MXPA01008006A - Stable non-aqueous single phase viscous vehicles and formulations utilizing such vehicles - Google Patents

Abstract

This invention relates to stable non-aqueous single phase viscous vehicles and to formulations utilizing such vehicles. The formulations comprise at least one beneficial agent uniformly suspended in the vehicle. The formulation is capable of being stored at temperatures ranging from cold to body temperature for long periods of time. The formulations are capable of being uniformly delivered from drug delivery systems at an exit shear rate of between about 1 to 1 x 107 reciprocal second.

Description

VISCOUS VEHICLES, SINGLE PHASE, NON-AQUEOUS, STABLE AND FORMULATIONS USING SUCH VEHICLES FIELD OF THE INVENTION This invention relates to viscous, biocompatible, single-phase, non-aqueous, stable vehicles capable of suspending beneficial agent and uniformly dispensing said agents at low flow rates, and more particularly, to mixed formulations. uniformly, stable, beneficial agents in viscous, biocompatible, single-phase, non-aqueous, stable vehicles.

References The following citations are referenced by the numbers in co hetes ([]) in the relevant portion of the specification. 1. Wang, et al., J. Parenteral Sci. Tech, 42: S4-S26 (1988). 2. Desai, et al., J. Am Chem. Soc, 116: 9420-9422 (1994). 3. Chang, et al., Pharm. Tech., 80-84 (January 1996). 4. Manning, et al., Pharm. Res., 6: 903-918 (1989). 5. Hageman, Drug Dev. Ind. Pharm, 14: 2047-2070 (1988). 6. Bell, et al., Biopolymers, 35: 201-209 (1995). 7. Zhang, et al., Pharm. Res. 12: 1447-1452 (1995). 8. Published PCT Application 98/00158 9 Published PCT Application 98/16250 10. Knepp, et al., Pharm. Res. 15 (7) 1090-1095 (1998) 11. Published application PCT 98/00157 12. Published application PCT 98/00152 13. US Patent 5,540,912 14. US Patent 5,571,525 15. US Patent 5,512,293 16. Published application of PCT 96/40049 17. Yu, et al., J. Pharm. Sci.85: 396-401 (1996) 18. Mitchell, U.S. Patent No. 5,411,951 (1995) 19. Brooks, et al., U.S. Patent No. 5,352,662 (1994) 20. Geller, L., U.S. Patent No. 3,869,549 (1975) 21. Larsen, et al., PCT publication no. WO95 / 34285 (1995). 22. Knepp, et al., J. Pharm. Sci. Tech, 50: 163-171 (1996). 23. U.S. Patent 5,614,221 24. U.S. Patent 4,594,108 25. U.S. Patent 5,300,302 26. U.S. Patent 4,588,614 U.S. Patent 4,310,516 28. U.S. Patent 5,635,213 29. EP 379,147 BACKGROUND OF THE INVENTION Peptides, polypeptides, proteins and other protein substances (eg, viruses, antibodies) collectively referred to herein as proteins, have great utility as pharmaceuticals in the prevention, treatment and diagnosis of diseases. The proteins are naturally active in aqueous environments, of this type, the preferred protein formulations have been in aqueous solutions. However, proteins are only marginally stable in aqueous solutions. Thus, protein pharmacists often have short shelf lives under ambient conditions or require refrigeration. In addition, many proteins have only limited solubility in aqueous solutions. Even when they are soluble at high concentrations, they are prone to aggregation and precipitation. Because proteins can easily degrade, the standard method for delivering such compounds has been daily injections. Proteins can be degraded via a variety of mechanisms, including deamidations of asparagine and glutamine; oxidation of methiopine and, to a lesser degree, tryptophan, tyrosine and histidine; hydrolysis of peptide bonds; disulfide bio exchange; and racemization of residues of amino acids q uirales [1 -7]. Water is a reagent in almost all of these deg radation routes. In addition, water acts as a plasticizer, which facilitates the unfolding and irreversible aggregation of proteins. Because water is a participant in almost all protein degradation pathways, the reduction of aqueous protein solution to a dry powder provides an alternative formulation methodology to enhance the stability of protein pharmacists. One approach to stabilize the proteins is to dry them using several techniques, including freeze drying, spray drying, lyophilization and drying. The dried proteins are stored as dry powders until their use is required.

