GB2328873A - The use of [3(R)-[2-(piperidin-4-yl)ethyl]-2-piperidone-1]acetyl-3(R)-methyl-ß-alanine for inhibiting blood platelet aggregation - Google Patents

Abstract

The aggregation of blood platelets in a non-fasting human patient is inhibited by the administration of a dose of between 0.1 mg/kg and 5.0 mg/kg of [3(R)-[2-(piperidin-4-yl)ethyl]-2-pipendone-1]acetyl-3(R)- methyl- #-alanine or a pharmaceutical acceptable salt thereof. [3(R)-[2-(piperidin-4-yl)ethyl]-2-piperidone-1]acetyl-3- (R)-methyl-#-alanine is a fibrinogen receptor antagonist and prevents fibrinogen from binding to the membrane bound glycoprotein complex GP IIa/IIIb. It is preferably administered as an oral formulation, such as a tablet or capsule. This treatment reduces the adverse risk of cardiovascular events in a non-fasting human and is used to prevent thrombus or embolism formation. The [3(R)-[2-(piperidin-4-yl)ethyl]-2-piperidone- 1] acetyl-3(R)-methyl-#-alanine may be administered in two daily doses (of between 0.1 to 5.0 mg/kg) in order to reduce the fibrinogen receptor antagonist plasma level variability in the patient. The treatment may be used in combination with other anticoagulant agents, thrombotytic agents or platelet anti-aggregation agents. A number of examples of tablet and capsule formulations are given as are a number of examples of administration regimes.

Description

TITLE OF THE INVENTION A METHOD FOR INHIBITING PLATELET AGGREGATION BACKGROUND OF THE INVENTION Platelet activation and aggregation are involved in unstable angina and acute myocardial infarction, in reocclusion following thrombolytic therapy and angioplasty, in transient ischemic attacks and in a variety of other vaso-occlusive disorders. When a blood vessel is damaged either by acute intervention such as angioplasty, or, more chronically, by the pathophysiological processes of atherosclerosis, platelets are activated to adhere to the disrupted surface and to each other. This activation, adherence and aggregation may lead to occlusive thrombus formation in the lumen of the blood vessel.

Antiplatelet therapy has been used in a wide variety of cardiovascular disease states and in conjunction with interventional therapy such as coronary artery or peripheral bypass grafting, cardiac valve replacement, and percutaneous transluminal coronary angioplasty (PTCA). Available drugs, such as aspirin and ticlopidine, have shown efficacy in syndromes involving vascular occlusion, presumably due to sustained inhibition of platelet function. However, the inhibitory effects of aspirin and ticlopidine are dependent upon the agonist which activates the platelet. For example, aspirin is effective in blocking platelet aggregation induced by agonists such as collagen that are dependent upon the cyclooxygenase pathway. It is, however, less effective against concentrations of thrombin which can act by cyclooxygenase independent pathways. Likewise, ticlopidine's inhibitory effects can be overcome by combinations of agonists. Thus, an efficacious inhibitor of platelet aggregation that acts independently of the agonist and the pathway activating the platelet could be an important therapeutic advance providing greater efficacy than aspirin or ticlopidine in a broader spectrum of thrombotic events.

GPIIb/IIIa Inhibitors The final obligatory step in platelet aggregation is the binding of fibrinogen to an activated membrane-bound glycoprotein complex, GP IIb/IIIa (aIIss3). Platelet activators such as thrombin, collagen, epinephrine or ADP, are generated as an outgrowth of tissue damage. During activation, GP IIb/IIIa undergoes changes in conformation that result in exposure of occult binding sites for fibrinogen. There are six putative recognition sites within fibrinogen for GP IIb/IIIa and thus fibrinogen can potentially act as a hexavalent ligand to crossing GP IIb/IIIa molecules on adjacent platelets. A deficiency in either fibrinogen or GP ITh/IlIa prevents normal platelet aggregation regardless of the agonist used to activate the platelets. Since the binding of fibrinogen to its platelet receptor is an obligatory component of normal aggregation, GP IIb/IIIa is an attractive target for an antithrombotic agent.

Oral fibrinogen receptor antagonists are readily absorbed when a patient consumes them on an empty stomach. However, it has been recently observed that absorption and bioavailability of oral fibrinogen receptor antagonists, when taken with food, may be reduced by the presence of food in the stomach.

Cook et al., The Journal of Pharmacology and Experimental Therapeutics, Vol. 278, No. 1, pp. 62-73 (1996) describes oral antithrombotic efficacy of [3(R)-[2-Piperidin-4-yl)ethyl]- 2-piperidone- 1]acetyl-3(R)-methyl-ss-alanine, and shows that the magnitudes and durations of inhibition of ADP-induced platelet aggregation elicited by the compound at a dose of 3.0 mg/kg appeared equivalent between fasting and non-fasting dogs at time points more than 6 hours after dosing. At initial timepoints after dosing (0-6 hours) inhibition shown exceeds the detectable limits of the assay, and food effect on the administered dose cannot be determined.

