COMPOUNDS AND COMPOSITIONS FOR THE TREATMENT OR PREVENTION OF PATHOLOGICAL CONDITIONS ASSOCIATED WITH EXCESS FIBRIN DEPOSITION AND/OR THROMBUS FORMATION

Abstract

There is herein provided an HDAC inhibitor, or a pharmaceutically acceptable salt thereof, as described in the description, for use in treating or preventing a pathological condition associated with excess fibrin deposition and/or thrombus formation, wherein said treatment comprises treating a patient with an HDAC inhibitor, or a pharmaceutically acceptable salt thereof, in a specific manner, and formulations for use or designed for use in such treatments.

Claims

1. A method of in treating or reducing the risk of developing a pathological condition associated with excess fibrin deposition and/or thrombus formation, the method comprising administering to a patient in need thereof at least one dose of an HDAC inhibitor, or a pharmaceutically acceptable salt thereof such that the maximum plasma concentration (Cmax) of the HDAC inhibitor, or a salt and/or metabolite thereof, in the patient occurs during a time period that is from four hours before to one hour after the maximum plasma concentration (Cmax) of PAI-1 in the patient, wherein the HDAC inhibitor is a compound selected from the list consisting of: (a) N-hydroxy-N′-phenyl-octanediamide or a pharmaceutically acceptable salt, hydrate, or solvate thereof; (b) {6-[(diethylamino) methyl]-naphthalen-2-yl}methyl[4-(hydroxycarbamoyl)phenyl]carbamate or a pharmaceutically acceptable salt, hydrate, or solvate thereof; (c) (2E)-3-[3-(anilinosulfonyl)phenyl]-N-hydroxy-acrylamide or a pharmaceutically acceptable salt, hydrate, or solvate thereof; (d) (E)-N-hydroxy-3-[4-[[2-(2-methyl-1H-indol-3-yl)-ethylamino]methyl]phenyl]prop-2-enamide or a pharmaceutically acceptable salt, hydrate, or solvate thereof; (e) 3-(dimethylaminomethyl)-N-[2-[4-(hydroxycarbamoyl)phenoxy]ethyl]-1-benzofuran-2-carboxamide or a pharmaceutically acceptable salt, hydrate, or solvate thereof; (f) N-hydroxy-2-(4-((((1-methyl-1H-indol-3-yl)methyl)amino)methyl)piperidin-1-yl)pyrimidine-5-carboxamide or a pharmaceutically acceptable salt, hydrate, or solvate thereof; (g) (2E)-3-{2-butyl-1-[2-(diethylamino)ethyl]-1H-benzimidazol-5-yl}-N-hydroxyacrylamide or a pharmaceutically acceptable salt, hydrate, or solvate thereof; (h) N-(2-aminophenyl)-4-[(4-pyridin-3-ylpyrimidin-2-ylamino)methyl]benzamide or a pharmaceutically acceptable salt, hydrate, or solvate thereof; and (i) CXD101 or a pharmaceutically acceptable salt, hydrate, or solvate thereof.

2. The method of claim 1, wherein the maximum plasma concentration (Cmax) of the HDAC inhibitor, or a salt and/or metabolite thereof, in the patient occurs during a time period that is from three hours before (e.g. two hours before) to one hour after the maximum plasma concentration (Cmax) of PAI-1 in the patient.

3. The method of claim 1, wherein the maximum plasma concentration (Cmax) of the HDAC inhibitor, or a salt and/or metabolite thereof, in the patient occurs during a time period that is from three hours before (e.g. two hours before) to the time of the maximum plasma concentration (Cmax) of PAI-1 in the patient.

4. The method of claim 1, wherein at the time when the patient experiences the maximum plasma concentration (Cmax) of PAI-1, the patient has a plasma concentration of the HDAC inhibitor, or a salt and/or metabolite thereof, that is at least within the therapeutic window for that HDAC inhibitor.

5. (canceled)

6. The method of claim 1, wherein the at least one dose a dose of the HDAC inhibitor, or a pharmaceutically acceptable salt thereof, is administered to a patient during a time period from about 20:00 hours to about 06:00 hours.

7. The method of claim 6, wherein the at least one dose of the HDAC inhibitor, or a pharmaceutically acceptable salt thereof, is administered to a patient during a time period from about 21:00 hours to about 05:00 hours (e.g. about 22:00 hours to about 04:00 hours).

