WO2000059480A1 - Lyophilized solid dosage forms and methods of making - Google Patents

Lyophilized solid dosage forms and methods of making Download PDF

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Publication number
WO2000059480A1
WO2000059480A1 PCT/US2000/009412 US0009412W WO0059480A1 WO 2000059480 A1 WO2000059480 A1 WO 2000059480A1 US 0009412 W US0009412 W US 0009412W WO 0059480 A1 WO0059480 A1 WO 0059480A1
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Prior art keywords
delivery agent
solution
delivery
solid dosage
agent
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PCT/US2000/009412
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French (fr)
Inventor
Michael M. Goldberg
Rajesh K. Agarwal
Kiran Chaudhary
Shingai Majuru
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Emisphere Technologies, Inc.
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Priority to AU42184/00A priority Critical patent/AU4218400A/en
Publication of WO2000059480A1 publication Critical patent/WO2000059480A1/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C231/00—Preparation of carboxylic acid amides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
    • A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
    • A61K47/183—Amino acids, e.g. glycine, EDTA or aspartame
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D265/00—Heterocyclic compounds containing six-membered rings having one nitrogen atom and one oxygen atom as the only ring hetero atoms
    • C07D265/04—1,3-Oxazines; Hydrogenated 1,3-oxazines
    • C07D265/12—1,3-Oxazines; Hydrogenated 1,3-oxazines condensed with carbocyclic rings or ring systems
    • C07D265/14—1,3-Oxazines; Hydrogenated 1,3-oxazines condensed with carbocyclic rings or ring systems condensed with one six-membered ring
    • C07D265/24—1,3-Oxazines; Hydrogenated 1,3-oxazines condensed with carbocyclic rings or ring systems condensed with one six-membered ring with hetero atoms directly attached in positions 2 and 4
    • C07D265/26—Two oxygen atoms, e.g. isatoic anhydride
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/20—Pills, tablets, discs, rods
    • A61K9/2004—Excipients; Inactive ingredients
    • A61K9/2013—Organic compounds, e.g. phospholipids, fats

Definitions

  • the present invention relates to a solid dosage form comprising a lyophilized powder obtained from a solution comprising a delivery agent and an active agent.
  • the solid dosage form is suitable for oral administration of active agents to animals. Methods of preparation and administration are also disclosed.
  • Solid dosage forms for delivering certain active agents are well known in the art.
  • a drug plus a diluent or excipient is milled to mix the powders to obtain a uniform mixture.
  • the mixture is then granulated by compression or is made into a wet dough-like mixture, which is then milled to make small granules.
  • the granules are dried in an oven or on a fluidized bed drive at temperatures of 40-80°C.
  • the dried granules are run through a sieve to obtain particles of a uniform size.
  • the granules are then put into capsules or compressed into tablets.
  • protein, polypeptide or peptide active agents such as calcitonin, insulin, human growth hormone and parathyroid hormone.
  • the process might denature the active agent, rendering it inactive and ineffective for its intended use. Therefore, protein, polypeptide or peptide drugs are not currently made in solid dosage forms .
  • a solid dosage form useful in the administration of active agents comprises a lyophilized powder obtained from lyophilization of a solution comprising a delivery agent and an active agent.
  • the solid dosage forms of the present invention have utility in the delivery of active agents to selected biological systems and in an increased bioavailability of the active agent compared to other dosage forms.
  • a method of preparing the solid dosage form is provided, the method comprising:
  • a method of administering a biologically-active agent to an animal in need of said agent comprising administering to the animal the solid dosage form is also provided.
  • the solid dosage forms of the present invention include an active agent and a delivery agent. These solid dosage forms may be used to deliver various active agents through various biological, chemical, and physical barriers and are particularly suited for delivering active agents which are subject to environmental degradation.
  • the delivery agent may be any compound that effects delivery of an active agent.
  • the delivery may be acylated amino acids, sulfonated amino acids, ketones or aldehydes of acylated or sulfonated amino acids, or polypeptides which include any of the foregoing.
  • the delivery agent units comprises:
  • R C ⁇ -C 2 alkyl, C 2 -C 2 alkenyl, C 3 -C ⁇ 0 cycloalkyl, C 3 - Cio cycloalkenyl, phenyl, naphthyl, (C ⁇ -C ⁇ 0 alkyDphenyl, (C 2 -Ci 0 alkenyl) phenyl, (C -C ⁇ 0 alkyl) naphthyl, (C 2 -C ⁇ 0 alkenyl) naphthyl , phenyl (Ci-Cio alkyl), phenyl (C 2 -C ⁇ 0 alkenyl), naphthyl (C ⁇ -C ⁇ 0 alkyl) and naphthyl (C 2 -C ⁇ 0 alkenyl) ;
  • R may be optionally substituted with C ⁇ -C ⁇ 0 alkyl, C 2 - Cio alkenyl, C ⁇ -C 4 alkoxy, -OH, -SH, -C0 2 R' (where R' is H, C ⁇ -C 4 alkyl or C 2 -C 4 alkenyl) , C 3 -C ⁇ 0 cycloalkyl, C 3 -C ⁇ 0 cycloalkenyl, heterocyclic having 3-10 ring atoms wherein the hetero atom is one or more atoms of N, 0, S or any combination thereof, aryl, (C 1 -C 1 0 alkyl) aryl, aryl(C ⁇ - C 1 0) alkyl, or any combination thereof;
  • R may be optionally interrupted by 0, N, S or any combination thereof; or a salt thereof .
  • the delivery agent comprises
  • R 2 is H, OH
  • the delivery agent comprises
  • the delivery agent comprises :
  • the delivery agent comprises
  • the delivery agents may be in the form of the carboxylic acid and/or their salts, including but not limited to sodium salts.
  • poly amino acids and peptides comprising one or more of these compound may be used.
  • amino acid is any carboxylic acid having at least one free amine group and includes naturally occurring and synthetic amino acids.
  • Poly amino acids are either peptides (which are two or more amino acids joined by a peptide bond) or are two or more amino acids linked by a bond formed by other groups which can be linked by, e.g., an ester or an anhydride linkage.
  • Peptides can vary in length from dipeptides with two amino acids to polypeptides with several hundred amino acids. See Chambers Biological Dictionary, editor Peter M.B. Walker, Cambridge, England: Chambers Cambridge, 1989, page 215.
  • One or more of the amino acids or peptide units may be acylated or sulfonated.
  • amino acid derivatives may be derived from amino acids and can be readily prepared from amino acids by methods within the skill of those in the art based upon the present disclosure and the methods described in WO96/30036, WO97/36480, US 5,643,957 and US 5,650,386.
  • the compounds may be prepared by reacting the single amino acid with the appropriate acylating or amine-modifying agent, which reacts with a free amino moiety present in the amino acid to form amides .
  • Protecting groups may be used to avoid unwanted side reactions as would be known to those skilled in the art . With regard to protecting groups, reference is made to T.W. Greene, Protecting Groups in Organic Synthesis, Wiley, New York (1981) , the disclosure of which is hereby incorporated by reference.
  • the compound may be purified by recrystallization or by fractionation on one or more solid chromatographic supports, alone or linked in tandem.
  • Suitable recrystallization solvent systems include, but are not limited to, acetonitrile, methanol, and tetrahydrofuran. Fractionation may be performed on a suitable chromatographic support such as alumina, using methanol/n- propanol mixtures as the mobile phase; reverse phase chromatography using trifluoroacetic acid/acetonitrile mixtures as the mobile phase; and ion exchange chromatography using water or an appropriate buffer as the mobile phase.
  • anion exchange chromatography preferably a 0-500 mM sodium chloride gradient is employed.
  • Active agents suitable for use in the present invention include biologically active agents and chemically active agents, including, but not limited to, pesticides, pharmacological agents, and therapeutic agents.
  • biologically or chemically active agents suitable for use in the present invention include, but are not limited to, proteins, polypeptides, peptides, and particularly small peptides; hormones, and particularly hormones which by themselves do not pass (or which pass only a fraction of the administered dose) through the gastro-intestinal mucosa and/or are susceptible to chemical cleavage by acids and enzymes in the gastro-intestinal tract; polysaccharides, and particularly mixtures of muco- polysaccharides; carbohydrates; lipids; or any combination thereof.
  • growth hormones including human growth hormones (hGH) , recombinant human growth hormones (rhGH) , bovine growth hormones, and porcine growth hormones; growth hormone-releasing hormones; interferons, including , ⁇ and ⁇ ; interleukin-l; insulin; insulin-like growth factor, including IGF-1; heparin, including unfractionated heparin, heparinoids, dermatans, chondroitins, low molecular weight heparin, very low molecular weight heparin and ultra low molecular weight heparin; calcitonin, including salmon, eel and human; erythropoietin; atrial naturetic factor; antigens; monoclonal antibodies; somatostatin; protease inhibitors; adrenocorticotropin, gonadotropin releasing hormone; oxytocin; leut
  • the solid dosage forms of the present invention comprise a delivery agent and one or more active agents and may optionally contain additives such as phosphate buffer salts, citric acid, glycols, or other dispersing agents. Stabilizing additives may be incorporated into the solution, preferably at a concentration ranging between about 0.1 and 20% (w/v) .
  • the solid dosage unit forms may also include one or more enzyme inhibitors.