SUMMARY OF THE INVENTION The present invention relates to a method for administering an amount of between about 0.1 mg/kg and about 5 mg/kg of a fibrinogen receptor antagonist to a non-fasting human patient in a non-fasting state wherein the fibrinogen receptor antagonist is [3(R)-[2-Piperidin-4- yl)ethyl]-2-piperidone-1]acetyl-3(R)-methyl-ss-alanine,

or a pharmaceutically acceptable salt thereof, to inhibit the aggregation of blood platelets The invention also includes a method for reducing the risk of adverse cardiovascular events in a non-fasting human at risk to adverse cardiovascular events comprising arlministering to the human patient a dose of [3(R)-[2-Piperidin-4-yl)ethyl]-2-piperidone-1]acetyl-3(R)- methyl-alanine or a pharmaceutically acceptable salt thereof, in an amount of between about 0.1 mg/kg and about 5 mg/kg.

The invention also includes a method for inhibiting the binding of fibrinogen to blood platelets in a non-fasting human, comprising administering to the mammal a dose of E3(R)-[2-PiperidinA- yl )ethyl)-2-piperidone- 1] acetyl-3 (R)-methyl-alanine or a pharmaceutically acceptable salt thereof, in an amount of between about 0.1 mg)kg and about 5 mgl:kg.

The invention also includes a method treating thrombus formation or embolism, or preventing thrombus or embolism a nonfasting human, which comprises administering to the human a dose of [3(R)-[2-Piperidin-4-yl)ethyl]-2-piperidone-1]acetyl-3(R)-methyl- alanine or a pharmaceutically acceptable salt thereof, in an amount of between about 0.1 mgig and about 5 mgAkg.

Preferably, the active agent administered in each of the above methods is [3(R)-[2-Piperidin-4-yl)ethyl]-2-piperidone-1]acetyl-3(R)- methyl-ss-alanine.

The invention is also a method for inhibiting the aggregation of blood platelets in a human, comprising administering to the human a fibrinogen receptor antagonist in an amount of between 0.1 mg/kg and about 5 mg/kg, wherein the fibrinogen receptor antagonist is C3(R)-[2 -Piperidin-4-yl)ethy1] -2-piperidone-l] acetyl-3(R)-methyl- P-alanine which provides reduced fibrinogen receptor antagonist plasma level variability.

The invention is also a method for inhibiting the aggregation of blood platelets in a human, comprising administering to the human a daily amount of a fibrinogen receptor antagonist in an amount of between 0.2 mg/kg and about 10 mg/kg, wherein the fibrinogen receptor antagonist is [3(R)-[2-Piperidin-4-yl)ethyl]-2 piperidone-1]acetyl-3(R)-methyl-ss-alanine which provides reduced fibrinogen receptor antagonist plasma level variability, and wherein the fibrinogen receptor antagonist is administered in two substantially equal doses each day.

The invention is useful for reducing the variability of fibrinogen receptor antagonist levels in the treated patient.

DETAILED DESCRIPTION OF THE INVENTION Hereinafter, it is understood that reference to [3(R)-[2 Piperidin-4-yl)ethyl]-2-piperidone- I]acetyl-3 (R)-methyl- -alanine, except for specific examples, includes both the active acid and salts thereof.

The method is specifically designed to inhibit platelet aggregation in non-fasting human patients. Thus, the method is particularly useful for reducing the risk of adverse cardiovascular events in such human patients at risk to adverse cardiovascular events.

[3(R)-[2-Piperidin-4-yl)ethyl]-2-piperidone- 1] acetyl-3(R)- methyl-ss-alanine plasma level variability is reduced when administered to a non-fasting human. The presence of food depresses the initial peak concentration that would otherwise result in the absence of food, and reduces plasma level variability over time to provide more uniform plamsa levels.

A non-fasting human patient is one who consumes food or beverage in quantities sufficient to deliver food or beverage to the stomach which eliminates physical conditions associated with a fasting state. Meals such as breakfast, lunch, and dinner which are exemplary of such foods and beverages are those which are composed of carbohydrates, fat and protein and representative of a breakfast, lunch or dinner typical of a normal balanced diet.

Administration to a nonfasting human occurs when the patient is in a nonfasting state. Such administration can be prior to a meal (e.g. immediately prior to, one-half hour prior to, one hour prior to a meal) during a meal, or following a meal (e.g. 2 hours after, 3 hours after) such that the patient is not in a fasting state.

In accordance with the invention, [3(R)-[2-Piperidin4- yl)ethyl]-2-piperidone- 1iacetyl-3(R)-methyl--alanine can be administered to the human patient in one oral composition or in several oral compositions. Preferably, the compound is administered two times each day. An extended release formulation, however, would also be suitable for administration.