8. The method of claim 6, wherein the at least one dose of the HDAC inhibitor, or a pharmaceutically acceptable salt thereof, is administered to a patient during a time period from about 02:00 hours to about 06:00 hours (e.g. about 03:00 hours to about 05:00 hours, such as about 04:00 hours).

9. The method of claim 1, wherein in treating or reducing the risk of developing a pathological condition associated with excess fibrin deposition and/or thrombus formation comprises administering a pharmaceutical composition comprising a dose of the HDAC inhibitor, or a pharmaceutically acceptable salt thereof, to a patient at a time and in a form such that substantially all of the HDAC inhibitor, or a pharmaceutically acceptable salt thereof, is released from the composition during a time period from about 02:00 hours to about 06:00 hours.

10. The method of claim 9, wherein treating or reducing the risk of developing a pathological condition associated with excess fibrin deposition and/or thrombus formation comprises administering a pharmaceutical composition comprising a therapeutically effective dose of the HDAC inhibitor, or a pharmaceutically acceptable salt thereof, to a patient at a time and in a form such that substantially all of the HDAC inhibitor, or a pharmaceutically acceptable salt thereof, is released from the composition during a time period from about 03:00 hours to about 05:00 hours (e.g. from about 04:00 hours to about 05:00 hours, such as at about 05:00 hours).

11. A method of in treating or reducing the risk of developing a pathological condition associated with excess fibrin deposition and/or thrombus formation in a patient, wherein said treatment comprises: (i) monitoring the plasma concentration of PAI-1 in the patient in order to determine the time at, or time period during which, the maximum plasma concentration of PAI-1 occurs; and (ii) administering at least one dose of the HDAC inhibitor, or a pharmaceutically acceptable salt thereof, to the patient such that the maximum plasma concentration (Cmax) of the HDAC inhibitor, or a salt and/or metabolite thereof, in the patent occurs during a time period that is from four hours before to one hour after the time at which, or time period during which, the maximum plasma concentration of PAI-1 occurs, wherein the HDAC inhibitor is a compound as defined in claim 1 selected from the list consisting of: (a) N-hydroxy-N′-phenyl-octanediamide or a pharmaceutically acceptable salt, hydrate, or solvate thereof; (b) {6-[(diethylamino) methyl]-naphthalen-2-yl}methyl[4-(hydroxycarbamoyl)phenyl]carbamate or a pharmaceutically acceptable salt, hydrate, or solvate thereof; (c) (2E)-3-[3-(anilinosulfonyl)phenyl]-N-hydroxy-acrylamide or a pharmaceutically acceptable salt, hydrate, or solvate thereof; (d) (E)-N-hydroxy-3-[4-[[2-(2-methyl-1H-indol-3-yl)ethylamino]methyl]phenyl]prop-2-enamide or a pharmaceutically acceptable salt, hydrate, or solvate thereof; (e) 3-(dimethylaminomethyl)-N-[2-[4-(hydroxycarbamoyl)phenoxy]ethyl]-1-benzofuran-2-carboxamide or a pharmaceutically acceptable salt, hydrate, or solvate thereof; (f) N-hydroxy-2-(4-((((1-methyl-1H-indol-3-yl)methyl)amino)methyl)piperidin-1-yl)pyrimidine-5-carboxamide or a pharmaceutically acceptable salt, hydrate, or solvate thereof; (g) (2E)-3-{2-butyl-1-[2-(diethylamino)ethyl]-1H-benzimidazol-5-yl}-N-hydroxyacrylamide or a pharmaceutically acceptable salt, hydrate, or solvate thereof; (h) N-(2-aminophenyl)-4-[(4-pyridin-3-ylpyrimidin-2-ylamino)methyl]benzamide or a pharmaceutically acceptable salt, hydrate, or solvate thereof; and (i) CXD101 or a pharmaceutically acceptable salt, hydrate, or solvate thereof.

12. The method of claim 11, wherein the maximum plasma concentration (Cmax) of the HDAC inhibitor, or a salt and/or metabolite thereof, in the patient occurs during a time period that is from three hours before (e.g. two hours before) to the time of the maximum plasma concentration (Cmax) of PAI-1 in the patient.

