Methods of treating hypercholesterolemia

Abstract

A method of reducing elevated plasma levels of cholesterol in a subject is disclosed. Also disclosed is a method of treating hypercholesterolemia in a subject. The method comprises administering to the subject an effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, in the absence of a cholesterol-lowering drug.

Description

[0001] Related applications

[0002] This application claims the benefit of U.S. Provisional Application No. 63 / 067,786, filed August 19, 2020, pursuant to 35 U.S. SC §119(e), the entire contents of which are incorporated herein by reference. Background Technology

[0003] High cholesterol is one of the many risk factors for heart attack and stroke. Poor diet and lack of exercise are common causes of high cholesterol. High cholesterol may also have underlying genetic causes, such as the common hypercholesterolemia (FH). Many cholesterol-lowering drugs are currently available, but they are not without risks or contraindications for certain conditions or other medications. These drugs include statins, fibrates, niacin, bile acid sequestrants (resins), phytosterols, or other compounds that prevent fat absorption, reduce cholesterol absorption, or target genes in cholesterol transport pathways. Due to the risks and contraindications associated with current cholesterol-lowering drugs, further medications are needed to lower cholesterol levels in patients. Summary of the Invention

[0004] Compound 1 has been found to be an inhibitor of multiple enzymes in the cholesterol biosynthesis pathway, including LBR / TM7SF2 and EBP, and its structure is shown below.

[0005]

[0006] 1-((6-(((1s,4s)-4-ethylcyclohexyl)oxy)naphth-2-yl)methyl)piperidin-4-carboxylic acid

[0007] Specifically, compound 1 reduced cholesterol levels in healthy human volunteers in a dose-dependent manner (Example 1) and induced the accumulation of 7-dehydrocholesterol (7-DHC). The accumulation of 7-DHC was also reproduced in rat OPCs treated with compound 1 (Example 2). Based on these findings, this paper discloses a method for reducing blood cholesterol levels in subjects in need.

[0008] One embodiment of the present invention is a method for reducing elevated plasma cholesterol levels in a subject, the method comprising administering an effective amount of compound 1 to the subject:

[0009]

[0010] Or its pharmaceutically acceptable salt.

[0011] Another embodiment of the present invention is a method for treating a subject suffering from hypercholesterolemia, the method comprising administering an effective amount of compound 1 to the subject:

[0012]

[0013] Or its pharmaceutically acceptable salt.

[0014] Another embodiment of the invention is an effective amount of compound 1 or a pharmaceutically acceptable salt thereof, used to reduce elevated plasma cholesterol levels in a subject.

[0015] Another embodiment of the invention is the use of compound 1 or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for reducing elevated cholesterol plasma levels in a subject.

[0016] Another embodiment of the invention is an effective amount of compound 1 or a pharmaceutically acceptable salt thereof, used to treat a subject suffering from hypercholesterolemia.

[0017] Another embodiment of the invention is the use of an effective amount of compound 1 or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for use in treating a subject suffering from hypercholesterolemia. Attached Figure Description

[0018] Figure 1 This study demonstrates the time course of decrease in circulating mean total cholesterol levels in healthy volunteers when administered daily with placebo, 10 mg, 30 mg, or 60 mg of compound 1 over a 28-day period.

[0019] Figure 2 This is a bar graph showing the predicted steady-state concentrations of circulating cholesterol levels under pharmacokinetic steady-state during treatment with compound 1 in subjects receiving placebo, 1 mg, 3 mg, 10 mg, 30 mg, or 60 mg of compound 1 daily. The predicted concentrations are derived from simulations based on data from three Phase I trials of compound 1.

[0020] Figure 3 This is a bar graph showing the changes in 7-DHC, cholesterol, and sterol levels in rat OPC cultures treated with compound 1.