Suitable oral compositions include tablets, compressed capsules, gel capsules containing (3(R)- [2-Piperidin-4-yl)ethylj-2- piperidone-liacetyl-3(R)-methyl-J3-alanine integrated into the gel matrix forming the gel capsule, gel capsules containing t3(R)-[2-Piperidin-4- yl)ethyl3-2-piperi done- 1]acetyl-3 (R)-methyl- 13-alanine particles, or gel capsules containing liquid [3(R)- E2.Piperidin-4-yl)ethyl]-2.piperidone- 1iacetyl-3(R)-methyl--alanine composition (each of which may include sustained release or timed release formulations).

Glycoprotein IIb/IIIa receptor antagonists and their pharmaceutically acceptable salts are useful in the present invention.

The term "pharmaceutically acceptable salts" means non-toxicsalts of the compounds which include, but are not limited to, acetate, bezizenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate, bromide, calcium edetate, camsylate, carbonate, chloride, clavulanate, citrate, dihydrochioride, edetate, edisylate, estolate, esylate, flanarate, gluceptate, gluconate, glutamate, glycollylarsanilate, hexylresorcinate, hydrabamine, hydrobromide, hydrochloride, hydroxynapthoate, iodide, isothionate, lactate, lactobionate, laurate, malate, maleate, mandelate, mesylate, methylbromide, methylnitrate, methylsulfate, mucate, napsylate, nitrate, oleate, oxalate, pamaote, palmitate, panthothenate, phosphate/diphosphate, polygalacturonate, salicylate, stearate, subacetate, succinate, tannate, tartrate, teoclate, tosylate, triethiodide, valerate.

Pharmaceutically effective amounts of [3(R)-[2-Piperidin-4 yl)ethyl]-2-piperidone-1]acetyl-3(R)-methyl-ss-alanine are suitable for use in the methods of the present invention. The term "pharmaceutically effective amount" means that amount of [3(R)-[2-Piperidin-4-yl)ethyl]-2 piperidone-1]acetyl-3(R)-methyl-ss-alanine that will elicit the biological or medical response of a tissue, system or animal that is being sought by a researcher or clinician.

The methods of the present invention are useful in combination with procedures for treating human patients with other anticoagulants (e.g. heparin, warfarin, thrombin inhibitors), thrombolytic agents (e.g. streptokinase and tissue plasminogen activator), and platelet antiaggregation agents (e.g. aspirin and dipyridamole).

The [3(R)-[2-Piperidin-4-yl)ethyl]-2-piperidone- 1]acetyl-3(R)- methyl-ss-alanine may be administered to human patients where prevention of thrombosis by inhibition of binding of fibrinogen to the platelet membrane glycoprotein complex IIb/IIIa receptor is desired.

The compound prevents or reduces the risk of adverse cardiovascular events such as death, acute coronary ischemic syndrome (e.g. myocardial infarction and unstable angina), stroke and amputation.

Administration is useful in surgery on peripheral arteries (arterial grafts, carotid endarterectomy), with stents, and in cardiovascular surgery where manipulation of arteries and organs, and/or the . interaction of platelets with artificial surfaces, leads to platelet aggregation and consumption. The aggregated platelets may form thrombi and thromboemboli. [3 (R)-[2-Piperidin-4-yl )ethyl]-2-piperidone- 1]acetyl-3(R)-methyl-ss-alanine may be administered to these surgical human patients to prevent the formation of thrombi and thromboemboli.

Other applications include prevention of platelet thrombosis, thromboembolism and reocclusion during and after thrombolytic therapy and prevention of platelet thrombosis, thromboembolism and reocclusion after angioplasty or coronary artery bypass procedures.

The dosage regimen utilizing [3(R)-F2-Piperidin-4-yl )ethyl]- 2-piperidone-1]acetyl-3(R)-methyl-ss-alanine is selected in accordance with a variety of factors including type, species, age, weight, sex and medical condition of the human patient; the severity of the condition to be treated; the route of administration; the renal and hepatic function of the human patient; and the particular compound or salt thereof employed.

An ordinarily skilled physician or veterinarian can readily determine and prescribe the effective amount of t3(R)-[2-Piperidin-4-yl)ethyl]-2- piperidone-13acetyl-3(R)-methyl-P-alanin required to prevent, counter, or arrest the progress of the condition.

Oral dosages of the present invention, when used for the indicated effects, will range between about 0.1 mg per kg of body weight (mgl) twice each day to about 5 mg/kg twice each day and preferably 0.1-3.0 mgAkg twice each day. For example, a typical 70 kg human patient would receive oral dosages ranging between about 7 mg twice each day and about 350 mg twice each day, preferably between about 7 mg twice each day and 210 mg twice each day. Suitable pharmaceutical oral compositions may contain, for example, 2 mg, 10 mg, 25 mg, 50 mg, 100 mg, 120 mg, 150 mg, and 300 mg, administered twice a day to maintain therapeutically effective levels of t3(R)-[2-Piperidin-4-yl)ethyl]- 2-piperidone-1]acetyl-3(R)-methyl-ss-alanine.