13. A method of treating or reducing the risk of developing a pathological condition associated with excess fibrin deposition and/or thrombus formation in a patient, wherein said treatment comprises: (i) monitoring the plasma concentration of PAI-1 in the patient in order to determine the time at, or time period during which, the maximum plasma concentration of PAI-1 occurs; and (ii) administering at least one dose of the HDAC inhibitor, or a pharmaceutically acceptable salt thereof, to the patient such that at the time when the patient experiences the maximum plasma concentration of PAI-1, the patient has a plasma concentration of the HDAC inhibitor, or a salt and/or metabolite thereof, that is within the therapeutic window for the relevant HDAC inhibitor, wherein the HDAC inhibitor is a compound selected from the list consisting of: (a) N-hydroxy-N′-phenyl-octanediamide or a pharmaceutically acceptable salt, hydrate, or solvate thereof; (b) {6-[(diethylamino) methyl]-naphthalen-2-yl}methyl[4-(hydroxycarbamoyl)phenyl]carbamate or a pharmaceutically acceptable salt, hydrate, or solvate thereof; (c) (2E)-3-[3-(anilinosulfonyl)phenyl]-N-hydroxy-acrylamide or a pharmaceutically acceptable salt, hydrate, or solvate thereof; (d) (E)-N-hydroxy-3-[4-[[2-(2-methyl-1H-indol-3-yl)ethylamino]methyl]phenyl]prop-2-enamide or a pharmaceutically acceptable salt, hydrate, or solvate thereof; (e) 3-(dimethylaminomethyl)-N-[2-[4-(hydroxycarbamoyl)phenoxy]ethyl]-1-benzofuran-2-carboxamide or a pharmaceutically acceptable salt, hydrate, or solvate thereof; (f) N-hydroxy-2-(4-((((1-methyl-1H-indol-3-yl)methyl)amino)methyl)piperidin-1-yl)pyrimidine-5-carboxamide or a pharmaceutically acceptable salt, hydrate, or solvate thereof; (g) (2E)-3-{2-butyl-1-[2-(diethylamino)ethyl]-1H-benzimidazol-5-yl}-N-hydroxyacrylamide or a pharmaceutically acceptable salt, hydrate, or solvate thereof; (h) N-(2-aminophenyl)-4-[(4-pyridin-3-ylpyrimidin-2-ylamino)methyl]benzamide or a pharmaceutically acceptable salt, hydrate, or solvate thereof; and (i) CXD101 or a pharmaceutically acceptable salt, hydrate, or solvate thereof.

14. (canceled)

15. The method of claim 1, wherein the HDAC inhibitor or a pharmaceutically acceptable salt thereof is administered: (i) as a single dose per 24 hour period (i.e. a single daily dose); and/or (ii) at a dose sufficient to achieve a reduction in PAI-1 plasma levels of at least about 20% (such as at least about 30%).

16. The method of claim 1, wherein the administration of the HDAC inhibitor or pharmaceutically acceptable salt thereof is in a manner such that the plasma concentration of the HDAC inhibitor, or a salt and/or metabolite thereof, during a 24 hour period mimics the plasma concentration of PAI-1 during the same period.

17. The method of claim 1, wherein the pathological condition associated with excess fibrin deposition and/or thrombus formation is selected from the group consisting of atherosclerosis, myocardial infarction, ischemic stroke, deep vein thrombosis, pulmonary embolism, disseminated intravascular coagulation, renal vascular disease and intermittent claudication.

18. The method of claim 1, wherein the pathological condition associated with excess fibrin deposition and/or thrombus formation is: (a) ischemic stroke, such as a major ischemic stroke and minor ischemic stroke; and/or (b) myocardial infarction.

19. The method of claim 1, wherein the patient is a human.

20. The method of claim 1, wherein the patient is at increased risk of developing a pathological condition associated with excess fibrin deposition and/or thrombus formation.

21. The method of claim 1, comprising administration of the HDAC inhibitor or pharmaceutically acceptable salt thereof in combination with aspirin, clopidogrel and/or ticagrelor.

22-27. (canceled)

Description

BRIEF DESCRIPTION OF THE FIGURES

[0510] FIG. 1 shows a schematic representation of the circadian rhythm (i.e. variation) of PAI-1 levels in an adult human during a typical 24 hour period. The lower curve represents the variation of PAI-1 levels in a normal (i.e. healthy) patient. The upper curve represents the variation in PAI-1 levels in a patient having increased levels of PAI-1 (e.g. patients with obesity and/or the metabolic syndrome). The y-axis represents arbitrary plasma levels and is abbreviated to illustrate the positively skewed distribution toward high plasma levels in obesity/metabolic syndrome. The x-axis represents clock time.

