MXPA98008786A - A pharmaceutical formulation containing a growth hormone, an amino acid and a non-ion detergent - Google Patents

Abstract

The invention relates to pharmaceutical formulations comprising a growth hormone, an amino acid selected from the group of Asp, Ile, Val, Leu or His, or a histidine derivative, or a peptide comprising at least one basic amino acid and at least one acidic amino acid, and a non-ionic detergent, for example, a polysorbate or a poloxamer. The formulation is synthesized against deamidation and aggregation. The formulation may be an aqueous formulation. Growth hormone can be a human growth hormone

Description

A PHARMACEUTICAL FORMULATION CONTAINING A HORMONE OF GROWTH, AN AMINO A NON-IONIC DETERGENT FIELD OF THE INVENTION The invention relates to new pharmaceutical formulations comprising a growth hormone, or a growth hormone derivative, and an amino acid selected from the group of Asp, He, Val, Leu, or His, or a histidine derivative, or a a peptide comprising at least one basic amino acid and at least one acidic amino acid, and a nonionic detergent, for example, a polysorbate or a poloxamer. The invention further relates to a method for producing such formulations and to a method for treatment using such formulations.

BACKGROUND OF THE INVENTION The growth hormones of man and common domestic animals are proteins of approximately 191 amino acids, synthesized and secreted from the anterior lope of the gland REF: 028598 pituitary. Human growth hormone consists of 191 amino acids. Growth hormone is a key hormone involved in the regulation of growth not only somatic, but also in the regulation of the metabolism of proteins, carbohydrates and lipids. The greatest effect of growth hormone is to promote growth. __ Organ systems affected by growth hormone include the skeleton, connective tissue, muscles, and viscera such as the liver, intestines, and kidneys. Until the development of recombinant technology and the cloning of the growth hormone gene that now gives rise to the production of, for example, human growth hormone (hGH) and Met-hGH in a On an industrial scale, human growth hormone could only be obtained by extracting the pituitary glands from human cadavers. Very limited supplies of growth hormone limited the use of growth hormone for the acceleration of longitudinal growth in childhood and puberty for the treatment of dwarfism, although it has been proposed for the inter alia treatment of short stature (due to the Growth hormone deficiency, normal short stature and Turner syndrome), growth hormone deficiency in adults, infertility, treatment of burns, healing sores, dystrophy, repair of bone fractures, osteoporosis, diffuse gastric bleeding, and pseudoarthrosis. In addition, growth hormone has been proposed to increase the growth rate of domestic animals or to decrease the proportion of fat. The pharmaceutical preparations of growth hormone tend to be unstable. Degradation products are generated such as deaminated or sulfoxidated products and dimer or polymer forms - especially in growth hormone solutions. The predominant chemical degradation reactions of hGH are 1) deamidation by direct hydrolysis or by means of a cyclic succinimide intermediate to form various amounts of L-asp-hGH, L-iso-asp-hGH, D-asp- hGH and D-iso-asp-hGH (ref 1-3), 2) oxidation of merionine residues at positions 14 and 125 (ref 4-9), and 3) decomposition of peptide bonds. The deamidation takes place especially in the Asn at position 149. The hGH is easily oxidized preferably at positions 14 and 125, especially in the solution (4-8), since the oxidation of hGH in the sulfoxides forming the solution is normally due to dissolved oxygen in the preparation. Now, it is not believed that these degradation products should have a biological, toxic or altered activity or binding properties of the receptor, but there is an indication to the effect that the stability of the conformation of the sulfoxides is reduced when compared to natural hGH . Other "degradation products" of growth hormone are the aggregation products such as dimers and polymers. Studies have been conducted to clarify the role of these forms in inducing an immune response with measurable amounts of antibodies to natural hGH. These studies have indicated that the aggregates of hGH are the primary cause of immunogenicity in patients.

