Classifications

• • 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
  • A61K9/141—Intimate drug-carrier mixtures characterised by the carrier, e.g. ordered mixtures, adsorbates, solid solutions, eutectica, co-dried, co-solubilised, co-kneaded, co-milled, co-ground products, co-precipitates, co-evaporates, co-extrudates, co-melts; Drug nanoparticles with adsorbed surface modifiers
  • A61K9/145—Intimate drug-carrier mixtures characterised by the carrier, e.g. ordered mixtures, adsorbates, solid solutions, eutectica, co-dried, co-solubilised, co-kneaded, co-milled, co-ground products, co-precipitates, co-evaporates, co-extrudates, co-melts; Drug nanoparticles with adsorbed surface modifiers with organic compounds
• • 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
  • A61K9/1694—Processes resulting in granules or microspheres of the matrix type containing more than 5% of excipient

Definitions

• the present invention relates to pharmaceutical formulations involving the inclusion of an active pharmaceutical ingredient ("API") in a pharmaceuticaUy- acceptable single crystal matrix. More particularly, the crystals contain growth- sector specific, oriented inclusions of active pharmaceutical ingredients which are isolated. The active pharmaceutical ingredients have higher stability and shelf-life, and can be delivered in conventional dosage forms.
• This invention has general application to active pharmaceutical ingredients, and in one aspect has particular application to biopharmaceuticals.
• biopharmaceuticals is used to refer to a subset of API's which are polymeric in nature, including for example, proteins, polypeptides, enzymes, immunoglobulins, polynucleic acids, and plasmids. Description of the Prior Art:
• compositions which are capable of maintaining the quality and efficacy of the API during storage and delivery.
• the loss of potency of an API is a critical concern in assuring that viable, effective drugs are delivered to patients. It is similarly desirable to have formulations which do not require special packaging or handling. FuVther, it remains a constant goal to provide active pharmaceutical ingredients in a form which facilitates their use by the consumer, such as through convenient dosage forms.
• the present invention addresses these and other issues concerning pharmaceutical compositions and formulations.
• biopharmaceuticals Although not limited to biopharmaceuticals, the usefulness of the present invention is well exemplified with respect to biopharmaceuticals, many of which demonstrate the problems encountered in prior-art approaches. Ensuring long-term stability and maintaining activity of biopharmaceuticals is a prevalent concern.
• the chemical complexity and conformational fragility of protein drugs make them highly susceptible to both physical and chemical instabilities and threaten their emergence into the marketplace. Denaturation, adsorption with container walls, aggregation, and precipitation can result from non-covalent interactions between a drug and its environment. Insulin, for instance, has been shown to adsorb onto the surfaces of glass and plastic containers, and to have interactions at air-water interfaces, leading to denaturation, aggregation and precipitation.
• HGH human growth hormone
• glucagon in solution has been shown to readily gel or aggregate when subjected to mechanical stress.
• researchers have distinguished nine major reaction mechanisms by which proteins degrade, including hydrolysis, imide formation, deamidation, isomerization, racemization, diketopiperazine formation, oxidation, disulfide exchange, and photodecomposition. The rates of these deleterious processes depend in large measure on the protein and its environment.
• the primary chemical degradation products of glucagon include oxidation of Met (27), deamidation of Gin (24), and acid-catalyzed hydrolysis at Asp (9), Asp (15) and Asp (21). HGH undergoes chemical decomposition via oxidation at Met (14) and deamidation at Asn (149).
• biopharmaceuticals One technique used in formulating biopharmaceuticals has been lyophilization of the biopharmaceutical solution in the presence of excipients, buffers and/or bulking agents.
• lyophilized preparations must typically be stored under refrigeration, a requirement which is neither technically nor economically feasible in many markets and inhibits flexibility of patient use.
• formulations of many biopharmaceuticals which would permit their storage at ambient temperatures. This would permit more rapid development of products, increasing flexibility in shipping, storing and carrying the drug products, and allowing introduction and use of such products in markets where refrigeration is too costly.
• the increased stabilization of biopharmaceuticals would naturally improve the general use of the biopharmaceuticals where shelf life is an important consideration, whether or not refrigeration or other concerns are at issue.
• aqueous solution containing a biopharmaceutical with a limited amount of excipient(s) is frozen and then dried under vacuum to produce solids of sufficient stability for storage and distribution.
• Excipients are added to prevent blow out of the product, to provide stability during lyophilization and/or dissolution, and to enhance compatibility for parenteral use.
• Various excipients used with lyophilization have included salts, metal ions, polyalcohols, surfactants, reducing agents, chelating agents, other proteins, amino acids, fatty acids, and phospholipids.
• excipients include mannitol, alanine, glycine, sorbitol, lactose, arginine, and maltose.
• the results obtained with such excipients have usually been inconsistent.
• Most lyophilized biopharmaceuticals are amorphous powders that have no specific structure, and as a result, the amount and location of the incorporated biopharmaceutical varies widely for the product particles. Also, they are typically readily dissolved, rendering them unsuitable for use as a sustained-release material. Further, there is no isolation of the pharmaceutical molecules from the environment or one another, leaving them susceptible to degradation by various mechanisms. Studies have shown that lyophilization of excipients can typically damage proteins rather than protect them.
• Crystallized pharmaceuticals have been used in some instances, but there have been inherent limitations. Some API's , e.g. insulin, can be crystallized themselves, and are useful in that form for administration to patients. However, the majority of biopharmaceuticals either do not crystallize or the crystallization is very difficult, particularly on a commercial scale. Further, crystallization procedures are limited to the use of pharmaceuticaUy-acceptable ingredients and process conditions that do not adversely affect the active pharmaceutical ingredient, thus further constraining the ability to obtain desired microcrystalline suspensions.
• United States Patent No. 5,075,291 describes a process for preparing a uniformly-dispersed, pharmaceutically-active material in a crystalline sugar alcohol matrix.
• this process requires the addition of the API into a molten sugar alcohol with considerable mechanical agitation.
• Many API's and virtually all biopharmaceuticals would not be stable in the extreme temperature of 110°C and the physical stresses of a high-shear vortex mixer used for agitation.
• the present invention does not require these extremes of temperature and physical agitation.
• the process of the present invention slowly includes the API into the growing crystal lattice in specific growth sectors, instead of homogeneous mixing and entrapping of the active pharmaceutical ingredient in a viscous melt.
• the present invention relates to pharmaceutical compositions comprising single crystals of a pharmaceuticaUy-acceptable crystal lattice component, and an active pharmaceutical ingredient different from and included within the crystal lattice component in a growth-sector specific orientation.
• the crystals are prepared using components and methods which yield crystals having suitable purity and efficacy for use in administering the API's to a patient.
• the crystals may be coated or combined with adjuvants such as excipients, diluents or carriers, and are preferably formulated into tablets, capsules, suspensions, and other conventional forms containing dosage amounts of the API's.
• the crystals are prepared as depot formulations which may be administered, as by subcutaneous injection or implantation, to provide a long-term payout or sustained release of the active pharmaceutical ingredient.
• the present invention further provides methods for preparing the crystals and for storing and administering the active pharmaceutical ingredient either in crystal form or upon reconstitution to a solution.
• Another object of the present invention is to provide compositions comprising API's included in single crystals to provide improved stability and shelf-life.
• the active pharmaceutical ingredients may therefore be stored for extended periods of time prior to use either as crystals or as reconstituted solutions. It is a further object of the present invention to provide single crystals with included API's to provide quick, delayed-release or sustained-release formulations for flexibility in pharmacokinetic profiles in delivery of the API's to patients.
