogues (e.g., tolbutamide, glyburide, glipizide, glimepiride, mitiglinide, meglitinide, nateglinide and repaglinide), α-glucosidase inhibitors (e.g., acarbose, voglibose, and miglitol), glucagon receptor antagonists (e.g., MK-3577, MK-0893, LY-2409021 and KT6-971), incretin mimetics such as GLP-1, GLP-1 analogues, GLP-1 derivatives and GLP-1 mimetics, GLP-1 receptor agonists (e.g., dulaglutide, semaglutide, albiglutide, exenatide, liraglutide, lixisenatide, taspoglutide, CJC-1131 and BIM-51077, including intranasal, transdermal and once-weekly formulations thereof), bile acid sequestrants (e.g., colestilan, colestimide,colesevelam hydrochloride, colestipol, dialkylaminoalkyl derivatives of cholestyramine and cross-linked dextran), acyl-CoA:cholesterol acyltransferase inhibitors (e.g., avasimibe), anti-obesity compounds, drugs intended for use in inflammatory conditions such as aspirin, nonsteroidal anti-inflammatory drugs or NSAIDs, glucocorticoids, and selective cyclooxygenase-2 or COX-2 inhibitors, glucokinase activators (GKAs) (e.g., AZD6370), inhibitors of type I 11β-hydroxysteroid dehydrogenase (e.g., those disclosed in U.S. Pat. No. 6,730,690 and LY-2523199), CETP inhibitors (e.g., anacetrapib, torcetrapib, and evacetrapib), fructose 1,6-bisphosphatase (e.g., those disclosed in U.S. Pat. Nos. 6,054,587, 6,110,903, 6,284,748, 6,399,782, and 6,489,476), inhibitors of acetyl-CoA carboxylase-1 or 2 (ACC1 or ACC2), AMP-activated protein kinase (AMPK) activators, other agonists of G-protein coupled receptors: (i) GPR-109, (ii) GPR-119 (e.g., MBX2982 and PSN821), and (iii) GPR-40 (e.g., TAK875), SSTR3 antagonists (e.g., those disclosed in WO 2009 / 001836), neuromedin U receptor agonists (e.g., neuromedin S (NMS)), including, but not limited to, those disclosed in WO 2009 / 042053), SCD modulators, GPR-105 antagonists (such as those disclosed in WO 2009 / 000087), SGLT inhibitors (such as ASP1941, SGLT-3, empagliflozin, canagliflozin, BI-10773, ertugliflozin, remogroflozin, TS-071, tofogliflozin, ipragliflozin, and LX-4211), inhibitors of acyl-coenzyme A: diacylglycerol acyltransferase 1 and 2 (DGAT-1 and DGAT-2), inhibitors of fatty acid synthase, inhibitors of acyl-coenzyme A: monoacylglycerol acyltransferase 1 and 2 (MGAT-1 and MGAT-2),Other agents useful for the treatment of the above-mentioned conditions or disorders include agonists of the TGR5 receptor (also known as GPBAR1, BG37, GPCR19, GPR131 and M-BAR), agonists of the TGR5 receptor (GPBAR1, BG37, GPCR19, GPR131 and M-BAR), ileal bile acid transporter inhibitors, PACAP, PACAP mimetics and PACAP receptor 3 agonists, PPAR agonists, protein tyrosine phosphatase-1B (PTP-1B) inhibitors, IL-1b antibodies (e.g., XOMA052 and canakinumab), bromocriptine methyl salt and its rapid release formulations, and bempedoic acid, as well as, where chemically possible, the free acid, free base and pharma- ceutically acceptable salt forms of the above additional pharmacologically active agents.

[0075] In one embodiment, the additional pharmacologically active agent is a statin, ezetimibe, bempedoic acid, any other cholesterol-lowering agent considered to be standard of care, or any combination thereof.

[0076] In one embodiment, the method of the present disclosure further comprises administering a statin drug. Thus, the compound of formula (I) is co-administered with at least one statin drug. The compound of formula (I) can be administered with the statin drug simultaneously or separately. This co-administration can include co-administration of the compound of formula (I) and the statin drug in the same oral dosage form, co-administration in separate dosage forms, and separate administration. That is, the compound of formula (I) and the statin drug can be formulated together in the same oral dosage form and administered at the same time. Alternatively, the compound of formula (I) and the statin drug can be co-administered, both of which are in separate formulations. In another alternative, the compound of formula (I) can be administered immediately after the administration of the statin drug, or vice versa. In some embodiments of the separate administration protocol, the compound of formula (I) and the statin drug are administered within minutes, hours, or days apart.

