Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
  • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
  • A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
  • A61K47/10—Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K38/00—Medicinal preparations containing peptides
  • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
  • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
  • A61K38/22—Hormones
  • A61K38/25—Growth hormone-releasing factor [GH-RF], i.e. somatoliberin
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K38/00—Medicinal preparations containing peptides
  • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
  • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
  • A61K38/22—Hormones
  • A61K38/27—Growth hormone [GH], i.e. somatotropin
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
  • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
  • A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
  • A61K47/14—Esters of carboxylic acids, e.g. fatty acid monoglycerides, medium-chain triglycerides, parabens or PEG fatty acid esters
• • 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
  • A61K9/0012—Galenical forms characterised by the site of application
  • A61K9/0014—Skin, i.e. galenical aspects of topical compositions
• • 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
  • A61K9/0012—Galenical forms characterised by the site of application
  • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
  • A61P15/14—Drugs for genital or sexual disorders; Contraceptives for lactation disorders, e.g. galactorrhoea

Definitions

• the present invention relates to novel compositions useful for the sustained-release of bioactive proteins. More particularly, the present invention relates to novel suspensions of a protein, more particularly somatotropins and growth hormone releasing factor, in triacetin and in polyethylene glycols. Also provided are methods of using these novel compositions for the sustained or prolonged release of the bioactive proteins.
• bovine somatotropin (bSt) is being produced by recombinant DNA technology. It is well recognized that lactating dairy cows administered recombinant bSt (rbSt) produce more milk compared with controls. Administration of rbSt in a sustained release vehicle will allow less frequent dosing which would be expected to result in a savings of time and labor to dairymen.
• Biocompatible oils have long been used as vehicles to provide sustained delivery of many drugs, including proteins and specifically somatotropins.
• Oleaginous vehicles also have the practical advantage of providing a low water content environment to the suspended, dehydrated protein, thus providing sufficient stability for long term storage of a ready-to-inject product.
• An alternative approach to stabilization involves the use of a non-aqueous, water miscible vehicle.
• a non-aqueous, water miscible vehicle e.g., propylene glycol, glycerin, dimethyl sulfoxide (DMSO), the polyethylene glycols, triacetin, n-methylpyrrolidone, 2-pyrrolidone, and N,N-dimethylacetamide.
• DMSO dimethyl sulfoxide
• these vehicles typically either do not form a suspension or, when mixture is possible, form a suspension not suitable for a commercial product. I have found an exception to these formulation difficulties in glyceryl triacetate (triacetin) and in the polyethylene glycols (PEG).
• Triacetin is a well known article of commerce which is used primarily in industrial applications.
• a number of pharmaceutical uses have been reported, e.g., as a solvent for preparing stable solution formulations of prostaglandin-like compounds (U.S. Patents 3,966,962, 4,301, 175, and 4,680,312), as an antifungal agent (U.S. Patent 3,070,497), as a solvent in tetracycline solutions (U.S. Patent 3,219,529), as a solvent in various topical formulations (GB 2150830, JP 63130541, and JP 59044311 (as a plasticizer in polymeric drug delivery systems); U.S.
• Patent 4,857,313 (as a co-polymer for transdermal delivery systems), and as a dispersing agent in the food industry for hydrophilic colloids such as gelatin (GB 1185340).
• Triacetin is included in the GRAS (generally regarded as safe) list compiled by the U.S. Food and Drug Administration. The Merck Index, 10th Edition, reports triacetin is soluble in water to the extent of about one part triacetin to 14 parts water.
• the polyethylene glycols are well known articles of commerce which have been put to a variety of applications.
• the PEGs are used as plasticizers in other polymeric systems, the high molecular weight PEGs as hydrophilic excipients in compressed tablets for oral drug delivery, and within osmotic minipumps. They are commonly used at low concentrations as dispersing agents in aqueous suspensions and as cosolvents with water or ethanol.
• United State Patent 4,041,155 discloses a composition of an aqueous formulation of 80% PEG 400 and somatostatin which reportedly sustained release of the active agent for 4 hours.
• European Patent Application 0 140 255 discloses a sustained release composition of interferon and a biodegradable carrier which is dispersed in a viscous solvent, e.g. polyethylene glycol. To my knowledge, there are no published reports of the use of a polyethylene glycol suspension for sustained release of a bioactive protein.
• Polyethylene glycols ( ⁇ -hydro- ⁇ -hydroxypoly(oxy-l,2-ethanediyl) have the general formula H(OCH 2 CH 2 ) radicalOH where n>4 and are included in the GRAS (generally regarded as safe) list compiled by the U.S. Food and Drug Administration.
• GB 1185340 reports the use of triacetin as a dispersing agent in the food industry for hydrophilic colloids, e.g. gelatin.