[0021] Figure 4 This is a diagram showing the biosynthetic pathways of cholesterol and sterols. Detailed Implementation

[0022] Compound 1 inhibits cholesterol biosynthesis, thus causing a decrease in plasma cholesterol levels, particularly peripheral cholesterol, in patients. Based on these results, this paper discloses methods for reducing elevated plasma cholesterol levels in subjects and treating hypercholesterolemia in subjects. According to the methods described herein, in some embodiments, based on animal data, compound 1 or a pharmaceutically acceptable salt thereof reduces peripheral cholesterol in humans but does not significantly reduce cholesterol in the central nervous system.

[0023] Subjects with hypercholesterolemia have elevated plasma cholesterol levels. "Elevated plasma cholesterol levels" are defined as plasma levels greater than 170 mg / dL, 200 mg / dL, 210 mg / dL, 220 mg / dL, 230 mg / dL, 240 mg / dL, 250 mg / dL, 260 mg / dL, 270 mg / dL, 300 mg / dL, 3300 mg / dL, or 3600 mg / dL.

[0024] The disclosed methods can be used to lower plasma cholesterol levels in diseases or disorders associated with or characterized by elevated blood cholesterol levels. "Diseases or disorders associated with or characterized by elevated blood cholesterol levels" refers to diseases or disorders where elevated blood cholesterol levels are a common or typical symptom. Examples include familial hypercholesterolemia, atherosclerosis, acute coronary syndrome (ACS), coronary artery disease, peripheral artery disease (PAD), cerebrovascular disease, cardiovascular disease caused by diabetes, macular degeneration, or congestive heart failure.

[0025] Compound 1 can be combined with an effective amount of another cholesterol-lowering drug in subjects with elevated plasma cholesterol levels or who have hypercholesterolemia. When co-administered with another drug that effectively treats hypercholesterolemia or lowers elevated plasma cholesterol levels, Compound 1 and the other drug can be administered simultaneously (in the same or different formulations) or at different times.

[0026] "Cholesterol-lowering drugs" are medications prescribed and / or administered to lower cholesterol levels in human patients with elevated cholesterol levels. Examples include statins, PCSK9 inhibitors, selective cholesterol absorption inhibitors, bile acid sequestrants, fibrates, or lipid-lowering therapies.

[0027] Statins are cholesterol-lowering drugs that work by inhibiting HMG-CoA reductase. Examples include atorvastatin. Fluvastatin (LESCOL) lovastatin Pitavastatin ), pravastatin ), rosuvastatin EZALLOR TM ) and simvastatin ).

[0028] PCSK9 inhibitors are cholesterol-lowering drugs that work by inhibiting the proprotein convertase subtilisin / kexin 9 serine protease. Examples include alicurumab and evolocumab.

[0029] Selective cholesterol absorption inhibitors are cholesterol-lowering drugs that work by inhibiting the absorption of cholesterol in the intestine. For example, ezetimibe... It is a selective cholesterol absorption inhibitor that works by inhibiting the transporter Niemann-Pick C-1-like 1 protein (NPC1L1).

[0030] Bile acid sequestrants are cholesterol-lowering drugs that work by binding to bile acids in the intestines and increasing their excretion in feces. This reduces the amount of bile acids returning to the liver and forces the liver to produce more bile acids to replace those lost in feces. To produce more bile acids, the liver converts more cholesterol into bile acids, thereby lowering blood cholesterol levels. Examples include cholestyramine. ), colestipol ) and colesevelam, ).

[0031] Fibrates (fibrates) are a class of drugs that lower blood triglyceride levels. Fibrates lower blood triglyceride levels by reducing the production of VLDL (volumes of triglycerides circulating in the blood) by the liver and by accelerating the removal of triglycerides from the blood. Fibrates also have a moderate effect on increasing blood HDL cholesterol levels. Examples of fibrates include gemfibrozil. ) and fenofibrate ).

[0032] Other cholesterol-lowering medications include fish oil, niacin (nicotinic acid cholesterol), and bempedioic acid. ) and probucol.