[0077] In one embodiment, the LDL-C level of the subject after treatment with a compound of formula (I) is reduced from the baseline level of LDL cholesterol before treatment with a compound of formula (I). In one embodiment, the LDL-C level of the subject after treatment with a compound of formula (I) is reduced by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, more than 50%, at least 60%, more than 60%, at least 65%, more than 65%, at least 70%, more than 70%, at least 75%, more than 75%, at least 80%, more than 80%, at least 85%, more than 85%, or at least 90% from the baseline level of LDL-C before treatment with a compound of formula (I). In one embodiment, the LDL-C level of the subject after treatment with a compound of formula (I) is reduced by more than 50% from the baseline level of LDL-C before treatment with a compound of formula (I). In one embodiment, the subject's LDL-C level after treatment with a compound of formula (I) is reduced by more than 60% from the baseline level of LDL-C before treatment with a compound of formula (I). In one embodiment, the subject's LDL-C level after treatment with a compound of formula (I) is reduced by more than 65% from the baseline level of LDL-C before treatment with a compound of formula (I). In one embodiment, the subject's LDL-C level after treatment with a compound of formula (I) is reduced by more than 70% from the baseline level of LDL-C before treatment with a compound of formula (I). In one embodiment, the subject's LDL-C level is reduced by more than 50% from the baseline level of LDL-C 14 days after treatment of the subject with a compound of formula (I). In one embodiment, the subject's LDL-C level is reduced by more than 60% from the baseline level of LDL-C 14 days after treatment of the subject with a compound of formula (I). In one embodiment, the subject's LDL-C level is reduced by more than 65% from the baseline level of LDL-C 14 days after treating the subject with a compound of formula (I). In one embodiment, the subject's LDL-C level is reduced by more than 70% from the baseline level of LDL-C 14 days after treating the subject with a compound of formula (I). Both baseline and post-treatment levels of LDL-C can be determined by standard laboratory tests used to measure blood cholesterol.

[0078] In one embodiment, the subject's LDL-C level after treatment with a compound of formula (I) is reduced by at least 50% from the baseline level of LDL-C before treatment with a compound of formula (I). In one embodiment, the subject's LDL-C level after treatment with a compound of formula (I) is reduced by at least 60% from the baseline level of LDL-C before treatment with a compound of formula (I). In one embodiment, the subject's LDL-C level after treatment with a compound of formula (I) is reduced by at least 65% from the baseline level of LDL-C before treatment with a compound of formula (I). In one embodiment, the subject's LDL-C level after treatment with a compound of formula (I) is reduced by at least 70% from the baseline level of LDL-C before treatment with a compound of formula (I). In one embodiment, the subject's LDL-C level is reduced by at least more than 50% from the baseline level of LDL-C 14 days after treatment of the subject with a compound of formula (I). In one embodiment, the subject's LDL-C level is reduced by at least 60% from the baseline level of LDL-C after 14 days of treating the subject with a compound of formula (I). In one embodiment, the subject's LDL-C level is reduced by at least 65% from the baseline level of LDL-C after 14 days of treating the subject with a compound of formula (I). In one embodiment, the subject's LDL-C level is reduced by at least 70% from the baseline level of LDL-C after 14 days of treating the subject with a compound of formula (I). Both baseline and post-treatment levels of LDL-C can be determined by standard laboratory tests used to measure blood cholesterol.

[0079] In one embodiment, the present invention relates to a method of reducing apolipoprotein B (apo B) levels in a subject in need thereof, comprising orally administering to the subject an amount of a compound of formula (I). In some embodiments, after treatment with a compound of formula (I), the apolipoprotein B (apo B) levels in the subject in need thereof are reduced from the baseline level of apo B before treatment with a compound of formula (I). In one embodiment, after treatment with a compound of formula (I), the apo B levels in the subject are reduced by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, more than 50%, at least 60%, more than 60%, at least 65%, more than 65%, at least 70%, more than 70%, at least 75%, more than 75%, at least 80%, more than 80%, at least 85%, more than 85%, or at least 90% from the baseline level of apo B before treatment with a compound of formula (I).