[0511] FIG. 2 shows the results of the in vitro release profile analysis as described in Example 7 herein below.

[0512] FIG. 3 provides an example of a release profile as may be provided by pharmaceutical formulations as described in the eighth aspect of the invention, compared to release profiles as may be provided by corresponding immediate release (IR) and extended release (ER) formulations.

EXAMPLES

[0513] The following examples are included to further illustrate the invention, although the skilled person will understand that the invention is in no way restricted to the specific aspects described therein.

Example 1—VPA and PAI-1

[0514] The effects of VPA on PAI-1 were analysed in two different proof-of-concept studies in healthy subjects as well as in patients with manifest atherosclerotic disease. The studies had a randomized cross-over design and PAI-1 levels were investigated before and after HDAC inhibition with valproic acid. PAI-1 plasma levels were measured in the morning at the first day of the study as well as at the end of the treatment period with VPA (see example 2 for details on the PAI-1 analysis).

[0515] In the first study, 10 healthy non-smoking white male subjects (with mean BMI of approximately 26), aged 50-70 years were included and treated with valproic acid 500 mg (Ergenyl Retard, Sanofi) twice daily during 14 days. Unexpectedly we detected a more than 50% reduction (from 22.2 to 10.8 ng/ml, p<0.05) in circulating plasma PAI-1 levels during mid-morning in comparison to the midmorning levels found before treatment with VPA.

[0516] In the second study, 16 non-smoking white male patients, aged 50-80 years with a history of a myocardial infarction were included. On top of their ordinary prescription (beta-blocker, ACE-inhibitor, statin, aspirin) they were treated with valproic acid 500 mg (Ergenyl Retard, Sanofi), twice daily during 28 days. In this study we detected a 45% reduction in circulating plasma PAI-1 levels (from 19.6 ng/ml to 11 ng/ml (p=0.01)), during midmorning.

Example 2—Intermediate Endpoint Study: Effects of Valproic Acid on In Vivo PAI-1 in Man

[0517] An intermediate endpoint proof-of-concept study is performed in patients with TIA/minor stroke investigated before and after treatment with Valproic acid. Valproic acid is administrated as an enteric-coated tablet with delayed absorption.

[0518] The study comprises 20 patients with TIA/minor stroke. Patients are investigated before and after oral treatment with 400 mg valproic acid once time daily at 11 pm for 2 weeks. Plasma PAI-1 levels and plasma concentrations of valproic acid is followed daily during the study period at the following time-points: 3 am, 6 am, 10 am, 16 pm, 22 pm PAI-1 levels are measured by commercially available ELISA-kits (Coaliza PAI-1, ChromogenixAB) and the plasma concentration of valproic acid an metabolites thereof is analyzed according to clinical routine at the Sahlgrenska University laboratory, Gothenburg, Sweden.

[0519] The plasma concentration of valproic acid is found to peak between 3 am and 6 am and thereafter declines to very low levels during the trough in PAI-1 concentrations. The peak in plasma valproic acid coincides with the peak level of plasma PAI-1 between 3 am and 6 am. The plasma concentration of valproic acid and plasma PAI-1 levels follow each other with a pronounced circadian elevation with its peak during the early morning hours. The plasma PAI-1 levels are lowered by approximately 30% after the treatment.

Example 3—Clinical Outcome Study in High-Risk Patients for Prevention of Recurrent Thromboembolic Events Using Valproic Acid

[0520] A clinical outcome study is performed in high-risk patients who have experienced a recent major atherothrombotic cardiovascular event (myocardial infarction or TIA/ischemic stroke) to investigate the preventive effect of valproic acid treatment on the risk for recurrent events. The annual risk for a recurrent atherothrombotic event in the investigated population is estimated to approximately 7%.

[0521] Patients are randomized in a parallel study design to receive double-blind oral treatment with 400 mg valproic acid (as in Example 2) or placebo once time daily at 11 pm, in addition to optimal conventional treatment. The event rate is monitored by Kaplan-Meyer statistics. The primary efficacy endpoint is the composite measure of either mortality, or non-fatal myocardial infarction or ischemic stroke. The study is event-driven to a total of 180 events.

[0522] The study is expected to show that long-term valproic acid treatment reduces this risk by approximately 30% in addition to that of conventional therapy, i.e. lowers the annual absolute event rate to approximately 5%. Thus, the study is expected to confirm the clinical efficacy and feasibility of using valproic acid for secondary prevention of cardiovascular disease.