• Another object of the present invention is to provide pharmaceutical delivery units including an amount of single crystals sufficient to provide a dosage amount of the included active pharmaceutical ingredient.
• the pharmaceutical delivery units include a quantity of crystals sufficient to provide a prolonged payout of the active pharmaceutical ingredient.
• the crystals may be coated or uncoated, and may be combined with various pharmaceutical adjuvants including excipients, diluents and carriers.
• a further object of the present invention is to provide methods for preparing compositions comprising single crystals with growth-sector specific inclusions of API's.
• Figure 1 is a photomicrograph illustrating fluorescence of a single crystal of green fluorescent protein in ⁇ -lactose monohydrate (1.8 (h) x 0.8 (w) x 0.5 (d) mm 3 ) with an idealized representation of habit.
• the sides of the crystal in the photomicrograph are bright due to internal reflection.
• Figure 2 is a graph of the fluorescence decay of the green fluorescent protein at 333°K in several environments: mixed crystal in ⁇ -lactose monohydrate (triangle), saturated lactose solution (square), and lyophilized ⁇ -lactose monohydrate (diamond).
• the present invention utilizes single-crystal matrix inclusion of active pharmaceutical ingredients ("API's”) to achieve advantageous storage and delivery of the API's.
• API's active pharmaceutical ingredients
• This invention has application to a wide range of API's to provide enhanced stability and/or delivery of the active pharmaceutical ingredients.
• the invention is particularly advantageous in providing greater stability over time and in providing alternative delivery and sustained release formulations to patients .
• the small molecule host crystals comprise a crystal lattice component which includes the API's in an oriented, growth-sector specific manner.
• the crystals and included API's are prepared to be pharmaceutically acceptable and pure, thereby being useful for administration to patients to be treated with the API's.
• the term "pharmaceuticaUy-acceptable" refers to sufficient quality to meet regulatory and compendial requirements for administration to humans and/or animals.
• the crystals provide a regular, predictable inclusion of the guest active pharmaceutical ingredient, and the crystals can consequently be used for obtaining a predetermined amount of the active pharmaceutical ingredient for delivery to a patient.
• the host crystal gradually dissolves upon contact with body tissue or fluids, and is therefore useful as a system for delivery of the active pharmaceutical ingredient into the body.
• the crystals and included active pharmaceutical ingredient may be reconstituted into a solution for administration to a patient.
• the active pharmaceutical ingredient molecules are generally isolated from one another and are insulated from the environment by the host crystal. This leads to reduced susceptibility of the API to degradation, and therefore enhanced stability and shelf-life. Also, the use of appropriate host crystal compounds, or selected dosage forms, permits the design of quick, delayed, or sustained-release formulations for delivery of the active pharmaceutical ingredient. Sustained- release formulations are particularly advantageous for treatment of chronic conditions as they provide a consistent amount of drug delivery over a long period of time to improve ease of use and patient compliance in administering the API.
• the crystals preferentially incorporate the active pharmaceutical ingredient on certain faces, thereby providing a growth-sector specific inclusion and orientation to the API's.
• growth-sector specific inclusion and orientation refers to the fact that the API molecules are included primarily at certain faces of the crystal matrix.
• the growth-sector specific inclusion and orientation can be determined by one skilled in the art, as demonstrated in the examples herein, by fluorescence microscopy and anisotropy measurements, single crystal desorption mass spectrometry, and autoradiography of 14 C-labeled material. In one embodiment, at least about
• 0.001% (on weight/weight (w/w) basis) of the pharmaceutical is included within specific faces of the crystal matrix, and in another embodiment at least about 0.1% (w/w) and up to about 10%.
• the crystal parameters including the particular crystal lattice component for a given API, the concentration of API, the use of crystal adjuvants, and the crystallization conditions, are selected to achieve the growth-sector specific inclusion and orientation of the API within the crystals.
• the method of the present invention broadly involves the including of the active pharmaceutical ingredient into the single crystal matrix formed from a pharmaceutically-acceptable crystal lattice component.
• the term "included” in the crystals refers to the active pharmaceutical ingredient being chemically adsorbed within the crystal lattice as the crystal is formed.
• This inclusion of the active pharmaceutical ingredient molecules is distinguished from crystallization of the API molecules with one another, and from simple and random entrapment of the API molecules by the formed crystal.
• the crystal product of the present invention is ordered, in contrast to the amorphous material produced by other approaches.
• the API is incorporated in the crystal in relation to its degree of affinity for the crystal lattice molecules.
• the crystal lattice component is therefore selected to be both chemically and physically compatible with the API such that the API is received by the crystal during formation, and remains stable and efficacious while within the crystal and upon release therefrom.
• the including of the active pharmaceutical ingredient involves combining the crystal lattice component, the active pharmaceutical ingredient and a pharmaceutically-acceptable adjuvant in a liquid state.
• the crystal lattice component is then crystallized under pharmaceutically-acceptable conditions to form the inventive crystals.
• one method uses spiking of the API into a saturated or supersaturated solution of the crystal lattice component in a suitable organic and/or aqueous solvent system.
• the saturated or supersaturated solution of the crystal lattice component may be spiked into the API solution.
• Other components may also be added to the solution, including compounds which facilitate or modify crystal growth or which are desired for incorporation in the final formulation.
• the solution may be seeded using any of a variety of conventional techniques.
• the solution is allowed to evaporate and/or equilibrate to cooler conditions for growth of the crystals.
• the crystals are then grown as the solvent is slowly evaporated away and/or the solution is cooled, with the evaporation and temperature gradient conditions being selected dependent on such factors as the solvent system and the desired crystal size.
• the crystals containing the active pharmaceutical ingredient are harvested from the remaining solution and are preferably washed to remove surface contamination. This procedure yields crystals which include the active pharmaceutical ingredient at a predictable concentration and facial orientation.
• crystals are grown under pharmaceutically-acceptable conditions.
• pharmaceutically-acceptable conditions refers to the use of crystal and API compounds which are pharmaceutically-pure, and for which such pharmaceutical purity is maintained in the final crystals.
• the crystal and API compounds are pharmaceutically pure, or have pharmaceutical purity, if they are of sufficient purity to be suitable for administration under applicable FDA or other administrative regulations regarding purity.
• pharmaceutically-acceptable conditions further refers to the use of crystallization conditions through which the API compounds retain pharmaceutical efficacy in the final crystals and upon subsequent administration to patients.
• the present invention readily allows the inclusion of API's by affinity with the small host molecules in the growing crystal lattice. This overcomes many of the limitations associated with previous approaches.
• the processing involved with preparing the present crystals does not expose the API's to harsh conditions, thereby substantially reducing or avoiding the possible degradation or disruption of the structural aspects of the API which could occur with prior art techniques.
• the inventive crystals have an added advantage in that they do not interfere with normal analytical methodologies used for characterizing the pharmaceutical product.
• the small host molecules can be easily separated on the basis of molecular size, which is not the case for prior art techniques which use polymers that interfere with analytical methodologies.
• the API molecules are incorporated into the host crystals typically at rates of at least about 0.001% (w/w), preferably at least about 0.1%, and more preferably about 1% to about 10% (w/w). Alternatively, the API molecules are included at rates of at least about 0.01 %, and as much as at least about 1 % (w/w).
• the limited molar concentration of the active pharmaceutical ingredient in the host crystals means that the active pharmaceutical ingredient molecules are generally isolated from one another in the crystals. Isolation of the API molecules is particularly advantageous for those molecules, such as certain biopharmaceuticals, which could otherwise react with one another (e.g., by polymerization) or the surrounding environment. The degree of isolation can be verified by those skilled in the art using atomic force microscopy or reaction fluorescence energy techniques.
