NO319827B1 - Use of cholesterol-lowering agent and method of preparation thereof, as well as pharmaceutical composition - Google Patents

Description

The present invention relates to the use of a cholesterol-reducing agent for the preparation of an oral dosage form. (E)-7-[4-{4-fluorophenyl)-6-isopropyl-2-[methyl(methyl-sulfonyl)amino]pyrimidin-5-yl]-(3R,5S)-3,5-dihydroxyhept-6 - an acid or a pharmaceutically acceptable salt thereof is used and is referred to as the "agent" and illustrated (as calcium salt) in formula (1) below. The invention further relates to methods for the production thereof, as well as pharmaceutical composition.

The agent is described in European Patent Application, Publication No. 0521471, and in Bioorganic and Medicial Chemistry, (1997), 5(2), 437-444 as an inhibitor of 3-hydroxy-3-methylglutaryl CoA reductase (HMG-CoA reductase) which is an essential rate-limiting enzyme in cholesterol biosynthesis. The agent is described to be useful for the treatment of hypercholesterolemia, hyperlipoproteinemia and atherosclerosis. HMG-CoA reductase inhibitors are the most commonly prescribed medication for the treatment of hypercholesterolemia. A number of HMG-CoA reductase inhibitors are marketed, ie lovastatin, pravastatin, simvastatin, fluvastatin, atorvastatin and cerivastatin and are collectively referred to as "statins".

Despite the advantages of statin therapy, less than optimal results may be achieved in patients due to the level of effectiveness and safety achieved by the recommended dosages of marketed statins. It is therefore important to find dosages of alternative statins that beneficially change lipid levels to a significantly higher degree than similar dosages of currently used statins and that have a similar or improved safety profile.

It has now surprisingly been discovered that when dosed orally to patients with hypercholesterolemia at particular dosages or in a particular dosage range, the agent reduces total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C) to an unexpected degree and without any significant negative side effects. When dosed at the same dosage ranges or in the same dosage range, the agent also modifies other lipoprotein levels (such as increasing high-density lipid cholesterol (HDL-C) levels, decreasing triglyceride (TG) levels, and decreasing apolipoprotein B-100 (Apo-B) levels) in an unexpected and beneficial degree without any significant negative side effects. Elevation of alanine aminotransferase (ALT) liver enzyme levels has been reported for other HMG-CoA reductase inhibitors. It has now surprisingly been discovered that when the agent is dosed at dosages or in dosage ranges discussed herein, clinically significant increases in these levels are less frequently observed.

One aspect of the present invention therefore comprises the use of the compound (E)-7-[4-(4-fluorophenyl)-6-isopropyl-2-[methyl-(methylsulfonyl)amino]pyrimidin-5-yl](3R,5S) -3,5-dihydroxyhept-6-ene acid, or a pharmaceutically acceptable salt thereof, for the preparation of an oral dosage form comprising 5 to 80 mg of the compound for use in reducing LDL-C levels by 40% or more, and/or reduction in total cholesterol levels by 30% or more, and/or reduction in triglyceride levels by 10% or more, and/or reduction in apolipoprotein B-100 levels by 30% or more, and/or increase in HLD-C levels by 5% or more in a human patient in need.

A further aspect of the present invention further comprises the use of the compound or a pharmaceutically acceptable salt thereof according to claim 1 for the preparation of an oral dosage form comprising 5 to 10 mg of the compound for use in reducing LDL-C levels by 40% or more, and/ or reduction in total cholesterol levels by 30% or more, and/or reduction in triglyceride levels by 10% or more, and/or reduction in apolipoprotein B-100 levels by 30% or more, and/or increase in HDL-C levels by 5 % or more in a human patient in need.

A further aspect of the present invention comprises the use of the compound or a pharmaceutically acceptable salt thereof according to claim 2 for the preparation of an oral dosage form comprising 5 to 10 mg of the compound for use in reducing LDL-C levels by 45% or more, in a human patient who needs it.

A further aspect of the present invention comprises the use of the compound or a pharmaceutically acceptable salt thereof according to claim 2 for the preparation of an oral dosage form comprising 5 to 10 mg of the compound for use in reducing total cholesterol levels by 35% or more in a human patient in need the.

A further aspect of the present invention comprises the use of the compound or a pharmaceutically acceptable salt thereof according to claim 2 for the preparation of an oral dosage form comprising 5 to 10 mg of the compound for use in reducing triglyceride levels by 10% or more in a human patient in need thereof .

