Abstract:
A unit dosage form, such as a capsule or the like for delivering drugs into the body in a circadian release fashion, is comprising of one or more populations of propranolol-containing particles (beads, pellets, granules, etc.). Each bead population exhibits a pre-designed rapid or sustained release profile with or without a predetermined lag time of 3 to 5 hours. Such a circadian rhythm release cardiovascular drug delivery system is designed to provide a plasma concentration—time profile, which varies according to physiological need during the day, i.e., mimicking the circadian rhythm and severity/manifestation of a cardiovascular disease, predicted based on pharmaco-kinetic and pharmaco-dynamic considerations and in vitro/in vivo correlations.

Description:
This application is a continuation-in-part application of U.S. application Ser. No. 10/334,052 filed Dec. 30, 2002, now abandoned, which is a continuation of U.S. application Ser. No. 09/971,167 filed Oct. 4, 2001, now U.S. Pat. No. 6,500,454. Each of the foregoing applications, each document cited or referenced in each of the foregoing applications and during the prosecution of each of the foregoing applications (“application cited documents”), each document referenced or cited in each of the application cited documents, each document cited or referenced in this application (“herein cited documents”) and each document cited or referenced in each of the herein cited documents are all incorporated herein by reference. 
    
    
     TECHNICAL FIELD 
     A major objective of chronotherapy for cardiovascular diseases is to deliver the drug in higher concentrations during the time of greatest need, typically during the early morning hours, and in lesser concentrations when the need is less, such as during the late evening and early sleep hours. This can be accomplished by administration of the release dosage form of the present invention, which relates to a controlled absorption of propranolol from dosage forms. The release dosage form of the present invention, which relates to a controlled absorption of propranolol from dosage form, comprises an assembly of Timed, Sustained Release (TSR) Beads, each of which is designed to release as a sustained release pulse after a predetermined delay (“time-controlled” drug delivery instead of “rate-controlled”) with resulting plasma concentration(s) of propranolol varying in a circadian rhythm fashion following administration of a single dosage form at bedtime, thereby minimizing potential risks of a cardiovascular disease, such as stroke, heart attack and myocardial infarction, decreasing systolic blood pressure, or reducing hypertension or beta-adrenergic stimulation, treating cardiac arrhythmia, hypertrophic subaortic stenosis or angina, or preventing migraine and thus enhancing patient compliance and therapeutic efficacy, while reducing cost of treatment. 
     BACKGROUND OF THE INVENTION 
     Many therapeutic agents are most effective when made available at a constant rate at or near the absorption site. The absorption of therapeutic agents thus made available generally results in desired plasma concentrations leading to maximum efficacy and minimum toxic side effects. Much effort has been devoted to developing sophisticated drug delivery systems, such as osmotic devices, for oral application. However, there are instances where maintaining a constant blood level of a drug is not desirable. For example, a “position-controlled” drug delivery system (e.g., treatment of colon disease or use of colon as an absorption site for peptide and protein based products) may prove to be more efficacious. A pulsatile delivery system is capable of providing one or more immediate release pulses at predetermined time points after a controlled lag time or at specific sites. However, there are only a few such orally applicable pulsatile release systems due to the potential limitation of the size or materials used for dosage forms. Ishino et al. disclose a dry-coated tablet form in Chemical Pharm. Bull. Vol. 40 (11), 3036-041 (1992). U.S. Pat. No. 4,851,229 to Magruder et al., U.S. Pat. No. 5,011,692 to Fujioka et al., U.S. Pat. No. 5,017,381 to Maruyama et al., U.S. Pat. No. 5,229,135 to Philippon et al., and U.S. Pat. No. 5,840,329 to Bai disclose preparation of pulsatile release systems. Some other devices are disclosed in U.S. Pat. No. 4,871,549 to Ueda et al. and U.S. Pat. Nos. 5,260,068; 5,260,069; and 5,508,040 to Chen. U.S. Pat. Nos. 5,229,135 and 5,567,441 both to Chen disclose a pulsatile release system consisting of pellets coated with delayed release or water insoluble polymeric membranes incorporating hydrophobic water insoluble agents or enteric polymers to alter membrane permeability. U.S. Pat. No. 5,837,284 to Mehta et al. discloses a dosage form which provides an immediate release dose of methylphenidate upon oral administration, followed by one or more additional doses spread over several hours. 