  • enzyme inhibitors include, but are not limited to, compounds such as actinonin or epiactinonin and derivatives thereof. Derivatives of these compounds are disclosed in US 5,206,384.
  • Other enzyme inhibitors include, but are not limited to, aprotinin (Trasylol) and Bowman-Birk inhibitor.
  • the amount of active agent is an amount effective to accomplish the purpose of the particular active agent for the target indication.
  • the amount of active agent in the compositions typically is a pharmacologically, biologically, therapeutically, or chemically effective amount. However, the amount can be less than that amount when the composition is used in a dosage unit form that contains a plurality of delivery agent/active agent compositions or a divided pharmacologically, biologically, therapeutically, or chemically effective amount.
  • the total effective amount can then be administered in cumulative units containing, in total, pharmacologically, biologically, therapeutically or chemically active amounts of biologically or pharmacologically active agent.
  • the total amount of active agent, and particularly biologically or chemically active agent, to be used can be determined by those skilled in the art. However, because the compositions may deliver active agents more efficiently than prior compositions, lower amounts of biologically or chemically active agents than those used in prior dosage unit forms or delivery systems can be administered to the subject, while still achieving the same blood levels and therapeutic effects.
  • Lyophilization techniques are well known in the art.
  • the delivery agent and active agent can be made up into solution and lyophilized to form a powder.
  • a solid dosage form can be a capsule, tablet or powder.
  • the powder may be in the form of a sachet that is mixed with a liquid and administered.
  • the solid dosage form may also be a topical delivery system, such as an ointment, cream or semi-solid.
  • the solid dosage form contemplated may include a sustained release or controlled release system.
  • the lyophilized powder may be packed into capsules, or pressed into tablets, used in powder form, or incorporated into an ointment, cream or semi-solid. Methods for forming solid dosage forms are well known in the art.
  • the presently disclosed solid dosage forms are suitable for delivering biologically and chemically active agents, particularly in oral, intranasal, sublingual, intraduodenal, buccal, rectal, vaginal, mucosal, pulmonary, transdermal, and intramuscular systems.
  • the amount of lyophilized material in the present solid dosage forms is a delivery effective amount and can be determined for any particular compound or biologically or chemically active agent by methods known to those skilled in the art.
  • Dosage unit forms can also include any of excipients, diluents, disintegrants, lubricants, plasticizers, colorants, flavorants, taste-masking agents, sugars, sweeteners, salts, and dosing vehicles, including, but not limited to, water, 1,2 -propane diol, ethanol, olive oil, or any combination thereof.
  • the dosage unit forms of the present invention are useful for administering biologically or chemically active agents to animals, including but not limited to poultry, such as chickens; mammals, such as cows, pigs, dogs, cats, primates, and particularly humans; and insects.
  • the system is particularly advantageous for delivering chemically or biologically active agents which would otherwise be destroyed or rendered less effective by conditions encountered before the active agent reaches its target zone (i.e. the area in which the active agent of the delivery composition is to be released) and within the body of the animal to which they are administered.
  • the solid dosage forms of the present invention are useful in orally administering active agents, especially those which are not ordinarily orally deliverable.
  • the active agent present in the dosage unit form is taken up into the circulation.
  • the bioavailability of the agent is readily assessed by measuring a known pharmacological activity in blood, e.g. an increase in blood clotting time caused by heparin, or a decrease in circulating calcium levels caused by calcitonin. Alternately, the circulating levels of the active agent itself can be measured directly.
  • the molecular level interactions of the delivery agent with the active agent is critical to effective delivery of the active agent.
  • the active agent is a protein
  • water forms a protective hydration layer around the protein when it is in solution. This layer interferes with the interactions of the active agent with the delivery agent. Lyophilization of the delivery agent/active agent solution may impact the hydration layer around the protein and allow for maximum interaction between the protein and the delivery agent, thus improving delivery. It is believed that lyophilization improves interactions for non-protein active agents as well by maximizing interactions with the delivery agent .
  • Example 1 Preparation of Delivery Agent 1 5-chlorosalicylamide (280 g, 1.6 mol) and acetonitrile (670 ml) were placed in a 5 liter, 4-neck, round bottomed, flask under a nitrogen atmosphere. To this stirred mixture, pyridine (161.3 g, 2.0 mol) was added over a period of 25 min. The reaction vessel was placed in an ice/water bath and portionwise addition of ethyl chloroformate was started. This addition continued over a period of one hour. When the addition was completed the ice/water bath was removed and the reaction mixture was allowed to come to room temperature. The reaction mixture was allowed to stir for an additional one hour at room temperature before the reaction vessel was reconfigured for distillation at atmospheric pressure.
  • reaction mixture was vacuum filtered and the filter cake was washed with ethyl alcohol (525 ml) .
  • Deionized water (525 ml) was slowly added to the stirred filtrate and a white solid precipitated.
  • An ice/water bath was placed around the reaction vessel and the slurry was cooled to 5 C. After stirring at this temperature for approximately 15 min the solids were recovered by vacuum filtration and the filter cake was washed first with ethanol (300 ml) and then with heptane (400 ml) .
  • the percent purity of Delivery Agent 1 was determined as follows. 0.9964 g of the free acid of Delivery Agent 1 was quantitatively dissolved in 40 ml reagent methanol . 2 ml of distilled water was added to this solution after the solids were dissolved. The solution was titrated in methanol with 0.33 N sodium hydroxide using a computer controlled burette (Hamilton automatic burette available from Hamilton (Reno, NV) ) . A glass electrode (computer controlled Orion model 525A pH meter available from VWR Scientific, S . Plainfield, NJ) was used to monitor the pH of the solution. The solution was stirred with a magnetic stirrer .
  • the volume of titrant to reach the second pH inflection point was 18.80 ml.
  • the inflection point determined by interpolation between the two data points where the second derivative of the pH plot changed from positive to negative, occurred at pH 11.3.
  • the purity of the free acid was determined using the following equation:
  • sCT/Delivery Agent 1 Na Salt Solution The aqueous solution of Delivery Agent 1 sodium salt was filtered through a sterile, 0.45 micron cellulose acetate, low protein binding membrane on a 150 ml Corning filter (available from VWR Scientific Product, S. Plainfield, NJ) . The pH of the solution was about 8.3. Dry sCT, stored at -70C, was brought to room temperature, 18.692 mg was weighed out and dissolved in 10 ml 0.1 M mono sodium phosphate buffer solution at pH 5, with gentle mixing. The sCT solution was added to the Delivery Agent 1 sodium salt solution with gentle mixing, taking precaution to avoid foaming or vortexing. 2c. Lyophilization of sCT/Delivery Agent 1 Na Salt Solution: Shelves of the lyophilizer (Genesis 25 LL-800 from The Virtis Company, Gardiner, NY) were prefrozen to -45C.
  • sCT/Delivery Agent 1 sodium salt solution was added to a 30cm x 18 cm stainless steel tray to give a cake thickness of about 0.48 cm.
  • Four clean, dry thermocouple probe tips were inserted into the solution such that the probe tip touched the solution level in the center. The probes were secured with clips to the side of the tray and the trays were loaded on to the precooled shelves .
  • the GPC2 was programmed for the cycle listed in Table 1.
  • the system cycle was terminated and the system vacuum was released.
  • the trays were carefully removed from the shelves and the lyophilized powder was transferred into amber HDPE NALGENE® bottles (from VWR Scientific) .
  • a powder with about 3% moisture content was obtained.
  • the powder was hand packed into hard gelatin capsules (size OEL/CS, from Capsugel, Division of Warner Lamber Co., Greenwood, SC) as needed.
  • the filled capsules and the lyophilized powder were stored in a closed container with dessicant.
  • Acetic anhydride 56.81 ml, 61.47 g, 0.6026 mol
  • 5- chlorosalicylic acid 100.00 g, 0.5794 mol
  • xylenes 200 ml
  • the flask was heated to reflux, the reaction mixture clearing to a yellow solution around 100°C.
  • Most of the volatile organics xylenes and acetic acid were distilled into the Dean-Stark trap (135-146°C) .
  • the sodium salt solution was prepared according to the method of Example 2a using 0.2 N NaOH solution. Percent purity was calculated to be 100% using 0.5038 g of delivery agent 1 and 16.06 ml 0.2 N NaOH. The sodium salt solution was prepared using 250 ml 0.2 N NaOH and 9.4585 g delivery agent 1 prepared as above. The solution was filtered through the 0.45 micron filter.
  • mice Male Sprague-Dawley rats weighing between 200-250g were fasted for 24 hours and were administered ketamine (44 mg/kg) and chlorpromazine (1.5 mg/kg) 15 minutes prior to dosing.
  • the rats were administered one of the following: (4a) orally, one capsule of 13 mg lyophilized powder (as prepared as in Example 2) with 0.5 ml water to flush the capsule down; (4b) orally, 1.0 ml/kg of a reconstituted aqueous solution of the lyophilized powder prepared in Example 2 ; (4c) orally, 1.0 ml/kg of "fresh", unlyophilized aqueous solution of Delivery Agent 1 sodium salt as prepared in Example 3 plus sCT; or (3d) subcutaneously, 5 ⁇ g/kg of sCT.
  • Doses (a) , (b) and (c) contained 50 mg/kg of Delivery agent 1 Na salt and 100 ⁇ g/kg of sCT. Doses for (a) are approximate because the animals were given one capsule filled with the stated amount of powder based on an average animal weight of 250 g, whereas actual animal weight varied. This is also the case in all later examples where a capsule is dosed.