Thus, it is desirable in a pharmaceutical formulation to avoid the formation of growth hormone aggregates since aggregate limes are capable of causing undesirable immunogenicity or altered half-life. The degradation kinetics depends on temperature, pH and various additives or auxiliaries in the formulation of hGH. Because of the instability, a growth hormone formulation is now freeze-dried and stored normally in the lyophilized form at 2-8 ° C until it is reconstituted for use in order to minimize degradation. The lyophilized pharmaceutical preparations comprising the hGH are reconstituted by the patient and then stored as a solution during use for a period of up to 30 days, during which some degradation will occur. It is now preferred to reconstitute the growth hormone as recently as possible before use and store and ship the preparation in a lyophilized state. The chain from the manufacturer to the pharmacy is able to handle the preparations at a low, controlled temperature of, for example 2-8 ° C which allows a shelf life of up to three years. Preferably, a lyophilized and then reconstituted preparation should be stable with the end user in a lyophilized state for about one month and additionally for one month in a reconstituted state in a storage device during the proposed period of use of a cartridge. As an alternative to lyophilized and reconstituted preparations, growth hormone can be formulated as a liquid formulation suitable for use in vials or in a storage system for self-medication. The widespread use of storage systems for self-medication and the expanded field of use requires a preparation which is stable for a sufficiently long time with the end user. Such stabilization is of great importance when transferring the administration of growth hormone from the clinics to the homes of the individuals to be treated where optimal storage conditions may not be available. A "ready-to-use" formulation further diminishes any of the handling problems in connection with reconstitution and thus represents a convenience for the patient. A dissolved, stable preparation comprising the growth hormone can be easily produced for use in the form of vials used by the patient in combination with conventional syringes or as cartridges that fit into the storage device used by the patient. In both cases, the patient can then avoid reconstitution of the preparation and, therefore, will not have to have a lyophilized preparation, a suitable excipient for reconstitution as well as the necessary skill and sterile equipment for sterile reconstitution of the preparation. Safety reasons also make it desirable to avoid reconstitution of a lyophilized preparation just prior to the use of the preparation. From the point of view of the manufacturers, it can be an advantage to avoid the step of lyophilization in the production of the growth hormone preparations. Freeze-drying is a costly and time-consuming process, and a "bottleneck" in production due to the limited capacity of the lyophilizer is also frequent.

In this way, there is a need for dissolved, more stable preparations of growth hormone in order to facilitate the handling to be performed by the patient. Thus, there is also a need to reduce the rate of degradation processes in order to allow the preparations of the dissolved hGH to be stable during shelf life and during the period of use of up to about one month.

Prior art Previous attempts to stabilize hGH have not been completely successful in preventing dimer formation. The problems associated with the formation of the dimer are observed for example in Becker, G.W., Biotechnology and Applied Biochemistry 9, 478 (1987). International Patent Publication No. WO 89/09614 and Australian Patent Application No. 30771/89 disclose a stable pharmaceutical formulation containing human growth hormone, glycine and mannitol. Such formulation shows improved stability during normal processing and storage in a lyophilized state as well as in the period of use after reconstitution. The European patent application, published No. 303 746, teaches that the animal growth hormone can be stabilized with various stabilizers to give a decreased formation of insolubles and the conservation of the soluble activity in aqueous environments, such stabilizers include certain polyols, amino acids, polymers of amino acids having a secondary group, loaded at a physiological pH, and choline salts. The polyols are selected from the group consisting of non-reducing sugars, sugar alcohols, sugar acids, pentaerythritol, lactose, water soluble dextrans and Ficoll.; the amino acids are selected from the group consisting of glycine, sarcosine, lysine or salts thereof, serine, arginine or salts thereof, betaine, N, N, -dimethylglycine, aspartic acid or salts thereof, glutamic acid or salts thereof; a polymer of an amino acid having a secondary group charged at a physiological pH can be selected from polylysine, polyaspartic acid, polyglutamic acid, polyarginine, polyhistidine, polyornithine and salts thereof; and the choline derivatives are selected from the group consisting of choline chloride, choline diacid citrate, choline bitartrate, choline bicarbonate, tricholine citrate, choline ascorbate, choline borate, choline gluconate, choline phosphate, (choline) sulfate and dicoline mucate. US Patent Specification No. 4,917,685 discloses a delivery device designed to be implanted, comprising stabilized growth hormone using the same stabilizers as those mentioned in European patent 303746. European patent application, published No. 374,120 publishes a stabilized formulation comprising a hGH and a polyol having three hydroxy groups. Glycerol and tris (hydroxymethyl) aminomethane are mentioned. In addition, the presence of histidine hydrochloride is reported as a buffer together with the polyol. The International Patent Publication No.

WO 93/12811 publishes stabilized formulations of growth hormone in the form of a lyophilized powder or an aqueous solution - comprising asparagine.