A serious disadvantage of drying proteins is that one would often like to use proteins in some kind of fluidizable form. A parenteral injection and the use of medication delivery devices for sustained drug delivery are two examples of applications where one would like to use proteins in a fluidizable form. For injection, the dried proteins must be reconstituted, add additional steps which are slow and where contamination may occur, and expose the protein to potentially destabilizing conditions [7]. For drug delivery devices, protein formulations must be stable for extended periods at body temperature and maintain their ability to flow over the expected life of the device. It has been shown that protein / peptide solution formulations in non-aqueous polar aprotic solvents, such as DMSO and DMF, are stable at elevated temperatures for periods of time [8]. However, such solvent-based formulations will not be useful for all proteins, because many proteins have low solubility in these solvents. While the solubility of the protein in the formulation is less, the greater the amount of solvent to be used for the delivery of a specific amount of protein, the low concentration solutions may be useful for Injections, but may not be useful for long-term delivery at low flow rates.Proteins have been formulated for delivery ^ using perfluorodeca lin [9, 10], methosiflurane [9], high concentrations in water [11], polyethylene glycol [12], PLGA [13,14], mixtures of ethylene vinyl acetate / polyvinylpyrrodone [15], PEG400 / povidone [16] .However, these formulations were not shown to retain a uniform suspension of protein in viscous vehicle over long periods. Biologically active compounds are degraded over time in aqueous solution.The carriers in which the proteins do not dissolve, but instead are suspended, can often offer improved chemical stability. Alternatively, it may be beneficial to suspend the beneficial agent in a carrier when the agent exhibits low solubility in the desired vehicle. However, the suspensions may have poor physical stability due to settlement and agglomeration of the suspended beneficial agent. Problems with non-aqueous carriers tend to be exacerbated as the concentration of the active compound increases. It has been investigated to disperse proteins or peptides in powder in lipid vehicles to produce formulations of sustained release, parenteral [17-21]. The vehicles used were either several vegetable oils (sesame, soy, peanuts, etc.) or synthetic (for example, M ig I ol) gelled with esters of fatty acids of aluminum, such as aluminum stearates (mono-, di- or tri-) or with a polyglycerol ester. Although theoretically these vehicles could prevent solution denaturation and could protect the drug from aqueous chemical degradation, the vehicles by themselves are unstable at higher temperatures. The storage of liquid vegetable oils at body temperatures results in the formation of reactive species, such as free fatty acids and peroxides (a process, which is accelerated by the presence of traces of various metallic ions, such as copper or iron). , which can be leached from some implantab devices is). These peroxides not only adversely affect protein stability [22], but would be toxic when delivered directly to, for example, the central nervous system of a human or animal. The sustained delivery of medications has many advantages. The use of implantable devices ensures the patient's docility, because the delivery device is tamper proof. With an insert of a device, instead of daily injections, there is reduced site irritation, fewer occupational hazards for practitioners, improved cost effectiveness through decreased costs of equipment for repeated injections, reduced hazards of waste disposal, and enhanced efficacy through the release controlled, or compared to a deposit injection. The use of implantable devices for sustained delivery of a wide variety of drugs or other beneficial agents is well known in the art; standard devices are described, for example, in US Pat. 5, 034,229; 5, 057, 31 8; 5, 11, 596; and 5, 782, 396. The description of each of these patents is incorporated herein by reference. For medication delivery plans, dosing durations of up to one year are not unusual. Charitable agents, which have only therapeutic delivery rates, are the main candidates for use in im plates. When the device is implanted or stored, the settlement of the beneficial agent can occur in a liquid form. This heterogeneity can adversely affect the concentration of the beneficial agent dispensed. The solution to this problem is the size of the transplanted charitable agent deposit. The implant deposits are generally in the order of 25-250 μl, but can be up to 25 ml. Viscous formulations have been prepared using two separate components to be mixed with medication in use [23], agents Freeze drying, lyophilization or spray drying of the active network. The drying process of the active ingredient includes additional vents, such as compounds, which are relatively unstable in aqueous solution, can be processed and filled in dosing containers, can be dried without elevated temperatures and then stored in the container. dry state, in which there are relatively few problems of stability.

The pharmaceutical formulations, in particular parenteral products, should be sterilized after being sealed in the final container and within as short a time as possible after the filling and sealing have been completed. (See, for example, Remington, Pharmaceutical Sciences, 5th ed. (1975)). Examples of sterilization techniques include thermal, or dry heat, aseptic and ionized radiation. Combinations of these sterilization procedures can also be used to produce a sterile product. There is a need to be able to deliver protein compositions to the body, which are stable at body temperatures over prolonged periods to allow long-term delivery of the protein. There is a need to be able to deliver concentrations of proteins that are effective. There is a need for a novel non-aqueous formulation, capable of homogeneously suspending proteins and dispensing such agents at body temperatures and low flow rates over prolonged periods.

Brief description of the invention The present invention provides viscous, biocompatible, non-aqueous, single-phase, stable vehicles capable of forming uniform suspensions with proteins. Viscous vehicle components comprise at least two of polymer, surfactant and? solvent. The proportions of the components will vary depending | of the molecular weight of the components and the desired viscosity of the final vehicle.

The proportions of currently preferred components are: polymer, about 5% up to about 60%; solvent, approximately 30% up to about 50%; and surfactant, approximately 5% up to about 20%. The present invention also provides stable formulations, in which the beneficial agents are uniformly suspended in viscous, biocompatible, non-aqueous, single-phase, stable vehicles. In particular, the beneficial agents are formulated in the viscous vehicles at concentrations of at least about 0.1%, depending on the potency of the beneficial agent. These stable formulations can be stored at the appropriate temperature for the beneficial agent, varying from cold to bodily temperature (approximately 37 ° C) for long periods (1 month to 1 year or more). In a preferred embodiment, the formulation comprises approximately 0. 1 to 50% (w / w) of beneficial agent, depending on the potency of the beneficial agent, the duration of treatment and the release rate for the drug delivery system. These formulations are especially useful in implantable delivery devices for long-term delivery (eg, 1 to 12 months or more) of beneficial agent at body temperature, of reference p approximately 37 ° C. Thus, the present invention also provides delivery of said proteins to the body over a prolonged period to allow long-term delivery of the protein at low flow rates of about 0.3 to 4 μl / day for approximately one period __tf_MilM_ for delivery of 6 months and preferably, 5 to 8 μl / c during a delivery period of 3 months. | In another aspect, the invention provides methods for preparing stable, non-aqueous, biocompatible formulations of a beneficial agent in a single-phase viscous vehicle. Preferred formulations comprise approximately 0.1 to 50% (w / w) of beneficial agent depending on the potency of the beneficial agent, the duration of treatment and the release speed of the delivery system. In yet a further aspect, the invention provides methods of treating a subject suffering from a condition, which can be alleviated by the administration of a beneficial agent, said methods comprising administering to said subject an effective amount of a stable non-aqueous formulation. , which comprises at least one beneficial agent uniformly suspended in a simple viscous vehicle. An additional aspect of the invention is that viscous, single-phase, non-aqueous vehicles containing beneficial agents are chemically and physically stable over a wide temperature range for extended periods. The beneficial agents in viscous vehicles are also chemically and physically stable over a wide! temperature range for long periods. In this way, these forms are advantageous, since they can be stored and stored at temperatures below, at or above the ambient temperature for a long period. They are also suitable for use in implantable delivery devices, in which the formulation must be stable at body temperature for prolonged periods. The formulations of the present invention also remain stable when delivered from injectable drug delivery systems. It has been shown that the beneficial agents exhibit zero order release rates when delivered from implantable drug delivery systems at very low flow rates over extended periods.

BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 shows the stability of hGH formulations of the present invention as determined at 37 ° C by reverse phase HPLC. Figure 2 shows the stability of hGH formulations of the present invention as determined at 37 ° C by size excision chromatography. Figure 3 shows the average release rate (μl / day) of 10% (w / w) of spray-dried lysozyme in formulations of the present invention. Figure 4 shows the average release rate (μl / day) of 10% (w / w) of hG H dried by vehicle spray of glycerol mono aurate / lauryl lactate / polyvinylpyrrolidone. Figure 5 shows the average release rate (μg / day) of 10% lysozyme in a lauryl alcohol / polyvinylpyrrolildone vehicle. Figure 6 shows the average release rate (μg / day) of 25% isozyme in a glyceryl monolaurate / lauryl lactate / polyvinylpyrrolidone vehicle.

Figure 7 shows the average release rate ¡(μg / day) of 33% lysozyme in a lauryl glycerol monolaurate / polyvinylpyrrolidone monolaurate vehicle. Figure 8 shows the average release rate (μg / day) of 45% lysozyme in a vehicle of glycerol monolaurate / lauryl lactate / poly ivinylpyrrole idone.

DETAILED DESCRIPTION OF THE INVENTION The present invention is led to the unexpected discovery that uniformly suspending beneficial agents in 1 viscous, biocompatible, single-phase, non-aqueous vehicles results in stable formulations, which can be delivered to body temperature over a prolonged period at low flow rates. The previously known formulations of beneficial agents, which are buffered aqueous or non-aqueous solutions, which may or may not contain excipients, do not provide formulations that can be uniformly dispensed at body temperatures at low flow rates over a prolonged period without exhibiting unacceptable amounts. of ag regation or degradation of the formulation. The formulations currently claimed stabilize beneficial agents and can be stored at the appropriate temperature for the beneficial agent. Temperatures can vary from cold (without exceeding 8 ° C) to corporal (approximately 37 ° C) for extended periods. These formulations are especially useful in implantable delivery devices for long-term delivery (eg, 1 to 1 2 months or more) of medicament at low flow rates and at body temperature, preferably about 37 ° C. Standard benefit agent formulations consist of dilute aqueous or non-aqueous solutions or suspensions. Drug stability is usually achieved by varying one or more of the following: pH, type of buffer, ionic strength, excipients (EDTA, ascorbic acid, etc.). For these formulations, degradation pathways requiring water (hydrolysis, deamidation, racemization) can not be fully stabilized. In the present invention, it was shown that beneficial agents formulated in viscous, single-phase, biocompatible, non-aqueous vehicles containing, for example, polyvinylpyrrolidone, vinyl acetate and / or polyoxyethylenepolyoxypropylene block copolymers are chemically and physically stable. The viscosity of the formulation will depend on a variety of criteria, including the potency and concentration of beneficial agent, and the process by which the formulation is prepared. The viscosity of the formulation can be chosen so that the desired amount of beneficial agent is delivered over the desired period. The invention also consists of viscous, biocompatible, single-phase, non-aqueous vehicles capable of uniformly suspending beneficial agents and formulations containing at least one beneficial agent uniformly suspended in said viscous vehicle. The invention also consists of formulations containing alomenes un! Beneficial agent uniformly suspended in a viscous vehicle, biocompatible, single phase, non-aqueous, said formulations are stable for a prolonged period at body temperatures, and are capable of delivering said beneficial agents uniformly at low flow rates. The discovery consists of the understanding that viscous, non-aqueous, stable vehicles improve the stability of the beneficial agents in a wide range of formulation conditions, including concentration, high temperatures and duration of stable formulation. , making it possible in this way, the delivery of beneficial agents in long-term implantable devices that would not otherwise be feasible.

Definitions As used herein, the following terms have the following meanings: The term "chemical stability" means that an acceptable percentage of degradation products produced by chemical routes, such as oxidation, deamidation or hydrolysis, is formed. In particular, a formulation is considered chemically stable if no more than about 35% decomposition products are formed after 2 months at 37 ° C. The term "physical stability" means that an acceptable percentage of aggregates (eg, dimers, trimers, and larger forms) is formed by the beneficial agent. For the formulation (viscous vehicle and beneficial agent) this term means that the formulation retains stability, ability to flow, the ability to uniformly dispense the beneficial agent. In particular, a form ulation is - ^ y ^ considered physically stable if no more than about 15% of ag is formed after two months at 37 ° C. The term "stable formulation" means that at least about 65% of chemically beneficial and physically stable agent remains after two months at 37 ° C (or equivalent conditions at a high temperature). Particularly preferred formulations are those which retain at least about 80% chemical beneficial agent and physically stable under these conditions. Especially preferred stable formulations are those which do not exhibit degradation after sterilizing irradiation (eg, ga mm a, beta or electron beam). The term "beneficial agent" means peptides, proteins, nucleotides, hormones, viruses, antibodies, etc. which comprise polymers of amino acid residues or nucleic acids. These beneficial agents are generally detectable in water and generally stable as a dry powder at elevated temperatures. The portions produced, synthetically, derived naturally or produced recombinantly, are included in this term. The term also includes lipoproteins and modified post-translational forms, for example, glycosylated proteins. Analogs, derivatives, agonists, antagonists and pharmaceutically acceptable salts of any of these are included in this term. The term also includes proteins and / or protein substances, which have D-amino acids, modified amino acids, derivatives or that do not occur naturally in the D or L configuration and / or peptom ine units as part of their structure. The term "protein" will be used in the present invention. The term also means that the beneficial agent is present in the solid state, for example, powder or crystalline. The term "excipient" means a more or less inert substance in a formulation that is added as a diluent or vehicle or to give shape or consistency. The excipients are distinguished from solvents, such as ETOH, which are used to dissolve drugs in formulations. The excipients include nonionic surfactants, such as polysorbates, which are used to solubilize medicaments in formulations; preservatives, such as benzyl alcohols or methyl or propyl parabens, which are used to prevent or inhibit microbial growth; chelating agents; flavoring agents; and other pharmaceutically acceptable formulation auxiliaries. I The term "viscous vehicle" means a vehicle with a viscosity in the range of approximately 1,000 to 10,000,000 poises. The term includes Newtonian and non-Newtorian materials. Vehicles with a viscosity of approximately 10,000 to 250,000 poises. The formulations of this invention can uniformly expel beneficial agents suspended in the viscous vehicle from implantable drug delivery devices. The formulations exhibit a cutting speed at the output of said devices of 1 z 1 x 10"7 reciprocal seconds, preferably an output cutting speed of 1 x 10" 2 to 1 x 105 reciprocal seconds.