• the Merck Index, 11th Edition, page 1204 lists a variety of uses for the PEGs, including, water-soluble lubricants, in food packaging, in hair and cosmetic preparations, and as an ointment and suppository base in pharmaceuticals.
• United State Patent 4,041,155 discloses a composition of an aqueous formulation of 80% PEG 400 and somatostatin which reportedly sustained release of the active agent for 4 hours.
• European Patent Application 0 140 255 discloses a composition for sustained release comprising interferon and a biodegradable carrier dispersed in a viscous solvent, e.g. polyethylene glycol.
• the present invention relates to compositions of proteins in triacetin or polyethylene glycol (PEG). More particularly, the invention provides prolonged release of hormones, most particularly growth hormone or somatotropin and growth hormone releasing factor.
• a particular embodiment of this aspect of the invention provides bovine somatotropin suspended in triacetin in a concentration of about five to about fifty percent (W/V).
• An alternative embodiment of this aspect of the invention provides bovine somatotropin suspended in PEG 400 in a concentration of about five to about fifty percent (w/v).
• compositions of the invention as sustained release vehicles, especially for subcutaneous and intramuscular injection.
• this aspect of the invention relates to the use of somatotropin suspended in triacetin or polyethylene glycol 400 to increase milk production in cows.
• this aspect of the invention relates to the use of the somatotropin suspended in triacetin or in PEG, in a concentration of about ten to about thirty percent for injection into cows to increase milk production.
• Polyethylene glycols ( ⁇ -hydro- ⁇ -hydroxypoly(oxy-l,2-ethanediyl) have the general formula H(OCH 2 CH 2 ) radicalOH where ⁇ >4.
• PEGs are assigned a numerical suffix based on the average value of n and, thus, include polymers having a range of molecular weights.
• a polymer having an average value for n of 4 has a range of molecular weights of 190- 210 is designated PEG 200; a polymer having an average range of values for n of 8.2-9.1 has a range of molecular weights of 380-420 and is designated PEG 400; a polymer having an average range of values for n of 12.5-13.9 has a range of molecular weights of 570-630 and is designated PEG 600.
• compositions of the invention are conveniently and easily made by dispersing or stirring the protein into the triacetin or polyethylene glycol (PEG).
• PEG compositions of the invention having values of 4 ⁇ n ⁇ 20 are preferred.
• the protein can be readily dispersed in the triacetin or PEG vehicle by mixing in a beaker or other suitable container until a homogenous dispersion is obtained. Alternatively, mixing may be effected by typical high-shear dispersion equipment. In addition, the compositions of the invention may be conveniently made and stored under aseptic conditions. The amount of protein to be added depends on, for example, the protein dose and volume to be administered, as well as by the viscosity, syringeability, and injectability of the final formulation. These and other factors are well known and adjustments easily made by a person skilled in the art of formulation technology.
• any protein or peptide the sustained or prolonged release of which is desired to be effectuated is suitable for use in the compositions of the invention.
• proteins or peptides which are unstable under the aqueous conditions encountered upon administration and are bioactive upon release into the circulatory system are preferred.
• Preferred proteins or peptides for the incorporation into the compositions of the invention are insulin and insulin-like growth factors, interferon, growth hormone releasing factor, interleukins, etc.
• Most preferred are the growth hormones or somatotropins, especially bovine and porcine somatotropin, and growth hormone releasing factor.
• the protein or peptide may be obtained from the natural tissue ("native") or produced by recombinant technology (“recombinant”) and includes proteins or peptides having modified or varied amino acid sequences.
• the essential feature is that the protein or peptide retain bioactivity in the species into which it is administered.
• the protein or peptide is to be added in a dry state, e.g., powder, cake, lyophilized, spray dried, granulated air dried, milled protein, etc., and with or without additives. Thus, it is preferred to obtain the protein or peptide in the appropriate physical state.
• compositions of the invention are used for the same purposes and by the same method as the prior art prolonged or sustained release protein compositions. Accordingly, dosage forms in accordance with the present invention are used for various purposes and dosage levels known in the art, e.g., for the prolonged release of bovine somatotropin by injection into bovine to effectuate increased milk yields and/or increases in lean tissue (U.S. Patents 5,013,713, and 4,977,140), for the prolonged release of insulin, ACTH, glucagon (U.S. Patent 2,902,408), LHRH (B.K. Davis, Experentia and Proc. Natl. Acad. Sci. (1972)), and cyclosporin (U.S. Patent 4,388,307).
• the triacetin and PEG formulations of the invention are suitable for use in both human and non-human animals.
• compositions of the invention provide a slow release of the active agent, i.e., protein or peptide, from a depot formulation. This is surprising indeed, in view of the relatively rapid exposure of the protein depot to the destabilizing aqueous environment.