[3(R)-[2-Piperidin-4-yl)ethyl]-2-piperidone-1]acetyl-3(R) methyl-J3-alanine can be administered in admixture with suitable pharmaceutical diluents, excipients or carriers (collectively referred to herein as "carrier" or "filler" materials) suitably selected with respect to the intended form of administration, that is, oral tablets and capsules and the like, and consistent with convention pharmaceutical practices.

In the oral tablet, for example, [3(R)-[2-Piperidin-4-yl)ethyl]-2-piperidone- 1)acetyl-3(R)-methyl-J3-alanine can be combined with an oral, non-toxic, pharmaceutically acceptable excipients such as inert fillers including lactose (e.g. spray dried from Foremost Foods Co., San Francisco, CA; extrafine crystalline EFK, DMV, Veghel, The Netherlands), starch, sucrose, glucose, mannitol, microcrystalline cellulose (e.g. Avicelt FMC Corp. Philadelphia, PA), sorbitol, calcium phosphate dihydrate (e.g. DiTab, Stauffer Chemical Co., Westport, CT), and the like; lubricants including sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and the like; and disintegrating agents including starch and starch derivatives, cellulose derivatives, alginates, microcrystalline cellulose, croscarmellose sodium, crosslinked polyvinyl pyrrolidone, and the like.

Coloring and flavoring agents can also be incorporated into the tablet when desired.

Suitable binders for formulating with the active ingredient [3(R)-[2-Piperidin-4-yl)ethyl]-2-piperidone- 1]acetyl-3 (Rmethyl- alanine are polymeric binders. These include polymeric materials such as pre gelatinized starch (Starch 1500, Colorcom, Inc., West Point, PA), nonpre-gelatinized starch, gelatin, natural and synthetic gums such as acacia, tragacanth or sodium alginate, hydroxymethylcellulose, hydroxypropylcellulose, polyethylene glycol, polyvinylpyrolidone, waxes and the like.

Granulating fluids useful for forming the granules used to make the tablets include aqueous compositions which are typically used to cause powdered pharmaceutical composition material to agglomerate, e.g. water, aqueous ethanol solutions, citric acid solutions, aqueous hydroxypropylcellulose solutions, aqueous polyvinyl pyrrolidinone solutions, and alkali metal citrate solutions, and any other granulating fluid which is typically used in wet granulation processes to form pharmaceutical granules. The fluids may be sprayed into the mixed powder at a given temperature while powder is mixed in a granulator such as a Fukae-Powtec granulator or other high shear granulator equivalent, e.g. from Baker Perkins, Fielder, etc.

Wet granules are dried in, for example, a suitable piece of equipment such as a forced air tray dryer at a temperature range of between ambient and about 800C, or a fluidized bed dryer. The granules are kept under these conditions for a period of time necessary to obtain acceptable residual moisture levels. Thus it is understood that other commercially available dryers would also be suitable for the practice of this invention provided that the above identified purpose is accomplished by the equipment.

Using sieving equipment the dried granules are sized into oversized and undersized granules. The oversized granules are blended with a suitable lubricant. The mixture of the oversized granules and lubricant is milled through a suitable piece of equipment such as a Comic) fitted with a 062R screen and impeller speed of 725 rpm. Milling reduces the particle size of the large granules to facilitate more uniform distribution during finished dosage form preparation.

The purpose of milling is to reduce the particle size of the large granules. Thus it is understood that other commercially available mills fitted with various size screens and run at different impeller speeds would also be suitable for the practice of this invention provided that the above-defined purpose is accomplished by the equipment.

The milled oversized mixture and the undersized granules are mixed in a suitable piece of equipment such as a twin shell blender for a period of time such as three minutes or until there is distribution of the lubricant throughout all granules. The granular mixture is then ready for final dosage form preparation on a tablet press. Thus it is understood that other commercially available blenders would also be suitable for the practice of this invention provided that the aboveidentified purpose is accomplished by the equipment.

Tablets may also be prepared using tablet compression techniques or capsule technology well known to persons skilled in the art.

(3(R)-[2-Piperidin4-yl)ethyl]-2-piperidone- 1)acetyl-3(R)- mplgl-ss-alsnine can also be administered in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles and multilamellar vesicles. Liposomes can be formed from a variety of phospholipids, such as cholesterol, stearylamine or phosphatidylcholines.

[3(R)-[2-Piperidin-4-yl)ethyl]-2-piperidone- 1]acetyl-3(R)- methyl-ss-alanine may also be delivered by the use of monoclonal antibodies as individual carriers to which the compound molecules are coupled. (3(R[2-Piperidin-41l )ethylj-2-piperidone- Iiacetyl-3(R)-methyl- p-alanine may also be coupled with soluble polymers as targetable drug carriers. Such polymers can include polyvinyl-pyrrolidone, pyran copolymer, polyhydroxy-propyl-methacrylamide-phenol, polyhydroxyethyl-aspartamide-phenol, or polyethyleneoxide-polylysine substituted with pannitoyl residues. Furthermore, active drug may be coupled to a class of biodegradable polymers useful in achieving controlled release of a drug, for example, polylactic acid, polyglycolic acid, copolymers of polylactic and polyglycolic acid, polyepsilon caprolactone, polyhydroxy butyric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates and cross linked or amphipathic block copolymers of hydrogels.