[0080] In one embodiment, the subject's non-high density lipoprotein cholesterol (non-HDL-C) levels after treatment with a compound of Formula (I) are reduced from the baseline level of non-HDL-C before treatment with a compound of Formula (I). In one embodiment, the subject's non-HDL-C levels after treatment with a compound of Formula (I) are reduced by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, more than 50%, at least 60%, more than 60%, at least 65%, more than 65%, at least 70%, more than 70%, at least 75%, more than 75%, at least 80%, more than 80%, at least 85%, more than 85%, or at least 90% from the baseline level of non-HDL-C before treatment with a compound of Formula (I).

[0081] Inhibition or antagonism of one or more of the PCSK9-related functional properties can be readily determined according to methodologies known in the art (see, e.g., Barak & Webb, 1981 J. Cell Biol. 90:595-604; Stephan & Yurachek, 1993 J. Lipid Res. 34:325330; and McNamara et al., 2006 Clinica Chimica Acta 369:158-167) and described herein. Inhibition or antagonism achieves a decrease in PCSK9 activity compared to the activity seen in the absence of an antagonist, or compared to the activity observed, for example, in the presence of a control antagonist of irrelevant specificity. Preferably, compounds of formula (I) antagonize functional PCSK9 to the point of at least a 10% reduction in a measured parameter, including but not limited to the activities disclosed herein, and more preferably to the point of at least a 20%, 30%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90% and 95% reduction in a measured parameter.

[0082] The compound of formula (I) is highly effective in lowering LDL cholesterol and is generally well tolerated after single and multiple oral doses in healthy volunteers. The compound of formula (I) reduces the level of free PCSK9 protein, which contributes to high LDL cholesterol, by more than 90% from baseline after treatment with a single dose of the compound of formula (I). After 14 days of oral administration once daily, the compound of formula (I) reduces blood LDL cholesterol by about 65% from baseline levels in participants already receiving moderate to high intensity statin basal therapy. These participants were already taking statin drugs to control their cholesterol levels. The compound of formula (I) can be a highly effective treatment for patients suffering from high cholesterol.

[0083] These examples are provided for the purpose of further illustration only and are not intended as limitations on the present disclosure.

[0084] [Example] [Example 1] Preparation of Compound 1 (Amorphous Chloride Salt) [ka]

[0085] A 50 L cylindrical reactor was charged with 0.5 L of acetonitrile (MeCN) followed by compound 4 (294.7 g, 206 mmol) at room temperature (methods for synthesizing starting material compound 4 are described in WO 2019 / 246349, see Example 1). An additional 2.4 L of MeCN was used to rinse all solids into the bottom of the reactor. Compound 5 (5-carboxy-N,N,N-trimethylpentan-1-aminium chloride, 47.5 g, 227 mmol) was added. An additional 2.0 L of MeCN was used to rinse all solids into the bottom of the reactor. N,N-diisopropylethylamine (iPr 2 NEt, 216 mL, 1236 mmol) was added and 0.5 L of MeCN was used to rinse the liquid into the bottom of the reactor. The reactor was charged with 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium-3-oxide hexafluorophosphate (HATU, 94 g, 247 mmol) and 0.5 L of MeCN was used to rinse all of the solids into the bottom of the reactor. After 3 hours at room temperature, isopropyl acetate (iPrOAc, 17.7 L) was added dropwise over 1 hour. The slurry was filtered and the wet cake was washed 3 times with 2.9 L of iPrOAc. N 2 The solid was dried under vacuum using a sweep to give 337 g of crude product.

[0086] Supercritical fluid chromatography (stationary phase: DIACEL DCpak P4VP [30 × 250 mm, 5 μm]; mobile phase: 45% modifier (0.25% NH in MeOH) 4 OH and 5% H 2 O) and 55% CO 2The crude product was purified using a 0.22 μm line filter. The fractions containing the product were concentrated using rotary evaporation. The residue after evaporation was dissolved in water (3.2 L) and 0.1 M aqueous HCl (1389 mL, 139 mmol) was added at room temperature (pH was measured to be 6 at the end of the addition using pH paper). The resulting solution was filtered through a 0.22 μm line filter and the filtrate was lyophilized to give 238 g of compound 1 (amorphous chloride salt).