  • the reconstituted solution for (b) was prepared by mixing 150 mg of the lyophilized powder prepared as in Example 2 in 3 ml of water, and was dosed at 1.0 ml/mg.
  • the "fresh" solution for (c) was prepared from unlyophilized material using 150 mg delivery agent 1 Na salt prepared in Example 3 in 3 ml water plus 150 ⁇ l sCT stock solution (2000 ⁇ l/ml prepared in 0. IM phospate buffer, pH adjusted to 4 with HCl and NaOH.
  • the "fresh” solution had a final concentration of 50 mg/ml delivery agent 1 Na salt and of 100 ⁇ g/ml sCT, and 1.0 ml/kg was dosed.
  • the subcutaneous doses were prepared by dissolving 2 mg sCT in 1 ml water. 5 ⁇ L of this solution was added to 995 ⁇ L water. This solution was dosed at 0.5 ml/kg. Blood samples were collected serially from the tail artery. Serum sCT was determined by testing with an EIA kit (Kit # EIAS-6003 from Peninsula Laboratories, Inc., San Carlos, CA) , modifying the standard protocol from the kit as follows: incubated with 50 ⁇ l peptide antibody for 2 hours with shaking in the dark, washed the plate, added serum and biotinylated peptide and diluted with 4 ml buffer, and shook overnight in the dark. Results are illustrated in Table 2, below.
  • rats were administered one of the following:
  • the reaction mixture was vacuum filtered and the filter cake was washed with ethyl alcohol.
  • the filter cake and filtrate were monitored by HPLC to determine where the product was. Most was washed into the filtrate, however, product was still present in the filter cake.
  • the filter cake was worked up to recover product to increase the final yield.
  • the filter cake was washed first with copious amounts of water, then with ethyl acetate.
  • the washes from the filter cake were separated and the ethyl acetate layer was next washed with 2x water, lx brine, then dried over sodium sulfate, isolated and concentrated in vacuo to recover more solids (solids B) . Water was added to the filtrate that had been isolated earlier and solids precipitated out.
  • Solids A and B were combined and transferred to a round bottom flask and 2N NaOH was added to the filtrate and heating was begun with stirring. The reaction was monitored by HPLC to determine when the reaction was done. The reaction was cooled to 25 C, stirred overnight, and concentrated in vacuo to remove excess ethanol . An ice/water bath was placed around the reaction vessel and the slurry was acidified. The solids were recovered by vacuum filtration and the filter cake was washed with water, dried and sent for NMR analysis.
  • the solids were isolated and transferred to an Erlenmeyer flask to be recrystallized.
  • the solids were recrystallized first with methanol/water . Solids formed and were washed into a Buchner funnel, more solids precipitated out in the filtrate and were also recovered. The first solids recovered after recrystalization had formed methyl ester. All the solids were combined, 2N NaOH was added and heated again to reflux to regain the free acid. Once the ester had disappeared, as determined by HPLC, acidification of the reaction to pH 4.7 caused solids to develop. The solids were isolated by filtration and combined with all the solids and recrystallized using a 1.5: 1.0 ratio of methanol to water.
  • Example 7 Lyophilization of sCT/Delivery Agent 2 Na salt
  • a lyophilized powder of sCT/Delivery Agent 2 sodium salt was prepared and packed into capsules.
  • 10.528 g delivery agent 2 as prepared in Example 6 was dissolved in 150 ml water. 4.72 ml ION NaOH was added. 21.0566 mg sCT was dissolved in 10 ml phosphate buffer, and this was added to the delivery agent solution. Water was added to make the volume up to 250 ml.
  • rats were administered orally one capsule of 13 mg lyophilized powder with 0.5 ml water to flush the capsule down with the approximate amounts of Delivery Agent 2 sodium salt and sCT as set forth in Table 4 below, where results are recorded.
  • ethylchloroformate can contain 0.1% phosgene and is extremely reactive with water. The reacton is highly exothermic and requires the use of a process chiller to moderate reaction temperature.
  • the reactor contents was agitated for 30 minutes at 10-14°C once the ethylchloroformate addition was complete. The reactor contents was heated to 85°C over 25 minutes, collecting all distillate into a receiver. The reactor contents was held at 85-94°C for approximately 6 hours, collecting all distilled material into a receiver. The reaction mixture was sampled and the conversion (>90%) monitored by HPLC. The conversion was found to be 99.9% after 6 hours. The reactor contents was cooled to 19°C over a one-hour period.
  • the contents was centrifuged and the filtrate sent into a second 200-gallon glass-lined reactor.
  • 119 L of ethanol (200 proof denatured with 0.5% toluene) was charged to the first 200-gallon reactor, warmed to 45°C and the filter cake washed with warm ethanol, adding to the reaction mixture in the second 200-gallon reactor.
  • the agitator was started on the second 200-gallon reactor.
  • the reactor contents was cooled to 29°C.
  • 120 L distilled water was slowly charged to the second reactor, with the water falling directly into the batch.
  • the reactor contents was cooled to 8°C.
  • the intermediate came out of solution and was held for 9.5 hours.
  • the resultant slurry was centrifuged.
  • the product precipitated out of solution The reaction mixture was adjusted to pH 2.0-4.0 with 50% NaOH solution (2L water, 2 kg NaOH) .
  • the reactor contents was cooled to 9-15°C.
  • the reactor slurry was centrifuged to isolate the intermediate.
  • 50 L purified water, USP was charged to a 200-gallon glass-lined reactor and this rinse was used to wash the centrifuge wet cake.
  • the wet cake was unloaded into double polyethylene bags placed inside a plastic drum.
  • the N- (5-chlorosalicyloyl) -8-aminocaprylic acid was dried under vacuum (27" Hg) at 68°C for 38 hours.
  • the dry cake was unloaded into double polyethylene bags placed inside a 55-gallon, steel unlined, open-head drums with a desiccant bag placed on top.
  • the dried isolated yield was 81 kg of N- (5-chlorosalicyloyl) -8-aminocaprylic acid.
  • Example 10 Lyophilization of sCT/Delivery Agent 1 Na Salt for Tableting
  • Example 2 The method of Example 2 was used to prepare lyophilized powder using delivery agent 1 from Example 9. 200 g of delivery agent 1 was used. The NaOH solution was made by dissolving 42 g of 100% NaOH into 2000 ml water. The slurry was stirred at room temperature, and vacuum filtered over the 0.45 micron filter. The delivery agent 1 Na salt solution pH was about 8.6. 200 mg sCT was used.
  • Tablets of the lyophilized powder prepared in Example 10 were prepared as follows.
  • Agent 1 Na Salt 100.2 30, 060.0
  • the Ac-Di-Sol® (croscarmellose sodium NF, PH.Eur., JPE, available from FMC Corporation, Pharmaceutical Division, Philadelphia, PA) and Cab-O-Sil® (fumed silica, available from Cabot Corporation, Tuscola, IL) were weighed and transferred to a mixing bottle. The bottle was then closed and secured to the arm of a sustained release apparatus set at 25 rotations per minute (RPM) . The apparatus was rotated for 5 minutes to mix. The lyophilized powder of Delivery Agent 1/sCT was then added to the AC-DI-SOL®/CAB-0-SIL® mixture geometrically with a two minute mixing cycle after each addition. Magnesium stearate was then added to the above mixture and then mixing was continued for five minutes.
  • RPM rotations per minute
  • Example 12 Oral Delivery of sCT/Delivery Agent 1 Na Salt in Rats - Tablets
  • Example 11 The tablets prepared in Example 11 were pulverized and hand packed into capsules at 13 mg/capsule. Untableted, lyophilized powder as prepared in Example 10 was hand packed into capsules at 13 mg/capsule. The capsules were dosed with 1 ml water to flush them down.
  • rats were administered orally one capsule with 1 ml water to flush the capsule down with the approximate amounts of Delivery Agent 1 Na salt and sCT as set forth in Table 6 below, where results are recorded.
  • Example 14 Lyophilization of sCT/Delivery Agent 1 Na salt
  • Delivery Agent 1 as prepared in Example 13 was formulated into a lyophilized powder with sCT as in Example 2 with 485 ml 0.2 N NaOH and 19.0072 g of Delivery Agent 1 in a steam bath. The final volume was 505 ml.
  • Four separate batches were prepared from: 187, 138, 74 and 160 ml delivery agent 2 Na salt solution plus 28, 48, 40 and 360 mg sCT, respectively.
  • the estimated amounts of delivery agent 2 Na salt were: 7, 5, 2.5 and 4.5 g, respectively.
  • rats were administered orally one capsule of 13 mg lyophilized powder using one of the four batches prepared in Example 14, with 1 ml water to flush the capsule down.
  • the approximate amounts of Delivery Agent 1 Na salt and sCT are set forth in Table 7 below, where results are recorded.

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Abstract

A solid dosage form useful in the administration of active agents is provided. The solid dosage form comprises a lyophilized powder obtained from a solution comprising a delivery agent and an active agent. The solid dosage forms of the present invention have utility in the delivery of active agents to selected biological systems and in an increased bioavailability of the active agent compared to other dosage forms. Methods of preparation and administration are also provided.