A further aspect of the present invention comprises the use of the compound or a pharmaceutically acceptable salt thereof according to claim 2 for the preparation of an oral dosage form comprising 5 to 10 mg of the compound for use in reducing apolipoprotein B-100 levels by 35% or more in a human patient who needs it.

A further aspect of the present invention comprises the use of the compound or a pharmaceutically acceptable salt thereof according to claim 2 for the preparation of an oral dosage form comprising 5 to 10 mg of the compound for use in increasing HLD-C levels by 8% or more in a human patient who needs it.

A preferred feature of the present invention comprises use according to any one of claims 1 to 7 where the oral dosage form is administered with a single dose once a day.

The present invention further relates to a pharmaceutical composition adapted for oral administration, characterized in that it comprises 5 to 80 mg of {E)-7-[4-(4-fluorophenyl)-6-isopropyl-2-[methyl(methylsulfonyl)amino]pyrimidine- 5-yl](3R,5S)-3,5-dihydroxy-hept-6-ene acid, or a pharmaceutically acceptable salt thereof, together with a pharmaceutically acceptable diluent or carrier.

When a dosage range of 5 to 80 mg per day, 10 to 80 mg per day, 20 to 80 mg per day, or 40 to 80 mg per day is referred to herein, other particular dosage ranges include (as appropriate) 10 to 80 mg per day, 10 to 60 mg per day, 10 to 40 mg per day, 5 to 40 mg per day, 5 to 20 mg per day, 10 to 20 mg per day, 20 to 60 mg per day, 20 tii 40 mg per day and 40 to 60 mg per day.

The present invention further relates to a method for a pharmaceutical composition according to claim 9, 10 or 11, characterized in that it comprises mixing the compound or a pharmaceutically acceptable salt thereof with a pharmaceutically acceptable diluent or a carrier.

Special pharmaceutical compositions comprise, for example, 5 mg, 10 mg, 20 mg, 40 mg and 80 mg of the agent together with a pharmaceutically acceptable excipient or diluent. The pharmaceutical compositions will be in the form of a conventional dosage unit form, for example, tablets or capsules. The compositions according to the invention can be obtained by conventional procedures using conventional pharmaceutical excipients, well known in the art. It is preferred that the agent is administered as a single dose once a day.

It is preferred that the agent is bis-[(E)-7-[4.(4)-fluorophenyl)-6-isopropyl-2-[methyl-(methylsulfonyl)amino]pyrimidin-5-yl](3R,5S) -3,5-dihydroxyhept-6-ene acid] calcium salt. When reference is made herein to a dose of (or a pharmaceutical composition comprising) 5 mg, 10 mg, 20 mg, 40 mg or 80 mg of the agent, this includes a dose (or pharmaceutical composition comprising) 5.2 mg , 10.4 mg, 20.8 mg, 41.6 mg and 83.2 mg of the calcium salt of formula 1. Such doses (or compositions) are intended to provide the equivalent of 5 mg, 10 mg, 20 mg, 40 mg and 80 mg of free acid form when dosed.

Pharmaceutical compositions

The following example illustrates pharmaceutical dosage forms suitable for use in the invention as defined herein:

Capsules containing 1, 2, 5 or 10 mg of agent can also be obtained using more or less lactose as needed to maintain a total fill weight of 105 mg.

Preferred formulations are those in which the agent is bis[(E)-7-[4-(4-fluorophenyl)-6-isopropyl-2-[methyl(methylsulfonyl)amino]pyrimidin-5-yl]-(3R,5S)- 3,5-dihydroxyhept-6-ene acid] calcium salt.

To illustrate the invention, a randomized, dose-response parallel-group study with bis[(E)-7-[4-(4-fluorophenyl)-6-isopropyl-2-[methyl(methylsulfonyl)-amino]pyrimidin-5-yl] (3R,5S)-3,5-dihydroxyhept-6-ene acid] calcium salt (referred to below as ZD4522) and atorvastatin (ATORV) in subjects with primary hyper-cholesterolemia performed.

Primary purposes

The primary objectives of this trial were to assess the dose-response relationship between the dose of ZD4522 and the percentage reduction of LDL-C from the baseline value with respect to placebo.

