WO2005072709A2 - Formulations for poorly soluble drugs - Google Patents

Formulations for poorly soluble drugs Download PDF

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Publication number
WO2005072709A2
WO2005072709A2 PCT/IL2005/000093 IL2005000093W WO2005072709A2 WO 2005072709 A2 WO2005072709 A2 WO 2005072709A2 IL 2005000093 W IL2005000093 W IL 2005000093W WO 2005072709 A2 WO2005072709 A2 WO 2005072709A2
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WO
WIPO (PCT)
Prior art keywords
drag
delivery system
beads
drag delivery
solvent
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Application number
PCT/IL2005/000093
Other languages
French (fr)
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WO2005072709A3 (en
Inventor
Shlomo Magdassi
Yoram Sela
Keren Cohen
Original Assignee
Bio Dar Ltd
Yissum Research Development Company Of The Hebrew University Of Jerusalem
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Application filed by Bio Dar Ltd, Yissum Research Development Company Of The Hebrew University Of Jerusalem filed Critical Bio Dar Ltd
Priority to EP05703136A priority Critical patent/EP1708682A2/en
Priority to US10/587,456 priority patent/US20080038333A1/en
Publication of WO2005072709A2 publication Critical patent/WO2005072709A2/en
Publication of WO2005072709A3 publication Critical patent/WO2005072709A3/en
Priority to IL177120A priority patent/IL177120A0/en
Priority to US12/843,958 priority patent/US20100291200A1/en

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    • 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
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
    • A61K9/1682—Processes
    • 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
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
    • A61K9/1605—Excipients; Inactive ingredients
    • A61K9/1629—Organic macromolecular compounds
    • A61K9/1635—Organic macromolecular compounds obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyvinyl pyrrolidone, poly(meth)acrylates
    • 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
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
    • A61K9/1605—Excipients; Inactive ingredients
    • A61K9/1629—Organic macromolecular compounds
    • A61K9/1652—Polysaccharides, e.g. alginate, cellulose derivatives; Cyclodextrin
    • 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
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
    • A61K9/1605—Excipients; Inactive ingredients
    • A61K9/1629—Organic macromolecular compounds
    • A61K9/1658—Proteins, e.g. albumin, gelatin