• the present invention has a particular application to guest-host systems in which the guest API molecules are reactive with one another, but in which these molecules are sufficiently isolated from one another in the crystals as to substantially prevent such interaction. Consequently, the invention provides containment of the API molecules in the solid state crystals and provides for the API to be comformationally stable.
• the method preferably involves preparing a mixture of crystals of substantially uniform size. This may include processing of the harvested crystals, such as by grinding or milling, to reduce the crystals to a substantially uniform size. Greater uniformity can be achieved by sorting the processed crystals, such as by sieving.
• a preferred method further includes obtaining crystals which have a substantially uniform concentration of pharmaceuticals, for example, about 1% (w/w) of pharmaceuticals, that do not vary between crystals by more than 10 percent.
• the method of the present invention may further include formulating the crystals into pharmaceutical preparations.
• the collected crystals may optionally be coated with a suitable composition.
• Coated or uncoated crystals may be blended with one or more pharmaceutically-acceptable adjuvants, such as excipients, diluents, carriers or mixtures thereof.
• the blended crystals and adjuvant(s) are then formulated into pharmaceutical delivery units.
• each unit includes a predetermined amount of the pharmaceutical.
• the crystals are combined in a delivery unit intended to deliver multiple or sustained dosing of the API over a period of time, such as by subcutaneous implantation of the delivery unit.
• a further aspect of the method of the present invention involves reconstituting the crystals to liquid form.
• the harvested crystals are dissolved in a suitable diluent for the crystal lattice component.
• the dissolution of the crystals releases the API from the crystals.
• the resulting solution may include other adjuvants, such as excipients, diluents or carriers, and the mixture is formulated under conventional procedures to desired delivery forms.
• the crystals are used to store the pharmaceutical for a period of time, such as at least one month, or at least one year, and the crystals are subsequently dissolved to use the active pharmaceutical ingredient.
• the present invention involves the use of any of a wide variety of pharmaceutically-acceptable host crystal systems that can incorporate API's in a growing crystal lattice.
• the crystal lattice component is selected to be compatible with the guest API, and to be suited to the use of the resulting formulation for storage and administration. Selection of the crystal lattice component will involve consideration of such factors as affinity for the API, crystal size distribution and morphology, and desired pharmaceutical concentration and delivery rate, as well as other factors well known in the art of pharmaceutical delivery systems.
• the crystal systems must consistently incorporate the guest active pharmaceutical ingredient in terms of concentration and placement within the crystal lattice.
• the crystals also must grow under conditions which will not degrade or otherwise adversely affect the viability of the active pharmaceutical ingredient.
• Preferred host crystal materials are those that have a high affinity for the included API. It appears that the oriented inclusion of the API's is related to the affinity between the crystal lattice component and the API. The affinity between these materials is therefore important in obtaining the desired inclusion of the API's, and also permits control of the inclusion based upon this affinity. For example, the concentration of the pharmaceutical in a crystal can be controlled by selecting the host component to have an affinity for the API which yields the desired inclusion rate. Also, mixtures of host materials, or of host materials and other excipients, can be used to provide an affinity yielding the desired inclusion level. In one aspect of the present invention, the API's are incorporated at levels of at least about 0.001% (w/w of guest ⁇ ost), more preferably at least about 0.1% (w/w).
• the preferred host crystal materials will also be very stable and readily crystallizable, and will maintain their "order" or crystal morphology when including a guest molecule, particularly large biomolecules.
• the use of particular host crystal components will also depend on such factors as how small or large the crystals can be produced and how readily they dissolve.
• it is desirable to have very small crystals e.g., pulmonary
• moderately sized crystals e.g., injectable
• very large crystals e.g., implantation and long term payout.
• the useful crystal sizes will therefore vary accordingly, ranging from submicron to millimeter sizes.
• the preferred crystals are in the order of 5-100 microns in size.
• the useful host crystal systems are therefore diverse, and include various small molecule crystal systems which meet the desired criteria.
• pharmaceutically-acceptable crystal lattice components include sugars, polyhydroxy alcohols, single and polyamino acids, vitamins, salts, metals, preservatives, aromatic compounds especially aromatic acids, purified natural products, and polymers.
• Preferred crystal lattice components include, for example, sucrose, lactose, trehalose, maltose, galactose, sorbose, mannitol, lactitol, sorbitol, glycine, alanine, lysine, arginine, ascorbic acid, nicotinamide, thiamine, adenine, pyridoxine hydrochloride, caffeic acid, vanillic acid, ferulic acid, benzoate, sorbate, methyl paraben, sodium ascorbate, sodium saccharin, and potassium citrate. Also, compatible mixtures of these materials are also useful, and can be selected to obtain the desired rate of inclusion of the pharmaceutical, or to achieve desired characteristics, such as dissolution rate and pharmacokinetic profile, for the product crystals.
• the crystal lattice components are selected to achieve the desired pharmacokinetics for the final crystals.
• the term "pharmacokinetics" is used to refer to the profile of the delivery of active pharmaceutical ingredient from the crystals into the circulatory system. This will depend primarily on the concentration of the active pharmaceutical ingredient in the crystals, as well as parameters of the active pharmaceutical ingredient itself. While given crystal lattice components will have associated inclusion and dissolution characteristics, these can be modified by including other crystal lattice components, other API's, or a variety of excipients. Thus, single crystals having two different, co-crystallized lattice components will typically be characterized by pharmacokinetic profiles different from crystals prepared with either of the crystal lattice components alone.
• the present invention involves the use of mixtures of crystals having different pharmacokinetics in order to achieve desired payout profiles.
• a pharmaceutical product can be obtained by combining two different types of crystals, one type of crystal using a first crystal lattice component characterized by a first pharmacokinetic profile, and the second type of crystal using a second crystal lattice component characterized by a second pharmacokinetic profile.
• the mixture of crystals will give a payout of API that is different from either of the individual payouts for the two crystal types.
• the included API's are similarly diverse, limited simply by the requirements of compatibility with the host crystal and the crystal growth conditions.
• the active pharmaceutical ingredient cannot be unacceptably degraded or otherwise adversely affected by the conditions under which the crystals are formed. Also, the active pharmaceutical ingredient should remain stable for an extended period of time while included within the host crystal, and pharmaceutically efficacious upon release from the crystal.
• API's useful in accordance with the present include: antibiotics (such as dirithryomycin, loracarbef, tilmicosin, vancomycin, tylosin, monensin), fluoxetine, raloxifene, olanzapine, and nizatidine.
• antibiotics such as dirithryomycin, loracarbef, tilmicosin, vancomycin, tylosin, monensin
• fluoxetine raloxifene
• olanzapine olanzapine
• nizatidine nizatidine
• Tissue Piasminogen Activator T-PA
• Intravenous injection- Lyophilized powder which is reconstituted with sterile water (supplied) to 1 mg/mL and results in a final pK of 7.3. Can not be reconstituted with preserved water due to precipitation.
• the 1 mg mL solution can be diluted 1 : 1 with 0.9% NaCl or D5W and help for 8 hou ⁇ at room temperature. TPA is incapable with preservatives.
• Expression System Natural source - human leukocytes which arc exposed to an avian vims in order to produce interferon.
• Glyc ⁇ protein single N-linked complex carbohydrate
• 166 amino acids with a predicted molecular weight of 22,500 daltons, human sequence, has a specific activity of 200 million units per mg protein.
• ⁇ Formulation Lyophilized product (stored refrigerated) which is reconstituted with 0-54% NaCl (supplied] to 0.25 mg/mL i__te.fa.on beta-lb, 12.5 mg mL human albumin, 12.5 mg/ml dextrose, and has a pH of approximately 7.3 (recon solution is stable for 3 hours). Injected subcutaneously every other day (chronic use).