The term "simple phase" means a solid, semi-solid or liquid homogenous system that is both physically and chemically uniform throughout, as determined by differential scanning calorimetry (DSC). The DSC scan should show a peak indicative of a simple phase. The term "biocompatible" means a property or characteristic 5 of a viscous vehicle to disintegrate or decompose, over a prolonged period, in response to the biological environment in the patient, by one or more physical or chemical degradation processes, for example, by action enzymatic, oxidation or reduction, hydrolysis (proteolysis), displacement, for example, ion exchange, or dissolution by solubilization, emulsion or micelle formation, and said material is then absorbed by the surrounding body and tissue or is otherwise dissipated therefrom. The term "polymer" includes polyesters, such as PLA (polylactic acid) [having an inherent viscosity in the range of approximately 0.5 to 2.0 iv] and PLGA (polylactic polyglycolic acid) [having an inherent viscosity in the range of about 0.5 to 2.0 iv], pyrrolidones, such as polyvinylpyrrolidone (having a molecular weight range of about 2,000 to 1,000,000), esters or ethers of unsaturated alcohols, such as vinyl acetate and polyoxyethylenepolyoxypropylene block copolymers (exhibiting a high viscosity at 37 ° C), such as Pluronic 105. The currently preferred polymer is polyvinylpyrrolidone. The term "solvent" includes carboxylic acid esters, such as lauryl lactate, polyhydric alcohols, such as glycerin, polymers of polyhydric alcohols, such as polyethylene glycol, (having -fi l? T - - ™ »> - * J - -. - • - - B? _Ué a molecular weight of about 200 to 600), fatty acids, such as oleic acid and octanoic acid, oils such as rici oil, propylene carbonate, lauryl alcohol or poly alcohol esters rich, such as triacetin acetate. The currently preferred one is lauryl lactate. The term "surfactant" includes polyhydric alcohol esters, such as glycerol monolaurate, ethoxylated castor oil, polysorbates, esters or ethers of saturated alcohols, such as myristyl lactate (Ceraphyl 50) and polyoxyethylenepolyoxypropylene block copolymers, such as Pluronic. Monolaurate and glycerol and polysorbates are currently preferred. The term "antioxidant" means a pharmaceutically acceptable auxiliary for stabilization of the beneficial agent against degradation, such as oxidation. Antioxidants include, but are not limited to, tocopherol (vitamin E), ascorbic acid, ascorbyl palmitate, butylated hydroxyanisole, butylated hydroxytoluene and propyl gallate. A preferred antioxidant depends on the solubility and efficiency of the antioxidant to protect against degradation or chemical change of the beneficial agent in the preferred vehicle. Currently, ascorbyl palm itato is preferred.

FORMULATION PREPARATION The present invention is directed to viscous, biocompatible, simple phase, nonaqueous, stable vehicles capable of suspending beneficial agents and uniformly dispensing said agents. ......... ____.._. beneficial agents at body temperatures at low flow rates over a prolonged period. The present invention is also directed to formulations containing suspended beneficial agents (uniformly in such viscous, biocompatible, single phase vehicles, which are stable for extended periods at body temperatures.) Examples of beneficial agents that can be formulated using the present invention, include those peptides or proteins which have biological activity or which can be used to treat a disease or other pathological condition.But they are not limited to, adrenocorticotropic hormone, angioensina I and II, atrial natriuretic peptide, bombesin, radicinina, calcitonin, cerebellína, dynorphin N, alpha and beta endorphin, endothelin, enkephalin, epidermal growth factor, fertirelin, releasing peptide follicular gonadotropin, galanin, glucagon, GLP-1, gonadorelin, gonadotropin, goserelin, releasing peptide growth hormone, histrelin, human growth hormone, insulin, interferons, leuprolide, LHRH, motilin, nafarerlína, neurotensin, oxytocin, relaxin, somatostatin, substance P, tumor necrosis factor, triptorelin, vasopressin, growth hormone, nerve growth factor, blood coagulation factors, ribozymes and antisense oligonucleotides. Analogs, derivatives, antagonists, agonists and pharmaceutically acceptable salts of the foregoing can also be used. The beneficial agents useful in the formulations and methods of the present invention can be used in the form of a salt, preferably a pharmaceutically acceptable salt. Useful salts are known to those skilled in the art and include salts with inorganic acids, organic acids, inorganic bases or organic bases. Beneficial agents that are not readily soluble in non-aqueous solvents are preferred for use in the present invention. One skilled in the art can easily determine which compounds will be useful on the basis of their solubility. The amount of beneficial agent may vary depending on the potency of the compound, the composition to be treated, the solubility of the compound, the expected dose and the duration of administration. (See, for example, Gilman, et al., The Pharmacological Basis of Therapeutics, 7th ed. (1990) and Remington, Pharmacological Sciences, 18th ed. (1990), the descriptions of which are incorporated herein by reference.) Unexpectedly, it has been found that using a viscous, biocompatible, single-phase, non-aqueous, stable vehicle increases the stability of the beneficial agent. For example, as seen in Figures 1 and 2, it was found that human growth hormone (hGH) was stable at 37 ° C over 12 weeks in polyvinylpyrrolidone / PEG formulations; Pluronic; and glycerol monolaurate / lauryl lactate / polyvinylpyrrolidone. Figure 1 shows the stability results using reverse phase HPLC. Figure 2 shows the stability results using size exclusion chromatography. In general, viscous, biocompatible, single-phase, nonaqueous, stable vehicles can be prepared by combining the dry ingredients (low moisture content) in a dry box or, under other dry conditions, and mixing them at elevated temperature, preferably about 40 to about 70 ° C, to allow them to liquefy. The liquid vehicle is allowed to cool to room temperature. Differential scanning calorimetry was used to verify that the vehicle was in the simple phase. The final moisture content of the viscous vehicle was < 2% . In general, the stable formulations of the present invention can be prepared by combining the carrier and benefit agent under dry conditions and mixing them under vacuum at elevated temperature, preferably about 40 to about 70 ° C, to disperse the beneficial agent uniformly through the vehicle. icle Allow the formulation to cool to room temperature. It has been found that drying the beneficial agent prior to formulation intensifies the stability of the formulation. It has also been found that adding antioxidants, such as tocopherol, ascorbic acid, ascorbyl palm itoate, butylated hydroxyanisole, butylated hydroxytoluene and propyl gallate, reduces the formation of degradation products (eg, unstable chemical intermediates *) during sterilization.