[0033] "Effective amount" means the amount of a drug that reduces one or more symptoms of a disease or condition and / or slows the progression of the disease or condition. Regarding Compound 1 used to treat hypercholesterolemia or lower elevated plasma cholesterol levels, "effective amount" includes the amount that lowers plasma cholesterol levels or otherwise reduces symptoms of a disease or condition associated with elevated plasma cholesterol levels. Examples include, but are not limited to, reductions in atherosclerosis, reductions in arterial wall stiffness, and reductions in isolated systolic hypertension. Exemplary effective amounts of Compound 1 for lowering elevated plasma cholesterol levels or treating hypercholesterolemia include, but are not limited to, 10 mg to 60 mg daily (or a pharmaceutically acceptable salt of Compound 1 equivalent to 10 mg to 60 mg of Compound 1), such as 10 mg daily, 30 mg daily, or 60 mg daily. Exemplary effective amounts of pharmaceutically acceptable salts of Compound 1 include, but are not limited to, amounts equivalent to 10 mg to 60 mg daily of Compound 1, such as amounts equivalent to 10 mg daily, 30 mg daily, or 60 mg daily of Compound 1. In some embodiments, the effective amount of compound 1 may be between 10 mg and 20 mg daily, between 20 mg and 30 mg daily, between 30 mg and 40 mg daily, between 40 mg and 50 mg daily, or between 50 mg and 60 mg daily. In some embodiments, the effective amount of a pharmaceutically acceptable salt of compound 1 may be an amount equivalent to 10 mg to 20 mg daily, 20 mg to 30 mg daily, 30 mg to 40 mg daily, 40 mg to 50 mg daily, or 50 mg to 60 mg daily of compound 1.

[0034] As used herein, when expressing a numerical range, it includes both endpoints. For example, amounts from 10 mg to 60 mg include both 10 mg and 60 mg. Similarly, amounts between 10 mg and 20 mg include both 10 mg and 20 mg.

[0035] "Subject" and "patient" are used interchangeably and refer to mammals in need of treatment, such as companion animals (e.g., dogs, cats), farm animals (e.g., cattle, pigs, horses, sheep, goats), and laboratory animals (e.g., rats, mice, guinea pigs). Typically, a subject is a person in need of treatment.

[0036] The synthesis and preparation of compound 1 and suitable formulations of compound 1 are described in U.S. Patent No. 9,340,527, the entire teachings of which are incorporated herein by reference.

[0037] In compound 1, the two substituents on the cyclohexyl group have a cis configuration relative to each other. When referring to the name or structure of compound 1, its stereochemical purity is at least 90 wt%, at least 95 wt%, at least 98 wt%, or at least 99 wt%. Stereochemical purity is the weight ratio of the cis-configured compound to the sum of the cis and trans-configured compounds.

[0038] The present invention is illustrated by the following embodiments, which are not intended to be limiting in any way.

[0039] example

[0040] Example 1 - As demonstrated by the accumulation of 7-DHC, compound 1 inhibits DHCR7 activity.

[0041] Forty-two healthy volunteers received Compound 1 QD for 28 days (or before early withdrawal). Six individual participants in each cohort received either 1 mg (cohort 1), 3 mg (cohort 2), 10 mg (cohort 3), 30 mg (cohort 4), a 60 mg loading dose and a 10 mg maintenance dose (cohort 5) of Compound 1, 60 mg (cohort 6), or a 90 mg loading dose and a 30 mg maintenance dose on days 1 and 2 (cohort 7). Fourteen participants also received a placebo; one participant in cohort 4 made a dosing error on day 19 and appeared to have received at least one 30 mg dose of Compound 1. Participants in cohorts 1 through 5 reported the following protocol deviations: they randomly drank water before dosing (not limited to 1 hour before and 1 hour after dosing) and ate food 30 minutes after dosing (not 4 hours after dosing). A total of 49 participants completed treatment, including 37 who received active treatment.

[0042] Participants received their first dose of study treatment (compound 1 or placebo) on day 1 and continued to receive the study treatment once daily until day 28. Participants remained at the clinic throughout the treatment period. Participants were discharged on day 29 after completing all assessments.