Definitions

  • the present invention generally concerns formulations for drugs, and more particularly formulations for poorly soluble drugs.
  • Solubility is defined as the concentration of the solute in a saturated solution.
  • the solubility of compounds varies in accordance with factors such as temperature, the type of solvent, the pH of the solution, and atmospheric pressure.
  • the solubility of drugs found in the US Pharmacopeia is expressed as the number of milliliters of solvent in which one gram of solute can dissolve. Where the exact solubility of various compounds cannot be precisely determined general quality terms are used to describe the solubility of a specific compound, typically with reference to other compounds. Solubility may also be expressed in terms of molarity, percentage, and molality.
  • drugs defined as “poorly soluble" are those that require more than 1 ml part of solvent per 10 mg of solute.
  • Some poorly soluble drugs are further limited by their intrinsic bioavailability for example due to extensive first pass metabolism by the liverok (first pass effect), or further limited due to various drug-drug interactions .
  • Usage of poorly soluble compounds has increased by 25% on average over the last five year period.
  • the increase in formulations containing poorly soluble compounds is attributed to factors associated with both the pharmaceutical and biotechnology sectors.
  • drugs are now more frequently designed by combinatorial chemistry in order to improve their distribution through various tissues in the body, increase their half life, and improve their therapeutic index (more potency with low concentrations).
  • newly developed drugs produced by combinatorial techniques are poorly soluble as during development, and in contrast to rational drug design, solubility was never a factor considered for their production.
  • Patent Application 20030215513 concerns release of substantially water insoluble nano-sized particles from a composition, by coating the pharmaceutical composition with a diffusion-control membranes that contains a multiplicity of pores and pore-forming substances. This establishes a diffusion gradient that enables mass-transport of nano-suspensions from the pharmaceutical composition through the pores, thereby resulting in a diffusion controlled release through the membrane.
  • U.S. Patent Application 20020106403 discloses a water insoluble drug, in a nanometer or micrometer particulate solid format, which is surface stabilized by a phospholipid, being dispersed throughout a bulking matrix. This construction can dissolve upon contact with aqueous environments, thereby releasing the water insoluble particulate solid in an unaggregated or un-agglomerated form.
  • the matrix is composed of water insoluble substance.
  • U.S. Patent No. 5,439,686 discloses compositions for in vivo delivery of water insoluble pharmaceutical agents, notably the anticancer drag taxol, wherein the active agent is solubilized in a biocompatible dispersing agent contained within a protein walled shell.
  • the protein walled shell can contain particles of the taxol itself.
  • U.S. Patent No. 6,387,409 discloses nano- or micro-sized particles of water insoluble, or of poorly soluble drugs, produced by a combination of natural and synthetic phospholipids and charge surface modifiers such as highly purified charge phospholipids, together with a block copolymer which are coated or adhered on to the surfaces of water insoluble compound particles.
  • the pore forming agent creating the porous matrix is typically a volatile liquid that is immiscible with the drag solvent, or alternatively, a volatile solid compound such as a volatile salt.
  • the resulting porous matrix has a faster rate of dissolution following administration to a patient as compared to a non porous matrix form of the drag.
  • Sustained, or controlled release drug delivery systems include any drug delivery system that achieves a slow release of a drag over an extended period of time.
  • the main aim of slow release systems is improved efficiency of treatment as a result of obtaining constant drug-blood levels, thus maintaining the desired therapeutic effect for extended periods of time. This results in reduction and elimination of fluctuations in blood levels, thus allowing better disease management.
  • Some controlled release systems were not developed for the main purpose of sustained release, but rather having been developed in order to improve the bioavailability of drags, due to their activity in isolating the drags from the environment, for example by protecting drags susceptible to enzymatic inactivation or bacterial decomposition by encapsulation in polymeric systems.
  • Microparticles containing poorly soluble drags and a polymer were prepared in order to overcome some technical problems of tabulating encountered during fo ⁇ nulations of medicaments with microparticles. In these formulations propranonol was the poorly soluble drag, and the polymer was ethylcellulose.
  • the polymer and the poorly soluble drags were mixed to form microspheres containing a drug-polymer mixture, which were subsequently entrapped within a chitosan or calcium alginate beads.
  • the beads contained initially a mixture of drugs and insoluble polymers, subsequently mixed with a soluble polymer.
  • the ionic characteristics of the polysaccharides of this delivery system allowed a pH-dependent release of the microparticles in the gastrointestinal tract Bodmeier et al. Pharmaceutical Research 6:5, 1989).
  • the present invention is based on the realization that particles of water insoluble or poorly soluble drags can have improved solubility, and hence improved bioavailability, if they are administered dispersed in a hydrophilic polymeric bead in the fonn of nanoparticles or microparticles of the drag.
  • the present invention concerns a drag delivery system comprising nanoparticles or microparticles of a poorly soluble drag dispersed in a polymeric bead containing essentially only of hydrophilic polymers (i.e. without hydrophobic polymers).
  • nanoparticle in the context of the drugs refers to particles which have the size of 3 nm to 900 nm, preferably 5 nm to 450 nm.
  • microparticle refers to particles which have the size of 1 to 500 micrometers.
  • the polymeric beads consist essentially of a single hydrophilic polymer, this being in contrast to the publication of Bodmeier et al. wherein the poorly soluble drag is first entrapped within an insoluble, hydrophobic polymer, and the obtained microparticles of the insoluble polymer and drag are then mixed with a soluble polymer- forming bead.
  • the beads of the present invention consist of drag nanoparticles essentially free of water insoluble polymer, while the single hydrophilic polymer serves as a former of porous bead, which prevents the increase in the size of the drag particle, and greatly simplifies the manner of production as will be explained hereinbelow.
  • the bead formation process by itself leads to fonnation of the drag nanoparticles, which are formed from a nanoemulsion, in a way that overcomes the problems associated with conventional methods for preparation of nanoparticles by solvent evaporation from submicron emulsions.
  • the beads themselves serve as the delivery system, having the ability of controlling the release of the nano/micro particles of the poorly soluble drugs therefrom.
  • the control can be achieved by the inherent polymeric structure of the bead, or by a combination of the bead skeleton polymers and polymeric additives, mainly water soluble polymers.
  • drug delivery system in the context of the present invention concerns active ingredient - i.e.
  • the drag delivery system in accordance with the invention may be used for subsequent preparation of dosage administration forms, for example, in the form of capsules (coated or uncoated), tablets (coated or uncoated), wherein the coating may be functional such as enteric coating, colonic delivery coating, chrono-therapeutic and controlled release coating, taste-masking coating and the like.
  • the dosage form may be suitable for any mode of administration such as oral, rectal, depo-administration, parenteral, subcutaneous, ocular, nasal, vaginal and the like.
  • polymer in accordance with the present invention shall be understood as referring both to a polymer composed of a single re-occurring building block (monomer) as well as to a polymer composed of two or more different polymeric units (co-polymer).
  • poorly soluble drug refers to a drag which is insoluble or poorly soluble in an aqueous solution, and typically this refers to a drag which has a solubility of less than 10 mg/ml, and preferably less than about 5 mg/ml in aqueous media at approximately physiological temperature and pH.
  • drug refers to chemical and biological molecules having therapeutic, diagnostic or prophylactic effects in vivo.
  • drag therefore may include food additives which have biological activity such as lycomene, lycopene and beta carotene.
  • Drugs contemplated for use in the system described herein include the following categories and examples of drugs and alternative forms of these drags such as alternative salt forms, free acid forms, free base fonns, prodrag forms and solvates e.g.
  • the drags may also include biological produced agents.such as proteins, protein f agments, peptides, nucleic acid sequences, oligonucleotides, glycoproteins as long as they are water insoluble Most preferable drags are simvastatine, statines, risperidone, carvedilol, carbamazepine, oxcarbazepine, zaleplon, galantamine, avandia, and poorly soluble anti psychotic, anti epileptic, anti parkinsonian and other indicated for CNS indications.
  • the polymeric bead may comprise at least one of a polysaccharide polymer, a protein, a synthetic polymer which may be either crosslinked or not crosslinked or mixtures thereof.
  • polysaccharide polymers are: alginates, chitosans, gellan gums, agarose, pectin, carrageenan.
  • proteins are: gelatins, albumins, lactalbumin.
  • synthetic polymers are polyacrylic acid, polyethylene glycol ("PEG"), polyvinyl pyrrolidone, polymethacrylates, polylysine, poloxamers, polyvinyl alcohol, polyethylene oxide, and polyethyoxazoline.
  • PEG polyethylene glycol
  • polyvinyl pyrrolidone polymethacrylates
  • polylysine poloxamers
  • polyvinyl alcohol polyethylene oxide
  • polyethyoxazoline polyethyoxazoline.
  • the nanoparticles or microparticles are in an amorphous state, which increases their solubility rate, and subsequent crystallization is prevented due to the presence of hydrophilic polymer and surfactants used in the process of production.
  • the drag delivery system may include externally added crosslinking agents, which are, for anionic polyssacharides and synthetic polymers, multivalent cations, such as calcium, magnesium, barium, ferrous, polycations and cupper salts.
  • crosslinking agents for anionic polyssacharides and synthetic polymers, multivalent cations, such as calcium, magnesium, barium, ferrous, polycations and cupper salts.
  • cationic polymers such as chitosan
  • a polyvalent anion such as tripolyphosphate or anionic polymers may be used.
  • the polymeric beads may also be formed by heating -cooling effects, such as formation of gelatin beads , which is obtained by dropwise addition of warm gelation solution into cold liquid, water or oil.
  • the drag delivery system including said externally added crosslinking agents further comprises a disintegrant which may be a chelator of the crosslinking cation, for example calcium or magnesium.
  • a disintegrant which may be a chelator of the crosslinking cation, for example calcium or magnesium.
  • Such chelators in contact with water, interact with the crosslinking agents, thus breaking the crosslinking of the polymeric bead and enhancing the disintegration of the bead.
  • disintegrants are EDTA, sodium citrate, citric acid, sodium dodecyl sulfate, phosphate salts and phosphate buffer saline.
  • the present invention concerns a drug delivery system comprising an active ingredient dispersed within a polymeric bead, wherein the polymer may be crosslinked , while the crosslinking is achieved (in case of sodium alginate, for example) by a multivalent cation such as calcium, magnesium, barium, ferrous or copper salts and wherein the drug delivery system further comprises as a disintegrate, a chelator of the multivalent cation.
  • the drag is a poorly soluble drug, more preferably in the form of a nano-particle, a micro-particle, most preferably in the form of a nanoparticle.
  • the present invention further concerns a method of producing the drug delivery system of the invention comprising: (i) providing poorly water soluble drug dissolved in organic volatile solvent, optionally in the presence of at least one surfactant; (ii) mixing the drag-containing solvent with an aqueous phase, optionally in the presence of at least one agent selected from surfactant, co-solvent and electrolyte, thereby producing an oil-in-water nanoemulsion or microemulsion; (iii) mixing the oil-in-water nano- or micro emulsion with water-soluble bead-forming polymers to produce a continuous phase of the emulsion which comprises the bead forming polymer; (iv) providing conditions enabling bead formation from the continuous phase of (iii); (v) drying of the beads, by evaporating the volatile organic solvent and the aqueous phase of the bead; thereby obtaining dry beads comprising in their matrix dispersed nanoparticles or microparticles of poorly water-soluble drags.
  • the beads containing the drug nanoparticles or microparticles obtained by the method of the invention may be formulated to fonn a suitable dosage form, for example they may be packed within a capsule or a tablet, optionally together with a disintegrant as will be explained herein bellow, thus providing a delivery system of th ⁇ poorly soluble drag.
  • polymeric additives may be added in order to control the drag release.
  • the poorly soluble drag is rendered in a nanoparticle form by consequent evaporation of the organic solvent and the water, thus the previously dissolved drug in the solvent droplets, becomes insoluble, and having a size similar to the initial size of the nanoemulsion droplets, and in most cases having a non-crystalline morphology.
  • each nanoemulsion droplet is dispersed within the crosslined polymeric network of the bead, there is no possibility for coalescence of emulsion droplets, and therefore there is no increase in the size of drag particles which are maintained in their original nanoparticle size.
  • the obtained drag nanoparticles are amorphous (not crystalline).
  • the surfactants in the nanoemulsion the nanoparticles remain in an amorphic structure that brings significant advantages for enhanced dissolution and bioavailability.
  • the processes described in this invention allow obtaining nanoparticles of drags, which otherwise, upon application of conventional solvent evaporation method, would have formed large crystals. It was surprisingly found that by performing the solvent evaporation process only after the beads are formed, the crystallization and increase of the size of the drag molecule could be prevented.
  • the solvent used in the method of the invention is an organic solvent that is volatile (at the concentration used ) i.e. has a relatively low boiling point, or can be removed under vacuum, and which is acceptable for administration to humans in trace amounts.
  • Representative solvents include, chloroform, chlorofluorocarbons, dichloromethane, dipropyl ether, diisopropyl ether, ethyl acetate, butyl acetate, methyl ethyl ketone (MEK), limonene, heptane, hexane, butanol, octane, pentane, toluene, 1,1,1-trichloroethane, 1,1,2-trichloroethylene, xylene, and combinations thereof.
  • the drug is dissolved in the volatile solvent to fonn a drag solution having a concentration of between 0.01 and 80% weight to volume (w/v).
  • the solvent in which the drag is dissolved may contain a co-solvent which is either miscible or immiscible with water.
  • co-solvents are: ethanol, isopropanol, pentanol THF, DMF, DMSO, propylene glycol, polyethylene glycol, glyme , diglyme , triglyme and the like.
  • suitable surfactants are: nonionic surfactants such as for example block copolymers, e.g.
  • Pluronic F 68 polyglycerol esters, alkyl glucosides ethoxylated sorbitan esters and ethoxylated sorbitan esters; ionic surfactants; and polymers such as polyvinyl alcohol, gelatin and BSA.
  • the surfactants are selected from molecules acceptable for pharmaceutical preparations, which are capable of yielding nanoemulsions or microemulsions.
  • the nanoemulsions can be formed by various methods, preferably by using a high pressure homogenization technology, or phase inversion methods (such as the PIT method) and the microemulsions are prepared by simple mixing of proper compositions of water, surfactants, solvents and co-solvents (microemulsions may form spontaneously, according the phase diagram of the compositions ).