• Cost of therapy is $13,140 (based on 025 mg/injection, dose every other day for 1 year; equals 46 mg protein).
• Formulation Comes in a lyophilized and a solution formulation.
• the lyophilized formulations when reconstituted with 0.9% benzyl alcohol (supplied) contains either 0.015, 0.025, 0.05, 0.90, or 0.125 mg/ml.
• Interferon alfa-2b 20 mg/ml glycine, 2.3 mg/ml sodium phosphate dibasic, 0-55 mg/ml sodium phosphate monobasic, and 1 mg/ml human albumin
• the solution formulations contain either 0.05, 0.114, or 0.1 5 mg/mL Interferon alfa-2b, 20 mg/ml glycine, 2.3 mg ml sodium phosphate dibasic, 0.55 mg/ml sodium phosphate monobasic, 1 mg/ml human albumin, 1.2 mg/mL methylparaben, and 0.12 mg/ml propyparaben.
• These formulations be injected intramusc ⁇ lar, subcutaneous, or intralesi ⁇ nal. All formulations and reconstituted products are stored at reftigerated temperatures.
• Multi-use and lyophilized formulation indented for intramuscular or subcutaneous administration contains either 0.015, 0.045, 0.090, 0.18 mg/mL Interferon alfa-2a, 9 mg/ml NaCl. 5 g/ l human albumin, and 3 mg/ml phenol.
• the lyophilized formulation reconstituted with 3 mL of supplied diluent (6 mg ml NaCl, .3 mg/ml phenol) consists of 0.03 mg/ml Interferon alfa-2*, 9 rag/ml NaCl, 1.67 mg/ml human albumin, and 3.3 mg/ml phenol.
• Cost of therapy is $59,200 (28mg protein over 1 year). Specific activity is 200 million international units per mg protein.
• Lyophilized product which is reconstituted with sterile water containing 0.3% m-cresoL 1.7% glycerin (supplied) to 2 mg/inL hGH and has a final pH of approximately 7.5, subcutaneous or intramuscular _uin_Jnistratio ⁇ .
• Each 5 mg lyophilized vial contains 5 mg hGH, 25 mg mannitol, 1.13 mg dibasic sodium phosphate, and 5 mg glycine.
• Strncture 191 amino acids, molecular weight of 22,125 daltons, sequence is identical to human pituitary- derived material.
• Lyophilized product which is reconstituted with bacteriostatic water (0.9% benzyl alcohol, supplied) to 5 mg/mL hGH and has a final pH of approximately 7.4, subcutaneous or ulcerramuscular administration.
• Each 5 mg lyophilized vial contains 5 g hGH, 45 mg mannitol, 1.7 mg sodium phosphates (0.4 mg monobasic and 1.3 mg dibasic), and 1.7 mg glycine.
• Expression System E. coli, expressed with a leading secretion signal precursor which directs the protein to the plasma membrane of the cell where the sequence is removed and the native protein is secreted into the periplas so that the protein if folded appropriately as it is synthesized
• Lyophilized product 212 units ghicocerebrosidase, 155 mg mannitol, 70 mg sodium citrate, and 0.53 mg polysotbate-80; stored refrigerated
• Lyophilized product 212 units ghicocerebrosidase, 155 mg mannitol, 70 mg sodium citrate, and 0.53 mg polysotbate-80; stored refrigerated
• 5.1 mL of sterile water final pH is approximately 6.1.
• the reconstituted material is combined with 100 to 200 mL of 0.9% NaCl and administered intravenously.
• Recombinant protein differs from human placental ghicocerebrosidase by a arginine substituted for histidine at position 495 and the glycosylation sites have been modified to terminate in mannose sugars (which are specifically recognized by endocytic carbohydrate receptors on macrophages, the cells that accumulate lipid in Gaucher disease).
• Orphan Drug sales > S100 million- Cost of therapy is $351,130 (1 year).
• Formulation Suspension (20 ⁇ g/mL hepatitis B surface antigen adsorbed onto 0.5 mg aluminum, 1:20,000 thimerosal, 9 mg/ml NaCl, 1.7 mg/ml sodium phosphates). IJcrO ⁇ uscular administration.
• Formulation contains no more than 5% yeast proteins.
• Formulation Suspension (lO ⁇ g/ml hepatitis B surface antigen adsorbed onto 0.5 mg aliiminuirj, 1:20, 00G thimerosal). Intramuscular administration.
• Formulation contains no more than 1% yeast proteins.
• Produet name Epogen or Epoetin alfa (Also sold under the name Procrit by Ortho Biotech but manufactured by A geo)
• GM-CSF (Granuloc te Macrophage-Colo ⁇ y Stimulating Factor)
• Formulation Lyophilized solution which feconsrJnned with sterile water (stored at refrigerated temperatures for ⁇ 6 hours) o ⁇ 0.9% benzyl alcohol (can be stored for ⁇ 20 days at refrigerated TABLE A temperatures) and administered intravenous.
• the lyophilized single use product contains either 0.25 mg/mL or 0.50 mg/mL GM-CSF, 40 mg/ml mannitol, 10 mg/ml sucrose, and 12 mg ml tromethamine (final pH is 7.4 +/- 03).
• the reconstituted solution is then diluted into a 0.9% NaCl bag for IV administration (note if final GM-CSF is below 0.01 mg/mL add human albumin to 0.1% to prevent adsorption to the IV bag.
• G-CSF G-CSF (Granulocyte Colony Stimulating Factor)
• the protein has an amino acid sequence identical to the human protein except for an additional N-terminal mcthionine (necessary for expression in E. coli).
• the human protein is glycosylated but the recombinant Neupogen is not
• Proleuldn has a molecular weight of 1 ,300 daltons and differs from the natural human protein (is not giycosylated, the N-terminal alanine is removed, and has a serine substituted for the free cysteine at position 125)
• Formulation Lyophilized formulation which is reconstituted with 0.9% benzyl alcohol (supplied) and aclministered intramuscular or subcutaneous.
• the lyophilized vial contains 5 mg Somatrem, 40 mg mannitol and 1.7 mg sodium phosphates (0.1 mg sodium phosphate monobasic and 1.6 mg sodium phosphate dibasic) and is reconstituted with 1 to 5 mL of 0.9% benzyl akohoL
• the lyophilized product is stored at refrigerated temperatures, the reconstituted product is good for 14 days at refrigerated temperatures.
• Inhalation solution (aerosol mist produced by a compressed air driven nebulizer system). Comes in a single-use 2 -5 toL ampule containing 1.0 mg/mL DNase, 0.15 mg/ ⁇ nl- calcium chloride dihydratc, and 8.77 mg/ml sodium chloride, at a pH of 6.3. The solution is stored at refrigerated temperatures and should not be exposed to light.
• M-CSF Macrophage-Colony Stimulating Factor
• Epoetin Beta (Erythropoietin)
• Human Serum Albumin Product name Produced by Indication: Human use, Date of approval: Formulation: Expression System: Refolding Conditions: Post-Transitional Modifications: Structure: Additional Information:
• Epoetin alfa Thousand Oaks, CA failure including patients on dialysis and not on (rSO) dialysis, and anemia in Retrovi ⁇ *treated HlV-infected patients ( _ne 1909); treatment of anemia caused by chemotherapy in patients with non-myaloid malignancies (April 1993); prevention of anemia associated with surgical blood loss, autoiogous blood __ ___ _ donation adjirvant (DecemberJ 996) _
• Epoetin alfa NJ failure including patients on dialysis and not on (rEPO) dialysis, and anemia in Retrovfr*trea_ed HlV-infeded patients (December 1990); treatment of anemia caused by dierno erapy in patients with non-myd ⁇ id malignancies (April 1993); prevention of anemia associated with surgical blood lots, autologcus blood donation adjuvant (December 1996) [PROCRIT was approved for marketing under Amgtn's epoetin alfa PLA.