     There is a well-established circadian variation in frequency of onset of cardiovascular events including ventricular arrhythmias, stroke, angina, and myocardial infarction. The peak frequency of such events is exhibited in the morning hours, theoretically in conjunction with the morning surge in systolic blood pressure and heart rate. There is also some evidence of a secondary peak in frequency of such events in the late afternoon or evening hours. Although there is some evidence which suggests that long acting propranolol may blunt the circadian variability of sudden cardiac death, it does not appear to attenuate early morning increases in blood pressure observed in hypertensive patients. Thus it would be physiologically advantageous to tailor plasma concentrations of propranolol to the typical circadian patterns of blood pressure and heart rate. 
     Chronotherapeutics is a means of proportioning plasma drug concentrations during a 24 hour period, relative to the biological rhythm determinates of disease activity. The objective of chronotherapy is to deliver the drug in higher concentrations during the time of greatest need, and in lesser concentrations when the need is less. The dosage forms disclosed in the prior arts above are not specifically designed to provide drug release profiles varying predictably in time over 24 hours, i.e., in a circadian rhythm fashion to effectively treat cardiovascular diseases. The dosage forms of the present invention, Propranolol Hydrochloride ER Capsules, 80, 120, and 160 mg), which are typically administered at bedtime, i.e., at about 10:00 PM, are novel formulations designed to provide reductions in blood pressure and heart rate over 24 hours, including optimal protection in the early morning hours when patients are most vulnerable to cardiovascular events. At steady state, blood levels of propranolol begin to increase approximately 4 hours after bedtime administration of these capsules and rise progressively over the early morning hours to reach peak plasma concentrations approximately 14 hours after dosing. These capsules produce peak plasma propranolol levels that rise slowly to attenuate the rapid increase in blood pressure and heart rate that precedes and follows waking. This increase is associated with circadian variation in catecholamine secretion and in rennin release. The rise in plasma propranolol concentration after dosing with these capsules parallel the circadian rise in morning blood pressure associated with target organ damage in patients with hypertensive and ischemic cardiovascular disease. 
     Propranolol [1-(isopropyl amino)-3-(1-naphthyloxy)-2-propanol] is a betaadrenergic blocking agent and as such is a competitive inhibitor of the effects of catecholamines at beta-adrenergic receptor sites. The principal effect of propranolol is to reduce cardiac activity by diminishing or preventing beta-adrenergic stimulation. By reducing the rate and force of contraction of the heart, and decreasing the rate of conduction of impulses through the conducting system, the response of the heart to stress and exercise is reduced. These properties are used in the treatment of angina in an effort to reduce the oxygen consumption and increase the exercise tolerance of the heart. Propranolol is also used in the treatment of cardiac arrhythmias to block adrenergic stimulation of cardiac pacemaker potentials. Propranolol is also beneficial in the long term treatment of hypertension. Other uses of propranolol are in the treatment of migraine and anxiety. 
     SUMMARY OF THE INVENTION 
     The preparation of the dosage form of the present invention is fully described in U.S. Pat. No. 6,500,454 assigned to Eurand Pharmaceuticals, Ltd., which is incorporated here by reference in its entirety. The dosage form, a hard gelatin capsule, is a timed, sustained release multi-particulate dosage form comprising a propranolol core having a first membrane of a sustained release polymer and a second membrane of a mixture of water insoluble polymer and an enteric polymer (2 nd  or outer coating), wherein the water insoluble polymer and the enteric polymer may be present at a weight ratio of from 10:1 to 1:2, and the total weight of the coatings is 10 to 60 weight % based on the total weight of the coated beads. In some cases depending on the type of drug release profile needed, an immediate release component may be included to provide a modified, timed, sustained release dosage form. When administered at bedtime, the dosage form comprising Timed, Sustained Release Capsule, commercially known as TSR Formulation XL, is designed to provide reductions in blood pressure and heart rate over 24 hours, including optimal protection in the early morning hours when patients are most vulnerable to cardiovascular events. At steady state, blood levels of propranolol begin to increase approximately 4 hours after bedtime administration of TSR Formulation XL and rise progressively over the early morning hours to reach peak plasma concentrations approximately 14 hours after dosing. TSR Formulation XL produces peak plasma propranolol levels that rise slowly to attenuate the rapid increase in blood pressure and heart rate that precedes and follows waking. This increase is associated with circadian variation in catecholamine secretion and in rennin release. The rise in plasma propranolol concentration after dosing with TSR Formulation XL parallels the circadian rise in morning blood pressure associated with target organ damage in patients with hypertensive and ischemic cardiovascular disease. 
    