[0087] [Example 2] Preparation of frozen Compound 2 (amorphous caprate salt) Porous anion exchange resin AG MP-1M (6 g, 100-200 mesh, chloride form) was loaded into a 60 mL funnel. The loaded resin was washed five times with 9 mL of a mixture of acetonitrile and water (1:1). The resin was washed twice with 200 mL of 1 M NaOH, then with 10 mL of water. The resin was transferred to a glass column and washed three times with 10 mL of water. The resin was then washed two times with 10 mL of EtOH, followed by five times with 9 mL of 1 M caprylic acid EtOH solution, followed by three times with 9 mL of EtOH. Compound 1 (0.3 g) was dissolved in 6 mL MeCN / water (1:1) and placed into the resin-packed column. The filtrate was collected in a 20 mL vial. The column was washed three times with 15 mL of a solution of MeCN and water (1:1) and the filtrate was collected in a 20 mL vial. Fractions containing the caprate salt of compound 2 were combined and concentrated to remove MeCN, and the desired amorphous compound 2 (0.29 g) was then isolated by lyophilization.

[0088] [Example 3] Preparation of freeze-dried compound 3 (amorphous acetate salt) Porous anion exchange resin AG MP-1M (6 g, 100-200 mesh, chloride form) was loaded into a 60 mL funnel. The loaded resin was washed five times with 9 mL of acetonitrile / water mixture (1:1 ratio). The resin was washed twice with 200 mL of 1 M NaOH, then 50 mL of 1 M AcOH in water. The resin was transferred to a 100 mL round bottom flask containing a solution of compound 1 (chloride salt, 0.3 g) in 6 mL of a 1:1 mixture of acetonitrile and water. An additional 18 mL of MeCN / water (1:1) was added. The mixture was aged at room temperature for 30 min, and the resulting mixture was transferred to a 60 mL funnel. The filtrate was collected and the resin was washed three times with 10 mL MeCN / water (1 / 1), and the filtrate was collected in a 20 mL vial. The fractions containing compound 3 were combined and concentrated to remove MeCN. The desired amorphous compound 3 (0.304 g) was then isolated by lyophilization of the solution.

[0089] [Example 4] Preparation of tablets containing a compound of formula (I) Sodium caprate (1.5 kg) and macrogol (499.9 g) were placed in a 10 L high shear granulator. The two components were dry mixed in a high shear granulator for 1 minute at an impeller speed of 183 rpm. During continuous mixing in the high shear granulator, water was added until the appropriate granulation was reached. The wet powder was milled in a cone mill with a 2.0 mm screen size, then transferred to a fluidized bed dryer, using an inlet temperature of 70° C., and dried until the specified loss on drying of the wet powder was reached (less than 3.00%). The dry powder was milled in a cone mill with a 1.0 mm screen size. The dry powder (1.275 kg) was then mixed in a 10 L diffusion blender with compound 1 (26.96 g), lactose (150.4 g) and silicon dioxide (22.58 g) at 920 revolutions. It was then milled in a cone mill with a 0.8 mm screen size. The mixed blend was then mixed with magnesium stearate (22.58 g) in a 10 L diffusion blender at 460 revolutions. The finally lubricated powder was compressed into tablets using a rotary tablet press with a target weight of 564.9 mg.

[0090] [Example 5] Dry-filled capsule manufacturing process Microcrystalline cellulose (179.7 g), sodium caprate (661 g), compound 1 (41 g) and silicon dioxide (8.996 g) were mixed using a 10 L diffusion blender at 375 revolutions. Magnesium stearate (4.498 g) was then added to the blender and mixed for 250 revolutions. The blend was then granulated by roller compaction using a roll pressure of 21 bar, a coarse screen of 2.0 mm and a fine screen of 1 mm. The roller compacted powder (761.9 g) was mixed with magnesium stearate (3.8 g) using a 5 L diffusion blender at 250 revolutions. The final lubricated powder was then manually encapsulated with a target fill weight of 490 mg.