• CanoBopm 1 Pha ⁇ r ⁇ a & Upjohn human short stature in children due to growth hormone somatropin Kalam*zDo, A ⁇ l growth deficiency (March 1995) (rONA origin) hormone for injection
• Prtrtropt ⁇ * Genentech human human growth hormone deficiency in children somatrem 5. San Ftan ⁇ sco, CA growth (October 1985) for injection hormone
• RetrraseTM Boehringer Mannheim tissue treatment of acute myocardial infarction (October 1996) reteplase Gaithet ⁇ urg,MD plasminogen Centocor factor Mah-m, PA
• ISIS 2922 Isis Pharmaceuticals antisense cytomegalovi ⁇ s retin ris Phase III fomivirsen Carlsbad, CA
• NEUPOCCN* A gen colony treatment and prevention of application
• PRO 542 Progenies Pharmaceuticals HIV infection Phase 1 Ta ⁇ ytown, NY
• DABsJW H ⁇ pkintmMA protein see also cancer, skin
• interleukin-10 Schering-Plough interleukin rheumatoid arthritis Phase
• ISIS 2302 Isis Pharmaceuticals antisense rheumatoid arthritis Phas d Carlsbad, CA (see also digestive, skin, transplantation) TABLE A
• CA-EPO Hoechst Marion Roussel erythropoierin anemia associated with Phased Kansas City, MO chronic renal failure Transkaryotic Therapies Cambridge, MA ogenate-N Bayer dotting hemophilia A Phase ! rFVIII Berkeley, CA factor
• Aastrom Aastrom B Kciences cellular cancer immunosuppression/ Phase II CeU ProdocMn A ⁇ Abor.Ml therapy blood and immune system System recovery for patients receiving stem and progenitor ablative chemotherapy cell expansion from bone mamow and umbilical cord Wood
• AfP-ScmTM Immunomedics MAb extent of disease staging of liver Phased teehnetiuffl-99m- Moms Wains, NJ and germ cell cancers FAb' fragment (germctl allogeneic 5yStemix cellular advanced leukemia, rymphoma, Phase 1 hematopoietic Palo Alto, CA therapy r ⁇ yelodysplastic syndromes stem cell tran .plantation
• DAHu.gamma 15- Chiron Viagene gene therapy disseminated Phase! iraruduced San Diego, CA malignant melanoma autologou* tumor cells; ITAT daniplestim Static growth factor mobilization of peripheral Phased! SkokictL blood stem DCb dendritic cell Denditon cellular advanced prostate cancer Phase 11/111 therapy Mountain Vew, CA therapy multiple myeloma Phase 1
• KSPPC-96 Antigenics heat shock melanoma, pancreatic, Phase! autologous Ne yorf ⁇ NY protein renal cell cancers tumor derived
• IDEOInBB IDEC Pharmaceuticals MAb non-Hodgkir ⁇ B-cell lymphoma Phase VII San Diego, CA
• IDEC-Y288 IDEC Pharmaceuticals MAb ⁇ on-Hodgldn's B-cell lymphoma Phase l/ll San Diego, CA TABLE A
• GM SF factor prophylaxis of chemotherapy-induced neutropenia in acute myelogenous leukemia
• LymphoScanTM Immunomedics MAb extent of disease staging of Phase III tech ⁇ etium-99m- Morris trains, NJ non-Hodgx 's B-cell lymphoma, bectumomab detection of residual disease (lymphoma) following radiatiorVchemotherapy
• OncoRad* ra CYTOCEN MAb targeted radiotherapy for prostate Phase d CYT-356.Y.90 Princeton, NJ malignancies
• Sigoslx* Ares-Serono and interleukin hematologkal conditions Phase l/H recombinant Serono Laboratories (r ⁇ yelodvsplastic syndromes, cancer) interleukl ⁇ -6 Norwell, MA (r-IL-6) TABLE A
• TBC CEA Therion Biologies vacant colorectal and lung cancers Phase VII (vaccinia virus Cambridge. MA expressing carcinoembryonic antigen)
• BetaRx-H VivoRx cellular insulin-dependent diabetes Phasel encapsulated Santa Monica, CA therapy human isles
• I51S2302 Is ⁇ Pharmaceuticals antisense Crohn's disease, ulcerative colitis Phased Carlsbad. C ⁇ (see also autoimmune, skin, transplantation)
• Tc-99m apcitide Londonderry NH anri-CD18 Genentech MAb acute myocardial infarction Phase humanized MAb 5. San Francisco, CA
• BioByPa ⁇ TM GenVec gene therapy cardiovascular disease Phase 1 therapeutic Rockville. MD including cardiac artery disease angiogenesis and peripheral vascular disease,
• VEC cardiac artery bypass grafts
• CPC-1 11 Cypros Pharmaceuticals cellular coronary bypass surgery Phase C-risbad.
• CA therapy (see also blood) factor Vila Corvas deep vein thrombosis, pulmonary Phase l inhibitors San Diego, CA embolism, unstable angina, myocardial infarction
• Bothel WA see also neurologic other) ⁇ legiflhTM COR Therapeutics percutaneous trar ⁇ himinal application eptifibatide S. San Francisco, CA coronary angioplasty. submitted (llh/llla platelet Schering-Plough unstable angina aggregation Madison, NJ inhibitor) acute myocardial infarction Phase! inter leu kin-10 Schering-Prough interleukin ischemic reperfua'on injury Phase ! (IL-10! Madison, NJ (see also ⁇ IDVHIV, autoimmune, digestive, neurologic respiratory, skin) lanoteplase Bristol-Myers Squibb t-P ⁇ acute myocardial infarction Phase 111 ftmcefon, NJ
• VEG 121 Scios growth cardiovascular disorders Phasel (vascular Mountain View, CA factor endodietial growth factor)
• hepatitis C Phase 111 l ⁇ tron* A Schering-Plough interferon relapsed hepatitis C application XebetolTM Madison, NJ submitted interferon alfa-2b (recombinant)/ naive hepatitis C (not previously Phase III ribavirin treated with interferon) hepatitis C (PEG-tntron A/Rebetol) Phase l
• LhADt* Ares-Serono and recombinam female infertility— foHicular support, Phase d/Ill recombinam Serono Laboratories fertility stimulation of follicular development human leuiinizing Norwel MA hormone hormone (r-hLH)
• FIBIA5T* Sdos growth stroke Phase d/Ill trafermin Mountain View, CA factor bee a o heart) Wyeth-Ayerst Laboratories Philadelphia, PA
• NeuroCeflf-HD Diacrin cellular Huntington's disease Phase l (cellular Charkstown, MA therapy completed transplantation Genzyme Tissue Repair therapy) Catrib ⁇ dg-, A
• Influenza rHAO Protein Sdences vacdne prevention of influenza Phase n Vacdne Maiden, CT influenza vaccine influenza virus Aviro ⁇ vacdne prevention of influenza Phase Id vaccine Mountain View, CA
• NCUPOCtN* A gen colony multilobar pneumonia, Phase IU
• PIV vaccine Vvyeth- derie continuous prevention of parainfluenza Phase l live, attenuated Vaccines & Pediatrics ceil line virus-mediated lower respiratory Philadelphia, PA vacdne disease in infants
• ISIS 2302 Isis Pharmaceuticals antisense psoriasis Phase d Carlsbad, CA (see also autoimmune, digestive, transplantation) k ⁇ rattnoeytf Human Genome Sciences growth wound healing Phase ! growth factor-2 Rodeville. MD fa ⁇ or (see also other) ( GF-2J
• IStS 2302 isis Pharmaceuticals antisense renal transplant rejection Phase d Carlsbad, CA (see also autoirnraurg, digestive, skin)
• biopharmaceuticals which include, for example, any proteins, polypeptides, enzymes, irnmunoglobulins, polynucleic acids, and plasmids or other biopolymers.