Example 4

[0523] To determine if a particular substance is an HDAC inhibitor (HDACi) as required in the present invention, screening for activity towards a panel of recombinant human HDAC enzymes HDAC1-11 may be performed. In such studies a dilution series of the compound is generated with ten steps of three-fold dilutions starting at 10 μM (e.g. 10 μM, 3 μM, 1 μM, 300 nM, 100 nM, 30 nM, 10 nM, 3 nM, 1 nM, 0.3 nM) and this is plotted in a dose-response curve to yield the IC.sub.50 value.

Example 5

[0524] Core tablets with a composition according to Table 1 were manufactured at a batch size of 200 g.

TABLE-US-00004 TABLE 1 Core tablet formulation. Component Amount, % w/w Sodium valproate 23.06 MCC 64.94 Copovidone 5 Croscarmellose sodium 5 Silica, colloidal anhydrous 1 Magnesium stearate 1

[0525] Sodium valproate was crushed in a mortar and sieved through a 0.50 mm screen. 46.1 g of the screened material was charged in a 1 L-vessel of Turbula T2F together with 129.9 g MCC, 10 g copovidone, 10 g croscarmellose sodium and 2 g silica. After mixing for 4 min at 32 rpm the mixture was sieved through a 0.50 mm screen and mixed for 4 min further. Magnesium stearate 2 g was roughly pre-mixed with a similar volume of the powder mixture in a steel vessel with a spoon and sieved through a 0.50 mm screen, added to the 1 L-vessel and mixed with the powder mixture for 2 min at 22 rpm. Tablets were compressed in 5 mm circular punch/die sets with normal cup depth in a rotary press (Fette 52i) at a main compression force of 2 kN. Tablet weight was approx. 65 mg and resistance to crushing approx. 5 kp.

Example 6

[0526] Core tablets according to Example 5 were coated with Eudragit® FS30D (aqueous dispersion 30%)/PlasACRYL™ T20 according to Table 2 using a Hüttlin Kugelcoater HKC005. The batch size was 50 g. The coating was performed with an air inlet temperature of 47° C., resulting in a product temperature of 28-29° C. The air flow was adjusted to achieve an appropriate fluidization of the tablets during the coating. The coating layer was applied to the core tablets so as to obtain a weight gain of 20%. After the coating, the tablets were cured at 40° C. for 2 hours.

TABLE-US-00005 TABLE 2 Coating spray suspension for Example 6 Ingredient Quantity, % w/w Eudragit ® FS30D 60.61 PlasACRYL ™ T20 9.09 Water 30.3

Example 7—In Vitro Release

[0527] The in vitro release profile of the compositions as prepared in Example 6 was analysed using USP dissolution Apparatus 2 (paddle) Ph. Eur. 2.9.3 (as described herein). The following conditions were used: temperature 37.0±0.5° C.; paddle speed 75 rpm. The samples were analyzed for valproic acid by HPLC using a Phenomenex Luna C18 column, 150×4.6 mm, particle size 5 μm, column temperature 40° C., mobile phase acetonitrile/phosphate buffer (pH 3.0) 1:1, flow rate 1 mL/min.

[0528] The level of release at particular time points and pH of solution was analysed. pH adjustments and sample pulls are described below. pH adjustments are performed immediately after sample pulls. Time points refer to total running time.

[0529] Stomach, pH 1

[0530] One tablet was added to a vessel containing 250 mL 0.1 M hydrochloric acid solution and the content was stirred for 1 hour and samples were pulled.

[0531] Small Intestine, pH 6.4

[0532] 181 mL of a solution of potassium phosphate buffer and potassium hydroxide was added to the vessel to give pH 6.4. Samples were pulled after 1.5 and 2.5 hours.

[0533] Ilium, pH 6.8

[0534] 69 mL of a solution of potassium phosphate buffer and potassium hydroxide was added to the vessel to give pH 6.8. Samples were pulled after 3 and 4 hours.

[0535] Terminal Ilium, pH 7.3

[0536] 253 mL of an aqueous potassium hydroxide solution was added to the vessel to give pH 7.3. Samples were pulled after 4.25, 4.5, 4.75, 5, 5.5 and 6 hours.

[0537] The release profile observed for the composition of Example 6 is shown in FIG. 2 as provided herein.