    
     
       BRIEF DESCRIPTION OF THE FIGURES 
       The invention will be described in further detail with reference to the accompanying Figures wherein: 
         FIG. 1  shows in vitro drug release profiles from Propranolol HCl TSR Capsules, 80, 120, and 160 mg, used in the pivotal clinical studies. 
         FIG. 2  shows mean propranolol concentration versus time plots for 80 mg, 120 mg and 160 mg dosage strengths of TSR Formulation following a single administration of the drug product. 
         FIG. 3 a    shows the mean propranolol plasma concentration over time (0-80 hours) during multiple dosing of either Inderal® LA 160 mg or TSR Formulation 160 mg (i.e., at steady state). 
         FIG. 3 b    shows the mean propranolol plasma concentration over time (24 hours) during multiple dosing of either Inderal® LA 160 mg or TSR (RelPro) Formulation 160 mg (i.e., at steady state). 
     
    
    
     DETAILED DESCRIPTION OF THE INVENTION 
     The active core of the novel dosage form of the present invention may comprise an inert particle or an acidic or alkaline buffer crystal, which is coated with a propranolol-containing film-forming formulation and preferably a water-soluble film forming composition to form a water-soluble/dispersible particle. Alternatively, the active core may be prepared by granulating and milling and/or by extrusion and spheronization of a polymer composition containing propranolol. Generally, the individual polymeric coating on the active core will be from 1 to 50% based on the weight of the coated particle. Those skilled in the art will be able to select an appropriate amount of propranolol for coating onto or incorporating into the core to achieve the desired dosage. In one embodiment, the inactive core may be a sugar sphere, a buffer crystal or an encapsulated buffer crystal, such as calcium carbonate, sodium bicarbonate, fumaric acid, tartaric acid, etc. Buffer crystals are useful to alter the microenvironment. 
     In accordance with one embodiment of the present invention, the water soluble/dispersible drug-containing particle is first coated with a water insoluble polymer (1 st  or inner coating), and further coated with a mixture of a water insoluble polymer and an enteric polymer (2 nd  or outer coating). The water insoluble polymer and enteric polymer may be present at a weight ratio of from 10:1 to 1:2, more preferably 2:1 to 1:1, and the total weight of the coatings is 10 to 60 weight % based on the total weight of the coated beads. The polymeric coatings typically contain plasticizers and may be applied from aqueous and/or solvent based systems. 
     The composition of the outer layer and the individual weights of the inner and outer layers of the polymeric membrane are optimized for achieving desired drug release profiles. The unit dosage form according to certain embodiments of the present invention may comprise an immediate release bead population which provides an immediate release component of propranolol to act as a bolus dose. 
     The invention also provides a method of making a timed, sustained release dosage form comprising the steps of: 
     1. preparing an active-containing core by coating an inert particle such as a nonpareil seed, an acidic buffer crystal or an alkaline buffer crystal, with propranolol and polymeric binder or by granulation and milling or by extrusion/spheronization to form an immediate release (IR) bead; 
     2. coating the core with a plasticized solution or suspension of a water insoluble polymer to form sustained release (SR) coated drug particle; 
     3. coating the SR coated particle with a mixture of plasticized water insoluble and enteric polymers to form a Timed Sustained Release (TSR) coated drug particle; and filling capsules with TSR particles to produce Timed, Sustained Release (TSR) capsules. 
     The release profile for TSR beads can be determined according to the following procedure: 
     Dissolution testing is conducted with a USP Apparatus  2  (Paddles at 50 rpm) using a two-stage dissolution medium (first 2 hours in 700 mL 0.1N HC1 at 37° C. followed by dissolution at pH=6.8 obtained by the addition of 200 mL of pH modifier). Drug release with time is determined by HPLC on samples pulled at selected intervals. 
     The TSR Beads prepared in accordance with present invention release not more than 20%, more preferably not more than 10%, and most preferably not more than 5% in 2 hours, about 5-35%, more preferably about 5-25%, and most preferably about 5-15% in 4 hours, about 10-60%, more preferably about 20-45%, and most preferably about 25-35% in 6 hours, about 4090%, more preferably about 50-80%, and most preferably about 55-70% in 10 hours, and not less than 60%, more preferably not less than 70%, and most preferably not less than 75% in 16 hours. 
     In accordance with the present invention, the desired release properties are obtained as a result of the different characteristics of the two coating layers. The inner layer membrane provides sustained or extended drug release over several hours, while the second or outer membrane provides a lag time of three to four hours. Typical release profiles for SR beads (ethylcellulose coated drug particle) and TSR beads when tested by the two-stage dissolution medium are provided in Table 1, below: 
     
       
         
               
               
               
             
               
               
               
             
               
               
               
             
           
               
                   
                 TABLE 1 
               
             
             
               
                   
                   
               
               
                   
                 SR Beads 
                 TSR Beads 
               
             
          
           
               
                 Time 
                 (% Propranolol Released) 
                   
               
               
                   
               
             
          
           
               
                  1 hr 
                 11.2 
                  0.0 
               
               
                  2 hr 
                 32.1 
                  0.1 
               
               
                  3 hr 
                 39.8 
                  1.1 
               
               
                  4 hr 
                 52.3 
                  8.6 
               
               
                  5 hr 
                 62.3 
                 18.3 
               
               
                  6 hr 
                 69.2 
                 27.4 
               
               
                  8 hr 
                 79.4 
                 44.5 
               
               
                 10 hr 
                 84.6 
                 58.4 
               
               
                 12 hr 
                 90.0 
                 68.8 
               
               
                 16 hr 
                 95.6 
                 90.0 
               
               
                   
               
             
          
         
       
     