[0091] [Example 6] Liquid-filled capsule manufacturing process A solution of Labrasol® ALF (caprylocaproyl macrogol-8 glyceride, 100 mL) and propylene glycol (50 mL) was prepared as solvent in a 250 mL bottle using a stir plate. Compound 1 (0.7747 g) was dissolved in solvent (49.7 mL) in a 125 mL bottle using a stir plate for 5 minutes, followed by sonication for 15 minutes. The final solution was filled into hard gelatin capsules to a target weight of 548 mg. The hard gelatin capsules were then manually sealed using 50% aqueous ethanol and inspected for leaks. The final hard gelatin capsules were encapsulated in enteric capsules that were manually sealed using 90% aqueous ethanol. A 50% aqueous ethanol solution was prepared in a 30 mL bottle by mixing 10.4 mL of ethanol (96%) and 9.6 mL of water using a stir plate for 15 minutes. A 90% aqueous ethanol solution was prepared in a 30 mL bottle by mixing 18.8 mL of ethanol (96%) with 1.2 mL of water using a stir plate for 15 min.

[0092] test Compound 1, which is chemically stable and resistant to gastrointestinal (GI) degradation: [ka] Amorphous chloride salts of compounds of formula (I), such as , have demonstrated picomolar binding affinity to human PCSK9. GI absorption of Compound 1 was improved by co-administration with permeation enhancers (Labrasol, sodium caprate) in rats and non-human primates. Pre-clinical Good Laboratory Practice (GLP) toxicity studies in rats and non-human primates support clinical development and were conducted using both subcutaneous (to achieve high forward exposure of Compound 1) and oral-arm (to assess local / GI tolerability). No adverse events were observed in these GLP toxicity studies up to and including the highest dose administered.

[0093] Safety Testing The pharmacokinetics, pharmacodynamics (reduction in free PCSK9 from baseline), and safety and tolerability of a single dose of the compound of formula (I) were studied in normal, healthy male volunteers aged 18-50 years. The objective of this study was to evaluate the safety and tolerability of a single dose of compound 1 (about 10 mg to about 300 mg), as well as the pharmacokinetics (PK) of compound 1. Additionally, this study tested the effect of permeation enhancer administration on PK, the effect of food on PK, and the effect of various capsule formulations on PK. The pharmacodynamic endpoint measured in this study was target engagement (% change in free PCSK9). For each panel in this study, participants were randomized to receive either compound 1 or placebo (PBO) in a 9:3 randomization scheme (n=9 compound 1:n=3 PBO). The baseline characteristics of the participants in this study are shown in Table 2.

[0094] [Table 2]

[0095] A single dose of Compound 1, the amorphous chloride salt of the compound of formula (I), was administered in a liquid-filled hard gelatin capsule. The capsules contained various strengths of Compound 1, or no Compound 1 (placebo), and a 2:1 mixture of the liquid permeation enhancer Labrasol® and propylene glycol, with varying amounts of Labrasol® up to 1800 mg. The capsules were encapsulated in enteric capsules (HPMC Vcaps® Enteric, Capsugel®, Lonza).

[0096] The study also evaluated a 40 mg / mL suspension of Compound 1 in a 2:1 ratio of OraBlend SF and propylene glycol without permeation enhancers administered via syringe / PO administration, and dry-filled enteric-coated capsules (HPMC Vcaps® Enteric, Capsugel®, Lonza) containing various strengths of Compound 1 and sodium caprate up to 1800 mg. The minimum dose of Compound 1 in the study was 10 mg, and the maximum dose administered was 300 mg. Compound 1 was well tolerated at doses up to 300 mg, with no deaths, serious adverse events, or clinically meaningful trends in clinical safety tests, vital signs, or ECGs as of study treatment. There were no deaths or severe adverse events (SAEs) in the study. Of the 60 total participants, 6 discontinued, 3 due to adverse events (maculopapular rash, concussion / trauma-related wounds, and back pain), 2 due to protocol violations, and 1 due to withdrawal due to conflict with the participant's job. Adverse Events (AEs) related to Compound 1 reported by the investigators in this study included abdominal discomfort, diarrhea, dyspepsia, headache, and maculopapular rash. All treatment-related AEs were mild / moderate except for one participant who had severe back pain not related to treatment.

[0097] Compound 1 (amorphous chloride salt of the compound of formula (I)) exhibited a dose-dependent increase in plasma exposure and a mean maximum reduction in free plasma PCSK9 levels from baseline of greater than 90% at all dose levels tested. See Figure 2 and Table 3 below.