Secondary purposes

Secondary objectives of this trial include:

to assess the effect of 10 and 80 mg doses of atorvastatin on LDL-C levels; and assess effects of ZD4522 and atorvastatin on HDL-C, TG, TC, apolipoprotein, A-1, apolipoprotein Lp(a), apolipoprotein B-100 levels and LDL-C levels and LDL-C (by indirect method); to assess the pharmacokinetics of oral doses of 1, 2.5, 5.10, 20 and 40 mg ZD4522 (capsule formulations) over a 6-week treatment period; and to assess the tolerability and safety of ZD4522 compared to placebo.

Experimental design

After a 6-week diet habituation period, the subjects were randomized to either atorvastatin doses (10 or 80 mg), administered open-label, or to placebo or 1 of 6 ZD4522 doses (administered blindly). Analyzes of the blinded portion of the trial pertained to the primary objective. Open-label atorvastatin groups were included to obtain additional data regarding initial doses and high doses of a proven cholesterol-lowering agent in this patient population.

Trial plan

A total of 10 visits were made to each clinic. Three of these took place during the placebo-controlled diet run-in period (weeks -6, -2 and -1), five during the treatment phase (week 0 = randomized visit, weeks +1, +2, +4 and +6) and two follow-up visits (weeks +8 and +10).

Number of individuals

The primary endpoint on which the sample size is based is percent reduction from baseline in LDL-C (LDL cholesterol) values. A sample size of 9 in each group would have 90% power to detect a 25% difference between two groups assuming a common standard deviation of 15% using a 2-group t-test with a 0.05 two-tailed significance level. This has been increased to 12 subjects per group to adjust for multiple comparisons of groups against placebo while preserving a power of at least 90% (based on simulations). This sample size also led to an estimate of the dose-response curve for percent reduction in LDL-C with a safety band width of less than 10% for most of the dose range.

Inclusion criteria

For inclusion in the diet run-up period, the subjects had to meet all of the following criteria: (1) fasting LDL cholesterol (>4.14, but <6.21 mmol/l; (2) fasting levels of triglycerides (<3.39 mmol/h) ;

(3) male, age 18 to 70 years. Female age 50 to 70 years.

(4) Absence of menstruation for more than two years and follicle stimulating hormone (FSH), luteinizing hormone (LH) and estradiol within post-menopausal

the area (when using local laboratory areas);

(5) body mass index < 30 kg/m<2>.

For inclusion in the randomized treatment phase of the trial, the subjects had to fulfill all of the following criteria: (6) fasting LDL cholesterol >4.14 mmol/l, but < 5.69 mmol/l from measurements taken in weeks -2 and -1 before randomisation, with lower value within 15% of the higher value; (7) fasting TG < 3.39 mmol/l in weeks -6, -2 and -1, before randomisation; (8) a food record determination score (FRR) < 15 at weeks -2 and -1, before randomization, to demonstrate suitability with the American Heart Association Step -1 diet.

Exclusion criteria

Any of the following was considered a criterion for exclusion from the trial: (1) subjects using cholesterol-lowering drugs (this therapy must have been stopped at least 4 weeks before the start of the diet run-in period. Subjects taking probucol should have stopped this 12

months before inclusion in the study).

(2) Background of severe or hypersensitivity reactions to others

HMG-CoA reductase inhibitors

(3) Individuals with active arterial disease such as unstable angina, myocardial infarction, transient ischemic attack (TIA), cerebrovascular accident (CVA), or coronary artery bypass grafting (CABG) within 6 months or

angioplasty within 3 months before inclusion in the study.

(4) Background with malignancy with the exception of individuals whose only malignancy has been basal or squamous cell carcinoma. Female individuals with a background including cervical dysplasia if not three consecutive

normal cervical smears have been recorded before incorporation.

(5) Uncontrolled hypertension (diastolic blood pressure > 95 mm Hg). Individuals who have taken thiazide diuretics and beta blockers will not be excluded provided they are maintained at a stable dose from 3 months prior

beginning study and during the course of the study.

(6) Diabetes mellitus and/or other metabolic endocrine disease known to be associated with changes in plasma lipid levels (uncontrolled hyperthyroidism defined as TSH>1.5 times ULN at week -6, individuals with

nephrotic syndrome).

(7) Known homozygous familial hypercholesterolemia or known type III hyperlipoproteinemia (familial dyspetalipoproteinemia).

(8) Use of concomitant medications indicated as follows:

No medication allowed

(9) Background with alcohol and/or drug abuse.