Methodology We have found that formulations of beneficial, non-aqueous, stable agents can be prepared by utilizing viscous vehicles by combining the ingredients for the viscous vehicle under dry conditions and by using the same ingredients. mix them at elevated temperature to allow them to liquefy and form a simple phase. Once the single-phase viscous vehicle is formed, the vehicle is allowed to cool to room temperature. The beneficial agent is added with mixing at elevated temperature; under vacuum to disperse it evenly in the viscous vehicle. We have tested these formulations of beneficial agents, for example, hGH formulations, for stability when subjected to tests of accelerated aging. The results show that these forms remained stable over long periods. We have tested formulations of beneficial agents, for example, human growth hormone and lysozyme, for stability by suspending them in a variety of viscous, single-phase, non-aqueous vehicles prepared in accordance with the present invention, subjecting them then to accelerated aging. at elevated temperatures. The stability of the formulations was measured. The results of these studies show that these formulations were stable under conditions that approximate or exceed storage for one year at 37 ° C. We have also tested agent formulations. charities prepared as described herein, for stability after 2.5 meg arads of gamma irradiation. The results show that these formulations remained chemically and physically stable after such irradiation.

The methods _a¿_ The following methods were used to perform the studies in the following Examples. 1 . Prepare protein powders Human growth hormone (obtained, for example, from BresaGen Lim ited, Adelaide, Australia) The active agent was reconstituted in deionized water. The solution containing the active agent was exchanged as a buffer using an Amicon Diaflo® ultrafiltration membrane (molecular weight cutoff of 1,000,000). The diafiltered active agent solution was spray-dried using a Yamato spray mini-dryer. The powder was collected in a collection container through a cyclone trap. All handling of the spray-dried powder took place in a • dry box evacuated with nitrogen. The generated dust was analyzed by particle size and distribution, moisture content, protein content and stability by reverse phase chromatography and exclusion per year. It is known that the conformation of some proteins can be stabilized by the addition of a sugar (such as sucrose or mannitol) or a polyol (such as ethylene glycol, glycerol, glucose and dextran). 2. Preparation of viscous vehicles We have found that viscous, biocompatible, single-phase, stable vehicles can be prepared by combining the ingredients and mixing them at elevated temperatures to allow them to liquefy and form a simple phase. A differential scanning calorimetry scan showed a peak, indicative of a simple phase. The mixing was completed under vacuum to remove trapped air burbtffas produced by the powders. The mixer was a dual-blade blade mixer (D. I.T.), Which runs at a speed of about 40 rpm. Higher speeds may be used but are not required. If a three-component viscous vehicle is prepared, the solvent portion of the vehicle was added to the heated bowl of the mixer first, followed by the surfactant. The polymer was added to the latter, and the ingredients were mixed until a solution resulted (single phase). The vacuum was applied during mixing to remove the air bubbles. The solution was dispensed from the bowl while it was at elevated temperature and allowed to cool to room temperature. Upon cooling, the vehicle exhibited increased viscosity. One and two component gels were made using the same process. 3. Preparation of beneficial agent formulations To prepare the formulation, the single phase viscous vehicle was heated and then mixed under vacuum with a heavy amount of beneficial agent. The beneficial agent and the single phase viscous vehicle were mixed in the same manner as the vehicle was prepared, using a dual-blade blade mixer (or other similar mixer). The mixing speed was between 40 and 120 rpm for about 15 minutes or until a uniform dispersion was obtained. The resulting mixture was removed from the mixer, sealed in a dry container, and allowed to cool to room temperature. 4. Preparation of Deposits Deposits of implantable drug delivery devices (as described in US Patent Application Serial No. 08 / 595,761, incorporated herein by reference) were filled with the appropriate hG H formulation. The formulation was filled in titanium tanks with a polymer plug blocking each end. The filled reservoir was then sealed in a poly-laminated bag and placed in a stability testing oven. It should be noted that the formulations in the tanks of these devices are completely isolated from the outside environment.

. Reverse phase HPLC (RP-HPLC) All stability samples of hGH were assayed for protein content and chemical stability by reverse phase chromatography (RP-H PLC). The analyzes were performed on a Hewlett Packard HP-1090 system with a refrigerated auto-sampler (4 ° C). The chromatographic conditions used are listed below.

TABLE 1 RP-HPLC chromatographic conditions Description Parameter Column J .T. Bker-C 1 8, 4.6x250 mm Flow rate 1 .0 ml / min Detection 214 nm Mobile phase A: 0.1% TFA in water B: 0. 1% TFA in acetonitrile Gradient time% A% B 0 65 35 5 50 50 45 35 65 50 30 70 55 35 35 A standard reference solution of hGH was prepared and its protein content was calculated from the absorbance measurement at 280 nm. Three dilutions of this solution were run, representing 80%, 1,005 and 120% of the expected concentration of hGH in the samples in duplicate at the beginning and at the end of each run, and were used to calculate the total protein content of the samples. m uestras 6. Size exclusion chromatography (S EC) All stability samples of hG H were assayed for protein content and degradation products of high molecular weight medium size exclusion chromatography. The analyzes were performed on a Hewlett Packard H P-1 090 system with a refrigerated auto-sampler (4 ° C). The chromatographic conditions used are listed below.