Description

LYOPHILIZED SOLID DOSAGE FORMS AND METHODS OF MAKING
Field of the Invention The present invention relates to a solid dosage form comprising a lyophilized powder obtained from a solution comprising a delivery agent and an active agent. The solid dosage form is suitable for oral administration of active agents to animals. Methods of preparation and administration are also disclosed.
Background of the Invention
Solid dosage forms for delivering certain active agents are well known in the art. A drug plus a diluent or excipient is milled to mix the powders to obtain a uniform mixture. The mixture is then granulated by compression or is made into a wet dough-like mixture, which is then milled to make small granules. The granules are dried in an oven or on a fluidized bed drive at temperatures of 40-80°C. The dried granules are run through a sieve to obtain particles of a uniform size. The granules are then put into capsules or compressed into tablets. The harsh conditions of this process are of concern with protein, polypeptide or peptide active agents, such as calcitonin, insulin, human growth hormone and parathyroid hormone. The process might denature the active agent, rendering it inactive and ineffective for its intended use. Therefore, protein, polypeptide or peptide drugs are not currently made in solid dosage forms .
Recently, delivery agents have been discovered that facilitate oral and other routes of delivery of certain active agents that were previously only deliverable via injection. See, e.g., US 5,629,020; US 5,643,957; US 5,650,386; US 5,714,167; US 5,773,647; US 5,863,944; and US 5, 866, 536.
It has been reported that lyophilization can be used to stabilize a solution of an active agent and a delivery agent a solid which can then be reconstituted and dosed as a solution. See "Effect of Lyophilization on the Stability and In Vivo Absorption of Growth Hormone when Using PODDS™ Technology", Chaudhary et al . , (abstract PDD 7403 in Pharmaceutical Research 12:9 page S-293 (September 1995) (poster presented Nov. 8, 1995 at AAPS Annual Meeting, Miami Beach, FL (November 4-9, 1995).
There is still a need to improve current delivery systems .
Summary of the Invention
A solid dosage form useful in the administration of active agents is provided. The solid dosage form comprises a lyophilized powder obtained from lyophilization of a solution comprising a delivery agent and an active agent. The solid dosage forms of the present invention have utility in the delivery of active agents to selected biological systems and in an increased bioavailability of the active agent compared to other dosage forms. A method of preparing the solid dosage form is provided, the method comprising:
-making a solution comprising a delivery agent and an active agent; -lyophilizing the solution to obtain a solid powder; and -incorporating the solid powder into a solid dosage form. A method of administering a biologically-active agent to an animal in need of said agent comprising administering to the animal the solid dosage form is also provided.
Detailed Description of the Invention
The solid dosage forms of the present invention include an active agent and a delivery agent. These solid dosage forms may be used to deliver various active agents through various biological, chemical, and physical barriers and are particularly suited for delivering active agents which are subject to environmental degradation.
Delivery Agents The delivery agent may be any compound that effects delivery of an active agent. The delivery may be acylated amino acids, sulfonated amino acids, ketones or aldehydes of acylated or sulfonated amino acids, or polypeptides which include any of the foregoing. Preferably, the delivery agent units comprises:
R-C02H wherein: R = Cι-C2 alkyl, C2-C2 alkenyl, C3-Cι0 cycloalkyl, C3- Cio cycloalkenyl, phenyl, naphthyl, (Cι-Cι0 alkyDphenyl, (C2-Ci0 alkenyl) phenyl, (C -Cι0 alkyl) naphthyl, (C2-Cι0 alkenyl) naphthyl , phenyl (Ci-Cio alkyl), phenyl (C2-Cι0 alkenyl), naphthyl (Cι-Cι0 alkyl) and naphthyl (C2-Cι0 alkenyl) ;
R may be optionally substituted with Cι-Cι0 alkyl, C2- Cio alkenyl, Cι-C4 alkoxy, -OH, -SH, -C02R' (where R' is H, Cι-C4 alkyl or C2-C4 alkenyl) , C3-Cι0 cycloalkyl, C3-Cι0 cycloalkenyl, heterocyclic having 3-10 ring atoms wherein the hetero atom is one or more atoms of N, 0, S or any combination thereof, aryl, (C1-C10 alkyl) aryl, aryl(Cι- C10) alkyl, or any combination thereof;
R may be optionally interrupted by 0, N, S or any combination thereof; or a salt thereof .
More preferably, the delivery agent comprises
Figure imgf000006_0001
wherein:
Ri is H, Cl, Br, FI, CH3, OH, OCH3, (CH2)mCH3 where m=l- 8;
R2 is H, OH;
R3 is COOH or salts thereof, NH2, OH, Cl , Br; Z is (CH2)n where n=l-ll, and may be substituted, un- substituted, linear or branched; or
Figure imgf000007_0001
where o=l-5, and the ring and/or chain may be substituted or unsubstituted and the chain may be linear or branched.
In one embodiment, the delivery agent comprises
Figure imgf000007_0002
wherein n = 1-15, or salts thereof including but not limited to sodium salts. More preferably, the delivery agent comprises :
Figure imgf000007_0003
Delivery Agent 1
Figure imgf000007_0004
Delivery Agent 2
or salts thereof, including but not limited to sodium salts .
In another embodiment, the delivery agent comprises
Figure imgf000008_0001
Delivery Agent 3
or salts thereof, including but not limited to sodium salts.
The delivery agents may be in the form of the carboxylic acid and/or their salts, including but not limited to sodium salts. In addition, poly amino acids and peptides comprising one or more of these compound may be used.
An amino acid is any carboxylic acid having at least one free amine group and includes naturally occurring and synthetic amino acids. Poly amino acids are either peptides (which are two or more amino acids joined by a peptide bond) or are two or more amino acids linked by a bond formed by other groups which can be linked by, e.g., an ester or an anhydride linkage. Peptides can vary in length from dipeptides with two amino acids to polypeptides with several hundred amino acids. See Chambers Biological Dictionary, editor Peter M.B. Walker, Cambridge, England: Chambers Cambridge, 1989, page 215. One or more of the amino acids or peptide units may be acylated or sulfonated. These amino acid derivatives may be derived from amino acids and can be readily prepared from amino acids by methods within the skill of those in the art based upon the present disclosure and the methods described in WO96/30036, WO97/36480, US 5,643,957 and US 5,650,386.
For example, the compounds may be prepared by reacting the single amino acid with the appropriate acylating or amine-modifying agent, which reacts with a free amino moiety present in the amino acid to form amides . Protecting groups may be used to avoid unwanted side reactions as would be known to those skilled in the art . With regard to protecting groups, reference is made to T.W. Greene, Protecting Groups in Organic Synthesis, Wiley, New York (1981) , the disclosure of which is hereby incorporated by reference.
The compound may be purified by recrystallization or by fractionation on one or more solid chromatographic supports, alone or linked in tandem. Suitable recrystallization solvent systems include, but are not limited to, acetonitrile, methanol, and tetrahydrofuran. Fractionation may be performed on a suitable chromatographic support such as alumina, using methanol/n- propanol mixtures as the mobile phase; reverse phase chromatography using trifluoroacetic acid/acetonitrile mixtures as the mobile phase; and ion exchange chromatography using water or an appropriate buffer as the mobile phase. When anion exchange chromatography is performed, preferably a 0-500 mM sodium chloride gradient is employed.
Active Agents Active agents suitable for use in the present invention include biologically active agents and chemically active agents, including, but not limited to, pesticides, pharmacological agents, and therapeutic agents. For example, biologically or chemically active agents suitable for use in the present invention include, but are not limited to, proteins, polypeptides, peptides, and particularly small peptides; hormones, and particularly hormones which by themselves do not pass (or which pass only a fraction of the administered dose) through the gastro-intestinal mucosa and/or are susceptible to chemical cleavage by acids and enzymes in the gastro-intestinal tract; polysaccharides, and particularly mixtures of muco- polysaccharides; carbohydrates; lipids; or any combination thereof. Further examples include, but are not limited to, the following, including synthetic, natural or recombinant sources thereof: growth hormones, including human growth hormones (hGH) , recombinant human growth hormones (rhGH) , bovine growth hormones, and porcine growth hormones; growth hormone-releasing hormones; interferons, including , β and γ; interleukin-l; insulin; insulin-like growth factor, including IGF-1; heparin, including unfractionated heparin, heparinoids, dermatans, chondroitins, low molecular weight heparin, very low molecular weight heparin and ultra low molecular weight heparin; calcitonin, including salmon, eel and human; erythropoietin; atrial naturetic factor; antigens; monoclonal antibodies; somatostatin; protease inhibitors; adrenocorticotropin, gonadotropin releasing hormone; oxytocin; leutinizing-hormone-releasing-hormone; follicle stimulating hormone; glucocerebrosidase; thrombopoietin; filgrastim; prostaglandins; cyclosporin; vasopressin; cromolyn sodium (sodium or disodium chromoglycate) ; vancomycin; desferrioxamine (DFO) ; parathyroid hormone (PTH) , including its fragments; antimicrobials, including anti-fungal agents; analogs, fragments, mimetics or polyethylene glycol (PEG) -modified derivatives of these compounds; or any combination thereof. In one embodiment, the active agent comprises a protein, polypeptide, or peptide; analogs, fragments, mimetics or polyethylene glycol (PEG) -modified derivatives thereof; or any combination thereof.