  • Additional exemplary surfactants which may be used include most physiologically acceptable emulsifiers, for instance egg lecithin or soya bean lecithin, or synthetic lecithins such as saturated synthetic lecithins, for example, dimyristoyl phosphatidyl choline, dipalmitoyl phosphatidyl choline or distearoyl phosphatidyl choline or unsaturated synthetic lecithins, such as dioleyl phosphatidyl choline or dilinoleyl phosphatidyl choline.
  • emulsifiers for instance egg lecithin or soya bean lecithin
  • synthetic lecithins such as saturated synthetic lecithins, for example, dimyristoyl phosphatidyl choline, dipalmitoyl phosphatidyl choline or distearoyl phosphatidyl choline or unsaturated synthetic lecithins, such as dioley
  • Surfactants also include salts of fatty acids, esters of fatty acids with polyoxyalkylene compounds like polyoxpropylene glycol and polyoxyethylene glycol; ethers of fatty alcohols with polyoxyalkylene glycols; esters of fatty acids with polyoxyalkylated sorbitan; soaps; glycerol- polyalkylene stearate; glycerol-polyoxyethylene ricinoleate; homo- and co- polymers of poryalkylene glycols; polyethoxylated soya-oil and castor oil as well as hydrogenated derivatives; ethers and esters of sucrose or other carbohydrates with fatty acids, fatty alcohols, these being optionally polyoxyalkylated; mono-, di- and tri-glycerides of saturated or imsaturated fatty acids, glycerides of soya-oil and sucrose.
  • Beads are formed by solidifying drops of solutions containing the bead forming polymers either by contact with a crosslinking agent (when the polymer can react with the crosslinking agent to form an insoluble polymeric structure), or by solidification, for examples while using a polymer such as gelatin, which forms a liquid solution at elevated temperature, and solidifies at room temperature.
  • a crosslinking agent when the polymer can react with the crosslinking agent to form an insoluble polymeric structure
  • a polymer such as gelatin
  • the crosslinking ions migrate into the interior part of the bead, and form a solid matrix throughout the whole bead.
  • the structure of the beads can be tailored by proper selection of the bead formation conditions (such as crosslinker concentration, duration of crosslinking , presence of various electrolytes etc.).
  • the size of the beads can be controlled by proper selection of the nozzle diameter and instrumentation from which the bead forming polymeric solution is ejected.
  • the volatile (organic solvent) is evaporated together with the aqueous phase, for example by application of vacuum or by lyophilization processes, or by simply drying at room temperature or in an oven at elevated temperatures, to obtain the dry beads containing in their matrix dispersed nanoparticles of the poorly soluble drag.
  • the beads are packed in a suitable pharmaceutical formulation such as gelatin capsule or solid tablet (containing conventional pharmaceutical excipients), and optionally containing agents which enhance the disintegration of the beads upon contact with body fluids.
  • Such disintegrators can be molecules capable of replacing the crosslinking agent, such as chelators of the crosslinking agents such as EDTA, citric acid, sodium citrate, or surfactants such as sodium dodecyl sulfate, phosphate salts or phosphate buffer saline.
  • chelators of the crosslinking agents such as EDTA, citric acid, sodium citrate, or surfactants such as sodium dodecyl sulfate, phosphate salts or phosphate buffer saline.
  • the crosslinkers such as calcium ions
  • Fig. 1A shows an electron microscope picture of a polymeric bead containing nanoparticles of simvastatine, prepared as described in Example 1 which are vacuum dried
  • Fig IB shows an electron microscope picture of a cross section of the polymeric bead shown in Fig. 1 A.
  • Fig. 1C shows an electron microscope picture of a polymeric bead containing nanoparticles of simvastatine, prepared as described in Example 1 which are air dried.
  • Fig ID shows an electron microscope picture of a cross section of the polymeric bead shown in Fig. 1C.
  • Fig. 2 shows the dissolution of two samples of beads of the invention containing simvastatine as compared to dissolution of commercial simvastatine.
  • Fig. 3 shows an electron microscope picture of simvastatine crystals after solvent evaporation carried out without using bead formation.
  • Fig. 4 shows electron microscope pictures of simvastatine nanoparticles after solvent evaporation from bead nanoemulsion systems.
  • Fig. 5 shows the effect of varying concentrations of phosphate buffer (pH ⁇ 6.8) on beads disintegration.
  • Fig. 6 shows the effect of varying concentrations of citrate buffer (pH ⁇ 6.8) on beads disintegration.
  • Fig. 7 shows the effect of various crosslinking ions at a concentration of 25 mM on beads disintegration.
  • Fig. 8 shows the effect of various crosslinking ions at a concentration of 100
  • Droplets size in the nano/microemulsion may be tailored by controlling volatile solvent type, co- solvent type, surfactants and co-surfactant concentration and type, by controlling the cycles in high-pressure homogenizer (in case high pressure homogenization is utilized to obtain the nanoemulsions ), o/w ratio and temperature.
  • Type and molecular weight of the polysaccharide e.g. Alginate, K-Carrageenan, Chitosan, Gellan gum, Agarose, Pectin etc, or synthetic polymers.
  • alginates e.g. different ratio of guluronic and mannuronic acids.
  • crosslinking agent also termed “gelling agent”
  • gelling agent ion solution
  • Matrix composition of material other than the bead forming polymer other materials may be added, such as Silica, HPMC, Lactose, sodium chloride etc., which affect the morphology, porosity, size, and shrinkage of beads upon drying, disintegration rate and hydrophobicity.
  • the size of the polysaccharide beads can be controlled by controlling nozzle size, frequency, amplitude, velocity, physical parameters.
  • the rate of disintegration may be controlled by adding a disintegrate such as EDTA, phosphate or citrate ions, and controlling the amount of the disintegrant.
  • a disintegrate such as EDTA, phosphate or citrate ions
  • 4% Alginate solution 16g of Alginic acid sodium salt (Sigma, low viscosity, 2% solution- 250cps) was dissolved in 400g distilled water (4% w/w), together with 0.4g of Bronopol (preserving material). The mixture was mixed on magnetic stirrer for about 48 hours and heated to about 37°C until complete dissolution.
  • 3.3584g of Simvastatine powder (Teva Pharmaceuticals, Israel) used as the poorly soluble drag was weighed and mixed with 80. Og toluene until complete dissolution of the drag is achieved. Final concentration of Simvastatine is 42mg/g toluene.
  • Tween 20 was weighed and dissolved in 160.26g distilled water saturated with toluene (filtered through 0.2 ⁇ m filter) . 4.97g Span 20 was weighed and mixed with the 40.23g solution of 42mg/g Simvastatine in toluene, and stirred about lOmin together. The organic phase was added carefully to the water phase and mixed for 5 min in an Ultra Turrax homogenizer at 8000 RPM. A coarse, homogeneous emulsion was obtained. This emulsion was introduced into a high pressure homogenizer (Stansted), and was circulated through the high-pressure-homogenizer twice at 17,000 psi. Z-average particles size of the resulting emulsion was 250-255nm.
  • Beads formation 95. lg of sodium alginate solution (4% w/w) and 3.8g of Silica 6 ⁇ A Frutarom) used to prevent shrinking upon drying, were mixed together for about 10 min by a magnetic stirrer until the silica was dispersed homogeneously in the alginate solution. Then 95. lg of the above o/w emulsion were added and stirred together until homogenous mixture was achieved.
  • the alginate- emulsion mixture was introduced into an Innotech encapsulator, and jetted into lOOmM CaCl 2 crosslinking solution.
  • the Innotech encapsulator allows tailoring the final size of the beads by selecting the proper instrument parameters.
  • the parameters were: Nozzle size - 300 ⁇ m. Voltage - 0.914 Kv. Amplitude - 3. Frequency -1550 Hz. Pressure —0.4 bar.
  • the beads were kept in the crosslinking solution for 30min. Then, the beads were rinsed with about 2 liters of distilled water, filtered and air dried in an oven, at temperature of about 35 ° C for 48 hours, in order to remove the water and the volatile solvent. The final result was dry beads in the size range of less than 1 mm in which nanoparticles of Simvastatine were dispersed, as verified by electron microscopy and shown in Fig. 1.
  • FIG. 1A shows an electron microscope picture of a polymeric bead containing nanoparticles of simvastatine, which was vacuum dried. A cross section of same bead is shown in Fig IB.
  • Fig. 1C shows an electron microscope picture of a polymeric bead containing nanoparticles of simvastatine, which was air dried. A cross section of same bead is shown in Fig ID.
  • EXAMPLE 2 Reduction of gelling time and gelling ion concentration.
  • Emulsification Oil in water emulsion 20% oil phase fraction, 80% aqeous phase fraction was prepared, containing 3% w/w total surfactant (mixture of Tween 20and Span 20, HLB 10) concentration. 3.7869g of Simvastatine powder (Teva Pharmaceuticals, Israel) used as the poorly soluble drug was weighed and mixed with 90. lg toluene until complete dissolution of the drag is achieved. Final concentration of Simvastatine is 42 mg/g toluene . 1.04g Tween 20 was weighed and dissolved in 160.54g distilled water saturated with toluene (filtered through 0.2 ⁇ filter) .
  • Beads formation 75.3 g of sodium alginate solution (4%w/w) and 3.0g of silica 6 ⁇ A(Fratarom) were mixed together for about 10 min by a magnetic stirrer until the silica was dispersed homogeneously in the alginate solution. Then 75.2g of the above o/w emulsion were added and stirred together until homogenous mixture was achieved.
  • the alginate- emulsion mixture was introduced into an Innotech encapsulator, and jetted into 25mM CaCl 2 crosslinking solution .
  • the Innotech encapsulator allows tailoring the final size of the beads by selecting the proper instrument parameters. In this example, the parameters were: Nozzle size - 300 ⁇ m. Voltage - 1.005 Kv.
  • the beads were kept in the crosslinking solution for lOmin. Then, the beads were rinsed with about 2 liters of distilled water, filtered and air dried in an oven, at temperature of about 35 C for 48 hours, in order to remove the water and the volatile solvent.
  • Hexaglycerol sesquistearate was weighed and dissolved in 160.28g distilled water saturated with toluene (filtered through 0.2 ⁇ m filter) .
  • 2.02g Hexaglycerol sesquistearate was weighed and mixed with the 40.46g solution of 42mg/g Simvastatine in toluene, and stirred about lOmin together.
  • the organic phase was added carefully to the water phase and mixed for 5 min in an Ultra Turrax homogenizer at 8000 RPM. A coarse, homogeneous emulsion was obtained. This emulsion was introduced into a high-pressure homogenizer (Stansted), and was circulated through the high-pressure-homogenizer twice at 17,000 psi. Z-average particles size of the resulting emulsion was 126-140nm.
  • Beads formation 75.2g of sodium alginate solution (4%w/w) and 3.0g of Silica 60A (Frutarom ) were mixed together for about 10 min by a magnetic stirrer until the silica was dispersed homogeneously in the alginate solution. Then 75.5g of the above o/w emulsion were added and stirred together until homogenous mixture was achieved.
  • the alginate- emulsion mixture was introduced into an Innotech encapsulator , and jetted into 25mM CaCl 2 crosslinking solution .
  • the Innotech encapsulator allows tailoring the final size of the beads by selecting the proper instrument parameters. In this example, the parameters were: Nozzle size - 300 ⁇ m.
  • Dissolution tests Dissolution test was performed to the dried beads and the results are shown in Fig. 2, where samples 2 and 3 are the beads of the invention compared to commercial simvastatine.
  • Dissolution test parameters Instrument: Caleva 7 ST , Test method: USP II at 75rpm
  • Dissolution medium Citarate Buffer 0.1M pH-6.8
  • Assay Procedure UV at 239nm.
  • Dissolution test shows (see Fig. 2) the advantage of the beads of the invention, which uses hydrophilic polymer beads containing dispersed nanoparticles of simvastatine (water insoluble drug) by solvent evaporation upon commercial simvastatine particles. The overall dissolution rate of the beads containing dispersed nanoparticles is much faster than that of commercial drag particles.
  • Example 4 Solvent evaporation of nanoemulsion in conventional way
  • solvent evaporation was performed to the nanoemulsion before beads formation. This experiment prove the necessity of solvent evaporation after the beads formation in order to prevent crystal formation and growing of the lipophilic drag.
  • Emulsificatioii Oil in water emulsion 20% oil phase fraction, 80% aqueous phase fraction was prepared, containing 3% (w/w) total surfactant (mixture of Tween 20 and Span 20, HLB T0) concentration.
  • Span 20 was weighed and mixed with the 20.56g solution of 41mg/g Simvastatine in toluene, and stirred about lOmin together. The organic phase was added carefully to the water phase and mixed for 5 min in an Ultra Turrax homogenizer at 8000 RPM. A coarse, homogeneous emulsion was obtained. This emulsion was introduced into a high pressure homogenizer (Stansted), and was circulated through the high-pressure-homogenizer twice at 17,000 psi. Z-average particles size of the resulting emulsion was 186-198nm.
  • the organic solvent toluene was evaporated with Rotavapor (R-114 BUCHI) from the emulsion to form a dispersion of lipophilic drag in water.
  • the organic solvent evaporation was performed in four steps, water was added up to the initial weight after each step. After several hours, it was found that huge large crystals (needles) (crystal size: 0.5-2mm) of the raw material were formed (see Fig. 3) that indicate the instability of the drug nanoparticles that was formed after evaporation, while the evaporation is performed not within the polymeric bead..
  • EXAMPLE 5 Disintegrant effect on the beads Alginate beads are insoluble in water or acidic media.
  • a disintegrant was included in the drag formulation, which contains the beads.
  • the effect of disintegrant is demonstrated by experiments in which the beads were immersed in liquid containing the disintegrant.
  • the beads disintegration measurements were performed using turbidimeter (HACH RATIO/XR).
  • the turbidity values represent the beads disintegration. It is expected that the disintegration will enhance the drug release in the system. It should be emphasize that the beads cannot disintegrate without the presence of suitable disintegrating agents.
  • phosphate buffer concentrations in the range of 0.05M-0.25M, on the beads disintegration rate.
  • 0.05M phosphate buffer the beads were slightly disintegrated while in 0.25M phosphate buffer the beads were completely disintegrated within 10 mins.
  • Fig. 6 demonstrates the influence of citrate buffer concentrations, in the range of 0.05M-0.25M, on the beads disintegration rate. The beads were completely disintegrated within lOmins in all tested concentrations (0.05M-0.25M) of citrate buffer.
  • the citrate buffer is more efficient disintegrating agent than phosphate buffer and it disintegrate the beads in lower concentration.
  • the obtained order is influenced by several parameters such as: the cation valence, the cationic radius, and the ability of the disintegrating agent to competitive on the cation against the alginate polymer. It was found that by proper selection of disintegrants (type and concentration) and crosslinking (type and concentration) we can control the release rate of the drag.
  • EXAMPLE 6 microemulsions Microemulsions were prepared by mixing, without any special equipment - of the solvent (which contains the pre-dissolved drag molecule), the surfactant, co- surfactant and water, at proper composition according to the phase diagram. Than, the obtained microemulsion was mixed with alginate solutions, which upon contact with 2% CaCl 2 solution formed beads in which the microemulsion droplets were dispersed within. The last stage was drying the beads, which lead to formation of drag nanoparticles (size 10-50 nm) dispersed within the bead.
  • Beads formation 2.5%Alginate (type LF10/60) solution was mixed with 25%) of microemulsion having the composition: 9.1% Brij 96V (polyoxyethylene 10 oleyl ether surfactant) 81.8% Ethanol/Waterl : 1 9.1%) Limonene/Triglymel:l which contains the dissolved drag.
  • 2.5%Alginate (type LF 10/60) solution was mixed with 25% microemulsion having the composition: 8%> SDS (dodecyl sodium sulfate surfactant) % Water% BuAc/2-Propanoll:l containing the dissolved drug.