     It is also possible that the TSR Capsule may optionally also contain a population of Immediate Release (IR) beads or particles to provide an immediate release component of active to act as a bolus dose in addition to the timed, sustained release of active provided by the TSR beads. These dosage forms provide a Modified Timed Sustained Release (MTSR) profile. 
     An aqueous or a pharmaceutically acceptable solvent medium may be used for preparing drug containing core particles. The type of film forming binder that is used to bind propranolol to the inert sugar sphere is not critical but usually water-soluble, alcohol-soluble or acetone/water soluble binders are used. Binders such as polyvinylpyrrolidone (PVP), polyethylene oxide, hydroxypropyl methylcellulose (HPMC), hydroxypropylcellulose (HPC), polysaccharides, such as dextran, and corn starch may be used at concentrations of from about 0.5 to 10 weight %. Propranolol may be present in the coating formulation in solution form or may be suspended at a solids content up to about 35 weight % depending on the viscosity of the coating formulation. 
     Dissolution rate controlling polymers suitable for incorporating in the formulation for producing granules by high shear or fluid bed granulation or by dry granulation include high molecular weight hydroxypropyl methylcellulose, hydroxypropyl cellulose, ethyl cellulose, sodium carboxymethyl cellulose, alginic acid, polymethylmethacrylate copolymers and polyvinyl acetate/crotonic acid copolymer or combinations thereof. Acidic buffers, which help maintain an acidic microenvironment within drug containing particles, include fumaric acid, tartaric acid, maleic acid, succinic acid and mixtures thereof. An acidic microenvironment helps dissolve basic drugs with poor solubility at the intestinal pHs and become available for absorption. Examples of alkaline buffers include sodium bicarbonate, calcium carbonate, and sodium dihydrogen phosphate. 
     Propranolol, a binder such as PVP, a buffer, a dissolution rate controlling polymer (if used), and optionally other pharmaceutically acceptable excipients are blended together in a high shear granulator such as Fielder or a fluid bed granulator such as Glatt GPCG 5 and granulated to form agglomerates by adding/spraying a granulating fluid such as water or alcohol and dried. The wet mass can be extruded and spheronized to produce spherical particles (beads) using an extruder/marumerizer. In these embodiments, the drug load could be as high as 90% by weight based on the total weight of the extruded/spheronized core. The blend can also be used to produce dry granules by slugging in a tablet press or a chilsonator, without the addition of any granulating fluid. 
     The active containing cores (beads, pellets or granular particles) thus obtained may be coated with one or two layers of polymers to obtain desired release profiles with or without a lag time. The inner layer membrane, which largely controls the rate of release following imbibition of water or body fluids into the core, comprises a water insoluble polymer, such as ethylcellulose, at a thickness of from 1 weight % up to 6 weight %, preferably from 1.5 to 4% and most preferably about 2%, depending on the solvent or latex suspension based coating formulation used. 
     The outer membrane, which largely controls the lag time of up to 6 hours, comprises an enteric polymer and a water insoluble polymer at a thickness of 10 to 60, preferably from 10 to 56 weight % based on the total weight of the coated beads. The ratio of water insoluble polymer to enteric polymer may vary from 10:1 to 1:2, preferably from 2:1 to 1:1. 
     Representative examples of water insoluble polymers useful in the invention include cellulose derivatives (e.g. ethylcellulose), polyvinyl acetate (Kollicoat SR30D from BASF), neutral copolymers based on ethyl acrylate and methylmethacrylate, copolymers of acrylic and methacrylic acid esters with quaternary ammonium groups, such as Eudragit NE, RS or RS30D, RL or RL30D and the like. 
     Representative examples of enteric polymers useful in the invention include esters of cellulose and its derivatives (cellulose acetate phthalate, hydroxypropyl methylcellulose phthalate, hydroxypropyl methylcellulose acetate succinate), polyvinyl acetate phthalate, pHsensitive methacrylic acid-methamethacrylate copolymers and shellac. These polymers may be used as a dry powder or an aqueous dispersion. Some commercially available materials that may be used are methacrylic acid copolymers sold under the trademark Eudragit (L100, S100, L30D) manufactured by Rhom Pharma, Cellacefate (cellulose acetate phthalate) from Eastman Chemical Co., Aquateric (cellulose acetate phthalate aqueous dispersion) from FMC Corp. and Aqoat (hydroxypropyl methylcellulose acetate succinate aqueous dispersion) from Shin Etsu K.K. 
     Both enteric and water insoluble polymers used in forming the membranes are usually plasticized. Representative examples of plasticizers that may be used to plasticize the membranes include triacetin, tributyl citrate, triethyl citrate, acetyl tri-n-butyl citrate diethyl phthalate, castor oil, dibutyl sebacate, acetylated monoglycerides and the like or mixtures thereof. The plasticizer may comprise about 3 to 30 wt. % and more typically about 10 to 25 wt. % based on the polymer. The type of plasticizer and its content depends on the polymer or polymers, nature of the coating system (e.g., aqueous or solvent based, solution or dispersion based and the total solids). 
     In general, it is desirable to prime the surface of the particle before applying the pulsatile release membrane coatings or to separate the different membrane layers by applying a thin hydroxypropyl methylcellulose (HPMC) (Opadry Clear) film. While HPMC is typically used, other primers such as hydroxypropylcellulose (HPC) can also be used. 