• biopharmaceuticals to be included in the crystal formulations of the present invention include the following: insulin, glucagon, Glucagon-Like Peptide- 1 (7-37)OH (GLP-1), human growth hormone, leptin, follicle-stimulating hormone (FSH), ribozyme, and analogs thereof .
• the API's useful with the present invention include those which themselves may form crystalline products, as well as those which do not.
• any proteins can be prepared as microcrystalline suspension products, but the results have frequently been unsatisfactory using existing technology.
• inclusion of these biomolecules into a host crystal system in accordance with the present invention overcomes this limitation on crystallization.
• the invention further finds utility even with API's that are readily crystallized, such as insulin.
• the incorporation of such API's into a single crystal lattice can be used to enhance stability or provide means of delivery that have different characteristics.
• Solvents for preparation of the saturated and supersaturated crystal lattice component include, but are not limited to, water, alcohols (e.g., ethanol, isopropanol), other organic solvents, acids, bases, and buffers.
• the crystals of the present invention are prepared to have a predetermined amount of active pharmaceutical ingredient.
• the desired amount of active pharmaceutical ingredient will depend on typical considerations, such as the effective amount of API used for administering to a patient.
• the concentration of API in the crystal is controlled, such as by previously described means, to yield crystals which are readily used in preparing pharmaceutical formulations for administration.
• the active pharmaceutical ingredient can be incorporated into the crystals at any of a wide variety of molar or weight percentages.
• Preferred percentages can be easily selected by a skilled artisan taking into account the usual considerations for later formulation of the desired pharmaceutical compositions, depending on the application, route of delivery, and desired pharmacological profile. Preferred percentages include, for example, concentrations of 0.01 - 1 weight percent. As used herein, all weight percentages are given as the percent based on the weight of the crystal including the crystal lattice component, the active pharmaceutical ingredient and any other components included within the crystals, unless stated otherwise.
• the crystals may be prepared at varying size distributions, similarly depending on the subsequent formulating to be done with the crystals, or on crystal growth parameters.
• the crystals may be harvested and then sorted directly to desired size ranges, or may first be processed, such as by grinding or milling, and then sorted such as by sieving.
• a desired amount of active pharmaceutical ingredient may be obtained simply by obtaining a determined weight of crystals containing the active pharmaceutical ingredient at a known weight concentration.
• the useful size or weight range of the crystals of the present invention accordingly varies widely, depending on such factors as the inclusion level of the active pharmaceutical ingredient, the dosage amount for the active pharmaceutical ingredient, and the method of delivery of the crystals.
• suitable crystals may have an average size distribution of 1 ⁇ m to 1 mm .
• the crystals of the present invention will typically be used in a formulation comprising a large number of crystals. It is a feature of the present invention that the active pharmaceutical ingredient is included within the crystal lattice component in a predictable, oriented fashion. This leads to a uniform concentration of the active pharmaceutical ingredient as a molar, and therefore weight, percentage of the crystals.
• a composition of crystals having a substantially uniform weight concentration of active pharmaceutical ingredient as between crystals.
• substantially uniform weight concentration refers to the fact that the weight concentration of active pharmaceutical ingredient in the various crystals is sufficiently uniform that an acceptably accurate weight of active pharmaceutical ingredient can be obtained based on the weight of the crystals and the average concentration of active pharmaceutical ingredient in such crystals.
• a composition of crystals in which the size distribution of active pharmaceutical ingredient does not vary between crystals by more than about 20 percent.
• alternate embodiments may be equally useful, including mixtures of different size crystals.
• a desired quantity of active pharmaceutical ingredient is then accurately obtained by measuring a weight amount of crystals which, given the concentration of active pharmaceutical ingredient, yields the selected weight of active pharmaceutical ingredient.
• the crystals and included API's are useful in the crystal form for both the stabilization and storage of the API and for the administration of the API to a patient.
• patient refers to either humans or non-humans, depending on the nature of the active pharmaceutical ingredient.
• the crystals may be used as such, and in one aspect of the present invention the crystals consist essentially of simply the crystal lattice component and the API. Alternatively, the crystals include the crystal lattice component and the API in combination with other pharmaceutically-acceptable adjuvants also contained within the crystals.
• the crystals of the present invention are preferably formulated as pharmaceutical materials for ultimate delivery in solid or liquid form.
• the crystals are typically formulated with common, compatible, pharmaceutically-acceptable adjuvants, such as excipients, diluents, carriers or mixtures thereof.
• pharmaceutically-acceptable adjuvants such as excipients, diluents, carriers or mixtures thereof.
• pharmaceutically-acceptable refers in this context to the excipients, diluents or carriers, as well as coatings or other components referred to elsewhere, being compatible with the other ingredients of the formulation and not deleterious to the recipient thereof.
• excipients, diluents, and carriers that are suitable for such dosage forms are well known in the art, and include the following: suspension additives such as tonicity modifiers, buffers, precipitants, and preservatives; fillers and extenders such as starch, lactose, dextrose, sucrose, sorbitol, mannitol, and silicic derivatives; binding agents such as carboxymethyl cellulose and other cellulose derivatives, alginates, gelatin, and polyvinyl pyrrolidone; moisturizing agents such as glycerol; disintegrating agents such as calcium carbonate and sodium bicarbonate; agents for retarding dissolution such as paraffin; resorption accelerators such as quaternary ammonium compounds; surface active agents such as cetyl alcohol and glycerol monostearate; adsorptive carriers such as kaolin and bentonite; and lubricants such as talc, calcium and magnesium stearate, and solid polyethyl glycols. Additionally
• the crystals may be coated to achieve various effects.
• the crystals are coated with the same crystal lattice component which forms the underlying crystal, but without the included API. This assures that the coating and the underlying crystal have compatibility.
• the coating is then applied at a thickness which provides the desired effect, such as further protection of the active pharmaceutical ingredient, bulking of the crystal for handling, and/or effecting a sustained or delayed release of the active pharmaceutical ingredient.
• the same effects can be accomplished by coating the crystals with other compatible coating compositions, such as those which are well known in the pharmaceutical coating art.
• the crystals can also be coated so as to release the active pharmaceutical ingredient only or preferably in a particular part of the intestinal tract or other route of administration, possibly over a period of time. This is accomplished, in known fashion, using coatings, envelopes, and protective matrices made, for example, from polymeric substances or waxes.
• the crystals and included API's may be packaged and administered to patients in discrete pharmaceutical dosage forms.
• the crystals may be used as such in solid form, or may be formulated into liquid solutions or suspensions prior to use.
• the compositions may accordingly be administered by various routes, for example, by the oral, rectal, vaginal, ocular, buccal, nasal, pulmonary, iontophoretic, topical or parenteral routes. Such compositions form part of the present invention and are prepared in manners well known in the pharmaceutical art.
• the API's of the present invention are effective over a varied dosage range.
• compositions are formulated in one embodiment as a unit dosage form.
• unit dosage form refers to physically discrete units, such as tablets, capsules, and suspensions in vials or cartridge/pen systems suitable as unitary dosages, particularly as unitary daily dosages.