• the present composition of the invention provide a water-miscible, non-aqueous vehicle which unexpectedly provides a redispersable suspension and, again unexpectedly, provides acceptably prolonged duration of release, while maintaining stability and ease of use similar to that of the prior art oil suspensions.
• compositions of the invention may optionally include preservatives, gelling agents, dispersing agents, stabilizers, etc., known in the art to further optimize the prolonged release of the protein or peptide from the depot or to further enhance the stability of the formulation. All chemicals used are reagent grade or better and are available from commercial vendors. Triacetin is obtained from Sigma Chemical Co. and can be sterilized using pharmaceutically acceptable methods, e.g., filtration, heat treatment, etc., prior to use. The various polyethylene glycols are widely available from commercial vendors. The PEGs also may be sterilized using pharmaceutically acceptable methods, e.g., filtration, heat treatment, etc., prior to use.
• Bioactive means a protein which has biological activity in the species of interest.
• W/V means weight protein in grams per unit volume of suspension as administered.
• Example 1 Somatotropin Triacetin Sustained Release Formulation 25.01 g of triacetin is added to 6.85 grams of bioactive recombinant bovine somatotropin (Somavubove, The Upjohn Company) and mixed with a spatula to result with a suspension containing 200 mg rbSt/ml. The formulation is designated SR-SbV TAC and is stored in a 20 ml stoppered vial until use.
• Example 2 Serum Somatotropin (Triacetin) Aliquots of the suspension made in Example 1 are removed for administration into non- pregnant non-lactating heifers.
• non-pregnant, non-lactating Holstein heifers are housed in dry lots in southeastern Michigan. Alfalfa/grass hay and water are available ad libitum; heifers also receive 15 kg corn silage per head per day. Heifers are assigned randomly to the following groups: (1) Control (no injection); (2) 150 mg SR-SbV/TAC (150 mg rbSt in triacetin); and (3) 300 mg SR-SbV/TAC (300 mg rbSt in triacetin).
• the formulations are injected subcutaneously over the rib, caudal to the left shoulder. Hair at injection site is clipped prior to injection.
• the formulation is easily injected with an 18 gauge X 2.5 cm needle. Injection volume is 0.7 ml for heifers receiving 150 mg rbSt in triacetin and 1.5 ml for heifers receiving 300 mg rbSt in triacetin.
• Blood is collected from the tail vein for 7 days following injection. Injections are given at time zero (0) and samples collected at 0, 6, 12, 18, 24, 30, 36, 48, 60, 72, 84, 96, 120, and 144 hours. Blood is allowed to clot, centrifuged, and serum decanted and stored at -20° until assay by radioimmunoassay following the procedure of G.D. Niswander, et ah, Endocrinol. 84:1166 (1969).
• Example 3 Somatotropin/PEG Sustained Release Formulation 21.8 g of PEG 400 (Sigma Chemical) is added to 6.00 grams of bioactive recombinant bovine somatotropin (rbSt) (Somavubove, The Upjohn Company) and mixed with a spatula to result with a suspension containing 200 mg rbSt ml. The formulation is designated SR- SbV/PEG and is stored in a 20 ml stoppered vial until use.
• Example 4 Serum Somatotropin (PEG) Aliquots of the suspension made in Example 3 are removed for administration into non- pregnant non-lactating heifers.
• non-pregnant, non-lactating Holstein heifers are housed in dry lots in southeastern Michigan. Alfalfa/grass hay and water are available ad libitum; heifers also receive 15 kg corn silage per head per day. Heifers are assigned randomly to the following groups: (1) Control (no injection); (2) 150 mg SR-SbV/PEG (150 mg rbSt in PEG 400); and (3) 300 mg SR-SbV/PEG (300 mg rbSt in PEG 400). The formulations are injected subcutaneously over the rib, caudal to the left shoulder.
• Blood is collected from the tail vein for 7 days following injection. Injections are given at time zero (0) and samples collected at 0, 6, 12, 18, 24, 30, 36, 48, 60, 72, 84, 96, 120, and 144 hours. Blood is allowed to clot, centrifuged, and serum decanted and stored at -20° until assay by radioimmunoassay following the procedure of G.D. Niswander, et al., Endocrinol. 84:1166 (1969).
• a least squares means (ng bSt/ml of serum).
• m Means with this superscript are not significantly (P>,05) different from overall control mean (3.7 ng bSt/ml serum).
• the control mean is the average concentration of bSt for all blood sampling times (through 144h).

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• 1993
  • 1993-08-23 EP EP93920157A patent/EP0661989B1/en not_active Expired - Lifetime
  • 1993-08-23 ES ES93920157T patent/ES2107051T3/es not_active Expired - Lifetime
  • 1993-08-23 AU AU50186/93A patent/AU5018693A/en not_active Abandoned
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  • 1993-08-23 JP JP50808394A patent/JP3756512B2/ja not_active Expired - Fee Related
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• 1995
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• 1997
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