     The membrane coatings can be applied to the core using any of the coating techniques commonly used in the pharmaceutical industry, but fluid bed coating is particularly useful. 
     The present invention is applied to multi-dose forms, i.e., drug products in the form of multi-particulate dosage forms (pellets, beads, granules or mini-tablets) or in other forms suitable for oral administration. 
     The following non-limiting examples illustrate the capsule dosage forms manufactured in accordance with the invention, which exhibit in vitro drug release profiles, similar to that predicted by performing modeling exercises, and in vivo plasma concentrations following circadian rhythm pharmaco-dynamic profile of angina attacks. Such dosage forms when administered at bed time, would enable maintaining drug plasma concentration at a level potentially beneficial in minimizing the occurrence of heart attacks in the early hours of the morning. 
     EXAMPLES 
     Modified Timed, Sustained Release (MTSR) capsules of Propranolol Hydrochloride may contain a mixture of two sets of beads: The first set is referred to as immediate release (IR) Beads and are designed to provide a loading dose by releasing all of the drug within the first hour, preferably within the first 30 minutes. The second set is referred to as Timed Sustained Release (TSR) Beads and are designed to release the remainder of the dose slowly over a period of 12-15 hours after a 3-5-hour lag time. The TSR Beads are produced by applying an inner layer of sustained release coating (with a dissolution rate controlling polymer such as ethylcellulose) (producing IntR Beads, intermediate release beads) and then an outer layer of pulse coating (with a blend of an enteric polymer such as HPMCP and a water-insoluble polymer such as ethylcellulose) on IR Beads. The two sets of beads (IR and TSR) when filled into capsule shells at an appropriate ratio will produce the target circadian rhythm release profile required for maintaining drug plasma concentrations at potentially beneficial levels in minimizing the occurrence of heart attacks. Alternatively, the capsules may comprise only the TSR Beads, the capsules containing only the TSR Beads are referred to as TSR Formulation. It is well known that the blood pressure begins to drop as the night advances, and consequently, only the Formulations containing only the TSR beads for oral administration are described below. 
     Example 1 
     Propranolol HCl (45.2 kg) was slowly added to an aqueous solution of polyvinylpyrrolidone (2.34 kg Povidone K-30) and mixed well. # 25-30 mesh sugar spheres (31.6 kg) were coated with the drug solution in a Glatt fluid bed granulator. The drug containing pellets were dried, and a seal coat of Opadry Clear (2% w/w) was first applied (batch size: 80.75 kg). The inner sustained release coating was applied to the active particles (73.7 kg) by spraying a solution of ethylcellulose and diethyl phthalate in 98/2 acetone/water. The outer coating of a blend of ethylcellulose and HPMCP plasticized with diethyl phthalate was sprayed onto the active particles having the inner coating to produce TSR Beads (batch size: 82.5 kg). These TSR Beads were filled into hard gelatin capsules using an MG capsule filling equipment to produce Propranolol hydrochloride TSR Capsules, 80, 120, and 160 mg. 
     These Propranolol TSR Capsules were tested for drug release profiles by the two-stage dissolution method, wherein capsules were dissoluted at pH 1.5 in 700 mL 0.1N HCl for two hours followed by testing at pH 6.8 in 900 mL obtained by adding 200 mL of concentrated buffer modifier.  FIG. 1  presents the drug release profiles from Propranolol Hydrochloride TSR Capsules, 80, 120, and 160 mg, used in the pivotal clinical studies. 
     Example 2 
     A double-blind, randomized, placebo-controlled, 4-period crossover study to assess the efficacy and dose proportionality of TSR Formulation 80 mg, 120 mg, and 160 mg in 39 healthy male and female subjects of ages 18 to 40 was carried out. In each study period a single dose of one strength of TSR Formulation or placebo was administered at approximately 10:00 PM. There was a minimum of 7 days between the administration of each dose of TSR Formulation or placebo. Efficacy was assessed by the mean change in post-exercise heart rate, heart rate pressure product, and systolic blood pressure from pre-dose to Hour 24 post-dose (trough) assessments. The secondary efficacy variable was the mean change in post-exercise heart rate from pre-dose to Hour 12. Post-exercise heart rate was used as a surrogate marker for antianginal efficacy. 
     Analyses for concentrations of total plasma propranolol (conjugated and unconjugated) on the blood samples collected were carried out using a validated high performance liquid chromatography (HPLC) method. Plasma samples were collected pre-dose and up to 72 hours post-dose. The limit of quantitation was 2 ng/mL.  FIG. 2  displays mean propranolol concentration versus time plots for 80 mg, 120 mg and 160 mg dosage strengths of TSR Formulation following a single administration of the drug product. Maximum plasma propranolol concentrations (C max ) occurred at about 12 hours post-dose for the 80 mg, 120 mg, and 160 mg dose groups. After 12 hours post-dose, plasma propranolol concentrations decreased at a consistent and similar rate in all dose groups. The absorption lag time (T lag ) and time to maximum drug concentration (T max ) were similar across all TSR Formulation dose groups. Table 2 summarizes the pharmacokinetic parameters for this study. 
     