TABLE 2 SEC chromatography conditions Description Parameter Column TSK-2000SWXL Flow rate 0.5 ml / min Detection 214 nm Mobile phase 25 mM sodium phosphate, 100 mM sodium chloride, pH 7.0 A reference standard solution of hGH was prepared and its protein content was calculated from the abnorbance measurement at 280 nm. Three dilutions of this solution were run, representing 80%, 1,005 and 1 20% of the expected concentration of hGH in the samples, by d uplicate at the start and at the end of each run, and were used to calculate the content of total protein of the samples. The amount of high molecular weight degradation products was calculated by area normalization. The following examples are offered to illustrate this invention and will not be construed in any way as limiting the scope of this invention.

EXAMPLE 1 Preparation of viscous, single-phase, non-aqueous vehicles Viscous, single-phase, non-aqueous vehicles can be prepared as follows and are shown in the table below A. Glycerol monolaurate (Danisco Ingredients, New Century, kansas ) (25 g) was dissolved in lauryl lactate (ISP Van Dyk I nc. J Bellevílle, NJ) (35 g) at 65 ° C. polyvinylpyrrolidone C30 was added (BASF, Mount Olive, NJ) (40 g) and the mixture was stirred at approximately 40 rpm in a dual-blade blade mixer (D.I.T) until a single phase was reached. The trapped air bubbles were removed by applying vacuum to the mixing chamber. The single phase vehicle was dispensed from the mixer and allowed to cool to room temperature. B. Glycerol monolaurate (Danisco I ngredients, New Century, Kansas) (25 g) was dissolved in lauryl lactate (ISP Van Dyk Inc., Bellevílle, NJ) (35 g) at 65 ° C. Polyvinylpyrrole idona C 1 7 was added (BASF, Mount Olive, NJ) (40 g) and the mixture was stirred at approximately 40 rpm in a dual-blade blade mixer (D. I .T.) Until a single phase was reached. Trapped air bubbles were removed when vacuum was applied to the mixing chamber. The single phase vehicle was dispensed from the mixer and allowed to cool to room temperature. C. Polyvinylpyrrolidone C30 (BASF, Mount Olive, NJ) (50 g) was dissolved in polyethylene glycol 400 (Carbide Unit) (50 g) at a temperature of approximately 65 ° C until a simple phase solution formed. The single phase vehicle was dispensed from the mixer and allowed to cool to room temperature. D. Polyvinylpyrrolidone C 1 7 (BA; SF \ Mount Olive, NJ) (50 g) was dissolved in polyethylene glycol 400 (Union Carbide) (50 g) at about 65 ° C until a simple phase solution formed. The single phase vehicle was dispensed from the mixer and allowed to cool to room temperature. E. Polyvinylpyrrolidone C 1 7 (BASF, Mount Olive, NJ) (50 g) was dissolved in castor oil (Spectrum, Gardena, CA) (50 g) at about 65 ° C until a simple phase solution formed. The single phase vehicle was dispensed from the mixer and allowed to cool to room temperature. F. Polyvinylpyrrolidone C 1 7 (BASF, Mount Olive, NJ) (50 g) was dissolved in octanoic acid (Spectrum, Gardena, CA) at about 65 ° C until a simple phase solution formed. The sim ples vehicle was left out of the mixer and allowed to cool to room temperature. G Polyvinylpyrrolidone C 1 7 (BASF, Mount Olive, NJ) (50 g) was dissolved in oleic acid (S pectrum, Gardena, CA) at about 65 ° C until a simple phase solution formed. The simple phase vehicle was dispensed from the mixer and allowed to cool to room temperature. H. Polyvinylpyrrolidone C 1 7 (BASF, Mount Olive, NJ) (35%) was dissolved in glycerin (Baker, NJ) (65%) at about 65 ° C until a simple phase solution formed. The simple phase vehicle was dispensed from the mixer and allowed to cool to room temperature. Cremophor EL (ethoxylated castor oil) (BASF, Mount Olive, NJ) (5%) was dissolved in castor oil (Spectrum, Gardena, CA) (70%) and added polyvinylpyrrolidone C 1 7 (BASF, Mount Olive , NJ) (25%) and dissolved upon mixing at approximately 40 rpm to form a single phase vehicle. The simple phase vehicle was dispensed from the mixer and allowed to cool to room temperature. Pluronic 105 (BASF, Mount Olive, NJ) was heated to about 65 ° C with mixing until melted. The simple phase vehicle was dispensed from the mixer and allowed to cool to room temperature.

TABLE 3 Proportions of components Component Viscosity at low Solvent Surfactant Polymer Proportion of cutting speed (poíses) PVP GML LL 53: 5: 42 PVP GML LL 55:10:35 50,000 PVP GML LL 50:15:35 7,000 PVP LA 60:40 PVP Ceraphyl 50 LA 60:10:30 PVP __. Oleic acid 50:50 30,000 PVP --- Octanoic acid 55:45 7,000 PVP Polysorbate 80 --- 50:50 PVP ___ PEG 400 50:50 PVP Castor oil --- 50:50 --- Pluronic 105 --- 100 1,000,000 PVP Glycerin 50:50 5,000 Where: GML = glycerol monolaurate LL = lauryl lactate PVP = polyvinylpyrrolidine C30 LA = lauryl alcohol PEG = polyethylene glycol 400 EXAMPLE 2 Preparation of hGH A. Preparation by spray drying Lyophilized hGH was reconstituted (BresaGen Limited, Adelaide, Australia) in 150 ml of deionized water. This stock solution contained 1050 mg of hGH. Buffer exchange was achieved using an Amicon Diaflo® ultrafiltration membrane (10,000 molecular weight cut). The ultrafiltration cell was connected to an auxiliary reservoir containing 5Mm of phosphate buffer (Ph 7). The volume of fluid in the cell, as well as the concentration of hGH, remained constant as the excipients were replaced by phosphate buffer. The diafiltered protein solution (concentration of protein in the solution of approximately 2%) was spray-dried using a Yamato spray mini-dryer. The settings of the spray dryer were as follows: suction pressure constantly adjusted to 1.3 kgf / c 2, inlet temperature 120 ° C, solution flow rate 2.5 (approximately 3 ml / min). The powder was collected in a collection vessel through a cyclone trap. All handling of the spray-dried powder took place in a dry box evacuated with nitrogen (% RH: 1-4%). The water content of the suspension vehicles is shown in the following table.