[0043] Blood was periodically drawn from each volunteer and immediately stored at -80°C. To measure the concentration of metabolites in human serum, the samples were centrifuged, and the resulting supernatant was used for further analysis.

[0044] Free oxygen sterols were extracted from the sample using methanol via a Biocrates Kit filter plate. The plate was pre-loaded with an internal standard mixture. Electrospray ionization (ESI) was performed using a SCIEX API 5500. Metabolite concentrations were determined using an AB SCIEX, Darmstadt, German instrument via UHPLC-MS / MS in positive mode with multiple reaction monitoring (MRM). Data were quantified using appropriate mass spectrometry software and input into Biocrates Met / DQ. TM The software will be used for further analysis.

[0045] Figure 1 The figure shows the mean circulating total cholesterol level in healthy volunteers, indicating a gradual, time-dependent, and dose-dependent decrease in total circulating cholesterol.

[0046] Based on these observations, a population pharmacokinetic / pharmacodynamic (PK / PD) model was developed using Monolix to describe circulating cholesterol concentration as a function of plasma concentration and exposure to compound 1. This model was developed based on cholesterol data from the aforementioned study (Study 1) and two other clinical studies (Study 2 and Study 3) conducted in healthy volunteers. In Study 2, 30 healthy volunteers received a single oral dose of compound 1, with 6 individual participants in each cohort receiving 3 mg (cohort 1), 10 mg (cohort 2), 30 mg (cohort 3), 60 mg (cohort 4), and 100 mg (cohort 5). Nine participants in the study also received a placebo. In Study 3, 8 healthy adult volunteers received a single 30 mg dose of compound 1.

[0047] In this model, circulating cholesterol concentrations decreased at all daily doses above 10 mg. Although data variability can affect predictive performance, the model demonstrates clear evidence of a dose-dependent decrease in circulating cholesterol concentration. This reduction in EC 50 It is approximately 3 μg / mL, which is close to the steady-state concentration of compound 1 at a daily dose of 60 mg.

[0048] In Study 1, healthy volunteers receiving a 60 mg dose experienced an average reduction of approximately 20% in total cholesterol, with the effect currently considered greater on the LDL fraction than on the HDL fraction. The model predicts that at a dose level of 60 mg QD, circulating cholesterol could be reduced by approximately 35%. Higher doses of Compound 1 could cause even greater reductions in circulating cholesterol levels, but may also lead to neutropenia. The reduction in circulating cholesterol is expected to occur within a timeframe (approximately 15 days) commensurate with the increase in steady-state concentrations of Compound 1 in plasma, after which circulating cholesterol levels are expected to remain stable during the dosing period. The model predicts that circulating cholesterol will return to baseline levels within approximately 30 days after discontinuation of Compound 1 treatment. The model also predicts that both the rate of reduction and recovery of circulating cholesterol levels will be limited by the accumulation and clearance rate of Compound 1 in plasma. Figure 2The figure shows the predicted steady-state concentration of circulating cholesterol during treatment with compound 1 in the dose range used in clinical studies.

[0049] Example 2 - Compound 1 induced the accumulation of 7-DHC in rat OPC, reduced sterols, and no change in cholesterol.