       
         
               
             
               
               
             
               
               
               
               
             
               
               
               
               
             
           
               
                 TABLE 2 
               
             
             
               
                   
               
               
                 Summary of Pharmacokinetic Parameters—Pharmacokinetic Population 
               
             
          
           
               
                   
                 TSR Formulation 
               
             
          
           
               
                   
                 80 mg 
                 120 mg 
                 160 mg 
               
               
                 Parameter/Statistic 
                 (N = 36) 
                 (N = 36) 
                 (N = 36) 
               
               
                   
               
             
          
           
               
                 AUC( 0–T )(ng · hr/mL) a   
                   
                   
                   
               
               
                 N 
                 36 
                 36 
                 36 
               
               
                 Mean 
                 1442.6 
                 2270.1 
                 3417.1 
               
               
                 SD 
                 844.19 
                 1230.62 
                 2047.40 
               
               
                 Median 
                 1220.8 
                 2102.2 
                 3047.2 
               
               
                 Minimum 
                 551.8 
                 594.3 
                 1211.1 
               
               
                 Maximum 
                 3792.1 
                 5928.7 
                 11596.2 
               
               
                 AUC( 0–∞ )(ng · hr/mL) 
               
               
                 N 
                 36 
                 36 
                 36 
               
               
                 Mean 
                 1568.5 
                 2412.6 
                 3589.0 
               
               
                 SD 
                 865.20 
                 1287.41 
                 2080.77 
               
               
                 Median 
                 1422.9 
                 2180.1 
                 3199.5 
               
               
                 Minimum 
                 605.1 
                 646.6 
                 1291.1 
               
               
                 Maximum 
                 3994.0 
                 6660.0 
                 12157.2 
               
               
                 C max  (ng/mL) 
               
               
                 N 
                 36 
                 36 
                 36 
               
               
                 Mean 
                 80.5 
                 130.0 
                 177.2 
               
               
                 SD 
                 31.53 
                 52.30 
                 70.62 
               
               
                 Median 
                 75.8 
                 117.0 
                 171.0 
               
               
                 Minimum 
                 30.1 
                 41.5 
                 72.4 
               
               
                 Maximum 
                 155.0 
                 249.0 
                 426.0 
               
               
                 t 1/2  (hr) 
               
               
                 N 
                 36 
                 36 
                 36 
               
               
                 Mean 
                 9.5 
                 8.5 
                 9.1 
               
               
                 SD 
                 5.05 
                 4.7 
                 4.35 
               
               
                 Median 
                 8.1 
                 7.1 
                 8.8 
               
               
                 Minimum 
                 3.3 
                 3.2 
                 2.8 
               
               
                 Maximum 
                 22.7 
                 20.2 
                 21.3 
               
               
                 T max  (hr) 
               
               
                 N 
                 36 
                 36 
                 36 
               
               
                 Mean 
                 12.2 
                 12.8 
                 12.8 
               
               
                 SD 
                 1.93 
                 2.46 
                 2.72 
               
               
                 Median 
                 12.0 
                 12.0 
                 12.0 
               
               
                 Minimum 
                 8.0 
                 10.0 
                 8.0 
               
               
                 Maximum 
                 20.0 
                 20.0 
                 24.0 
               
               
                 T lag  (hr) 
               
               
                 N 
                 36 
                 36 
                 36 
               
               
                 Mean 
                 2.2 
                 2.1 
                 1.9 
               
               
                 SD 
                 1.05 
                 0.89 
                 0.67 
               
               
                 Median 
                 2.0 
                 2.0 
                 2.0 
               
               
                 Minimum 
                 0.0 
                 0.0 
                 0.0 
               
               
                 Maximum 
                 4.0 
                 4.0 
                 4.0 
               
               
                 K el  (1/hr) 
               
               
                 N 
                 36 
                 36 
                 36 
               
               
                 Mean 
                 0.0965 
                 0.1076 
                 0.0958 
               
               
                 SD 
                 0.05090 
                 0.05384 
                 0.05125 
               
               
                 Median 
                 0.0862 
                 0.0987 
                 0.0788 
               
               
                 Minimum 
                 0.0305 
                 0.0343 
                 0.0326 
               
               
                 Maximum 
                 0.2107 
                 0.2193 
                 0.2517 
               
               
                   
               
               
                   a t is the last time point with measurable propranolol concentration 
               
             
          
         
       