Delivery systems The solid dosage forms of the present invention comprise a delivery agent and one or more active agents and may optionally contain additives such as phosphate buffer salts, citric acid, glycols, or other dispersing agents. Stabilizing additives may be incorporated into the solution, preferably at a concentration ranging between about 0.1 and 20% (w/v) .
The solid dosage unit forms may also include one or more enzyme inhibitors. Such enzyme inhibitors include, but are not limited to, compounds such as actinonin or epiactinonin and derivatives thereof. Derivatives of these compounds are disclosed in US 5,206,384. Other enzyme inhibitors include, but are not limited to, aprotinin (Trasylol) and Bowman-Birk inhibitor.
The amount of active agent is an amount effective to accomplish the purpose of the particular active agent for the target indication. The amount of active agent in the compositions typically is a pharmacologically, biologically, therapeutically, or chemically effective amount. However, the amount can be less than that amount when the composition is used in a dosage unit form that contains a plurality of delivery agent/active agent compositions or a divided pharmacologically, biologically, therapeutically, or chemically effective amount. The total effective amount can then be administered in cumulative units containing, in total, pharmacologically, biologically, therapeutically or chemically active amounts of biologically or pharmacologically active agent. The total amount of active agent, and particularly biologically or chemically active agent, to be used can be determined by those skilled in the art. However, because the compositions may deliver active agents more efficiently than prior compositions, lower amounts of biologically or chemically active agents than those used in prior dosage unit forms or delivery systems can be administered to the subject, while still achieving the same blood levels and therapeutic effects.
Lyophilization
Lyophilization techniques are well known in the art. The delivery agent and active agent can be made up into solution and lyophilized to form a powder.
Solid Dosage Form
A solid dosage form can be a capsule, tablet or powder. The powder may be in the form of a sachet that is mixed with a liquid and administered. The solid dosage form may also be a topical delivery system, such as an ointment, cream or semi-solid. The solid dosage form contemplated may include a sustained release or controlled release system.
The lyophilized powder may be packed into capsules, or pressed into tablets, used in powder form, or incorporated into an ointment, cream or semi-solid. Methods for forming solid dosage forms are well known in the art.
The presently disclosed solid dosage forms are suitable for delivering biologically and chemically active agents, particularly in oral, intranasal, sublingual, intraduodenal, buccal, rectal, vaginal, mucosal, pulmonary, transdermal, and intramuscular systems. The amount of lyophilized material in the present solid dosage forms is a delivery effective amount and can be determined for any particular compound or biologically or chemically active agent by methods known to those skilled in the art.
Dosage unit forms can also include any of excipients, diluents, disintegrants, lubricants, plasticizers, colorants, flavorants, taste-masking agents, sugars, sweeteners, salts, and dosing vehicles, including, but not limited to, water, 1,2 -propane diol, ethanol, olive oil, or any combination thereof.
The dosage unit forms of the present invention are useful for administering biologically or chemically active agents to animals, including but not limited to poultry, such as chickens; mammals, such as cows, pigs, dogs, cats, primates, and particularly humans; and insects. The system is particularly advantageous for delivering chemically or biologically active agents which would otherwise be destroyed or rendered less effective by conditions encountered before the active agent reaches its target zone (i.e. the area in which the active agent of the delivery composition is to be released) and within the body of the animal to which they are administered. Particularly, the solid dosage forms of the present invention are useful in orally administering active agents, especially those which are not ordinarily orally deliverable.
Following administration, the active agent present in the dosage unit form is taken up into the circulation. The bioavailability of the agent is readily assessed by measuring a known pharmacological activity in blood, e.g. an increase in blood clotting time caused by heparin, or a decrease in circulating calcium levels caused by calcitonin. Alternately, the circulating levels of the active agent itself can be measured directly.
While not wishing to be bound by theory, it is believed that the molecular level interactions of the delivery agent with the active agent is critical to effective delivery of the active agent. For example, where the active agent is a protein, water forms a protective hydration layer around the protein when it is in solution. This layer interferes with the interactions of the active agent with the delivery agent. Lyophilization of the delivery agent/active agent solution may impact the hydration layer around the protein and allow for maximum interaction between the protein and the delivery agent, thus improving delivery. It is believed that lyophilization improves interactions for non-protein active agents as well by maximizing interactions with the delivery agent .
Examples
The following examples illustrate the invention without limitation.
Example 1: Preparation of Delivery Agent 1 5-chlorosalicylamide (280 g, 1.6 mol) and acetonitrile (670 ml) were placed in a 5 liter, 4-neck, round bottomed, flask under a nitrogen atmosphere. To this stirred mixture, pyridine (161.3 g, 2.0 mol) was added over a period of 25 min. The reaction vessel was placed in an ice/water bath and portionwise addition of ethyl chloroformate was started. This addition continued over a period of one hour. When the addition was completed the ice/water bath was removed and the reaction mixture was allowed to come to room temperature. The reaction mixture was allowed to stir for an additional one hour at room temperature before the reaction vessel was reconfigured for distillation at atmospheric pressure. The distillation that followed yielded 257.2 g of distillate at a head temperature of 78 C. 500 ml of deionized water was added to the reaction mixture that remained in the flask and the resulting slurry was vacuum filtered. The filter cake was washed with 200 ml deionized water and was allowed to dry overnight in vacuo at room temperature. 313.6 g (97.3%) of 6-chloro carsalam was isolated after drying. An additional batch was made using this same method and yielded 44.5 g 6- chloro-2H-l, 3-benzoxazine-2 , 4 (3H) -dione . Sodium carbonate (I94.0g, 1.8 mol) was added to 5 liter, 4-neck, round bottomed, flask containing 6-chloro- 2H-1, 3-benzoxazine-2 ,4 (3H) -dione (323. lg, 1.6 mol) and dimethylacetamide (970 ml). Ethyl-8-bromooctanoate (459.0 g, 1.8 mol) was added in one portion to the stirring reaction mixture. The atmospheric pressure in the reaction vessel was reduced (550 mm Hg) and heating of the reaction mixture was started. The reaction temperature was maintained at 70 C for approximately 5 hours before heating and vacuum were discontinued and the reaction mixture was allowed to cool to room temperature overnight . The reaction mixture was vacuum filtered and the filter cake was washed with ethyl alcohol (525 ml) . Deionized water (525 ml) was slowly added to the stirred filtrate and a white solid precipitated. An ice/water bath was placed around the reaction vessel and the slurry was cooled to 5 C. After stirring at this temperature for approximately 15 min the solids were recovered by vacuum filtration and the filter cake was washed first with ethanol (300 ml) and then with heptane (400 ml) . After drying overnight at room temperature in vacuo, 598.4 g (99.5%) of ethyl 8- (6-chloro- 2H-1, 3 -Benzoxazine-2, 4 (3H) -dionyl) octanoate was obtained. An additional 66.6 g of ethyl 8- (6-chloro-2H-l, 3-
Benzoxazine-2, 4 (3H) -dionyl) octanoate was made by this same method.
Ethyl 8- (6-chloro-2H-l, 3 -benzoxazine-2 , 4 (3H) - dionyl) octanoate (641 g, 1.7 mol) and ethyl alcohol (3200 ml) were added to a 22 liter, five neck flask. In a separate 5 liter flask NaOH (288.5 g, 7.2 mol) was dissolved in deionized water (3850 ml) . This mixture was added to the reaction mixture contained in the 22 liter flask. A temperature increase to 40 C was noted. Heating of the reaction mixture was started and when the reaction temperature had increased to 50 C it was noted that all of the solids in the reaction mixture had dissolved. A temperature of 50 C was maintained in the reaction mixture for a period of 1.5 hr. The reaction flask was then set up for vacuum distillation. 2200 mis of distillate were collected at a vapor temperature of 55 C (10 mm Hg) before the distillation was discontinued. The reaction flask was then placed in an ice/water bath and concentrated HC1 (752 ml) was added over a period of 45 min. During this addition the reaction mixture was noted to have thickened somewhat and an additional 4 liters of deionized water was added to aid the stirring of the reaction mixture. The reaction mixture was then vacuum filtered and the filter cake washed with 3 liters of deionized water. After drying in vacuo at room temperature 456.7 g (83.5%) of N- (5- chlorosalicyloyl) -8-aminocaprylic acid was isolated. Example 2 : Lyophilization of Salmon Calcitonin
(sCT) /Delivery Agent 1 Sodium (Na) Salt
2a. Preparation of Delivery Agent 1 Sodium Salt Solution:
The percent purity of Delivery Agent 1 was determined as follows. 0.9964 g of the free acid of Delivery Agent 1 was quantitatively dissolved in 40 ml reagent methanol . 2 ml of distilled water was added to this solution after the solids were dissolved. The solution was titrated in methanol with 0.33 N sodium hydroxide using a computer controlled burette (Hamilton automatic burette available from Hamilton (Reno, NV) ) . A glass electrode (computer controlled Orion model 525A pH meter available from VWR Scientific, S . Plainfield, NJ) was used to monitor the pH of the solution. The solution was stirred with a magnetic stirrer .