Therapeutic Treatment The method for administering [3(R)-[2-Piperidin-4-yl)ethyl] 2piperidone- Iacetyl-3(R)-methyl- p-alanine are useful for treating human patients where inhibition of human or mammalian platelet aggregation or adhesion is desired. It is useful in surgery on peripheral arteries (arterial grafts, carotid endaterectomy) and in cardiovascular surgery where manipulation of arteries and organs, and/or the interaction of platelets with artificial surfaces, leads to platelet aggregation and potential formation of thrombi and thromboemboli.

Methods of the invention may be used to prevent the formation of thrombi and thromboemboli.

The present invention can be demonstrated in a study of non-fasting human patients with acute coronary ischemic syndromes who are undergoing early coronary revascularization with percutaneous coronary angioplasty or atherectomy. Because of unstable plaque with thrombus, percutaneous revascularization procedures in these human patients carry with them considerable higher morbidity than procedures performed in human patients with stable coronary disease. All human patients receive heparin (a standard PTCA regimen, weight adjusted in lighter human patients) and aspirin.

Heparin is discontinued after completion of the procedure and sheaths removed when the heparin-effect has dissipated. The [3(RS[2-Piperidin- 4-yl)ethyl]-2-piperidone- 1]acetyl-3(R)-methyl-ss-alanine composition is administered on a daily basis to human patients who are evaluated at 30 days for acute coronary ischemic syndrome and the need for follow-up intervention associated with acute coronary ischemic syndrome, including coronary artery bypass grafting, repeat percutaneous intervention for acute ischemia, and insertion of a coronary endovascular stent.

EXAMPLE 1 Non-fasting human patients with acute coronary ischemic syndromes receive coronary revascularization with angioplasty.

Aspirin is administered in a dose of 325 mg at least two hours before angioplasty, and daily thereafter. Heparin is given intravenously in an initial bolus dose of 7,000 to 10,000 units followed by incremental bolus doses of up to 3000 units at 15-minute intervals. The goal is to keep the activated clotting time between 300 and 350 seconds during the operation.

Heparin is continued by constant infusion for at least 12 hours to maintain the activated partial-thromboplastin time at 1.5 to 2.5 times the control value. Aspirin is required at discharge in a dose of 325 mg per day.

Patients then receive an oral tablet containing 50 mg of [3(:R)-[2-Piperidin-4-yl)ethyl]-2-piperidone- 1]acetyl-3(R)-methyl-alanine twice each day. Tablets are administered on a daily basis to non-fasting human patients in order to inhibit platelet aggregation.

Patients are monitored 30 days following initiation of [3(R) [2-Piperidin-4-yl)ethyl] -2-piperidone 1]acetyl-3(R)-methyl-fralaiine therapy, and show reduction in acute coronary ischemic syndrome after 30 days.

EXAMPLE 2 Tablet Prenaration Tablets containing 50 mg of [3(R)-[2-Piperidin-4-yl)ethyl]-2 piperidone-1Jacetyl-3(R)-methyl--alanine are prepared as illustrated below: Ingredient m g [3(R)-[2-Piperidin-4-yl)ethyl]-2-piperidone- 1]acetyl-3(R)- 50.0 methyl- P-alanine Microcrystalline cellulose 200.0 Modified food corn starch 8.5 Magnesium stearate 1.5 [3(R)-[2-Piperidin-4-yl)ethyl]-2-piperidone-1]acetyl-3(R)- methyl-ss-alanine, cellulose, and a portion of the corn starch are mixed and granulated to 10% corn starch paste. The resulting granulation is sieved, dried and blended with the remainder of the corn starch and the magnesium stearate. The resulting granulation is then compressed into tablets.

EXAMPLE3 Capsule Preparation Capsules containing [3(R )-[2-Piperidin-4-yl )ethyl]-2- piperidone-1]acetyl-3(R)-methyl-ss-alanine are prepared as illustrated below: Ingredient m g [3(R)-[2-Piperidin-4-yl)ethyl]-2-piperidone- ljacetyl-3(R)- 50.0 methyl-ss-alanine Microcrystalline cellulose 42.2 Dicalcium phosphate 42.2 Cro scarmellose sodium 0.1 Magnesium stearate 0.5 Hydroxypropyl methyl cellulose phthalate 10 [3(R)-[2-Piperidin-4-yl)ethyl]-2-piperidone-1]acetyl-3(R)- methyl-ss-alanine, microcrystalline cellulose, disodium phosphate and croscarmellose sodium are mixed in a blender. The resulting mixture is granulated and dried prior to milling. Milled granules are lubricated and filled into a standard pharmaceutical capsule, having a narrow dimension of 4 mm, which is then coated with hydroxypropyl methyl cellulose phthalate.