     
     Linear regression analyses on untransformed pharmacokinetic parameters showed dose proportionality and a linear dose-response relationship between the TSR Formulation dosage strengths and the pharmacokinetic parameters AUC 0-T  (area under the concentration-time curve to the last measurable time point calculated by the linear trapezoidal rule), AUC 0-∞  (are under the concentration-time curve to infinity), and C max . Results of linear regression and ANOVA after log-transformation and dose-normalization confirmed a dose proportional relationship between AUC 0-T , AUC 0-∞ , and C max  and TSR Formulation 80 mg, 120 mg, and 160 mg doses. 
     Regarding efficacy, maximum decreases in systolic blood pressure were noted at 8-10 hours post-dose in all treatment groups. Maximum decreases in diastolic blood pressure were noted at 8-10 hours post-dose in the TSR Formulation 120 mg and placebo groups and at 14 hours post-dose in the TSR Formulation 80 mg and 160 mg groups. Maximum decreases in heart rate were observed at 10 hours post-dose in all treatment groups. Mean changes in systolic and diastolic blood pressure and heart rate showed increases from pre-dose values at 72 hours post-dose in all treatment groups. No subject experienced postural hypotension during the study. 
     Only the TSR Formulation 80 mg dose group showed a statistically significant decrease in the adjusted mean change in exercise-induced heart rate from baseline at 24 hours post-dose, relative to placebo (primary end point) and in 12-hour post-exercise heart rate (secondary end point). No dose response was observed for heart rate. Statistically significant changes, relative to placebo, were observed for two primary end points: hour 24 heart rate-pressure product seen for the lowest and highest doses and hour 24 heart rate at the lowest dose of TSR Formulation; these data support the efficacy of TSR formulations in the prevention and treatment of angina. Demonstration of blunting of exercise response to beta blockade for the 12-hour post p.m. dosing exercise challenge also supports the efficacy of the TSR Formulation in protection against morning ischemic events. 
     Example 3 
     A randomized, open-label, two-period crossover study to evaluate the single and multiple dose bioavailability and safety of TSR Formulation 160 mg compared to Inderal® LA 160 mg capsules was carried out. The primary objective of this study was to evaluate the relative bioavailability of TSR Formulation 160 mg to Inderal® LA 160 mg as assessed by an ANOVA on AUCs after single and multiple dose administrations. The secondary objective was to explore the pharmacokinetics of TSR Formulation and Inderal® LA after single and multiple dose administrations. Safety and tolerability were measured by evaluating adverse events, laboratory values, physical examinations, electrocardiograms, and vital signs. 
     Subjects were healthy, adult, males between 18 and 45 years of age who met the study eligibility criteria. In Period  1  of the study, following a 4-hour fasting period on Day  1 , subjects received a single dose of the study drug (TSR Formulation or Inderal® LA) between 9:30 and 10:30 PM. Serial blood samples for plasma total propranolol determinations were collected for 72 hours. Subjects received a daily dose of their Period  1  drug on Days  4  to  8  and 24-hour trough blood samples were collected. A seven-day washout period followed, and the same procedures were followed for Period  2  with the other study drug as determined by the sequence to which the subjects were randomized. Analyses for concentrations of total plasma propranolol (conjugated and unconjugated) on the blood samples collected were carried out using a validated high performance liquid chromatography (HPLC) method. The limit of quantitation was 2 ng/mL. 
     Plots of the mean propranolol plasma concentration over time following a single dose of 160 mg propranolol TSR Formulation or 160 mg Inderal® LA are presented in  FIG. 3 a   . Following a single dose of TSR Formulation 160 mg there was a delayed release of propranolol for 2 to 4 hours, whereas release of propranolol following a single dose of Inderal® LA 160 mg was almost immediate. Within 3 hours after administration of Inderal® LA, mean plasma propranolol concentrations increased to approximately 50% of the mean maximum propranolol concentration. In contrast, following administration of TSR Formulation, 50% of the mean maximum plasma propranolol concentration was not reached until approximately 7 hours post administration. Between 10 hr and 12 hr post dose, mean plasma concentrations in the Inderal® LA group remained relatively constant (range 122.8 ng/mL to 128.2 ng/mL) while mean plasma concentrations in the TSR Formulation group steadily increased (range 146.9 ng/mL to 174.8 ng/mL). The time to maximum plasma levels of propranolol (T max ) was 10.5 hr and 11.9 hr for subjects administered Inderal® LA and TSR Formulation, respectively. The C max  values were 149.1 ng/mL for Inderal® LA and 186.5 ng/mL for TSR Formulation, respectively. At 24 hr post-dose, the mean plasma concentrations were 51.8 ng/mL and 67.3 ng/mL in the Inderal® LA and TSR Formulation groups, respectively. After 24 hr post-dose, plasma concentrations decreased at a consistent and similar rate in both groups. The elimination half-life (t 1/2 ) was similar for both formulations. 
     A summary of the single dose pharmacokinetic parameters of AUC 0-T , AUC 0-∞  and C max  following an Analysis of Variance (ANOVA) is given in Table 3. The adjusted mean C max  values were 131.0 ng/mL and 171.3 ng/mL for subjects administered Inderal® LA and TSR Formulation, respectively. 
     
       
         
               
             
               
               
               
               
             
           
               
                 TABLE 3 
               
             
             
               
                   
               
               
                 Summary of Analysis of Variance on 
               
               
                 Single Dose Pharmacokinetic Parameters 
               
             
          
           
               
                   
                   
                   
                 Ratio (TSR 
               
               
                   
                   
                   
                 Formulation/ 
               
               
                   
                   
                   
                 Inderal ® 
               
               
                 Parameter/ 
                 Inderal ® LA 
                 TSR Formulation 
                 LA) 
               
               
                 Statistic 1   
                 160 mg (N = 35) 
                 160 mg (N = 35) 
                 (N = 35) 
               
               
                   
               
               
                 AUC 0–T   
                   
                   
                   
               
               
                 (ng · hr/mL) 
               
               
                 Adjusted Mean 
                 2588.7 
                 2943.9 
                 1.1 
               
               
                 90% CI 
                 (2399.77, 2792.44) 
                 (2729.06, 3175.62) 
                 (1.02, 1.27) 
               
               
                 AUC 0–∞   
               
               
                 (ng · hr/mL) 
               
               
                 Adjusted Mean 
                 2604.8 
                 2962.7 
                 1.1 
               
               
                 90% CI 
                 (2415.90, 2808.43) 
                 (2747.90, 3194.37) 
                 (1.02, 1.27) 
               
               
                 C max  (ng/mL) 
               
               
                 Adjusted Mean 
                  131.0 
                  171.3 
                 1.3 
               
               
                 90% CI 
                 (117.52, 146.12) 
                 (153.66, 191.04) 
                 (1.12, 1.53) 
               
               
                   
               
               
                   1 Adjusted mean and 90% CI calculated from ANOVA on log-transformed parameter with subject, period, sequence, and treatment as fixed effects. Adjusted means and CIs displayed have been transformed from the log to the arithmetic scale. 
               