The volume of titrant to reach the second pH inflection point was 18.80 ml. The inflection point, determined by interpolation between the two data points where the second derivative of the pH plot changed from positive to negative, occurred at pH 11.3. The purity of the free acid was determined using the following equation:
% purity = 100 x ml x Normality x MolWt 1000 x Equivalents x SampleWt wherein ml = milliliters of titrant Normality = Normality of Sodium hydroxide Mol Wt = molecular weight of the Delivery Agent 1 free acid (313.78) Equivalents = 2 (the free acid is dibasic) SampleWt = weight of the free acid sample being titrated
Purity was found to be 97.0%
9.3458 g delivery Agent 1 powder was weighed out. The amount of 0.33 N sodium hydroxide needed to have a sodium hydroxide to free acid molar ratio of 1.6 was calculated using the following equation: ml NaOH = freeAcidWt x %purity x 1000 x 1.6 313.78 x 100 x Normality wherein
FreeAcidWt = weight of free acid in formulated sample %purity = % purity of Delivery Agent 1 Normality = Normality of Sodium hydroxide ml NaOH = amount of NaOH needed
Delivery Agent 1 and 153.3 ml 0.33 N sodium hydroxide was mixed in a Pyrex bottle. The resulting slurry was warmed in a steam bath to 60-80C. The warm slurry became a clear solution in about 15 minutes with occasional stirring. The solution was cooled to room temperature. The pH of this solution was 8.1.
2b. Preparation of sCT/Delivery Agent 1 Na Salt Solution: The aqueous solution of Delivery Agent 1 sodium salt was filtered through a sterile, 0.45 micron cellulose acetate, low protein binding membrane on a 150 ml Corning filter (available from VWR Scientific Product, S. Plainfield, NJ) . The pH of the solution was about 8.3. Dry sCT, stored at -70C, was brought to room temperature, 18.692 mg was weighed out and dissolved in 10 ml 0.1 M mono sodium phosphate buffer solution at pH 5, with gentle mixing. The sCT solution was added to the Delivery Agent 1 sodium salt solution with gentle mixing, taking precaution to avoid foaming or vortexing. 2c. Lyophilization of sCT/Delivery Agent 1 Na Salt Solution: Shelves of the lyophilizer (Genesis 25 LL-800 from The Virtis Company, Gardiner, NY) were prefrozen to -45C.
Approximately 260 ml sCT/Delivery Agent 1 sodium salt solution was added to a 30cm x 18 cm stainless steel tray to give a cake thickness of about 0.48 cm. Four clean, dry thermocouple probe tips were inserted into the solution such that the probe tip touched the solution level in the center. The probes were secured with clips to the side of the tray and the trays were loaded on to the precooled shelves .
The GPC2 was programmed for the cycle listed in Table 1.
Table 1: Lyophilization Process Cycle
Figure imgf000019_0001
When the lyophilization cycle was completed, the system cycle was terminated and the system vacuum was released. The trays were carefully removed from the shelves and the lyophilized powder was transferred into amber HDPE NALGENE® bottles (from VWR Scientific) . Using the above cycle for lyophilization, a powder with about 3% moisture content was obtained. The powder was hand packed into hard gelatin capsules (size OEL/CS, from Capsugel, Division of Warner Lamber Co., Greenwood, SC) as needed. The filled capsules and the lyophilized powder were stored in a closed container with dessicant.
Example 3 : Preparation of sCT/Delivery Agent 1 Na Salt "Fresh" Solution (unlyophilized)
Acetic anhydride (56.81 ml, 61.47 g, 0.6026 mol), 5- chlorosalicylic acid (100.00 g, 0.5794 mol), and xylenes (200 ml) were added to a 500 ml, three-neck flask fitted with a magnetic stir bar, a thermometer, and a Dean-Stark trap with condenser. The flask was heated to reflux, the reaction mixture clearing to a yellow solution around 100°C. Most of the volatile organics (xylenes and acetic acid) were distilled into the Dean-Stark trap (135-146°C) . Distillation was continued for another hour, during which the pot temperature slowly rose to 190°C and the distillate slowed to a trickle to drive over any more solvent . Approximately 250 ml solvent was collected. The residue was cooled below 100 °C and dioxane was added. The reaction "set up" since it had cooled, however, in the next step heating will be applied again to make it more fluid.
A 2N sodium hydroxide (222.85 ml, 0.4457 mol) and 8- aminocaprylic acid (70.96 g, 0.4457 mol) solution was added to the solution of oligo (5-chloroasalicylic acid) (0.5794 mol) in dioxane. The reaction mixture was heated to 90°C for 5.5 hours, then shut off overnight and then restarted in the morning to heat to reflux (after restarting the heating the reaction was monitored at which time the reaction was determined to have finished, by HPLC) . The reaction mixture was cooled to 40°C. The dioxane was stripped off in vacuo. The residue was taken up in 2N sodium hydroxide and acidified. The material did not solidify. And was taken up in ethyl acetate and extracted (2x 100ml) to remove excess dioxane. The ethyl acetate layer was dried over sodium sulfate and concentrated in vacuo. The easily filtered solids were collected by filtration. The remaining material was taken up in 2N NaOH. The pH was adjusted to 4.3 to selectively isolate product from starting material. Once at pH 4.3 the solids were filtered off and then recrystallized in ethanol/water 1:1 any insoluble material was hot filtered out first. All the solids which were collected were combined and recrystallized from ethanol/water to give 52.06 g of the free acid product as a white solid.
The sodium salt solution was prepared according to the method of Example 2a using 0.2 N NaOH solution. Percent purity was calculated to be 100% using 0.5038 g of delivery agent 1 and 16.06 ml 0.2 N NaOH. The sodium salt solution was prepared using 250 ml 0.2 N NaOH and 9.4585 g delivery agent 1 prepared as above. The solution was filtered through the 0.45 micron filter.
Example : Oral Delivery of sCT/Delivery Agent 1 Na Salt in Rats:
Male Sprague-Dawley rats weighing between 200-250g were fasted for 24 hours and were administered ketamine (44 mg/kg) and chlorpromazine (1.5 mg/kg) 15 minutes prior to dosing. The rats were administered one of the following: (4a) orally, one capsule of 13 mg lyophilized powder (as prepared as in Example 2) with 0.5 ml water to flush the capsule down; (4b) orally, 1.0 ml/kg of a reconstituted aqueous solution of the lyophilized powder prepared in Example 2 ; (4c) orally, 1.0 ml/kg of "fresh", unlyophilized aqueous solution of Delivery Agent 1 sodium salt as prepared in Example 3 plus sCT; or (3d) subcutaneously, 5 μg/kg of sCT.
Doses (a) , (b) and (c) contained 50 mg/kg of Delivery agent 1 Na salt and 100 μg/kg of sCT. Doses for (a) are approximate because the animals were given one capsule filled with the stated amount of powder based on an average animal weight of 250 g, whereas actual animal weight varied. This is also the case in all later examples where a capsule is dosed.
The reconstituted solution for (b) was prepared by mixing 150 mg of the lyophilized powder prepared as in Example 2 in 3 ml of water, and was dosed at 1.0 ml/mg. The "fresh" solution for (c) was prepared from unlyophilized material using 150 mg delivery agent 1 Na salt prepared in Example 3 in 3 ml water plus 150 μl sCT stock solution (2000 μl/ml prepared in 0. IM phospate buffer, pH adjusted to 4 with HCl and NaOH. The "fresh" solution had a final concentration of 50 mg/ml delivery agent 1 Na salt and of 100 μg/ml sCT, and 1.0 ml/kg was dosed.
The subcutaneous doses were prepared by dissolving 2 mg sCT in 1 ml water. 5 μL of this solution was added to 995 μL water. This solution was dosed at 0.5 ml/kg. Blood samples were collected serially from the tail artery. Serum sCT was determined by testing with an EIA kit (Kit # EIAS-6003 from Peninsula Laboratories, Inc., San Carlos, CA) , modifying the standard protocol from the kit as follows: incubated with 50 μl peptide antibody for 2 hours with shaking in the dark, washed the plate, added serum and biotinylated peptide and diluted with 4 ml buffer, and shook overnight in the dark. Results are illustrated in Table 2, below.
Figure imgf000023_0001
approximate dose due to variations in animal weight
Example 5: Oral Delivery of sCT/Delivery Agent 1 Na Salt in Rats
According to the method of Example 4, rats were administered one of the following:
(5a) orally, one capsule of 13 mg lyophilized powder with 1 ml water to flush the capsule down;
(5b) orally, one capsule of 6.5 mg lyophilized powder with
1 ml water to flush the capsule down; (5c) orally, one capsule of 3.25 mg lyophilized powder with
1 ml water to flush the capsule down; (5d) subcutaneously 5 μg/kg of sCT.
Approximate amounts of delivery agent and sCT, as well s the results, are provided in Table 3, below.
Figure imgf000024_0001
* approximate dose due to variations in animal weight
Example 6 : Preparation of Delivery Agent 2
Sodium carbonate (30g, 0.2835 mol) was added to 500 ml 3-neck, round bottomed, flask containing 6-chloro-2H-l, 3- benzoxazine-2 , 4 (3H) -dione (prepared as in Example 1, paragraph 1) (50g, 0.2532 mol) and dimethylacetamide (75 ml). Methyl-4-bromobutyrate (45.83 g, 0.2532 mol) was added in one portion to the stirring reaction mixture, and heating of the reaction mixture was started. The reaction temperature was maintained at 70 C and allowed to heat overnight. Heating was discontinued, and the reaction mixture was allowed to cool to room temperature.