• Each discrete unit contains a predetermined quantity of active pharmaceutical material calculated to produce the desired effect, e.g., a prophylactic or therapeutic effect.
• the amount of active pharmaceutical ingredient contained in a given dosage unit can be varied depending on the manner of delivering the crystals.
• a single dosage unit in tablet form may contain 1/4, 1/3, 1/2 or 1 times the unit dose for the active pharmaceutical ingredient, according to which 1 to 4 tablets would be administered to achieve a unit dose of the active pharmaceutical ingredient.
• a pharmaceutical product in dosage form comprising a pharmaceutical delivery unit including a dosage amount of active pharmaceutical ingredient.
• the API is contained within the crystal lattice component, and a sufficient amount of crystals is included within the delivery unit to constitute the dosage amount of the API.
• the dosage amount of pharmaceutical may be obtained by provision of one or more crystals of the present invention.
• One form of the product consists essentially of a dosage amount of the crystals.
• the pharmaceutical product consists of the dosage amount of the crystals.
• the ultimate delivery forms may include, for example, tablets, soft and hard gelatin capsules, pellets, granules, marumes, lozenges, sachets, cachets, elixirs, suspensions, ointments, suppositories, injection solutions and suspensions, nonpareils, spheres and sterile packaged powders.
• the crystals may be coated or uncoated, and may be combined with various pharmaceutical adjuvants, including excipients, diluents and carriers, as already described.
• One preferred form of the pharmaceutical product consists essentially of the crystals, and an alternate form consists of the crystals and the pharmaceutically-acceptable adjuvants.
• the delivery forms are prepared by conventional techniques such as disclosed in Remington's Pharmaceutical Sciences, 19th Edition, Mack Publishing Company, Easton, PA (1995), which is incorporated herein by reference, or other treatises available to the skilled artisan.
• Compressed tablets for example, are prepared by well-known means which are conventional in the art.
• the tablets may be prepared by wet or dry granulation methods or by direct compression, and may be produced by any of a wide variety of tabletting machines. Tablet formulations usually incorporate diluents, binders, lubricants and disintegrators, as well as the crystals with included API's.
• Typical diluents include, for example, various types of starch, lactose, mannitol, kaolin, calcium phosphate or sulfate, inorganic salts such as sodium chloride, and powdered sugar. Powdered cellulose derivatives are also useful.
• Typical tablet binders are substances such as starch, gelatin, and sugars such as lactose, fructose, glucose and the like. Natural and synthetic gums are also convenient, including acacia, alginates, methylcellulose, polyvinylpyrrolidine and the like. Polyethylene glycol, ethylcellulose and waxes can also serve as binders.
• Certain solid pharmaceutical dosage forms of the present invention may be coated in conventional fashion with a wide variety of materials utilizing various processes.
• the products of the present invention may be sugar coated or film coated in accordance with well-known techniques.
• the coatings serve an aesthetic purpose as well as a practical one. Coatings can mask an unpleasant taste or odor, can increase ease of ingestion by the patient, and can serve to improve the ultimate appearance of the dosage form. Similarly, coatings can protect the product from the effects of air, moisture and light, can improve product identification, and can facilitate handling in packaging and fill lines during manufacture.
• Various adjuvants may be included in the coating formulations as is well known in the art. These include, for example, permeability enhancers, plasticizers, antitacking agents and the like.
• permeability enhancers include, for example, plasticizers, antitacking agents and the like.
• a discussion of coating techniques and adjuvants is presented in United States Patent No. 5,015,480, issued to Childers et al. on May 14, 1991, the pertinent portions of which are hereby incorporated herein by reference. Further information pertinent to coating processes and equipment may be obtained from Remington's Pharmaceutical Sciences, supra. Tablets are often coated with sugar as a flavorant and sealant, or with film- forming protecting agents to modify the dissolution properties of the tablet.
• the compounds may also be formulated as chewable tablets by using large amounts of pleasant-tasting substances such as mannitol in the formulation, as is now well- established practice.
• Instantly dissolving tablet-like formulations are also now frequently used to assure that the subject consumes the dosage form, and to avoid the difficulty in swallowing solid objects that bothers some subjects.
• a lubricant is used in a tablet formulation to prevent the tablet and punches from sticking in the die of the tabletting machine.
• the lubricant is chosen from such slippery solids as talc, magnesium and calcium stearate, stearic acid and hydrogenated vegetable oils.
• Tablet disintegrators are substances which swell when wetted to break up the tablet and release the crystals. They include starches, clays, celluloses, algins and gums. More particularly, corn and potato starches, methylcellulose, agar, bentonite, wood cellulose, powdered natural sponge, cation-exchange resins, alginic acid, guar gum, citrus pulp and carboxymethylcellulose, for example, may be used, as well as sodium lauryl sulfate.
• Enteric formulations are used to protect crystals and the included API's from the strongly acidic contents of the stomach. Such formulations are created by coating a solid dosage form with a film of a polymer which is insoluble in acidic environments, and soluble in basic environments. Exemplary films are cellulose acetate phthalate, polyvinyl acetate phthalate, hydroxypropyl methylcellulose phthalate and hydroxypropyl methylcellulose acetate succinate.
• the crystals with included API's may similarly be formulated into capsules for administration.
• Such capsules are prepared utilizing conventional encapsulating methods.
• a general method of manufacture involves preparing the crystals for use in capsules, such as by milling the crystals to a suitable size.
• the crystals are blended with desired excipients, diluents or carriers, and the resulting mixture is filled into suitably-sized capsules, typically hard gelatin capsules, using conventional capsule-filling machines.
• the usual diluents include inert powdered substances such as starch of many different kinds, powdered cellulose, especially crystalline and microcrystalline cellulose, sugars such as fructose, mannitol and sucrose, grain flours and similar edible powders.
• Cocoa butter is a traditional suppository base, which may be modified by addition of waxes to raise its melting point slightly.
• Water- miscible suppository bases comprising, particularly, polyethylene glycols of various molecular weights are also in wide use.
• the crystals can also be similarly formulated as elixirs or suspensions for convenient oral administration or for parenteral administration, for instance by intramuscular, subcutaneous or intravenous routes.
• the inventive crystals enable the design of sustained-release formulations based upon various factors to yield both the desired amount of active pharmaceutical ingredient and the desired pharmacokinetic profile for delivery of the active pharmaceutical ingredient.
• the choice of the crystal component and the process used to grow the crystals of excipient host and guest active pharmaceutical ingredient can be selected and/or modified to adjust parameters such as the delivery rate of the active pharmaceutical ingredient upon use of the formulation.
• the active pharmaceutical ingredient is incorporated into the crystal matrix at a selected rate, typically as only a small weight percentage of the overall crystal. This permits moderate and uniform rates of release.
• Various approaches may be used to accomplish a delayed or sustained release of active pharmaceutical ingredient from the crystals.
• the crystals of the desired size are combined with a compatible preservative and the mixture is injected subcutaneously or surgically implanted to provide a prolonged payout as the crystals dissolve as a result of contact with the surrounding body tissue and fluid.
• the concentration of the active pharmaceutical ingredient in the crystals is reduced in order to effect a sustained release over time.
• larger crystals may be used to provide for more prolonged payout of the active pharmaceutical ingredient.
• coatings on the crystals are used to affect the rate of release of the active pharmaceutical ingredient. Such coatings may comprise the same crystal lattice component but without the included active pharmaceutical ingredient, as well as other coating compositions useful for this purpose.
• the crystals of the present invention can be used to isolate and/or store the active pharmaceutical ingredient for later reconstitution into solution.