Abstract

The present invention provides a drug delivery system comprising nanoparticles or microparticles of a water poorly soluble drug dispersed in a polymeric bead containing essentially only of hydrophilic polymers (i.e. without hydrophobic polymers). The present invention further provides a method of producing the drug delivery system of the invention.

Description

FORMULATIONS FOR POORLY SOLUBLE DRUGS
FIELD OF THE INVENTION The present invention generally concerns formulations for drugs, and more particularly formulations for poorly soluble drugs.
BACKGROUND OF THE INVENTION Solubility is defined as the concentration of the solute in a saturated solution. The solubility of compounds varies in accordance with factors such as temperature, the type of solvent, the pH of the solution, and atmospheric pressure. The solubility of drugs found in the US Pharmacopeia is expressed as the number of milliliters of solvent in which one gram of solute can dissolve. Where the exact solubility of various compounds cannot be precisely determined general quality terms are used to describe the solubility of a specific compound, typically with reference to other compounds. Solubility may also be expressed in terms of molarity, percentage, and molality. Typically, drugs defined as "poorly soluble " are those that require more than 1 ml part of solvent per 10 mg of solute. Some poorly soluble drugs are further limited by their intrinsic bioavailability for example due to extensive first pass metabolism by the liverok (first pass effect), or further limited due to various drug-drug interactions . Usage of poorly soluble compounds has increased by 25% on average over the last five year period. The increase in formulations containing poorly soluble compounds is attributed to factors associated with both the pharmaceutical and biotechnology sectors. For example, within the pharmaceutical sector, drugs are now more frequently designed by combinatorial chemistry in order to improve their distribution through various tissues in the body, increase their half life, and improve their therapeutic index (more potency with low concentrations). Sometimes newly developed drugs produced by combinatorial techniques are poorly soluble as during development, and in contrast to rational drug design, solubility was never a factor considered for their production. In the biotechnology field, compounds, such as peptides, nucleic acid sequences, monoclonal antibodies, etc. resulting from biotechnological development are also typically poorly soluble. There are several different approaches to solve the problem of solubility of poorly soluble drugs. These include traditional solubilizing approaches using a combination of solvents, surfactants and co-solvents, various sophisticated dispersion systems, as well as novel technologies, including micronization, complexation and liposomal delivery. One approach directed to delivery and release of poorly soluble drags is their formulation as nano sized particles/crystals. U.S. Patent Application 20030215513 concerns release of substantially water insoluble nano-sized particles from a composition, by coating the pharmaceutical composition with a diffusion-control membranes that contains a multiplicity of pores and pore-forming substances. This establishes a diffusion gradient that enables mass-transport of nano-suspensions from the pharmaceutical composition through the pores, thereby resulting in a diffusion controlled release through the membrane. U.S. Patent Application 20020106403 discloses a water insoluble drug, in a nanometer or micrometer particulate solid format, which is surface stabilized by a phospholipid, being dispersed throughout a bulking matrix. This construction can dissolve upon contact with aqueous environments, thereby releasing the water insoluble particulate solid in an unaggregated or un-agglomerated form. Typically, the matrix is composed of water insoluble substance. U.S. Patent No. 5,439,686 discloses compositions for in vivo delivery of water insoluble pharmaceutical agents, notably the anticancer drag taxol, wherein the active agent is solubilized in a biocompatible dispersing agent contained within a protein walled shell. By another alternative, the protein walled shell can contain particles of the taxol itself. U.S. Patent No. 6,387,409 discloses nano- or micro-sized particles of water insoluble, or of poorly soluble drugs, produced by a combination of natural and synthetic phospholipids and charge surface modifiers such as highly purified charge phospholipids, together with a block copolymer which are coated or adhered on to the surfaces of water insoluble compound particles. These constructs enable the formation and stabilization of submicron and micron sized compound particles stabilized by the charge phospholipids which provides electrostatic stabilization; and stabilized by the block copolymer to provide steric stabilization. Such constructs prevent the particles from aggregation and flocculation. International Patent Application WO 9725028 concerns controlled release beads which comprise a core of insoluble drags, and a layer of furosemide dispersed in a hydrophilic polymer and a membrane which regulates the release of the furosemide in a controlled manner. U.S. Patent No. 6,645,528 concerns poorly soluble drags provided in a porous matrix form which enhances the dissolution of the drug in an aqueous media. The pore forming agent creating the porous matrix is typically a volatile liquid that is immiscible with the drag solvent, or alternatively, a volatile solid compound such as a volatile salt. The resulting porous matrix has a faster rate of dissolution following administration to a patient as compared to a non porous matrix form of the drag. Sustained, or controlled release drug delivery systems, include any drug delivery system that achieves a slow release of a drag over an extended period of time. The main aim of slow release systems is improved efficiency of treatment as a result of obtaining constant drug-blood levels, thus maintaining the desired therapeutic effect for extended periods of time. This results in reduction and elimination of fluctuations in blood levels, thus allowing better disease management. Some controlled release systems were not developed for the main purpose of sustained release, but rather having been developed in order to improve the bioavailability of drags, due to their activity in isolating the drags from the environment, for example by protecting drags susceptible to enzymatic inactivation or bacterial decomposition by encapsulation in polymeric systems. Microparticles containing poorly soluble drags and a polymer were prepared in order to overcome some technical problems of tabulating encountered during foπnulations of medicaments with microparticles. In these formulations propranonol was the poorly soluble drag, and the polymer was ethylcellulose. Together, the polymer and the poorly soluble drags were mixed to form microspheres containing a drug-polymer mixture, which were subsequently entrapped within a chitosan or calcium alginate beads. Thus the beads contained initially a mixture of drugs and insoluble polymers, subsequently mixed with a soluble polymer. The ionic characteristics of the polysaccharides of this delivery system allowed a pH-dependent release of the microparticles in the gastrointestinal tract Bodmeier et al. Pharmaceutical Research 6:5, 1989).
SUMMARY OF THE INVENTION The present invention is based on the realization that particles of water insoluble or poorly soluble drags can have improved solubility, and hence improved bioavailability, if they are administered dispersed in a hydrophilic polymeric bead in the fonn of nanoparticles or microparticles of the drag. Thus, by one aspect the present invention concerns a drag delivery system comprising nanoparticles or microparticles of a poorly soluble drag dispersed in a polymeric bead containing essentially only of hydrophilic polymers (i.e. without hydrophobic polymers). The tenn "nanoparticle " in the context of the drugs refers to particles which have the size of 3 nm to 900 nm, preferably 5 nm to 450 nm. Similarly, the term "microparticle" refers to particles which have the size of 1 to 500 micrometers. By a preferred embodiment, the polymeric beads consist essentially of a single hydrophilic polymer, this being in contrast to the publication of Bodmeier et al. wherein the poorly soluble drag is first entrapped within an insoluble, hydrophobic polymer, and the obtained microparticles of the insoluble polymer and drag are then mixed with a soluble polymer- forming bead. Therefore, by Bodmeier publication one obtains drag molecules entrapped within a water insoluble polymeric matrix, which leads to decreased solubility of the drag, and that would cause a decreased bioavailability. Against this, the beads of the present invention consist of drag nanoparticles essentially free of water insoluble polymer, while the single hydrophilic polymer serves as a former of porous bead, which prevents the increase in the size of the drag particle, and greatly simplifies the manner of production as will be explained hereinbelow. In addition, in accordance with one preferred embodiment of the invention, the bead formation process by itself leads to fonnation of the drag nanoparticles, which are formed from a nanoemulsion, in a way that overcomes the problems associated with conventional methods for preparation of nanoparticles by solvent evaporation from submicron emulsions. The beads themselves serve as the delivery system, having the ability of controlling the release of the nano/micro particles of the poorly soluble drugs therefrom. The control can be achieved by the inherent polymeric structure of the bead, or by a combination of the bead skeleton polymers and polymeric additives, mainly water soluble polymers. The term "drug delivery system " in the context of the present invention concerns active ingredient - i.e. the drag - in its carrier matrix. The drag delivery system in accordance with the invention may be used for subsequent preparation of dosage administration forms, for example, in the form of capsules (coated or uncoated), tablets (coated or uncoated), wherein the coating may be functional such as enteric coating, colonic delivery coating, chrono-therapeutic and controlled release coating, taste-masking coating and the like. The dosage form may be suitable for any mode of administration such as oral, rectal, depo-administration, parenteral, subcutaneous, ocular, nasal, vaginal and the like. The term "polymer" in accordance with the present invention shall be understood as referring both to a polymer composed of a single re-occurring building block (monomer) as well as to a polymer composed of two or more different polymeric units (co-polymer). The term "poorly soluble drug" refers to a drag which is insoluble or poorly soluble in an aqueous solution, and typically this refers to a drag which has a solubility of less than 10 mg/ml, and preferably less than about 5 mg/ml in aqueous media at approximately physiological temperature and pH. As used herein, the term "drug" refers to chemical and biological molecules having therapeutic, diagnostic or prophylactic effects in vivo. The term "drag" therefore may include food additives which have biological activity such as lycomene, lycopene and beta carotene. Drugs contemplated for use in the system described herein include the following categories and examples of drugs and alternative forms of these drags such as alternative salt forms, free acid forms, free base fonns, prodrag forms and solvates e.g. hydrates: Accupril (Quinapril), Accutane (Isotretinoin), Actos (Pioglitazone), AeroBid (Flunisolide), Agenerase (Amprenavir), Akinetron (Biperiden), Allegra (Fexofenadine), Aromasin (Exernestane), Asacol (Mesalamine), Atacand (Candesartan cilexetil), Avandia (Rosiglitazone), Azmacort (Triamcinolone), Biaxin (Clarithromycin), Camptosar (Irinotecan), Cefzon (Cfdinir), Celebrex (Celecoxib), Claritin (Loratadine), Clinoril (Sulindac), Cordarone (Amiodarone HCL), Diovan (Valsartan), Duragesic (Fentanyl citrate), DynaCirc (Isra apine), Elmiron (Pentosan polysulfate sodium), Elconon/Nasonex (Mometasone), Epogen/Procrit (EPO), Estratest (Methyltestosterone), Evista (Raloxifene hydrochloride), Fareston (Toremifene citrate), Flomax (Tamsulosin hydrochloride), Follistirn (Follitropin beta), Halcion (Triazolam), Hismanal (Astemizole), Hydergine LC (Ergoloid mesylates), Imodium (Loperamide), Invirase (Saquinavir), Lipitor (Atorvastatin Calcium), Luvox (Fluvoxamine), Mevacor (Lovastatin), Neoral and Sandimmune (Cyclosporine), Nitorol-R/Frandol (Isosorbide dinitrate), Noroxin (Norfloxacin), Norvir (Ritonavir), Pepcid (Famotidine), Platinol-AQ (Cisplatin), Plavix (Clopidrogel bisulfate), Plendil (Felodipine), Pletal (Cilostazol), Pulmicort TurbuhalerRhinocort (Budesonide). The drags may also include biological produced agents.such as proteins, protein f agments, peptides, nucleic acid sequences, oligonucleotides, glycoproteins as long as they are water insoluble Most preferable drags are simvastatine, statines, risperidone, carvedilol, carbamazepine, oxcarbazepine, zaleplon, galantamine, avandia, and poorly soluble anti psychotic, anti epileptic, anti parkinsonian and other indicated for CNS indications. The polymeric bead may comprise at least one of a polysaccharide polymer, a protein, a synthetic polymer which may be either crosslinked or not crosslinked or mixtures thereof. Examples of polysaccharide polymers are: alginates, chitosans, gellan gums, agarose, pectin, carrageenan. Examples of proteins are: gelatins, albumins, lactalbumin. Examples of synthetic polymers are polyacrylic acid, polyethylene glycol ("PEG"), polyvinyl pyrrolidone, polymethacrylates, polylysine, poloxamers, polyvinyl alcohol, polyethylene oxide, and polyethyoxazoline. Preferably, in accordance with the present invention, the nanoparticles or microparticles are in an amorphous state, which increases their solubility rate, and subsequent crystallization is prevented due to the presence of hydrophilic polymer and surfactants used in the process of production. Still more preferably, in accordance with the invention, the drag delivery system may include externally added crosslinking agents, which are, for anionic