ExAMPLE4 Compositions containing 2, 25 or 150 mg of [3(R)-[2-Piperidin-4-yl)ethyl]- 2-piperidone-1]acetyl-3(R)-methyl-ss-alanine Tablets containing 2, 25 or 150 mg respectively, of the compound r3(R)-r2-Piperidin4-yl)ethyl]-2-piperidone-1]acetyl-3(R)- methyl-I3-alanine are prepared as illustrated below.

Tngredients Amount (mg) [3(R)-[2-Piperidin-4-yl)ethyl)- 2.0 25.0 150 2-piperidone- 1]acetyl-3(RS methyl-ss-alanine Starch pre gelatinized NF 45.6 34.05 204.3 1500 Avicel PH 101 45.6 34.05 204.3 (Microcrystalline cellulose) Croscarmellose sodium NF 6.0 6.0 36.0 Talc 0.6 0.6 3.6 Magnesium stearate 0.3 0.3 1.8 Absolute ethanol 7.5 Cil 7.5 Cll 45K11 Water (purified) 22.5 p1 22.5 il 135 l [3(R)-[2-Piperidin-4-yl)ethyl]-2-piperidone- 1J acetyl-3(R)- methyl-ss-alanine, starch, microcrystalline cellulose, one-half quantity of croscarmellose, microcrystalline cellulose, starch pre gelatinized, magnesium stearate are mixed and placed in a clean Fukae-Powtec granulator or other high shear granulator equivalent e.g., from Baker Perkins, Fielder, etc., and mixed for one minute. Ethanol and water are mixed in a beaker and then added over a period of 1.5 minutes while the powder mixture is being mixed. The resulting mixture is then granulated for 7 minutes and thereafter collected. The granules are dried at 470C in a tray dryer for 3 hours. The dry granules are then milled and mixed with the remaining one-half of croscarmellose.

Magnesium stearate and talc are mixed and then added to granulecroscarmellose mixture in a V blender and mixed for 5 minutes. The blended material is then pressed into tablets using an F Press tablet machine or other tablet press.

Tablet hardness evaluation, using standard tablet hardness evaluation procedure, showed that the formed tablet having 2 mg of active ingredient had a hardness of 8.8 kp, and that the formed tablet having 10 mg active ingredient had a hardness of 9 kp.

EXAMPLE 5 Compositions containing 2, 25 or 150 mg of [3(R)-[2-Piperidin-4-yl)ethyl]- 2-niperidone-1]acetyl-3(R)-methyl-ss-alanine The same compositions described in Example 1 are made according to the same procedure except tray drying is replaced with fluidized bed drying.

EXAMPLE 6 Composition containing 50 mg [3(R)-[2-Piperidin-4-yl)ethyl]-2-piperidone- llacetvl-3(R)-methvl- 13.alanine A tablet containing 50 mg [3(R)-[2-Piperidin-4-yl)ethylj-2- piperidone-1]acetyl-3(R)-methyl-ss-alanine and the following additional ingredients was prepared following the procedure described in Example 1.

Ingredient Amount img) [3(R)-[2-Piperidin-4-yl )ethyl]-2-piperidone- 50.0 liacetyl-3(R)-methyl-13-alanine Starch pre gelatinized NF 1500 68.1 Avicel PH 101 (Microcrystalline cellulose) 68.1 Croscarmellose sodium NF 12.0 Talc 1.2 Magnesium s evaluation procedure, showed that the formed tablet had a hardness of 8 kp.

A material balance study of the tablets prepared in Example 2 shows that essentially 100% of the amount of [3(R)-[2-Piperidin-4 yl)ethyli-2-piperidone-1]acetyl-3(R)-methyl--alanine added to the granulator was usefillly incorporated into the finished product (i.e., no loss of [3(R)-[2-Piperidin-4-yl)ethyl]-2-piperidone-1]acetyl-3(R)-methyl-ss- alanine).

EXAMPLE 7 Composition containing 100 mg [3(R)-[2-Piperidin-4-yl)ethyl]-2 niperidone- 1lacetvl-3(R)-methvl-alanine A tablet containing 100 mg [3(R)-[2-Piperidin-4-yl)ethyl]-2 piperidone-1]acetyl-3(R)-methyl-ss-alanine and the following additional ingredients was prepared following the procedure described in Example 1.

Ingredient Amount (m) [3(R)-[2-Piperidin-4-yl )ethyl]-2-piperidone- 100.0 1]acetyl-3(R)-methyl-ss-alanine Starch pre gelatinized NF 1500 136.2 Avicel PH 101 (Microcrystalline cellulose) 136.2 Croscarmellose sodium NF 24.0 Talc 2.4 Magnesium stearate 1.2 Absolute ethanol 30 Water (purified) 90.0 p11 Croscarmellose was added in two steps as described in Example 1 except that amounts of 12 mg each rather than 3 mg each were added. The formulation was studied for stability and shown to be stable.

EXAMPLE8 A non-fasting human patient is treated with any of the tablets or capsules described in examples 2-7 by orally administering the tablets or capsules to the human patients. Pharmacologically effective delivery of [3(R)-[2-Piperidin-4-yl)ethyl]-2-piperidone- 1]acetyl-3(R)- methyl-13-alanine is achieved and platelet aggregation is inhibited.