The reaction mixture was vacuum filtered and the filter cake was washed with ethyl alcohol. The filter cake and filtrate were monitored by HPLC to determine where the product was. Most was washed into the filtrate, however, product was still present in the filter cake. The filter cake was worked up to recover product to increase the final yield. The filter cake was washed first with copious amounts of water, then with ethyl acetate. The washes from the filter cake were separated and the ethyl acetate layer was next washed with 2x water, lx brine, then dried over sodium sulfate, isolated and concentrated in vacuo to recover more solids (solids B) . Water was added to the filtrate that had been isolated earlier and solids precipitated out. Those solids were isolated (solids A). Solids A and B were combined and transferred to a round bottom flask and 2N NaOH was added to the filtrate and heating was begun with stirring. The reaction was monitored by HPLC to determine when the reaction was done. The reaction was cooled to 25 C, stirred overnight, and concentrated in vacuo to remove excess ethanol . An ice/water bath was placed around the reaction vessel and the slurry was acidified. The solids were recovered by vacuum filtration and the filter cake was washed with water, dried and sent for NMR analysis.
The solids were isolated and transferred to an Erlenmeyer flask to be recrystallized. The solids were recrystallized first with methanol/water . Solids formed and were washed into a Buchner funnel, more solids precipitated out in the filtrate and were also recovered. The first solids recovered after recrystalization had formed methyl ester. All the solids were combined, 2N NaOH was added and heated again to reflux to regain the free acid. Once the ester had disappeared, as determined by HPLC, acidification of the reaction to pH 4.7 caused solids to develop. The solids were isolated by filtration and combined with all the solids and recrystallized using a 1.5: 1.0 ratio of methanol to water. White solids precipitated out overnight and were isolated and dried to give 23.48g of N- (5-chlorosalicyloyl) -4 aminobutyric acid at a 36% yield. It was later determined that the filter cake should have first been washed with excess ethyl alcohol to avoid having the product remain in the filter cake. From that point, the filtrate and 2N NaOH could be heated with stirring, cooled to 25C and concentrated in vacuo to remove excess ethanol; in an ice/water bath, the slurry acidified to pH 4.7; the solids recovered by vacuum filtration and the filter cake washed with water; solids isolated and recrystallized.
Example 7 : Lyophilization of sCT/Delivery Agent 2 Na salt
Following the procedure in Example 2, a lyophilized powder of sCT/Delivery Agent 2 sodium salt was prepared and packed into capsules. 10.528 g delivery agent 2 as prepared in Example 6 was dissolved in 150 ml water. 4.72 ml ION NaOH was added. 21.0566 mg sCT was dissolved in 10 ml phosphate buffer, and this was added to the delivery agent solution. Water was added to make the volume up to 250 ml.
Example 8: Oral Delivery of sCT/Delivery Agent 2 Na Salt in Rats
According to the method of Example 4, with the exception that the standard protocol for the EIA kit was followed, rats were administered orally one capsule of 13 mg lyophilized powder with 0.5 ml water to flush the capsule down with the approximate amounts of Delivery Agent 2 sodium salt and sCT as set forth in Table 4 below, where results are recorded.
Figure imgf000027_0001
* approximate dose due to variations in animal weight
Example 9 : Preparation of Delivery Agent 1 for Tableting
To a clean, dry, 200 gallon glass-lined reactor 178 L of dry acetonitrile was added. The agitator was set to 100-125 RPM and the reactor contents was cooled to 9°C. 74 kg of 5-chloro salicylamide (available from Polycarbon Industries, Leominster, MA) was charged to the reactor and the charging port was closed. 47 L of dry pyridine was charged to the reactor. The slurry was cooled to 9°C prior to proceeding. Cooling was applied to the reactor condenser and valve overheads were set for total reflux. Over 2 hours, 49.7 kg of ethylchloroformate was charged to the 200 gallon reactor while maintaining the batch temperature at 14°C. (Note that ethylchloroformate can contain 0.1% phosgene and is extremely reactive with water. The reacton is highly exothermic and requires the use of a process chiller to moderate reaction temperature.) The reactor contents was agitated for 30 minutes at 10-14°C once the ethylchloroformate addition was complete. The reactor contents was heated to 85°C over 25 minutes, collecting all distillate into a receiver. The reactor contents was held at 85-94°C for approximately 6 hours, collecting all distilled material into a receiver. The reaction mixture was sampled and the conversion (>90%) monitored by HPLC. The conversion was found to be 99.9% after 6 hours. The reactor contents was cooled to 19°C over a one-hour period. 134 L of deionized water was charged to the reactor. A precipitate formed immediately. The reactor contents was cooled to 5°C and agitated for 10.5 hours. The product continued to crystallize out of solution. The reactor slurry was centrifuged. 55 L of deionized water was charged to the 200-gallon, glass-lined reactor and the centrifuge wet cake was washed. The intermediate was dried under full vacuum (28" Hg) and 58°C for 19.5 hours. The yield was 82.6 kg 6-chloro-2H-l, 3 -benzoxazine-2 , 4 (3H) - dione . This intermediate was packaged and stored so that it was not exposed to water.
In the next preparation, absolutely no water can be tolerated in the steps up to the point where distilled water is added. 222 L of dry dimethylacetamide was charged to a dry 200 gallon glass-lined reactor. The reactor agitator was set to 100-125 RPM. Cooling was applied to the condenser and valve reactor overheads were set for distillation. 41.6 kg of dry anhydrous sodium carbonate was charged to the reactor and the reactor charging port was closed. Caution was used due to some off-gassing and a slight exotherm. 77.5 kg of dry 6-chloro-2H-l, 3-benzoxazine- 2 , 4 (3H) -dione was charged to the reactor. Quickly, 88 kg of dry ethyl-8-bromooctanoate was charged to the reactor. 22-24 inches of vacuum was applied and the reactor temperature was raised to 65-75°C. The reactor temperature was maintained and the contents was watched for foaming. The reactor mixture was sampled and monitored for conversion by monitoring for the disappearance of the bromo ester in the reaction mixture by GC . The reaction was complete (0.6% bromo ester was found) after 7 hours. The vacuum was broken and the reactor contents cooled to 45-
50°C. The contents was centrifuged and the filtrate sent into a second 200-gallon glass-lined reactor. 119 L of ethanol (200 proof denatured with 0.5% toluene) was charged to the first 200-gallon reactor, warmed to 45°C and the filter cake washed with warm ethanol, adding to the reaction mixture in the second 200-gallon reactor. The agitator was started on the second 200-gallon reactor. The reactor contents was cooled to 29°C. 120 L distilled water was slowly charged to the second reactor, with the water falling directly into the batch. The reactor contents was cooled to 8°C. The intermediate came out of solution and was held for 9.5 hours. The resultant slurry was centrifuged. 70 L ethanol was charged to the reactor, cooled to 8°C and the centrifuge cake was washed. The wet cake was unloaded into double polyethylene bags placed inside a paper lined drum. The yield was 123.5 kg ethyl 8- (6-chloro-2H-l, 3 -benzoxazine-2 , 4 (3H) -dionyl) octanoate .
400 1 purified water, USP and 45.4 kg NaOH pellets was charged to a 200 gallon glass-lined reactor and the agitator was set to 100-125 RPM. 123.5 kg of the ethyl 8- (6-chloro-2H-l, 3 -benzoxazine-2 , 4 (3H) -dionyl) octanoate wet cake was charged to the reactor. The charging port was closed. Cooling water applied to the condenser and the valve reactor overheads were set for atmospheric distillation. The reactor contents was heated to 98°C and conversion monitored by HLPC . Initially (approximately 40 minutes) the reactor refluxed at 68°C, however, as the ethanol was removed (over 3 hours) by distillation the reactor temperature rose to 98°C. The starting material disappeared, as determined by HPLC, at approximately 4 hours. The reactor contents was cooled to 27°C. 150 L purified water, USP was charged to an adjacent 200 gallon glass-lined reactor and the agitator was set to 100-125 RPM. 104 L concentrated (12M) hydrochloric acid was charged to the reactor and cooled to 24°C. The saponified reaction mixture was slowly (over 5 hours) charged to the 200-gallon glass-lined reactor. The material (45 L and 45 L) was split into 2 reactors (200 gallons each) because of carbon dioxide evolution. The product precipitated out of solution. The reaction mixture was adjusted to pH 2.0-4.0 with 50% NaOH solution (2L water, 2 kg NaOH) . The reactor contents was cooled to 9-15°C. The intermediate crystallized out of solution over approximately 9 hours. The reactor slurry was centrifuged to isolate the intermediate. 50 L purified water, USP was charged to a 200-gallon glass-lined reactor and this rinse was used to wash the centrifuge wet cake. The wet cake was unloaded into double polyethylene bags placed inside a plastic drum. The N- (5-chlorosalicyloyl) -8-aminocaprylic acid was dried under vacuum (27" Hg) at 68°C for 38 hours. The dry cake was unloaded into double polyethylene bags placed inside a 55-gallon, steel unlined, open-head drums with a desiccant bag placed on top. The dried isolated yield was 81 kg of N- (5-chlorosalicyloyl) -8-aminocaprylic acid.