polyssacharides and synthetic polymers, multivalent cations, such as calcium, magnesium, barium, ferrous, polycations and cupper salts. For cationic polymers, such as chitosan, a polyvalent anion such as tripolyphosphate or anionic polymers may be used. It sould be noted that the polymeric beads may also be formed by heating -cooling effects, such as formation of gelatin beads , which is obtained by dropwise addition of warm gelation solution into cold liquid, water or oil. Still more preferably, the drag delivery system including said externally added crosslinking agents, further comprises a disintegrant which may be a chelator of the crosslinking cation, for example calcium or magnesium. Such chelators, in contact with water, interact with the crosslinking agents, thus breaking the crosslinking of the polymeric bead and enhancing the disintegration of the bead. Examples of disintegrants are EDTA, sodium citrate, citric acid, sodium dodecyl sulfate, phosphate salts and phosphate buffer saline. By using a disintegrate mixed with the polymer bead in the delivery system of the invention, it is possible on the one hand to improve the solubility of the poorly soluble drags by using the drag in the form of nanoparticles, and on the other hand to obtain rapid disintegration of the bead, for example in the gastrointestinal tract, in such a way that the drug nanoparticles are in close contact with the dissolution medium, without any barrier that could be formed by the crosslinked polymer. Such a construct which is unusual for polymeric beads, which typically are constructed without a disintegrant for sustained-release purposes, which results in drag particles that remain entrapped in the beads' core leading to slower dissolution rate and consequently to reduced bioavailability. Thus the present invention concerns a drug delivery system comprising an active ingredient dispersed within a polymeric bead, wherein the polymer may be crosslinked , while the crosslinking is achieved (in case of sodium alginate, for example) by a multivalent cation such as calcium, magnesium, barium, ferrous or copper salts and wherein the drug delivery system further comprises as a disintegrate, a chelator of the multivalent cation. Preferably, the drag is a poorly soluble drug, more preferably in the form of a nano-particle, a micro-particle, most preferably in the form of a nanoparticle. The present invention further concerns a method of producing the drug delivery system of the invention comprising: (i) providing poorly water soluble drug dissolved in organic volatile solvent, optionally in the presence of at least one surfactant; (ii) mixing the drag-containing solvent with an aqueous phase, optionally in the presence of at least one agent selected from surfactant, co-solvent and electrolyte, thereby producing an oil-in-water nanoemulsion or microemulsion; (iii) mixing the oil-in-water nano- or micro emulsion with water-soluble bead-forming polymers to produce a continuous phase of the emulsion which comprises the bead forming polymer; (iv) providing conditions enabling bead formation from the continuous phase of (iii); (v) drying of the beads, by evaporating the volatile organic solvent and the aqueous phase of the bead; thereby obtaining dry beads comprising in their matrix dispersed nanoparticles or microparticles of poorly water-soluble drags. The beads containing the drug nanoparticles or microparticles obtained by the method of the invention may be formulated to fonn a suitable dosage form, for example they may be packed within a capsule or a tablet, optionally together with a disintegrant as will be explained herein bellow, thus providing a delivery system of th© poorly soluble drag. Alternatively polymeric additives may be added in order to control the drag release. The poorly soluble drag is rendered in a nanoparticle form by consequent evaporation of the organic solvent and the water, thus the previously dissolved drug in the solvent droplets, becomes insoluble, and having a size similar to the initial size of the nanoemulsion droplets, and in most cases having a non-crystalline morphology. Since each nanoemulsion droplet is dispersed within the crosslined polymeric network of the bead, there is no possibility for coalescence of emulsion droplets, and therefore there is no increase in the size of drag particles which are maintained in their original nanoparticle size. In addition, since the evaporation of the solvent is rapid, and perfonned within a viscous, crosslinked polymeric network (which becomes more viscous as evaporation proceeds), the obtained drag nanoparticles are amorphous (not crystalline). Furthermore, due to the presence of the surfactants in the nanoemulsion the nanoparticles remain in an amorphic structure that brings significant advantages for enhanced dissolution and bioavailability. As will be shown in the examples, the processes described in this invention allow obtaining nanoparticles of drags, which otherwise, upon application of conventional solvent evaporation method, would have formed large crystals. It was surprisingly found that by performing the solvent evaporation process only after the beads are formed, the crystallization and increase of the size of the drag molecule could be prevented. The solvent used in the method of the invention is an organic solvent that is volatile (at the concentration used ) i.e. has a relatively low boiling point, or can be removed under vacuum, and which is acceptable for administration to humans in trace amounts. Representative solvents include, chloroform, chlorofluorocarbons, dichloromethane, dipropyl ether, diisopropyl ether, ethyl acetate, butyl acetate, methyl ethyl ketone (MEK), limonene, heptane, hexane, butanol, octane, pentane, toluene, 1,1,1-trichloroethane, 1,1,2-trichloroethylene, xylene, and combinations thereof. In general, the drug is dissolved in the volatile solvent to fonn a drag solution having a concentration of between 0.01 and 80% weight to volume (w/v). Alternatively, the solvent in which the drag is dissolved may contain a co-solvent which is either miscible or immiscible with water. Examples for co-solvents are: ethanol, isopropanol, pentanol THF, DMF, DMSO, propylene glycol, polyethylene glycol, glyme , diglyme , triglyme and the like. Examples of suitable surfactants are: nonionic surfactants such as for example block copolymers, e.g. Pluronic F 68, polyglycerol esters, alkyl glucosides ethoxylated sorbitan esters and ethoxylated sorbitan esters; ionic surfactants; and polymers such as polyvinyl alcohol, gelatin and BSA. The surfactants are selected from molecules acceptable for pharmaceutical preparations, which are capable of yielding nanoemulsions or microemulsions. The nanoemulsions can be formed by various methods, preferably by using a high pressure homogenization technology, or phase inversion methods (such as the PIT method) and the microemulsions are prepared by simple mixing of proper compositions of water, surfactants, solvents and co-solvents (microemulsions may form spontaneously, according the phase diagram of the compositions ). Additional exemplary surfactants which may be used include most physiologically acceptable emulsifiers, for instance egg lecithin or soya bean lecithin, or synthetic lecithins such as saturated synthetic lecithins, for example, dimyristoyl phosphatidyl choline, dipalmitoyl phosphatidyl choline or distearoyl phosphatidyl choline or unsaturated synthetic lecithins, such as dioleyl phosphatidyl choline or dilinoleyl phosphatidyl choline. Surfactants also include salts of fatty acids, esters of fatty acids with polyoxyalkylene compounds like polyoxpropylene glycol and polyoxyethylene glycol; ethers of fatty alcohols with polyoxyalkylene glycols; esters of fatty acids with polyoxyalkylated sorbitan; soaps; glycerol- polyalkylene stearate; glycerol-polyoxyethylene ricinoleate; homo- and co- polymers of poryalkylene glycols; polyethoxylated soya-oil and castor oil as well as hydrogenated derivatives; ethers and esters of sucrose or other carbohydrates with fatty acids, fatty alcohols, these being optionally polyoxyalkylated; mono-, di- and tri-glycerides of saturated or imsaturated fatty acids, glycerides of soya-oil and sucrose. Beads are formed by solidifying drops of solutions containing the bead forming polymers either by contact with a crosslinking agent (when the polymer can react with the crosslinking agent to form an insoluble polymeric structure), or by solidification, for examples while using a polymer such as gelatin, which forms a liquid solution at elevated temperature, and solidifies at room temperature. Thus, while the bead forming solution is added as small droplets through a suitable orifice, into a crosslinking solution or simply in a cold environment in case of temperature induced bead formation, immediate crosslinking (similar to solidification ) of the external part of the bead occurs, and therefore the external part of the droplets becomes solid. Upon further exposure to the crosslinking solution, the crosslinking ions migrate into the interior part of the bead, and form a solid matrix throughout the whole bead. The structure of the beads (porosity, rigidity etc.) can be tailored by proper selection of the bead formation conditions (such as crosslinker concentration, duration of crosslinking , presence of various electrolytes etc.). The size of the beads can be controlled by proper selection of the nozzle diameter and instrumentation from which the bead forming polymeric solution is ejected. Finally, as a last stage, the volatile (organic solvent) is evaporated together with the aqueous phase, for example by application of vacuum or by lyophilization processes, or by simply drying at room temperature or in an oven at elevated temperatures, to obtain the dry beads containing in their matrix dispersed nanoparticles of the poorly soluble drag. At the last preparation step, the beads are packed in a suitable pharmaceutical formulation such as gelatin capsule or solid tablet (containing conventional pharmaceutical excipients), and optionally containing agents which enhance the disintegration of the beads upon contact with body fluids. Such disintegrators can be molecules capable of replacing the crosslinking agent, such as chelators of the crosslinking agents such as EDTA, citric acid, sodium citrate, or surfactants such as sodium dodecyl sulfate, phosphate salts or phosphate buffer saline. Thus, when the polymeric beads are placed in an aqueous medium (such as in the gastrointestinal tract) water activates the disintegrating agent, causing it to chelate (for example in case the disintegrant is a chelator ) the crosslinkers (such as calcium ions), thereby disintegrating the beads and speeding up the release of the drag therefrom. Agents which modify the release, such as polymers may be added to the pharmaceutical dosage forms as well for decreasing rather then increasing, the release rate. Polymeric bead properties can be tailored to meet various requirements for proper drug dissolution as will be explained below.
BRIEF DESCRIPTION OF THE DRAWINGS In order to understand the invention and to see how it may be carried out in practice, some preferred embodiments will now be described, by way of non- limiting examples only, with reference to the accompanying drawings, in which: Fig. 1A shows an electron microscope picture of a polymeric bead containing nanoparticles of simvastatine, prepared as described in Example 1 which are vacuum dried; Fig IB shows an electron microscope picture of a cross section of the polymeric bead shown in Fig. 1 A. Fig. 1C shows an electron microscope picture of a polymeric bead containing nanoparticles of simvastatine, prepared as described in Example 1 which are air dried. Fig ID shows an electron microscope picture of a cross section of the polymeric bead shown in Fig. 1C. Fig. 2 shows the dissolution of two samples of beads of the invention containing simvastatine as compared to dissolution of commercial simvastatine. Fig. 3 shows an electron microscope picture of simvastatine crystals after solvent evaporation carried out without using bead formation. Fig. 4 shows electron microscope pictures of simvastatine nanoparticles after solvent evaporation from bead nanoemulsion systems. Fig. 5 shows the effect of varying concentrations of phosphate buffer (pH ~ 6.8) on beads disintegration. Fig. 6 shows the effect of varying concentrations of citrate buffer (pH ~ 6.8) on beads disintegration. Fig. 7 shows the effect of various crosslinking ions at a concentration of 25 mM on beads disintegration. Fig. 8 shows the effect of various crosslinking ions at a concentration of 100 mM on beads disintegration.
DETAILED DESCRIPTION OF THE INVENTION
Tailoring of the polymeric bead parameters: The following parameters may be varied when designing the drag delivery system of the present invention: 1) Droplets size in the nano/microemulsion may be tailored by controlling volatile solvent type, co- solvent type, surfactants and co-surfactant concentration and type, by controlling the cycles in high-pressure homogenizer (in case high pressure homogenization is utilized to obtain the nanoemulsions ), o/w ratio and temperature. 2) Type and molecular weight of the polysaccharide, (e.g. Alginate, K-Carrageenan, Chitosan, Gellan gum, Agarose, Pectin etc,) or synthetic polymers.
3) Structure of alginates (e.g. different ratio of guluronic and mannuronic acids).
4) Type and concentration of the crosslinking agent (also termed "gelling agent") ion solution (cation: Ca+2, Ba+2, AL+3,Fe+2, Cu+2, poly(amino acids) etc., and non-crosslinking ion (and Na+).
5) Crosslinking duration.
6) Matrix composition of material other than the bead forming polymer: other materials may be added, such as Silica, HPMC, Lactose, sodium chloride etc., which affect the morphology, porosity, size, and shrinkage of beads upon drying, disintegration rate and hydrophobicity.
7) The size of the polysaccharide beads can be controlled by controlling nozzle size, frequency, amplitude, velocity, physical parameters.
8) The rate of disintegration may be controlled by adding a disintegrate such as EDTA, phosphate or citrate ions, and controlling the amount of the disintegrant.
EXAMPLE 1: Solutions preparation:
4% Alginate solution: 16g of Alginic acid sodium salt (Sigma, low viscosity, 2% solution- 250cps) was dissolved in 400g distilled water (4% w/w), together with 0.4g of Bronopol (preserving material). The mixture was mixed on magnetic stirrer for about 48 hours and heated to about 37°C until complete dissolution.
lOOmM CaCl? solution (crosslinking agent) 14.8g of Dihydrate Calcium Chloride (Merck) was dissolved in lOOOg distilled water.
1. Emulsification Oil in water emulsion 20% oil phase fraction , 80% aqueous phase fraction was prepared, containing 3% w/w total surfactant (mixture of Tween 20, commercial name ofethoxylated sorbitan mono-laurate and Span 20, commercial name ofsorbitan monolaurate HLB=10) concentration. 3.3584g of Simvastatine powder (Teva Pharmaceuticals, Israel) used as the poorly soluble drag was weighed and mixed with 80. Og toluene until complete dissolution of the drag is achieved. Final concentration of Simvastatine is 42mg/g toluene. 1.02g Tween 20 was weighed and dissolved in 160.26g distilled water saturated with toluene (filtered through 0.2μm filter) . 4.97g Span 20 was weighed and mixed with the 40.23g solution of 42mg/g Simvastatine in toluene, and stirred about lOmin together. The organic phase was added carefully to the water phase and mixed for 5 min in an Ultra Turrax homogenizer at 8000 RPM. A coarse, homogeneous emulsion was obtained. This emulsion was introduced into a high pressure homogenizer (Stansted), and was circulated through the high-pressure-homogenizer twice at 17,000 psi. Z-average particles size of the resulting emulsion was 250-255nm.
2. Beads formation: 95. lg of sodium alginate solution (4% w/w) and 3.8g of Silica 6θA Frutarom) used to prevent shrinking upon drying, were mixed together for about 10 min by a magnetic stirrer until the silica was dispersed homogeneously in the alginate solution. Then 95. lg of the above o/w emulsion were added and stirred together until homogenous mixture was achieved. The alginate- emulsion mixture was introduced into an Innotech encapsulator, and jetted into lOOmM CaCl2 crosslinking solution. The Innotech encapsulator allows tailoring the final size of the beads by selecting the proper instrument parameters. In this example, the parameters were: Nozzle size - 300μm. Voltage - 0.914 Kv. Amplitude - 3. Frequency -1550 Hz. Pressure —0.4 bar. The beads were kept in the crosslinking solution for 30min. Then, the beads were rinsed with about 2 liters of distilled water, filtered and air dried in an oven, at temperature of about 35°C for 48 hours, in order to remove the water and the volatile solvent. The final result was dry beads in the size range of less than 1 mm in which nanoparticles of Simvastatine were dispersed, as verified by electron microscopy and shown in Fig. 1. Fig. 1A shows an electron microscope picture of a polymeric bead containing nanoparticles of simvastatine, which was vacuum dried. A cross section of same bead is shown in Fig IB. Fig. 1C shows an electron microscope picture of a polymeric bead containing nanoparticles of simvastatine, which was air dried. A cross section of same bead is shown in Fig ID. EXAMPLE 2: Reduction of gelling time and gelling ion concentration.
Solutions preparations:
4% Alginate solution: Was prepared as described in Example 1.
25mM CaCl? solution (crosslinking agent) 3.7g of Dihydrate Calcium Chloride (Merck) was dissolved in lOOOg distilled water.
1. Emulsification Oil in water emulsion 20% oil phase fraction, 80% aqeous phase fraction was prepared, containing 3% w/w total surfactant (mixture of Tween 20and Span 20, HLB=10) concentration. 3.7869g of Simvastatine powder (Teva Pharmaceuticals, Israel) used as the poorly soluble drug was weighed and mixed with 90. lg toluene until complete dissolution of the drag is achieved. Final concentration of Simvastatine is 42 mg/g toluene . 1.04g Tween 20 was weighed and dissolved in 160.54g distilled water saturated with toluene (filtered through 0.2 μ filter) . 4.97g span 20 was weighed and mixed with the 40.55g solution of 42mg/g Simvastatine in toluene, and stirred about lOmin together. The organic phase was added carefully to the water phase and mixed for 5 min in an Ultra Turrax homogenizer at 8000 RPM. A coarse, homogeneous emulsion was obtained. This emulsion was introduced into a high pressure homogenizer (Stansted), and was circulated through the high-pressure-homogenizer twice at 17,000 psi. Z-average particles size of the resulting emulsion was 194 -210nm. 2. Beads formation: 75.3 g of sodium alginate solution (4%w/w) and 3.0g of silica 6θA(Fratarom) were mixed together for about 10 min by a magnetic stirrer until the silica was dispersed homogeneously in the alginate solution. Then 75.2g of the above o/w emulsion were added and stirred together until homogenous mixture was achieved. The alginate- emulsion mixture was introduced into an Innotech encapsulator, and jetted into 25mM CaCl2 crosslinking solution . The Innotech encapsulator allows tailoring the final size of the beads by selecting the proper instrument parameters. In this example, the parameters were: Nozzle size - 300μm. Voltage - 1.005 Kv. Amplitude - 3. Frequency -1527 Hz. Pressure -0.3 bar. The beads were kept in the crosslinking solution for lOmin. Then, the beads were rinsed with about 2 liters of distilled water, filtered and air dried in an oven, at temperature of about 35 C for 48 hours, in order to remove the water and the volatile solvent.
EXAMPLE 3: Alteration of surfactant
Solutions preparations:
4% Alginate solution: Was prepared as described in Example 1.
25mM CaCl? solution (crosslinking agent) - Was prepared as described in Example 2. 1. Emulsification Oil in water emulsion 20% oil phase fraction , 80% aqeous phase fraction was prepared, containing 3% (w/w) total surfactant (Hexaglycerol sesquistearate, SY-GLYSTER SS-5S, SAKAMOTO YAKUHIN KOGYO CO., LTD. HLB=9.9) concentration. 3.7807g of Simvastatine powder (Teva Pharmaceuticals, Israel), used as the poorly soluble drug was weighed and mixed with 90. lg toluene until complete dissolution of the drag is achieved. Final concentration of Simvastatine is 42mg/g toluene . 4.02g Hexaglycerol sesquistearate was weighed and dissolved in 160.28g distilled water saturated with toluene (filtered through 0.2μm filter) . 2.02g Hexaglycerol sesquistearate was weighed and mixed with the 40.46g solution of 42mg/g Simvastatine in toluene, and stirred about lOmin together. The organic phase was added carefully to the water phase and mixed for 5 min in an Ultra Turrax homogenizer at 8000 RPM. A coarse, homogeneous emulsion was obtained. This emulsion was introduced into a high-pressure homogenizer (Stansted), and was circulated through the high-pressure-homogenizer twice at 17,000 psi. Z-average particles size of the resulting emulsion was 126-140nm.
2. Beads formation: 75.2g of sodium alginate solution (4%w/w) and 3.0g of Silica 60A (Frutarom ) were mixed together for about 10 min by a magnetic stirrer until the silica was dispersed homogeneously in the alginate solution. Then 75.5g of the above o/w emulsion were added and stirred together until homogenous mixture was achieved. The alginate- emulsion mixture was introduced into an Innotech encapsulator , and jetted into 25mM CaCl2 crosslinking solution . The Innotech encapsulator allows tailoring the final size of the beads by selecting the proper instrument parameters. In this example, the parameters were: Nozzle size - 300μm. Voltage - 1.005 Kv. Amplitude - 3. Frequency -1527 Hz. Pressure -0.3 bar. The beads were kept in the crosslinking solution for lOmin. Then, the beads were rinsed with about 2 liters of distilled water, filtered and air dried in an oven, at temperature of about 35 C for 48 hours, in order to remove the water and the volatile solvent.
Dissolution tests Dissolution test was performed to the dried beads and the results are shown in Fig. 2, where samples 2 and 3 are the beads of the invention compared to commercial simvastatine. Dissolution test parameters: Instrument: Caleva 7 ST , Test method: USP II at 75rpm Dissolution medium: Citarate Buffer 0.1M pH-6.8 Assay Procedure: UV at 239nm. Dissolution test shows (see Fig. 2) the advantage of the beads of the invention, which uses hydrophilic polymer beads containing dispersed nanoparticles of simvastatine (water insoluble drug) by solvent evaporation upon commercial simvastatine particles. The overall dissolution rate of the beads containing dispersed nanoparticles is much faster than that of commercial drag particles. Using beads nanoparticles system enable tailoring of release kinetics. The dried resulting beads can be inserted to capsules or compressed to tablets. Example 4: Solvent evaporation of nanoemulsion in conventional way In this example solvent evaporation was performed to the nanoemulsion before beads formation. This experiment prove the necessity of solvent evaporation after the beads formation in order to prevent crystal formation and growing of the lipophilic drag.
1. Emulsificatioii Oil in water emulsion 20% oil phase fraction, 80% aqueous phase fraction was prepared, containing 3% (w/w) total surfactant (mixture of Tween 20 and Span 20, HLB=T0) concentration. 2.523 lg of Simvastatine powder (Teva Pharmaceuticals, Israel) used as the poorly soluble drag was weighed and mixed with 61.7g toluene until complete dissolution of the drug is achieved. Final concentration of Simvastatine is 41mg/g toluene. 0.5 lg Tween 20 was weighed and dissolved in 80.26g distilled water saturated with toluene (filtered through 0.2μm filter). 2.49g Span 20 was weighed and mixed with the 20.56g solution of 41mg/g Simvastatine in toluene, and stirred about lOmin together. The organic phase was added carefully to the water phase and mixed for 5 min in an Ultra Turrax homogenizer at 8000 RPM. A coarse, homogeneous emulsion was obtained. This emulsion was introduced into a high pressure homogenizer (Stansted), and was circulated through the high-pressure-homogenizer twice at 17,000 psi. Z-average particles size of the resulting emulsion was 186-198nm. The organic solvent (toluene) was evaporated with Rotavapor (R-114 BUCHI) from the emulsion to form a dispersion of lipophilic drag in water. The organic solvent evaporation was performed in four steps, water was added up to the initial weight after each step. After several hours, it was found that huge large crystals (needles) (crystal size: 0.5-2mm) of the raw material were formed (see Fig. 3) that indicate the instability of the drug nanoparticles that was formed after evaporation, while the evaporation is performed not within the polymeric bead.. Against this, when the solvent evaporation was performed after the beads formation, the simvastatine remain as nanoparticles while performing the evaporation without beads forms large crystals of simvastatine (see Fig. 4). These experiments prove the necessity of solvent evaporation after the beads formation in order to prevent forming and growing of the drag crystals, which significantly reduce the bioavailability of the poorly soluble drag.
EXAMPLE 5: Disintegrant effect on the beads Alginate beads are insoluble in water or acidic media. In order to enable the disintegration of the drug uptake, a disintegrant was included in the drag formulation, which contains the beads. The effect of disintegrant is demonstrated by experiments in which the beads were immersed in liquid containing the disintegrant. The beads disintegration measurements were performed using turbidimeter (HACH RATIO/XR). The turbidity values represent the beads disintegration. It is expected that the disintegration will enhance the drug release in the system. It should be emphasize that the beads cannot disintegrate without the presence of suitable disintegrating agents. Fig. 5 demonstrates the influence of phosphate buffer concentrations, in the range of 0.05M-0.25M, on the beads disintegration rate. In 0.05M phosphate buffer the beads were slightly disintegrated while in 0.25M phosphate buffer the beads were completely disintegrated within 10 mins. Fig. 6 demonstrates the influence of citrate buffer concentrations, in the range of 0.05M-0.25M, on the beads disintegration rate. The beads were completely disintegrated within lOmins in all tested concentrations (0.05M-0.25M) of citrate buffer. The citrate buffer is more efficient disintegrating agent than phosphate buffer and it disintegrate the beads in lower concentration. In addition to the examination of disintegrating agents (which is in the external phase) on the beads disintegration, the influence of various crosslinking ions (Ca+2, Ba+2, Fe+3, Zn +2 and Co+2) in two different concentrations (which are added in the bead formation process) on the beads disintegration was determined. Figs. 7 and 8 demonstrate the influence of different crosslinking cation on the beads disintegration. It was found that the beads disintegration depends on the crosslinking ion according to the following order: Ca > Zn > Fe > Co > Ba . The obtained order is influenced by several parameters such as: the cation valence, the cationic radius, and the ability of the disintegrating agent to competitive on the cation against the alginate polymer. It was found that by proper selection of disintegrants (type and concentration) and crosslinking (type and concentration) we can control the release rate of the drag.
EXAMPLE 6: microemulsions Microemulsions were prepared by mixing, without any special equipment - of the solvent (which contains the pre-dissolved drag molecule), the surfactant, co- surfactant and water, at proper composition according to the phase diagram. Than, the obtained microemulsion was mixed with alginate solutions, which upon contact with 2% CaCl2 solution formed beads in which the microemulsion droplets were dispersed within. The last stage was drying the beads, which lead to formation of drag nanoparticles (size 10-50 nm) dispersed within the bead. Beads formation: 2.5%Alginate (type LF10/60) solution was mixed with 25%) of microemulsion having the composition: 9.1% Brij 96V (polyoxyethylene 10 oleyl ether surfactant) 81.8% Ethanol/Waterl : 1 9.1%) Limonene/Triglymel:l which contains the dissolved drag. In an alternative procedure: 2.5%Alginate (type LF 10/60) solution was mixed with 25% microemulsion having the composition: 8%> SDS (dodecyl sodium sulfate surfactant) % Water% BuAc/2-Propanoll:l containing the dissolved drug.