(10) Individuals with active liver disease or hepatic dysfunction as defined by

elevation in the liver enzymes (ALT, AST, yGT, ALP) or bilirubin >= 1.5

times ULN at any time before radomization at visit 4.

(11) Individuals with serum creatinine > 180 jamol/l before randomisation.

(12) Participation in another study less than three months before inclusion in the diet run-in period.

(13) Serum CK > 3 times ULN in weeks -6 and -1 (visits 1 and 3).

(14) Clinically significant ophthalmic abnormalities, simple refractive errors will be allowed. (15) Individuals who have received randomized medication cannot be returned to

this study.

(16) Individuals with serious or unstable medical or physiological conditions

which according to the researcher will destroy the safety of the patient or

successful participation in the study.

(17) Individuals who have accepted hormone replacement therapy (HRT).

(18) Individuals who have received digoxin and/or coumarin anticoagulants.

Experimental methods

Summary

The subjects had to take the study medication once per day and no less than three hours after the evening meal. At visits 1, 2, 3, 4, 6, 7, 8, 9, and 10, subjects reported to the clinic after fasting for 12 hours (water allowed) and blood was drawn to determine fasting lipids. The individuals submitted a urine sample, and blood samples for hematology and clinical biochemistry assessment. When lipid and clinical biochemistry measurements fell at the same visit, only one sample was taken. At visits 4, 6, 7 and 8, pharmacodynamics samples were collected. Weight, blood pressure and heart rate were measured and recorded at all visits. At visits 1 and 10, an ECG was performed.

Applied Analytical Laboratory is certified for standardization of lipid analyzes as specified by the standardization program of the Centers for Disease Control and Prevention and the National Heart, Lung and Blood Institute.

Analysis

(a) Lipid

Concentrations of the following lipids were determined LDL-C, HDL-C. TG, TC, apo A-I, apo A-I I, Lp(a), apo B, LDL-C (by Friedwald equation and quantification method).

(b) Biochemistry

Concentrations of the following were determined: AST, ALT, CK<1>, yGT, ALP and total bilirubin, sodium, potassium, calcium, chloride, phosphate, bicarbonate creatinine, fasting blood glucose and serum thyroxine (T4) and thyroid stimulating hormone (TSH).

If not surgically sterilized women had postmenopausal status confirmed by measurement of FSH, LH and estradiol at visit 1.

Hematology

Whole blood counts were performed and included the following; erythrocyte count, hemoglobin, hematocrit, leukocyte cell count, platelets, red cell distribution width, percentage differential leukocyte count (including percentage of large unstained cells), mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration. A coagulation screening was performed only at visits 1 and 10.

Urinalysis

On visits 1, 3, 4, 5, 6, 7, 8, 9 and 10, urinalyses were performed on 10 ml mid-stream urine samples and tested clinically follow-up using a lab stick/dip stick: pH, glucose, blood, ketones, proteins and bilirubin.

Results

1 Study strength average % reduction of LDL-C in week 6 (Friedewald method)

2 Study strength Lipid parameter (average % change from baseline in week 6)

Note: in 2, a positive value indicates an average percent increase of lipid parameter level and a negative value indicates an average percent decrease of lipid parameter level.

4. Effects in each study strength ("arm") on percentage change in total cholesterol from the baseline are given in fig. 1. Dosing began at baseline after an initial diet performed for a period of six weeks, according to the clinical protocol above. Dosing stopped six weeks after baseline. 5. Effects in each study strength on percentage of LDL cholesterol (Friedewald equation calculation) percentage change from baseline is given in fig. 2. Dosing began at baseline after an initial diet performed for a period of six weeks according to the above clinical protocol. Dosing stopped six weeks after baseline. 6. Effects on each study arm on HDL cholesterol percentage change from baseline are given in fig. 3. Dosing began at baseline after an initial diet performed for a period of six weeks, according to the above clinical protocol. Dosing stopped six weeks after baseline. 7. Effects of each study strength on Apolipoprotein B-100 percentage change from baseline are given in fig.4. Dosing began at baseline after an initial diet performed for a period of six weeks, as the clinical protocol above.

Dosing stopped six weeks after baseline.