[0098] Pharmacokinetic results are shown in Figure 1. The study also demonstrated that permeation enhancers improved absorption, as evidenced by increased Cmax and AUC0-24 (see Figure 2). The PK of Compound 1 in the presence of permeation enhancers Labrasol® and sodium caprate was similar (as shown in Figure 2). The study also demonstrated that food consumed 30 minutes prior to dosing resulted in lower plasma exposure compared to the fasted state, and food consumed 30 minutes after dosing had negligible effect on plasma exposure (see Figure 2).

[0099] As can be seen in FIG. 4, administration of Compound 1 is associated with a decrease in plasma levels of free PCSK9 protein, which contributes to over 90% of elevated LDL cholesterol compared to baseline levels.

[0100] [Table 3]

[0101] LDL cholesterol lowering test Dosing of Compound 1 to achieve a target LDL-C reduction of >50% was evaluated using a multiple-dose study in male and female participants aged 18-65 years receiving statin background therapy to control blood cholesterol. Participants' baseline mean LDL-C was approximately 87 mg / dL, with 85% of participants receiving moderate or high-intensity statins. Either placebo or Compound 1 was administered once daily for 14 days in the morning after an overnight fast. After receiving the daily dose, participants were provided with a standard take-home meal and allowed to consume it for only 30 minutes. Plasma lipids (total cholesterol, LDL-C, HDL-C and TG) were measured in addition to vital signs, ECGs and standard safety monitoring, including clinical safety tests. LDL-C was monitored as part of a safety laboratory.

[0102] A starting dose of 20 mg compound 1 and 360 mg sodium caprate was associated with a mean reduction in plasma LDL-C of approximately 62%. 10 mg compound 1 and 360 mg sodium caprate was the next dose tested. 10 mg compound 1 and 360 mg sodium caprate was associated with a mean reduction in LDL-C of approximately 64%. The third dose level was also 10 mg compound 1, but the formulation contained 180 mg sodium caprate. The 10 mg compound 1 and 180 mg sodium caprate dose was associated with a mean reduction in LDL-C of approximately 60%. PK from this dose was similar to that of the 10 mg compound 1 and 360 mg sodium caprate dose, supporting the similarity of LDL-C reduction. All formulations containing either sodium caprylate alone (placebo) or both sodium caprate and Compound 1 were in the form of dry-filled enteric-coated capsules (HPMC Vcaps® Enteric, Capsugel®, Lonza).

[0103] Blood LDL cholesterol levels were measured pre-dose and on days 3, 7, 14, 15, and 21 post-dose using standard clinical laboratory procedures. The results of this study are presented in Figure 3. As shown in Figure 3, the maximum reduction in LDL cholesterol observed at the 10 mg and 20 mg doses is in the range of LDL-C reductions observed with the anti-PCSK9 monoclonal antibodies Repatha and Praluent, as reported in a Phase III cardiovascular study and a Phase III lipid study of the anti-PCSK9 siRNA Inclisiran. See Repatha cardiovascular outcomes trial FOURIER reporting 59% reduction in LDL-C, N Engl J Med 2017 May 4,376(18):1713-1722; Praluent cardiovascular outcomes trial ODYSSEY reporting 59% reduction in LDL-C, N Engl J Med 2018,379:2097-2107; and Inclisiran phase 3 lipid trials reporting 49-52% reduction in LDL-C, N Engl J Med 2020,382:1507-1519. In contrast, placebo-treated participants experienced less than a 5% reduction in LDL-C from baseline.

[0104] Hard gelatin capsules containing 5 mg of Compound 1 and 180 mg of sodium caprate were administered to male and female participants taking statins to control cholesterol in a separate study. No % LDL-C reduction was observed in the study reported above (less than 50% reduction from baseline).

Claims

1. In the manufacture of a medicament for a method of treating hypercholesterolemia in a subject in need thereof, Compound 2 shown below: 【Chemistry 1】 said method comprising orally administering to said subject a therapeutically effective amount of Compound 2 in an oral dosage form further comprising sodium caprate; The use wherein the therapeutically effective amount of compound 2 administered is 5 mg to 300 mg.

2. The use according to claim 1, wherein the therapeutically effective amount of compound 2 administered is 10 mg to 30 mg.

3. 3. The use according to claim 1 or 2, wherein the therapeutically effective amount of compound 2 administered is 10 mg, 12.5 mg, 15 mg, 17.5 mg, 18 mg or 20 mg.

4. 3. The use according to claim 1 or 2, wherein the therapeutically effective amount of compound 2 administered is 10 mg or 20 mg.