Example 10 : Lyophilization of sCT/Delivery Agent 1 Na Salt for Tableting
The method of Example 2 was used to prepare lyophilized powder using delivery agent 1 from Example 9. 200 g of delivery agent 1 was used. The NaOH solution was made by dissolving 42 g of 100% NaOH into 2000 ml water. The slurry was stirred at room temperature, and vacuum filtered over the 0.45 micron filter. The delivery agent 1 Na salt solution pH was about 8.6. 200 mg sCT was used.
Example 11: Preparation of sCT/Delivery Agent 1 Na Salt Tablets
Tablets of the lyophilized powder prepared in Example 10 were prepared as follows.
An instrumented Carver press (Model C, available from Carver, Wabash, Indiana) was used for tablet compression. The die used was 0.245" in diameter. The top punch was flat- faced, bevel-edged and 0.245" in diameter while the bottom punch was flat-faced, scored, bevel-edged and 0.245" in diameter. The press was capable of measuring the upper and lower punch force as well as the displacement of the upper punch. A formula for direct compression was designed as shown in Table 5 below: Table 5 :
Material mg/tablet mg/300 tablet batch
Lyophilized powder of SCT/Delivery
Agent 1 Na Salt 100.2 30, 060.0
AC-DI-SOL® 2.004 601.2
Magnesium Stearate 0.511 153.3
CAB-O-SIL® 0.205 61.5
Total weight (mg) 102.92 30,876.0
The Ac-Di-Sol® (croscarmellose sodium NF, PH.Eur., JPE, available from FMC Corporation, Pharmaceutical Division, Philadelphia, PA) and Cab-O-Sil® (fumed silica, available from Cabot Corporation, Tuscola, IL) were weighed and transferred to a mixing bottle. The bottle was then closed and secured to the arm of a sustained release apparatus set at 25 rotations per minute (RPM) . The apparatus was rotated for 5 minutes to mix. The lyophilized powder of Delivery Agent 1/sCT was then added to the AC-DI-SOL®/CAB-0-SIL® mixture geometrically with a two minute mixing cycle after each addition. Magnesium stearate was then added to the above mixture and then mixing was continued for five minutes.
Approximately 103 mg of the above powder was then transferred to the die containing the lower punch. The powder was pressed down into the die using the upper punch. The upper punch was inserted and the punch die assembly was mounted onto the press . The compression was then performed. The upper punch was used to push the tablet out of the die . Example 12 : Oral Delivery of sCT/Delivery Agent 1 Na Salt in Rats - Tablets
The tablets prepared in Example 11 were pulverized and hand packed into capsules at 13 mg/capsule. Untableted, lyophilized powder as prepared in Example 10 was hand packed into capsules at 13 mg/capsule. The capsules were dosed with 1 ml water to flush them down.
Following the procedure of Example 4, with the exception that the standard protocol for the EIA kit was followed instead of the modified version, rats were administered orally one capsule with 1 ml water to flush the capsule down with the approximate amounts of Delivery Agent 1 Na salt and sCT as set forth in Table 6 below, where results are recorded.
Figure imgf000033_0001
* approximate dose due to variations in animal weight
Example 13 : Preparation of Delivery Agent 1
Delivery Agent 1 was made under similar conditions as in Example 9 in a laboratory environment . Example 14: Lyophilization of sCT/Delivery Agent 1 Na salt
Delivery Agent 1 as prepared in Example 13 was formulated into a lyophilized powder with sCT as in Example 2 with 485 ml 0.2 N NaOH and 19.0072 g of Delivery Agent 1 in a steam bath. The final volume was 505 ml. Four separate batches were prepared from: 187, 138, 74 and 160 ml delivery agent 2 Na salt solution plus 28, 48, 40 and 360 mg sCT, respectively. The estimated amounts of delivery agent 2 Na salt were: 7, 5, 2.5 and 4.5 g, respectively.
Example 15: Oral Delivery of sCT/Delivery Agent 1 Na Salt in Rats
According to the method of Example 4 , with the exception that the standard protocol for the EIA kit was followed instead of the modified version, rats were administered orally one capsule of 13 mg lyophilized powder using one of the four batches prepared in Example 14, with 1 ml water to flush the capsule down. The approximate amounts of Delivery Agent 1 Na salt and sCT are set forth in Table 7 below, where results are recorded.
* approximate dose due to variations in animal weight
The above mentioned patents, applications, test methods, and publications are hereby incorporated by reference in their entirety.
Many variations of the present invention will suggest themselves to those skilled in the art in light of the above detailed description. All such obvious variations are within the fully intended scope of the appended claims

Claims

What is claimed is :
1. A solid dosage form comprising a lyophilized powder obtained from a solution comprising a delivery agent and an active agent.
2. A method of making a solid dosage form comprising:
-making a solution comprising a delivery agent and an active agent; -lyophilizing the solution to obtain a solid powder; and
-incorporating the solid powder into a solid dosage form.
3. A method of administering a biologically-active agent to an animal in need of said agent comprising administering to the animal the solid dosage form of claim 1.
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WO2004012772A1 (en) * 2002-08-01 2004-02-12 Novartis Ag Oral administration of calcitonin
US6960355B2 (en) 1996-03-29 2005-11-01 Emisphere Technologies, Inc. Compounds and compositions for delivering active agents
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US6991798B1 (en) 1998-08-07 2006-01-31 Emisphere Technologies, Inc. Compounds and compositions for delivering active agents
WO2006072070A2 (en) 2004-12-29 2006-07-06 Emisphere Technologies, Inc. Pharmaceutical formulations of gallium salts
US7151191B2 (en) 2000-01-13 2006-12-19 Emisphere Technologies, Inc. Compounds and compositions for delivering active agents
US7186414B2 (en) 1998-08-07 2007-03-06 Emisphere Technologies, Inc Compounds and compositions for delivering active agents
US7351741B2 (en) 2000-06-29 2008-04-01 Emisphere Technologies, Inc. Compounds and compositions for delivering active agents
US7553872B2 (en) 1997-02-07 2009-06-30 Emisphere Technologies, Inc. Compounds and compositions for delivering active agents
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US8410052B2 (en) 2007-03-02 2013-04-02 Novartis Ag Oral administration of a calcitonin
US8927015B2 (en) 2006-04-12 2015-01-06 Emisphere Technologies, Inc. Formulations for delivering insulin
US8975227B2 (en) 2005-07-15 2015-03-10 Emisphere Technologies, Inc. Intraoral dosage forms of glucagon
US9364502B2 (en) 2006-06-28 2016-06-14 Emisphere Technologies, Inc. Gallium nitrate formulations
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Cited By (22)

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US6610329B2 (en) 1993-04-22 2003-08-26 Emisphere Technologies Inc. Compositions for the delivery of antigens
US6461643B2 (en) 1993-04-22 2002-10-08 Emisphere Technologies, Inc. Oral drug delivery compositions and methods
US6960355B2 (en) 1996-03-29 2005-11-01 Emisphere Technologies, Inc. Compounds and compositions for delivering active agents
US7417022B2 (en) 1996-03-29 2008-08-26 Mhr Institutional Partners Iia Lp Compounds and compositions for delivering active agents
US7553872B2 (en) 1997-02-07 2009-06-30 Emisphere Technologies, Inc. Compounds and compositions for delivering active agents
US7186414B2 (en) 1998-08-07 2007-03-06 Emisphere Technologies, Inc Compounds and compositions for delivering active agents
US6991798B1 (en) 1998-08-07 2006-01-31 Emisphere Technologies, Inc. Compounds and compositions for delivering active agents
US7151191B2 (en) 2000-01-13 2006-12-19 Emisphere Technologies, Inc. Compounds and compositions for delivering active agents
US7351741B2 (en) 2000-06-29 2008-04-01 Emisphere Technologies, Inc. Compounds and compositions for delivering active agents
EP1341526B1 (en) * 2000-12-06 2011-06-22 Novartis AG Pharmaceutical compositions for the oral delivery of calcitonin
WO2004012772A1 (en) * 2002-08-01 2004-02-12 Novartis Ag Oral administration of calcitonin
AU2003260346B2 (en) * 2002-08-01 2007-12-13 Novartis Ag Oral administration of calcitonin
US7569539B2 (en) 2002-08-01 2009-08-04 Novartis Ag Oral administration of calcitonin
WO2005112937A1 (en) 2004-05-19 2005-12-01 Emisphere Technologies, Inc. Acyclovir formulations
US9498487B2 (en) 2004-05-19 2016-11-22 Emisphere Technologies, Inc. Topical cromolyn formulations
WO2006072070A2 (en) 2004-12-29 2006-07-06 Emisphere Technologies, Inc. Pharmaceutical formulations of gallium salts
US8110547B2 (en) 2005-01-12 2012-02-07 Emisphere Technologies, Inc. Compositions for buccal delivery of parathyroid hormone
US8975227B2 (en) 2005-07-15 2015-03-10 Emisphere Technologies, Inc. Intraoral dosage forms of glucagon
US8927015B2 (en) 2006-04-12 2015-01-06 Emisphere Technologies, Inc. Formulations for delivering insulin
US9364502B2 (en) 2006-06-28 2016-06-14 Emisphere Technologies, Inc. Gallium nitrate formulations
US8410052B2 (en) 2007-03-02 2013-04-02 Novartis Ag Oral administration of a calcitonin
WO2011017346A2 (en) 2009-08-03 2011-02-10 Emisphere Technologies, Inc. Fast-acting naproxen composition with reduced gastrointestinal effects

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