8. Effects on each study arm on triglyceride percentage change from baseline are indicated in fig. 5. Dosing began at baseline after an initial diet round for a period of six weeks according to the clinical protocol above. Dosing stopped six weeks after baseline. It is to be noted that alternative or further trials of shorter or longer duration may be carried out with the agent to demonstrate the present invention such as other randomized double blind, possibly placebo controlled, trials with selected doses of the agent and selected doses of commercially available strains (for e.g. atorvastatin, pravastatin, simvastatin) as comparative subjects in individuals with hypercholesterolemia with etler without a background of CHD or PVD or with risk factors thereof that measure changes in LDL-C and other lipid or lipoprotein fractions and safety of the agent with regard to e.g. serum transaminase levels.

Claims (12)

1. Use of the compound (E)-7-[4-(4-fluorophenyl)-6-isopropyl-2-[methyl-(methylsulfonyl)amino]pyrimidin-5-yl](3R,5S)-3,5- dihydroxyhept-6-ene acid, or a pharmaceutically acceptable salt thereof, for the preparation of an oral dosage form comprising 5 to 80 mg of the compound for use in reducing LDL-C levels by 40% or more, and/or reducing total cholesterol levels by 30% or more, and/or reduction in triglyceride levels by 10% or more, and/or reduction in apolipoprotein B-100 levels by 30% or more, and/or increase in HLD-C levels by 5% or more in a human patient who needs it. 2. Use of the compound or a pharmaceutically acceptable salt thereof according to claim 1 for the preparation of an oral dosage form comprising 5 to 10 mg of the compound for use in reducing LDL-C levels by 40% or more, and/or reducing total cholesterol levels by 30% or more, and/or reduction in triglyceride levels by 10% or more, and/or reduction in apolipoprotein B-100 levels by 30% or more, and/or increase in HDL-C levels by 5% or more in a human patient who needs it. 3. Use of the compound or a pharmaceutically acceptable salt thereof according to claim 2 for the preparation of an oral dosage form comprising 5 to 10 mg of the compound for use in reducing LDL-C levels by 45% or more, in a human patient in need thereof. 4. Use of the compound or a pharmaceutically acceptable salt thereof according to claim 2 for the preparation of an oral dosage form comprising 5 tiMO mg of the compound for use in reducing total cholesterol levels by 35% or more in a human patient in need thereof. 5. Use of the compound or a pharmaceutically acceptable salt thereof according to claim 2 for the preparation of an oral dosage form comprising 5 to 10 mg of the compound for use in reducing triglyceride levels by 10% or more in a human patient in need. 6. Use of the compound or a pharmaceutically acceptable salt thereof according to claim 2 for the preparation of an oral dosage form comprising 5 to 10 mg of the compound for use in reducing apolipoprotein B-100 levels by 35% or more in a human patient in need thereof. 7. Use of the compound or a pharmaceutically acceptable salt thereof according to claim 2 for the preparation of an oral dosage form comprising 5 to 10 mg of the compound for use in increasing HLD-C levels by 8% or more in a human patient in need thereof. 8. Use according to any one of claims 1 to 7, wherein the oral dosage form is administered in a single dose once a day. 9. Pharmaceutical composition adapted for oral administration, characterized in that it comprises 5 to 80 mg of (E)-7-[4-(4-fluorophenyl)-6-isopropyl-2-[methyl(methylsulfonyl)amino]pyrimidin-5-yl ](3R,5S)-3,5-dihydroxy-hept-6-ene acid, or a pharmaceutically acceptable salt thereof, together with a pharmaceutically acceptable diluent or carrier. 10. Pharmaceutical composition adapted for oral administration according to claim 9, characterized in that it comprises 5 to 10 mg of (E)-7-[4-(4-fluorophenyl)-6-isopropyl-2-[methyl(methylsulfonyl)amino]pyrimidine- 5-yl](3R,5S)-3,5-dihydroxy-hept-6-ene acid, or a pharmaceutically acceptable salt thereof, together with a pharmaceutically acceptable diluent or carrier. 11. Pharmaceutical composition according to claim 9 or 10, characterized in that it comprises 5.2 to 10.4 mg of the calcium salt of (E)-7-[4-(4-fluorophenyl)-6-isopropyl-2-[methyl(methylsulfonyl)amino]pyrimidin-5-yl](3R ,5S)-3,5-dihydroxyhept-6-one acid, together with a pharmaceutically acceptable diluent or carrier. 12. Process for producing a pharmaceutical composition according to claim 9, 10 or 11, characterized in that it comprises mixing the compound or a pharmaceutically acceptable salt thereof with a pharmaceutically acceptable diluent or a carrier.