EXAMPLE 9 AdminisStion of [3(R)-[2-Piperidin-4-yl)ethyl]-2-piperidone-13acetyl- 3(Rmethvl-J3-alanine to fasting and non-fasting humans Subjects received a single oral dose of study drug on four occasions. Treatments were administered as follows: Treatment A: Subjects were required to fast from all food (and liquid except water) from midnight the night before until 2 hours postdose.

Subjects were given the appropriate dose of study drug between 8 and 10 AM with at least 150 mL of water. The exact time of dosing was recorded on the CRF. Additionally, liquid (as water and/or apple juice) was consumed after the 120-minute phlebotomy. A standard lunch was provided to all subjects after the 2-hour posttreatment blood samples were collected. A standard dinner was provided at approximately 9 hours postdose.

Treatment B: Subjects were required to fast from all food and liquids (except water) from midnight the preceding night to approximately 20 minutes prior to dosing. At approximately 20 minutes prior to dosing, subjects consumed a standard breakfast. Drug was administered between 8 and 10 AM with 150 mL of water and within 5 minutes of completing the breakfast. Water and/or apple juice was permitted after the 120-minute phlebotomy. No further food was consumed for a least 4 hours from dosing. A standard lunch was provided to all subjects after the 4-hour posttreatment blood samples were collected. A standard dinner was provided at 9 hours postdose. Meals were preferably eaten during a 20-minute period.

Treatment C: Subjects were required to fast from all food (and liquid except water) from midnight the night before dosing. Between 7 and 8 AM, subjects were given a standard breakfast. Breakfast was consumed within 20 minutes. Subjects were given the appropriate dose of study drug with at least 150 mL of water 2 hours after completing the breakfast between 9 and 10 AM. A standard lunch was provided to all subjects after the 2-hour posttreatment blood samples were collected. A standard dinner was provided at 9 hours postdose.

Treatment D: Subjects were admitted in the afternoon and provided a standard dinner between 5 and 7 PM. After the meal, subjects fasted until the following morning. At 2 hours after completing the meal, drug was administered. Water and/or apple juice was permitted after the 120minute phlebotomy. A standard breakfast and lunch was provided to subjects the following day at -7 to 9 AM and 11 AM to 1 PM, respectively.

For these treatments, the dose given was 100 mg [3(R)-[2- Piperidin-4-yl)ethyl]-2-piperidone-1]acetyl-3(R)-methyl-ss-alanine (n=12) or matching placebo (n=2). All meals (i.e., breakfast, lunch, and dinner) were standardized, and the contents were the same in each period. As shown in the attached table, each meal was composed of carbohydrates, fat and protein and representative of a breakfast, lunch or dinner typical of a normal balanced diet.

Standard Meals During Studv for Each Treatment Penod Food Item Intake Amount Meal Bread-Whole Wheat-Firm-Enriched 1.0 Slice Breakfast Egg Beaters-No Cholesterol 4.0 Ounce Breakfast (Fleischman) Bacon-Pork-Broiled/Pan 1.0 Slice Breakfast Fried/Roasted Cereal (Cheerios-USDA) 1.0 Cup Breakfast Juice-Orange-Frozen Concentrate- 0.5 Cup Brealiast Dilute Mi -2% FatbNonfat Milk Solids 1.0 Cup Breakfast Added Margarine (I Can't Believe Its Not 1.0 Serving Breakfast Butter) Jam/Preserves-Regular 1.0 Tbsp Breakfast Beef-Pot Roast-Chuck-Arm Cut- 3.0 Ounce Lunch Cooked Potato-Mashed from Dehydrated- 0.6 Cup Lunch With Carrots-Frozen-Boiled-Drained 0.5 Cup Lunch Bread-'White-Firm-Enriched 1.0 Slice Lunch Pineapple-Canned-Juice Pack- 0.5 Cup Lunch USDA Gravy-Brown 0.3 Cup Lunch Gelatin Dessert-Prepared 0.5 Cup Lunch Milk-2% Fat-Nonfat Milk Solids 1.0 Cup Lunch Added Pie-Cherry-Comm Prep From 3.0 Ounce Lunch Frozen Margarine (I Can't Believe Its Not 1.0 Serving Lunch Butter) Food Item Intake Amount Meal 6.0 Ounce Dinner Baked Macaroni and Cheese Fish-Cod-Scrod-Atlantic-Baked or 4.0 Ounce Dinner Broiled Peas-Green-Frozen-Boiled-Drained 0.5 Cup Dinner Bread-White-Firm-Enriched 1.0 Slice Dinner Pears-Canned-Juice Pack 0.5 Cup Dinner Milk-2% Fat Nonfat Mild Solids 1.0 Cup Dinner Added Salad-Coleslaw 0.5 Cup Dinner Margarine (I Can't Believe Its Not 1.0 Serving Dinner Butter) There was a washout interval of 5 to 7 days between doses.