Claims

CLAIMS:
1. A drag delivery system comprising nanoparticles or microparticles of a poorly soluble drag dispersed in a polymeric hydrophilic bead.
2. A drug according to Claim 1, wherein the polymeric bead consists essentially of a single species of hydrophilic polymer.
3. A drag delivery system according to Claim 2, wherein the polymeric bead is selected from: a polysaccharide polymer, a synthetic polymer, and a protein.
4. A drag delivery system according to Claim 3, wherein the polysaccharide is selected from: alginates, chitosans, gellan gum, agarose, pectin and carrageenan.
5. A drag delivery system according to Claim 3, wherein the synthetic polymer is polyacrylic acid sodium salt
6. A drag delivery system according to Claim 3, wherein the protein is selected from gelatins, albumins and lactalbumins.
7. A drag delivery system according to any one of the preceding claims, wherein the poorly soluble drag is selected from: simvastatine, statines, risperidone, carvedilol, carbamazepine, oxcarbazepine, zaleplon, galantamine, anti Alzheimer, anti epileptic, anti parkinsonian, and other used for CNS indications.
8. A drag delivery system according to Claim 1, wherein the nanoparticles are in an amorphous, non crystalline state which enhances dissolution of the drag.
9. A drag delivery system according to Claim 1, further comprising a crosslinker.
10. A drag delivery system according to Claim 9, wherein the crosslinker is a multivalent cation.
11. A drag delivery system according to claim 10, wherein the multivalent cation is selected from: calcium, barium, iron, magnesium and copper.
12. A drag delivery system according to Claim 1, further comprising a disintegrate mixed with the beads.
13. A drag delivery system according to Claim 11 and 12, wherein the disintegrate is capable of breaking the crosslinking by replacing or chelation of the crosslinking multivalent cation.
14. A drag delivery system according to Claim 11 and 13, wherein the disintegrate is a calcium chelator.
15. A drag delivery system according to Claim 14, wherein the calcium chelator is selected from EDTA, sodium citrate and citric acid.
16. A drag delivery system according to Claim 14, wherein the disintegrant is selected from sodium dodecyl sulfate, and phosphate buffer saline.
17. A drag delivery system according to claim lwherein the beads are formed without a cross linker.
18. A drag delivery system according to claim 1 wherein the beads are gelatin beads.
19. A drag delivery system according to any one of the preceding claims in a dosage form selected from: coated capsules, non-coated capsules, hard gelatin capsules, coated and non-coated tablets, suppositories, liquid suspensions for oral or parenteral administration and formulations for topical applications.
20. A drag delivery system according to Claim 19 in the form of coated hard gelatin capsules or coated tablets wherein the coating is selected from enteric coating, coating for colonic delivery and taste masking coating.
21. A drug delivery system comprising an active ingredient dispersed within a crosslinked polymeric bead wherein the crosslinking is by a cation selected from calcium, iron, magnesium and copper and wherein the drag delivery system further comprises as a disintegrant a chelator of calcium.
22. A drag delivery system according to Claim 21, wherein the active ingredient is a poorly soluble drag.
23. A drag delivery system according to Claim 21, wherein the active ingredient is a food additive.
24. A drag delivery system according to Claim 22, wherein the poorly soluble drag is in the form of nanoparticles.
25. A method for producing the drag delivery system of Claim 1 , comprising: (i) providing poorly water soluble drag dissolved in organic volatile solvent or mixture of organic volatile solvent with co-solvent that is either miscible or immiscible with water, optionally in • the presence of at least one surfactant; (ii) mixing the drag containing solvent with an aqueous phase comprising at least one surfactant and optionally co-solvent and other emulsification aids at such conditions in which an oil-in-water nanoemulsion or microemulsion is foraied; (iii) mixing the oil-in-water nanoemulsion or microemulsion with water-soluble bead fonning polymers to produce a continuous phase of the emulsion which is capable of forming a bead; (iv) providing conditions enabling bead fonnation from the continuous phase of (iii) containing nano- microemulsion droplets; (v) optionally evaporating the volatile organic solvent and the water, thereby obtaining dry beads containing in the polymeric bead dispersed nanoparticles of poorly water soluble drags.
26. A method according to Claim 25, wherein the mixing of the poorly water soluble drag in an organic solvent occurs in the presence of at least one surfactant.
27. A method according to Claim 25, wherein the drag containing solvent is mixed within an aqueous phase containing a surfactant, the aqueous phase further containing a co-surfactant and/or co-solvent, and/or electrolytes.
28. A method according to Claim 25, wherein the volatile organic solvent is selected from: toluene, butyl acetate, ethyl acetate, methylene chloride, chloroform, limonene and their mixtures.
29. A method according to Claim 25, wherein the evaporation is carried out by application of air, vacuum, by a lyophilizing process, or by drying under normal room conditions.
30. A method according to Claim 25, wherein the nanoemulsion is prepared by homogenization by a high pressure homogenizer or by a phase inversion method.
31. A method according to Claim 25, wherein the microemulsion is formed spontaneously by proper selection of the surfactants, solvent, co-solvent and co- surfactants.
32. A method according to Claim 25, wherein at step (iv) the beads are incubated under suitable conditions and for suitable periods of time, with external crosslinking agents.
33. A method according to Claim 32, wherein the polymer is an anionic polymer and external crosslinkers are multivalent cations selected from calcium, magnesium, copper, iron, barium and salts of these cations.
34. A method according to Claim 32, wherein the polymer is a cation polymer and external crosslinkers are polyvalent anions selected from polyanions or sodium tripolyphosphate.
35. A method for producing a pharmaceutical composition comprising packing the beads obtained in Claim 25 within a capsule or tablet.
36. A method according to Claim 35, wherein disintegrator is added to the dry beads prior to packing the beads in a capsule or tablet.
37. A method according to Claim 36, wherein the disintegrator is selected from chelators and molecules capable of replacing the crosslinking ions.
38. A method according to Claim 37, wherein the disintegrants are selected from: sodium citrate, citric acid EDTA, sodium dodecyl sulphate, phosphate salts, and phosphate buffer saline.
39. A method according to claim 35 further comprising adding other polymers internally to the beads to improved permeability, disintegration or release properties and hence dissolution properties of the drug.
40. A method according to claim 35 further comprising adding other polymers and surfactants externally to the beads to improved permeability, disintegration or release properties and hence dissolution properties of the drag.
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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2006061827A1 (en) * 2004-12-06 2006-06-15 Nutralease Ltd. Microemulsion comprising carbamazepine having solubility
WO2007133758A1 (en) * 2006-05-15 2007-11-22 Physical Pharmaceutica, Llc Composition and improved method for preparation of small particles
WO2007144894A1 (en) * 2006-06-15 2007-12-21 Yissum Research Development Company Of The Hebrew University Of Jerusalem Hydrocolloid carrier beads with inert filler material
WO2008032328A2 (en) 2006-09-14 2008-03-20 Yissum Research Development Company Of The Hebrew University Of Jerusalem Pesticide nanoparticles obtained from microemulsions and nanoemulsions
WO2008032327A2 (en) * 2006-09-14 2008-03-20 Yissum Research Development Company Of The Hebrew University Of Jerusalem Organic nanoparticles obtained from microemulsions by solvent evaporation
WO2008006715A3 (en) * 2006-07-13 2008-05-08 Unilever Plc Improvements relating to pharmaceutical compositions
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WO2010026993A1 (en) 2008-09-03 2010-03-11 武田薬品工業株式会社 Method for improving absorbability of preparation, and preparation having improved absorbability
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EP2266546A1 (en) 2009-06-08 2010-12-29 Advancell Advanced in Vitro Cell Technologies,S.A. Process for the preparation of colloidal systems for the delivery of active compounds
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WO2011063775A2 (en) 2009-11-25 2011-06-03 Zentiva, K.S. Pectin complexes of sartans and pharmaceutical compositions based thereon
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US9018193B2 (en) 2010-09-13 2015-04-28 Bev-Rx, Inc. Aqueous drug delivery system
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US11865206B2 (en) 2018-09-05 2024-01-09 Emcure Pharmaceuticals Ltd Stable ready-to-use carmustine pharmaceutical composition