• the crystals may be stored for extended periods of time prior to reconstitution in view of the added stability accorded the API's by the encompassing crystal lattice component.
• the crystals are then combined with pharmaceutically-acceptable excipients, diluents or carriers to prepare the solutions for subsequent administration.
• the crystals are readily dissolved or suspended in appropriate diluents, which may be selected, for example, from the list previously provided with regard to diluents used to initially prepare the crystals.
• Such solutions of dissolved crystals provide the active pharmaceutical ingredient free of the previously encompassing crystal lattice component.
• the solutions are useful, for example, for oral administration, parenteral use, or as suppositories.
• the crystals may be formulated in a pharmaceutically-acceptable diluent such as physiological saline (0.9%), 5% dextrose, Ringer's solution, and the like, along with other additives to reduce the solubility of the crystals in suspension.
• the resulting pharmaceutical formulations provide an active pharmaceutical ingredient which is included within the host crystal and has enhanced stability and shelf-life.
• the present invention therefore satisfies the desire to provide certain pharmaceuticals having an acceptable, room-temperature shelf-life.
• the desired shelf-life can be as little as one month, or may be at least one year, two years or more.
• the pharmaceutical molecules are generally isolated from one another and from the environment by the surrounding crystal lattice.
• the containment of the API in the solid crystal lattice also fixes the conformational orientation. This eliminates most of the potential degradation mechanisms, such as polymerization, oxidation, deamidation and proteolysis, that could otherwise reduce the stability of the pharmaceutical.
• Methods demonstrating stability include but are not limited to high- performance liquid chromatography for purity and potency, FT-IR for secondary structure, in-vitro and in-vivo bioassays, and pharmacokinetic profiles.
• the crystals of the present invention are readily prepared and are useful in containing the included API in an isolated, oriented position within the lattice.
• the utility of the present invention is demonstrated in the following examples, which are illustrative in nature, and are not to be considered limiting of the scope of the present invention.
• green fluorescent protein was incorporated into deionized ⁇ -lactose monohydrate.
• GFP was selected because it is known to fluoresce only in its native conformation. Upon denaturation, the interior of the ⁇ -barrel of the molecule is exposed and the fluorescence of the p-hydroxybenzylideneimidazolinone chromophore is rapidly quenched.
• Typical crystal growth conditions involved the addition of 8 volumes of an approximately 1 mg/mL (approximately 37 ⁇ mole) solution of GFP in 10 mM tris-HCl, pH8 and 10 mM EDTA to 100 volumes of a supersaturated aqueous solution (approximately 1.15 M) of deionized ⁇ -lactose monohydrate. The mixed solution was allowed to stand for 3-4 days at room temperature in a 24-well plate.
• Crystals were harvested between 1-3 days and displayed a hatchet morphology as shown in Figure 1 with a broad base (010) further bounded by ⁇ 100 ⁇ , ⁇ 110 ⁇ , ⁇ 1- 10 ⁇ , and ⁇ 0-11 ⁇ . Small (0-10) and ⁇ 1-50 ⁇ faces are also occasionally present. When illuminated with a long wavelength UV lamp, the crystals exhibited a bright green fluorescence localized within a sharply defined pyramid corresponding to the (010) growth sector. This indicates that GFP is selectively recognized and overgrown by the (010) face in preference to the others. More importantly, it is evidence that the GFP is in its native conformation. The level of GFP to lactose is approximately 0.008% (w/w).
• GFP fluorescence intensity was measured as a function of time and temperature in three environments: saturated aqueous ⁇ -lactose solution, lyophilized ⁇ -lactose, and crystalline ⁇ -lactose monohydrate. As shown in Figure 2, both the solution and lyophilized preparations lost nearly half of the fluorescence intensity at 333°K within one hour. The crystal showed no change at 333°K or even 343°K.
• Example 2 To investigate the potential for incorporation of a biopharmaceutical into crystals of biocompatible excipients, studies were conducted using rhodamine- labeled glandular glucagon and lactose.
• the rhodamine label was used to facilitate the visualization of glucagon in the host crystals.
• Typical crystal growth conditions involved the addition of 5 volumes of a supersaturated solution of deionized ⁇ -lactose monohydrate to 1 volume of an approximately 1.5 mg/mL (approximately 300 to 400 ⁇ mole) of rhodamine-labeled glucagon in purified water.
• the mixed solution was allowed to stand at room temperature in a 24-well plate. Crystals were harvested between 1-3 days and displayed a hatchet morphology with a broad base. With the rhodamine label, glucagon inclusion was visible in the crystals as a well-defined pyramid corresponding to the (010) growth sector.
• the level of inclusion was determined to be approximately 0.1% (w/w).
• In-vitro dissolution experiments were performed on the glucagon/lactose crystals to evaluate potential for in-vivo, sustained-release pharmacokinetics. The release of rhodamine-labeled glucagon into solution was followed by fluorescence spectroscopy. In a typical experiment, 1-2 crystals were added to 100 microliters of phophate buffered saline solution at room temperature and the increase in fluorescence of the solution was monitored over time. The release of glucagon from the dissolving crystals was generally complete after 24-48 hours depending on crystal size and was linear until the last few hours of dissolution.
• Example 3 To demonstrate the universality of this technology for incorporation of a diversity of biopharmaceuticals into crystals of biocompatible excipients, studies were conducted using biosynthetic human insulin and insulin analogs, V8-GLP-l(7-37)OH, a glucagon-like insulinotropic peptide- 1 analog, exendin, and human growth hormone in deionized ⁇ -lactose monohydrate or phthalic acid.
• V8-GLP Information regarding V8-GLP is available in United States Patent No. 5,705,483, issued to Galloway and Hoffman on January 6, 1998, which patent is hereby incorporated herein in its entirety.
• exendin see, e.g., R. Goke, H.C. Fehmann, T. Linn, H.
• Exendin- 4 is a High Potency Agonist and Truncated Exendin-(9-39)-amide an Antagonist at the Glucagon-like Peptide l-(7-36)-amide Receptor of Insulin-secreting Beta- cells," J. Biol. Chem. 1993, Sep 15, 268(26), pp. 19650-5, which reference is hereby incorporated herein in its entirety.
• Typical crystal growth conditions involved the addition of 1 volume of an approximately 10 mg/mL rhodamine- or Texas red-labeled peptide or protein in 0.1M phosphate-buffered saline solution (PBS, pH7.4) to 10 volumes of a supersaturated ⁇ -lactose solution or phthalic acid solution.
• PBS phosphate-buffered saline solution
• Supersaturated solutions of purified ⁇ -lactose were obtained by adding 0.41 grams of ⁇ -lactose to 1 mL of purified water, allowing to dissolve in a 50-70°C water bath, and cooling to room temperature.
• Supersaturated solutions of phthalic acid were prepared by adding 0.05 grams of phthalic acid to 1 mL of either 70/30 (v/v) water/acetonitrile or 90/10 water/ethanol, allowing to dissolve in a 50-70°C water bath, and cooling to room temperature. Larger volumes of supersaturated solutions are obtained by using the same solute-to-solvent ratio.
• the shape of the crystals formed was dependent on the solvent system used for the phthalic acid.
• the crystals formed with phthalic acid in water/ethanol were long, petal-shaped clusters.
• the crystals formed with water/ethanol were smaller and rhombic.
• Crystals of labeled-insulin/lactose were dissolved in PBS and analyzed by HPLC.
• the level of insulin inclusion was determined to be approximately 0.1%. This process is scalable from 100 ⁇ L to several liters.
• the larger volume crystallizations were performed using glass beakers, or other appropriate large containers, covered with watch glasses.

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