Subjects who received placebo in Period 1 also received placebo in each subsequent period. Therefore, the allocation schedule assigned subjects to either [3 (R [2-Piperidin4-yl )ethyl]- 2-piperidone- ijacetyl- 3(Rmethyi- p-alanine or placebo and to dosing conditions with regard to food intake.

Safety was assessed by measurement of blood pressure and heart rate and by careful questioning for adverse events. Plasma and urine was collected for [3(R)-[2-Piperidin-4-yl)ethyl]-2-piperidone- 1]acetyl-3(R)-methyl-ss-alanine concentrations.

For all human patients studied, no drugs (including aspirin, any NSAIDs or drugs that affect platelet function, or an anticoagulant) were taken within 14 days of the start of the study. No drugs were taken during the study unless required to treat a serious adverse effect. Subjects were allowed to consume a normal balanced diet except when treatment was administered as described in Treatments A, B, C, and D above. Strenuous physical exercise was avoided throughout the duration of the study. Subjects refrained from all alcoholic beverages from 48 hours before until 24 hours or longer after treatment if any pharmacological effects were still present following each treatment.

Subjects drank no caffeine-containing beverages beginning 24 hours prior to dosing and throughout the duration of admission. Smoking was not permitted during periods of admission.

The results of the study showed that a 100 mg dose of [3(R) [2-Piperidin-4-yl)ethyl]-2-piperidone- 1]acetyl-3(R)-methyl- 13-alanine in non-fasting human subjects provided drug plasma level concentrations in the subjects with minimal variability compared to dosing in a fasting human. The following table shows plasma level concentration in nglml over time.

Claims (10)

WHAT IS CLAIMED IS:

1. A method for inhibiting the aggregation of blood platelets in a non-fasting human, comprising treating the non-fasting human with a dose of [3 (R)-[2-Piperidin-4-yl )ethyl] -2-piperidone- liacetyl- 3(R)-methyl-ss-alanine or a pharmaceutically acceptable salt thereof, in an amount of between about 0.1 mg/kg and about 5 mg/kg.

2. A method of Claim 1 wherein the composition comprises [3(R)-[2-Piperidin-4-yl)ethyl]-2-piperidone- 1]acetyl-3(R)- methyl-ss-alanine.

3. A method for reducing the risk of adverse cardiovascular events in a non-fasting human at risk for adverse cardiovascular events comprising administering to the human patient a dose of [3(R)-[2-Piperidin-4-yl)ethyl]-2-piperidone-1]acetyl-3(R)-methyl-ss- alanine or a pharmaceutically acceptable salt thereof, in an amount of between about 0.1 mg/kg and about 5 mg/kg.

4. A method of Claim 3 wherein the composition comprises [3(R)-[2-Piperidin-4-yl)ethyl]-2-piperidone-1]acetyl-3(R)- methyl-ss-alanine.

5. A method for inhibiting the binding of fibrinogen to blood platelets in a non-fasting human, comprising administering to the non-fasting human a dose of [3(R)-[2-Piperidin-4-yl)ethyl]-2-piperidone- 1] acetyl-3(R)-methyl- P-alanine or a pharmaceutically acceptable salt thereof, in an amount of between about 0.1 mg/kg and about 5 mg/kg.

6. A method of Claim 5 wherein the composition comprises [3(R)-[2-Piperidin-4-yl)ethyl]-2-piperidone- 1] acetyl-3(R)- methyl-P-alanine.

7. A method for treating thrombus formation or embolism, or preventing thrombus or embolism in a non-fasting human, which comprises administering to the human a dose of [3(R)-[2 Piperidin.4.yl)ethyl]-2-piperidone- 1] acetyl-3(R)-methyl--alanine or a pharmaceutically acceptable salt thereof, in an amount of between about 0.1 mg/kg and about 5 mg/kg.

8. A method of Claim 7 wherein the composition comprises [3(R)-[2-Piperidin-4-yl)ethyl]-2-piperidone-1]acetyl-3(R)- methyl-j3-alanine.

9. A method for inhibiting the aggregation of blood platelets in a human, comprising administering to the human a fibrinogen receptor antagonist in an amount of between 0.1 mg/kg and about 5 mgAg, wherein the fibrinogen receptor antagonist is [3(R)-[2 Piperidin-4-yl)ethyl]-2-piperidone- 1)acetyl-3 (R)-methyl--alanine which provides reduced fibrinogen receptor antagonist plasma level variability.

10. A method for inhibiting the aggregation of blood platelets in a human, comprising administering to the human a daily amount of a fibrinogen receptor antagonist in an amount of between 0.2 mg/kg and about 10 mg/kg, wherein the fibrinogen receptor antagonist is [3(R)-[2-Piperidin-4-yl)ethyl]-2-piperidone-1]acetyl-3(R)-methyl-ss- alanine which provides reduced fibrinogen receptor antagonist plasma level variability, and wherein the fibrinogen receptor antagonist is administered in two substantially equal doses each day.