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* Cited by examiner, † Cited by third party
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Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1997025028A1 (en) * 1996-01-05 1997-07-17 Astra Aktiebolag (Publ) Pharmaceutical controlled release beads comprising furosemide and a formulation containing said controlled release beads
WO2000071079A2 (en) * 1999-05-21 2000-11-30 American Bioscience, Inc. Protein stabilized pharmacologically active agents, methods for the preparation thereof and methods for the use thereof
WO2001030145A1 (en) * 1999-10-22 2001-05-03 3M Innovative Properties Company Active material within hydrogel microbeads
US6238705B1 (en) * 1995-07-28 2001-05-29 Advanced Poymer Systems, Inc. Bioerodible porous compositions
US20020119125A1 (en) * 1999-06-16 2002-08-29 Yilong Xue Microencapsulated pheochromocyte of ox adrenal medulla as medicine for curing pain
WO2003018186A1 (en) * 2001-08-23 2003-03-06 Bio-Dar Ltd. Stable coated microcapsules
WO2005030257A2 (en) * 2003-09-30 2005-04-07 Solubest Ltd. Water soluble nanoparticles inclusion complexes

Family Cites Families (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5985354A (en) * 1995-06-07 1999-11-16 Brown University Research Foundation Preparation of multiwall polymeric microcapsules from hydrophilic polymers
US6471994B1 (en) * 1995-01-09 2002-10-29 Edward Mendell Co., Inc. Pharmaceutical excipient having improved compressibility
US8137684B2 (en) * 1996-10-01 2012-03-20 Abraxis Bioscience, Llc Formulations of pharmacological agents, methods for the preparation thereof and methods for the use thereof
US5904927A (en) * 1997-03-14 1999-05-18 Northeastern University Drug delivery using pH-sensitive semi-interpenetrating network hydrogels
US6146671A (en) * 1999-05-08 2000-11-14 Kemin Industries, Inc. Method and protecting heat-or oxygen-labile compounds to preserve activity and bioavailability
US6495161B1 (en) * 1999-03-09 2002-12-17 Vivorx, Inc. Cytoprotective biocompatible containment systems for biologically active materials and methods of making same
US20030157171A1 (en) * 2000-07-07 2003-08-21 Esteban Chornet Drug delivery system for poorly water soluble drugs
GB2388581A (en) * 2003-08-22 2003-11-19 Danisco Coated aqueous beads
AU2005219443A1 (en) * 2004-03-03 2005-09-15 Spherics, Inc. Polymeric drug delivery system for hydrophobic drugs

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6238705B1 (en) * 1995-07-28 2001-05-29 Advanced Poymer Systems, Inc. Bioerodible porous compositions
WO1997025028A1 (en) * 1996-01-05 1997-07-17 Astra Aktiebolag (Publ) Pharmaceutical controlled release beads comprising furosemide and a formulation containing said controlled release beads
WO2000071079A2 (en) * 1999-05-21 2000-11-30 American Bioscience, Inc. Protein stabilized pharmacologically active agents, methods for the preparation thereof and methods for the use thereof
US20020119125A1 (en) * 1999-06-16 2002-08-29 Yilong Xue Microencapsulated pheochromocyte of ox adrenal medulla as medicine for curing pain
WO2001030145A1 (en) * 1999-10-22 2001-05-03 3M Innovative Properties Company Active material within hydrogel microbeads
WO2003018186A1 (en) * 2001-08-23 2003-03-06 Bio-Dar Ltd. Stable coated microcapsules
WO2005030257A2 (en) * 2003-09-30 2005-04-07 Solubest Ltd. Water soluble nanoparticles inclusion complexes

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See also references of EP1708682A2 *

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WO2006061827A1 (en) * 2004-12-06 2006-06-15 Nutralease Ltd. Microemulsion comprising carbamazepine having solubility
WO2007133758A1 (en) * 2006-05-15 2007-11-22 Physical Pharmaceutica, Llc Composition and improved method for preparation of small particles
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US8932634B2 (en) 2006-06-15 2015-01-13 Yissum Research Development Company Of The Hebrew University Of Jerusalem Ltd Hydrocolloid carrier beads with inert filler material
US8821932B2 (en) 2006-07-13 2014-09-02 David John Duncalf Pharmaceutical compositions
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US9060937B2 (en) 2006-07-13 2015-06-23 David John Duncalf Pharmaceutical compositions
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US10568860B2 (en) 2006-08-30 2020-02-25 Kowa Co., Ltd. Pharmaceutical composition containing statin-encapsulated nanoparticle
KR101455413B1 (en) * 2006-08-30 2014-10-27 고쿠리쓰다이가쿠호진 규슈다이가쿠 Pharmaceutical composition containing statin-encapsulated nanoparticle
WO2008032327A3 (en) * 2006-09-14 2008-05-02 Yissum Res Dev Co Organic nanoparticles obtained from microemulsions by solvent evaporation
US9095133B2 (en) 2006-09-14 2015-08-04 Yissum Research Development Company Of The Hebrew University Of Jerusalem Pesticide nanoparticles obtained from microemulsions and nanoemulsions
US20110021592A1 (en) * 2006-09-14 2011-01-27 Shlomo Magdassi Organic nanoparticles obtained from microemulsions by solvent evaporation
WO2008032327A2 (en) * 2006-09-14 2008-03-20 Yissum Research Development Company Of The Hebrew University Of Jerusalem Organic nanoparticles obtained from microemulsions by solvent evaporation
WO2008032328A2 (en) 2006-09-14 2008-03-20 Yissum Research Development Company Of The Hebrew University Of Jerusalem Pesticide nanoparticles obtained from microemulsions and nanoemulsions
CN101322712B (en) * 2007-06-14 2010-10-13 沈阳市万嘉生物技术研究所 Alprostadil nano granule formulation and preparation thereof
WO2009095448A1 (en) * 2008-01-30 2009-08-06 Lek Pharmaceuticals D.D. Preparation of nanoparticles by using a vibrating nozzle device
EP2085080A1 (en) * 2008-01-30 2009-08-05 LEK Pharmaceuticals D.D. Preparation of nanoparticles by using a vibrating nozzle device
WO2010026993A1 (en) 2008-09-03 2010-03-11 武田薬品工業株式会社 Method for improving absorbability of preparation, and preparation having improved absorbability
US9089152B2 (en) 2009-04-13 2015-07-28 TEAGASC—Agriculture and Food Development Authority Method of producing microbeads
WO2010119041A3 (en) * 2009-04-13 2011-01-13 Agriculture And Food Development Authority (Teagasc) Method of producing microbeads
EP2266546A1 (en) 2009-06-08 2010-12-29 Advancell Advanced in Vitro Cell Technologies,S.A. Process for the preparation of colloidal systems for the delivery of active compounds
WO2011063774A2 (en) 2009-11-25 2011-06-03 Zentiva, K.S. Pectin complexes of steroids and pharmaceutical compositions based thereon
WO2011063775A2 (en) 2009-11-25 2011-06-03 Zentiva, K.S. Pectin complexes of sartans and pharmaceutical compositions based thereon
WO2011136376A1 (en) 2010-04-30 2011-11-03 武田薬品工業株式会社 Enteric tablet
WO2011136373A1 (en) 2010-04-30 2011-11-03 武田薬品工業株式会社 Enteric tablet
US9018193B2 (en) 2010-09-13 2015-04-28 Bev-Rx, Inc. Aqueous drug delivery system
US9789191B2 (en) 2010-09-13 2017-10-17 Solixa Technologies, Inc. Aqueous drug delivery system
WO2012130567A1 (en) * 2011-03-29 2012-10-04 Beta-Cell Nv Method for encapsulated therapeutic products and uses thereof
CN106999429A (en) * 2014-11-19 2017-08-01 Fmc有限公司 Nanoparticle suspension agent formulation
EP3220894A4 (en) * 2014-11-19 2018-06-06 FMC Corporation Nanosuspension formulation
CN106999429B (en) * 2014-11-19 2020-09-18 杜邦营养美国有限公司 Nanosuspension formulations
EP3474826A4 (en) * 2016-06-27 2020-03-04 Tamarisk Technologies Group, LLC Pharmaceutical preparation for delivery of peptides and proteins
WO2019193477A1 (en) * 2018-04-05 2019-10-10 Emcure Pharmaceuticals Limited Carmustine formulation
US11865206B2 (en) 2018-09-05 2024-01-09 Emcure Pharmaceuticals Ltd Stable ready-to-use carmustine pharmaceutical composition

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WO2005072709A3 (en) 2006-04-20
US20080038333A1 (en) 2008-02-14
IL160095A0 (en) 2004-06-20

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