5. 3. The use according to claim 1 or 2, wherein the oral dosage form comprises 180 mg of sodium caprate.

6. 3. The use of claim 1 or 2, wherein compound 2 is administered in a single oral dosage form administered once daily for at least 14 days.

7. 3. The use of claim 1 or 2, wherein the subject is currently being treated or has previously been treated with statin therapy.

8. 3. The use of claim 1 or 2, wherein the subject's level of LDL cholesterol after treatment is reduced from the subject's baseline level of LDL cholesterol before treatment.

9. 9. The use of claim 8, wherein the level of LDL cholesterol in the subject after treatment is reduced by more than 50% from the baseline level of LDL cholesterol before treatment.

10. The use according to claim 1 or 2, wherein the subject is a human.

11. In the manufacture of a medicament for a method of lowering LDL-C in a subject in need thereof, Compound 2 shown below: 【Transformation 5】 said method comprising orally administering to said subject a therapeutically effective amount of Compound 2 in an oral dosage form further comprising sodium caprate; The use wherein the therapeutically effective amount of compound 2 is 5 mg to 300 mg.

12. In the manufacture of a medicament for a method of treating atherosclerotic cardiovascular disease in a subject in need thereof, Compound 2 shown below: 【Transformation 6】 said method comprising orally administering to said subject a therapeutically effective amount of Compound 2 in an oral dosage form further comprising sodium caprate; The use wherein the therapeutically effective amount of compound 2 is 5 mg to 300 mg.

13. In the manufacture of a medicament for a method of inhibiting PCSK9 activity in a subject in need of treatment, 【Transformation 5】 said method comprising orally administering to said subject a therapeutically effective amount of Compound 2 in an oral dosage form further comprising sodium caprate; The use wherein the therapeutically effective amount of compound 2 is 5 mg to 300 mg.

14. The use according to claim 1 or 2, wherein the subject in need of administration fasts for about 30 minutes prior to administration of Compound 2.

15. Compound 2 shown below: 【Transformation 8】 and sodium caprate.

16. 16. The pharmaceutical composition of claim 15, further comprising a diluent selected from polyethylene glycol, microcrystalline cellulose, mannitol, starch, dicalcium phosphate, calcium carbonate, sodium carbonate, lactose, or a combination thereof.

17. 17. The pharmaceutical composition of claim 16, wherein the diluent is selected from microcrystalline cellulose, lactose, or macrogol (PEG 4000).

18. 17. The pharmaceutical composition according to claim 15 or 16, wherein the pharmaceutical composition is in tablet form.

19. 17. The pharmaceutical composition according to claim 15 or 16, wherein the pharmaceutical composition is in capsule form.

20. The pharmaceutical composition comprises: a) 1% to 7% by weight of compound 2 based on the total weight of the pharmaceutical composition; b) 1% to 75% by weight of sodium caprate based on the total weight of the pharmaceutical composition; c) at least one diluent; Including, d) Glidants and / or lubricants 16. The pharmaceutical composition of claim 15, which may comprise:

21. The pharmaceutical composition comprises: a) 2% to 6% by weight of compound 2 based on the total weight of the pharmaceutical composition; b) 18% to 74% by weight of sodium caprate based on the total weight of the pharmaceutical composition; c) at least one diluent selected from PEG 4000, microcrystalline cellulose, and lactose; d) 0% to 3% by weight of a lubricant, which is silicon dioxide, based on the total weight of the pharmaceutical composition; e) 0% to 2% by weight of a lubricant, which is magnesium stearate, based on the total weight of the pharmaceutical composition; Including, f) at least one disintegrant 16. The pharmaceutical composition of claim 15, which may comprise:

22. The use according to any one of claims 11 to 13, wherein the subject is a human.

23. The use according to any one of claims 11 to 13, wherein the therapeutically effective amount corresponds to 20 mg of compound 2 in free form.

24. 17. The pharmaceutical composition according to claim 15 or 16, comprising compound 2 in an amount equivalent to 20 mg in free form.

25. 17. The pharmaceutical composition of claim 15 or 16, comprising 22 mg, 22.5 mg, 23 mg, 23.5 mg, 24 mg or 24.5 mg of Compound 2.

26. 17. A pharmaceutical composition according to claim 15 or 16 for use in the treatment of hypercholesterolemia.

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