[0050] Enriched oligodendrocyte populations from female Sprague-Dawley day 2 (P2) rats were grown in cultures. Briefly, the forebrain was dissected and placed in Hank's buffered salt solution (HBSS) (Life Technologies). Tissue was cut into 1 mm fragments and incubated at 37°C for 15 min in 0.01% trypsin and 10 μg / ml DNase. The dissociated cells were plated into poly-D-lysine (PDL)-coated T75 tissue culture flasks and grown at 37°C for 10 days in Durbecco's Modified Eagle Medium (DMEM) with 20% fetal bovine serum (Life Technologies). Oligodendrocyte precursors (A2B5+) were collected overnight at 37°C with shaking at 200 rpm, resulting in a 95% pure cell population. Cultures were maintained in a defined growth medium (high glucose DMEM, 0.1% BSA, 50 μg / ml Apo-transferrin, 5 μg / ml insulin, 30 nM sodium selenite, 10 nM biotin, and hydrocortisone) containing 10 ng / ml fibroblast growth factor / platelet-derived growth factor (FGF / PDGF) (Peprotech) for 2–3 days. To assess the ability of compound 1 to promote the differentiation of rat A2B5+ progenitor cells into mature myelin-basic protein-positive (MBP+) myelinated oligodendrocytes, A2B5+ cells were plated in free FGF / PDGF growth medium supplemented with 10 ng / ml CNTF and 15 nM T3 onto 10 cm PDL-coated culture plates and immediately treated with compound 1. Cell clumps were collected at 24 and 72 hours of culture and stored at -80°C. Cell clumps were then transported to Metabolon (Morrisville, NC, USA) and maintained at -80°C during transport and storage until processing. Cell clump samples were extracted with methanol for 2 minutes under vigorous shaking (Glen Mills GenoGrinder 2000) to precipitate proteins and dissociate small molecules bound to proteins or trapped in the precipitated protein matrix, followed by centrifugation to recover chemically distinct metabolites. The resulting extracts were then aliquoted and analyzed on Metabolon's HD4 platform. Several types of quality control samples were applied during sample preparation and analysis, including recovery standards added before extraction, technical repeats using pools composed of each experimental sample, process, and solvent blank, and an additional mixture of QC standards for quality assessment and filtration of failed samples.

[0051] Metabolon data showed that, compared with the medium control at the same time point, compound 1 treatment increased the accumulation of the DHCR7 substrate 7-DHC in the culture and decreased or trended a decrease in the accumulation of DHCR7 products sterols and cholesterol. At 24 hours, compound 1-treated OPC cultures showed 8.3-fold increases in 7-DHC (p = 2.5e-5, q = 0.005), 0.44-fold increases in sterols (p = 0.002, q = 0.15), and 0.64-fold increases in cholesterol (p = 0.018, q = 0.15). By 72 hours, compound 1-treated OPC cultures showed 12.86-fold increases in 7-DHC (p = 8.1e-8, q = 2.6e-5), 0.4-fold increases in sterols (p = 0.001, q = 0.022), with no significant change in cholesterol. See also Figure 3 .

[0052] Data showed that although compound 1 was shown to lower peripheral cholesterol levels in humans, it did not lower cholesterol levels in rat OPC.

Claims

1. Effective amount of compound 1: ； The use of its pharmaceutically acceptable salts in the preparation of a medicine for reducing elevated cholesterol plasma levels in a subject.

2. Effective amount of compound 1: ； The use of its pharmaceutically acceptable salts in the preparation of a medicament for treating a subject with hypercholesterolemia.

3. The use as described in claim 1, wherein the elevated plasma cholesterol level is associated with familial hypercholesterolemia, atherosclerosis, acute coronary syndrome (ACS), coronary artery disease, peripheral artery disease (PAD), cerebrovascular disease, cardiovascular disease caused by diabetes, macular degeneration, or congestive heart failure.

4. Compound 1: ； The use of a pharmaceutically acceptable salt or combination of a cholesterol-lowering drug in the preparation of a drug for reducing elevated plasma cholesterol levels in a subject.

5. Compound 1: ； The use of a pharmaceutically acceptable combination of salts or cholesterol-lowering drugs in the preparation of a medicament for treating a subject with hypercholesterolemia.

6. The use as claimed in claim 4, wherein the elevated plasma cholesterol level is associated with familial hypercholesterolemia, atherosclerosis, acute coronary syndrome (ACS), coronary artery disease, peripheral artery disease (PAD), cerebrovascular disease, cardiovascular disease caused by diabetes, macular degeneration, or congestive heart failure.

7. The use as described in any one of claims 4-6, wherein the cholesterol-lowering drug is a statin, a PCSK9 inhibitor, a selective cholesterol absorption inhibitor, a bile acid sequestrant, or a fibrate.

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