TABLE 4 WATER CONTENT OF SUSPENSION VEHICLES Vehicle Water content Water content of vehicle vehicle at T 0 in 12 weeks at 37 ° C% w / w% w / w Pluronic 105 0.25 0.4 GML / LL / PVP 1.5 1.3 RRP / PEG 2.0 2.0 Where: GML = glycerol monoaurate LL = lauryl lactate PVP = polyvinylpyrrolidine C30 PEG = polyethylene glycol 400 EXAMPLE 3 Preparation of hGH formulation A portion of the single phase viscous vehicle was weighed (9 g) and heated to 60 ° C. HGH was added (BresaGen Limited, Adelaide, Australia) (1 g) to the vehicle and mixed for 15 minutes. The mixing was completed under vacuum to remove air bubbles added from the powder.

Approximately 10 mg of the spray dried hGH powder was weighed (the content of hGH in the powder was recalculated based on the determined water and salt content) and mixed with 100 μl of the vehicle at 55-65 ° C (3 samples per each vehicle). Special care was taken while mixing the powder in the suspension vehicle to achieve maximum uniform particle dispersion in the vehicle. 1 All the steps were done in a dry box. The resulting suspension was dissolved with 10 ml of buffer and release rate and analyzed by size exclusion and reverse phase chromatography. HGH spray-dried powder was used as a control.

TABLE 5 Stability of suspensions of hGH at 37 ° C as measured by size exclusion chromatography Time Dust Dried by Suspension of Suspension of Suspension weeks spray-80 ° C PVP / PEG 400 GML / LL / PVP Pluronic 105% LS % LS% LS% LS 0 96 ± 1 88 ± 6 92 ± 2 87 ± 7 1 99 ± 8 81 ± 2 94 ± 3 93 ± 3 2 99 ± 3 83 ± 1 97 ± 1 94 ± 1 3 97 ± 1 84 ± 2 95 + 2 95 + 3 4 95 + 2 82 + 8 94 ± 4 93 ± 5 7 95 + 4 76 ± 3 93 ± 4 88 ± 2 1 2 97 ± 4 79 ± 3 97 ± 1 95 ± 6 Each data represents the average ± the relative standard deviation of three individual samples taken from three separate bottles.

TABLE 6 Stability of suspensions of hGH at 37 ° C as measured by reverse phase chromatography Time Dust Dried by Suspension of Suspension of Week Suspension aspersion-80 ° C PVP / PEG 400 GML / LL / PVP Pluronic 105% LS% LS% LS% LS 0 104 ± 1 99 ± 3 99 ± 2 89 ± 7 1 104 ± 8 75 + 2 98 ± 3 96 ± 6 2 104 ± 4 73 ± 3 95 ± 1 96 ± 1 3 104 ± 2 78 ± 4 97 + 3 97 ± 4 4 100 + 2 74 ± 10 93 ± 4 96 + 4 7 108 + 5 72 ± 4 96 ± 2 94 + 2 9 102 ± 3 66 ± 3 92 ± 3 93 ± 2 12 101 ± 2 66 ± 1 89 + 2 92 + 5 Each data represents the average ± the relative standard deviation of three individual samples taken from three separate bottles.

EJ EM PLO 4 Preparation of deposits Release rate profiles The deposition systems of drug delivery devices (as described in US patent application Ser. No. 08/595, 761, incorporated herein by reference) were each assembled with an osmotic motor, piston and speed control membrane. The tanks were filled with the appropriate amount of viscous vehicle formulation and plugged with a flow plug. The systems were placed in a water bath at 37 ° C and allowed to form for a prolonged period. The material released was sampled twice a week. Assays for released material were completed using reverse phase HPLC. The resulting concentrations of beneficial agent for each system were converted to free capacity per day. The beneficial agent was found to have a zero order release from the implantable drug delivery device, as shown in Figures 3 to 8.

EJ EM P LO 5 Stability of hGH in viscous, non-aqueous vehicle formulations The formulations of 10% w / w of hGH in vehicle were prepared as described and placed in flasks. The formulations were subjected to accelerated aging by storing them at elevated temperatures and times shown in the table below in a temperature controlled oven.

TABLE 7 Vehicle Time (h) Temperature% LS per SEC% LS by RP-HPLC Pluronic 105"5 50 ° C ~ 98 + 3 101 ± 3 Pluronic 105 1 50 ° C 98 ± 3 101 + 4 Pluronic 105 2 50 ° C 100 + 1 1¡02 + 3 Pluronic 105 4 50 ° C 101 + 3 105 + 3 GML / LL / PVP 0 65 ° C 99 ± 3 101 + 3 GML / LL / PVP 1 65 ° C 93 ± 6 97 ± 6 GML / LL / PVP 2 65 ° C 91 + 5 95 ± 5 GML / LL / PVP 4 65 ° C 95 ± 3 98 + 3 Each data represents the mean ± the relative standard deviation of three individual samples taken from three separate bottles. The results, presented in the following table, demonstrate that these formulations are capable of maintaining the stability of hGH in each case. In each case, at least 70% hGH was retained.