               
                 CI: Confidence Interval 
               
             
          
         
       
     
       FIG. 3 b    displays the mean propranolol plasma concentration over time during multiple dosing of either Inderal® LA 160 mg or TSR Formulation 160 mg (i.e., at steady state). Following multiple doses of TSR Formulation 160 mg, there was a delayed release of propranolol for 2 to 6 hours, whereas the release of propranolol for Inderal® LA occurred after a slight delay of less than 1 hour. At steady state, the mean maximum plasma propranolol concentrations (C max SS ) were 229.9 ng/mL and 248.3 ng/mL for subjects administered Inderal® LA and TSR Formulation, respectively. Mean minimum plasma propranolol concentrations (C min ) at steady state were 56.3 ng/mL and 58.9 ng/mL for subjects administered Inderal® LA and TSR Formulation, respectively. The mean time to observed maximum propranolol concentration at steady state (T max SS ) was approximately 2 hours greater for TSR Formulation (12.6 hr) than for Inderal® LA (10.9 hr). Similarly, the mean time to the observed minimum plasma propranolol concentration at steady state (T min ) was approximately 2 hours greater for TSR Formulation (7.3 hr) than for Inderal® LA (5.1 hr). Steady state plasma propranolol concentrations for both TSR Formulation and Inderal® LA were attained after 2 days of dosing during the multiple dosing period. Both formulations showed similar accumulation at steady state and propranolol concentrations fluctuated in a similar manner over the dosing interval. 
     Table 4 describes a summary of the results of the ANOVA performed on the pharmacokinetic parameters AUC o-TSS , C AV SS , R ac SS , and Fi SS . 
     
       
         
               
             
               
               
               
               
             
           
               
                 TABLE 4 
               
             
             
               
                   
               
               
                 Summary of Analysis of Variance on 
               
               
                 Steady State Pharmacokinetic Parameters 
               
             
          
           
               
                   
                   
                   
                 Ratio 
               
               
                   
                   
                   
                 (Formulation 
               
               
                   
                   
                   
                 TSR/ 
               
               
                   
                   
                   
                 Inderal ® 
               
               
                 Parameter/ 
                 Inderal ® LA 
                 Formulation TSR 
                 LA) 
               
               
                 Statistic 1   
                 160 mg (N = 35) 
                 160 mg (N = 35) 
                 (N = 35) 
               
               
                   
               
               
                 AUC 0–τSS   
                   
                   
                   
               
               
                 (ng · hr/mL) 
               
               
                 Adjusted Mean 
                 3115.0  
                 3325.6  
                 1.1 
               
               
                 90% CI 
                 (2928.62, 3313.28) 
                 (3126.56, 3537.22) 
                  (.98, 1.16) 
               
               
                 C AV SS  (ng/mL) 
               
               
                 Adjusted Mean 
                 128.7 
                 137.5 
                 1.1 
               
               
                 90% CI 
                 (121.02, 136.92) 
                 (129.22, 146.20) 
                  (.98, 1.17) 
               
               
                 R ac SS   
               
               
                 (AUC 0–τSS / 
               
               
                 AUC 0–τ DAY 1 ) 
               
               
                 Adjusted Mean 
                  1.7 
                  1.7 
                 1.0 
               
               
                 90% CI 
                 (1.54, 1.94) 
                 (1.47, 1.86) 
                 (0.81, 1.13) 
               
               
                 Fi SS  (C max  − 
               
               
                 C min /C av ) 
               
               
                 Adjusted Mean 
                 114.3 
                 126.7 
                 1.1 
               
               
                 90% CI 
                 (105.38, 123.93) 
                 (116.87, 137.45) 
                 (0.99, 1.24) 
               
               
                   
               
               
                   1 Adjusted mean and 90% CI calculated from ANOVA on log-transformed parameter with subject, period, sequence, and treatment as fixed effects. Adjusted means and CIs displayed have been transformed from the log to the arithmetic scale. 
               
               
                 CI: Confidence Interval 
               
             
          
         
       
     
     Example 4 
     A randomized, double-blind, parallel, placebo-controlled, multicenter trial to study the efficacy, safety and steady state pharmacokinetics of 80 mg, 120 mg, 160 mg, and 640 mg TSR Formulation in patients with essential hypertension was carried out. The primary objective of this study was to assess the efficacy of the TSR Formulation in subjects with essential hypertension by evaluating the mean change from Baseline to Week  8  in morning sitting diastolic pressure. Subjects were randomized to one of the five double-blind treatment groups: placebo, TSR Formulation 80 mg/day, TSR Formulation 120 mg/day, TSR Formulation 160 mg/day, and TSR Formulation 640 mg/day taken once daily before bedtime. During Week  1  and Week  2  subjects were up titrated to the appropriate dose for 6 weeks, and then they were down titrated for the last two weeks of the study. 
     Morning sitting diastolic pressure, the measure of primary efficacy, decreased from Baseline to Endpoint for the placebo and all four TSR Formulation groups. Statistically significant differences in the magnitude of the decrease between the placebo group and the 120 mg, 160 mg, and 640 mg TSR Formulation groups were observed. Statistical trends were observed for the 80 mg TSR Formulation dose group. These results demonstrate that the TSR Formulation, administered once daily at bedtime in dosage strengths of 80 mg to 640 mg propranolol is an effective antihypertensive agent.