TABLE 8 Retrieval of hGH from non-aqueous suspensions Vehicle% LS by RP-HPLC% by HPLC exclusion by size PVP / PEG 400 99 + 3% 88 + 6% GML / LL / PVP 99 ± 2% 92 ± 2% Pluronic 105 89 ± 7% 87 ± 7% "? Each data represents the average ± the relative standard deviation of three individual samples taken from three separate bottles. % LS or% brand strength = (measured protein content '+ theoretical protein content) x 1 00% Modification of the modes described above to perform various embodiments of this invention will be apparent to those of skill in the art, following the teachings of this invention as set forth herein. The examples described above are not limiting, but simply exemplary of this invention, the scope of which is defined by the following claims.

Claims (38)

1. RE IVI N DI CATIONS 1 A viscous vehicle, biocompatible, simple phase, non-aqueous, stable, able to suspend beneficial agents and homogeneously dispense said beneficial agent over a prolonged period at body temperature and at low flow rates.
2. 2. The vehicle of claim 1, comprising two components selected from the group consisting of solvent, surfactant and polymer, wherein the two components are not of the same type.
3. 3. The vehicle of claim 1, comprising at least two components selected from the group consisting of solvent, surfactant and polymer, wherein the components are not of the same type.
4. 4. The vehicle of claim 1, which comprises three components selected from the group consisting of solvent, surfactant and polymer, wherein the components are not of the same type.
5. 5. The vehicle of claim 2 or 4, wherein said solvent is selected from the group of carboxylic acid esters, polyhydric alcohols, polyhydric alcohol polymers, fatty acids, oils, propylene carbonate, lauryl alcohol and esters of polyhydric alcohols.
6. 6. The vehicle of claim 2 or 4, wherein said surfactant is selected from the group of polyhydric alcohol esters, non-ethoxy side oil, polysorbates, esters or ethers of saturated alcohols and polyoxyethylenepolyoxypropylene block copolymers. .
7. 7. The vehicle of claim 2 or 4, wherein said polymer is selected from the group of polyesters, pyrrolidones, esters, esters of unsaturated alcohols, and polyoxyethylenepolyoxypropylene block copolymers. The vehicle of claim 2, wherein the proportions of the components are in the range of 40:60 to 60:40. The vehicle of claim 4, wherein the proportions of the components are in the range of about 30% to about 50% for solvent, about 5% to about 20% for surfactant, and about '5% to about 60% for polymer. The vehicle of claim 4, wherein the polymer is polyvinyl pyrrolidone, the surfactant is gml, and the solvent is lactate laudel. 11. The vehicle of claim 4, wherein the polymer is polyvinyl pyrrolidone, the surfactant is polysorbate and the solvent is lauyl lactate. 12. The vehicle of claim 1, which comprises an antioxidant. 13. The vehicle of claim 12, wherein said antioxidant is selected from the group consisting of tocopherol, ascorbic acid, ascorbyl palmitate, butylated hydroxyanisole, butylated hydroxytoluene and propyl gallate. 14. A stable, non-aqueous, viscous protein formulation comprising: a) at least one beneficial agent, and b) a viscous, biocompatible, single-phase, non-aqueous vehicle, said formulation is capable of being uniformly dispensed over a period of time. prolonged at a low flow rate. A non-aqueous formulation comprising at least one beneficial agent uniformly suspended in a viscous, biocompatible, single-phase, non-aqueous vehicle, said formulation can be delivered from an implantable drug delivery system, so that the cutting speed of the formulation is between about 1 and 1 x 1 0"7 reciprocal seconds 16. The formulation of claim 14, wherein said formulation is stable at body temperature for prolonged periods. 14, which comprises at least about 0. 1% (w / w) of beneficial agent 1 8. The formulation of claim 14, which comprises at least about 10% (w / w) of beneficial agent. 19. The formulation of claim 14, wherein said beneficial agent is selected from the group consisting of peptide, protein, nucleotide, hormone, virus or antibody. ation 1 9, wherein said beneficial agent is a protein. twenty-one . The formulation of claim 14, which is stable at 65 ° C for at least about 2 months. 22. The formulation of claim 1, which is stable at 37 ° C for at least about 3 months. 23. The formulation of claim 14, which is stable at 37 ° C for at least about one year. The formulation of claim 1, which is adapted for use in an implantable drug delivery device. The formulation of claim 14, wherein said carrier is selected from the group consisting of solvent, surfactant and polymer. 26. The formulation of claim 14, wherein said carrier comprises an antioxidant. 27. The formulation of claim 14, comprising a beneficial agent, which has been dried at a low moisture content before incorporation into said formulation. 28. The formulation of claim 14, which is stable after sterilization. 29. A method for preparing the single-phase, stable, viscous vehicle of claim 1, comprising the steps of (1) mixing the carriers at elevated temperature under dry conditions to allow them to liquefy, and (2) ) allow the liquid from step (1) to cool to room temperature. 30. A method for preparing the stable formulation of claim 14, which comprises combining the single-phase viscous vehicle and beneficial agent under dry conditions and mixing them under vacuum at elevated temperature to uniformly disperse the beneficial agent in the vehicle and allow the formulation to cool to room temperature. 31 The method of claim 30, wherein at least about 0. 1% (w / w) of beneficial agent is suspended in said vehicle. 32. The method of claim 30, wherein at least about 10% (w / w) of beneficial agent is suspended in said vehicle. 33. A method for treating a subject suffering from a condition, which can be alleviated by the administration of a beneficial agent, comprising administering to said subject a therapeutically effective amount of the formulation of claim 14. 34. The method of claim 33, wherein said administration is parenteral administration. 35. The method of claim 33, wherein said administration is continuous long-term administration. 36. The method of claim 33, wherein said administration is achieved by the use of an implantable drug delivery system. 37. The method of claim 33, wherein said day administration continues for a period selected from the group consisting of about 3 months, about 6 months and about 12 months. 38. The method of claim 37, wherein said daily administration is achieved using an im plantable drug delivery system.