International Patent Publication No. WO 93/12812 publishes stabilized formulations of growth hormone in the form of a lyophilized powder or an aqueous solution comprising histidine. In such formulations, deamidation is reduced to 25-30% as compared to a corresponding formulation of growth hormone comprising a phosphate buffer. International Patent Publication No. WO 96/11703 publishes stabilized formulations of growth hormone comprising isoleucine. International Patent Publication No. WO 96/11702 publishes stabilized formulations of growth hormone comprising valine. International Patent Publication No. WO 96/11704 publishes stabilized formulations of growth hormone comprising leucine. International Patent Publication No. WO 93/19776 discloses protein formulations comprising growth hormone comprising citrate as a buffer substance which is more stable than formulations comprising a phosphate buffer. The formulations may also comprise amino acids such as glycine and alanine and / or mannitol or other sugar alcohols and / or glycerol and / or other carbohydrates and optionally a preservative such as benzyl alcohol. International Patent Publication No. WO 94/03198 discloses a stable, aqueous formulation containing human growth hormone, a buffer, a non-ionic surfactant, and optionally, a neutral salt, mannitol, or, a preservative. The buffer can be histidine although citrate is preferred. The application publishes a preferred formulation containing citrate as a buffer, and natrium chloride and polysorbate 20 (Tween 20) for stabilization. This combination stabilizes the aqueous formulation against aggregation but gives rise to a considered amount of deamidated growth hormones. International Patent Publication No. WO 95/35116 publishes a lyophilized, stable formulation containing sucrose and, optionally, mannitol.

DESCRIPTION OF THE INVENTION It has now surprisingly been found that a pharmaceutical formulation comprising the growth hormone, or a growth hormone derivative, and an amino acid selected from the group of Asp, He, Val or Leu, histidine, or a histidine derivative, or a peptide comprising at least one residue of a basic amino acid and at least one residue of an acidic amino acid, and a nonionic detergent, wherein the amount of the peptide, or Asp, He, Val, Leu or His, or a derivative of histidine is about 0.01 to about 10 mg per mg of GH, shows a high stability against deamidation and aggregation. The stability of the product allows the storage and shipment thereof in a lyophilized state or in the form of a dissolved or redissolved formulation. The peptide comprising at least one residue of a basic amino acid and at least one residue of an acidic amino acid to be used according to the present invention can be a peptide comprising up to 20 amino acid residues, preferably from 3 to 10 residues of amino acid, more preferably from 3 to 6 amino acid residues such as 3 or 4 amino acid residues (for example, Lys-Gly-Asp-Ser). According to one aspect of the invention, the basic amino acid residues of the shorter peptides are separated by 1 or 2 amino acid residues. The peptides preferably comprise naturally occurring alpha amino acid residues. The amino acid (s) _ ^ can (n) be the amino acid (s) 1 or d or a mixture thereof. "The acidic amino acid residues" are for example, Glu or Asp, and "the basic amino acid residues" are for example, Lys or Arg. The histidine derivatives to be used according to the present invention may be amides and esters of histidine such as the methyl or ethyl ester, dipeptides having the His-X or X-His sequence, when X is a naturally-occurring amino acid residue, and analogues or His derivatives such as imidazole, des-amino-His or poly-His. In a preferred aspect, the invention relates to a pharmaceutical formulation comprising growth hormone (GH) or a growth hormone derivative and an amino acid selected from the group of Asp, He, Val or Leu, and a non-ionic detergent. , wherein the amount of the amino acid selected from the group of Asp, He, Val or Leu is from about 0.01 to about 10 mg per mg of GH. In another preferred aspect, the invention relates to a pharmaceutical formulation comprising growth hormone (GH) or a growth hormone derivative and histidine, or a histidine derivative, or a peptide comprising at least one residue of a basic amino acid and at least one residue of an acidic amino acid, and a nonionic detergent, wherein the amount of the peptide or histidine or a histidine derivative is from about 0.01 to about 10 mg per mg of GH. In a preferred embodiment of the above aspect, the amount of the selected peptide, or amino acid, or histidine, or histidine derivative, is from about 0.05 to about 0.5 mg per mg of GH, more preferably from about 0.05 to about 1 mg per mg of GH, even more preferred from about 0.05 to about 0.7 mg per mg of GH, and even more preferred from about 0.05 to about 0.5 mg per mg of GH. In a preferred embodiment of the above aspect, the non-ionic detergent is selected from a polysorbate or a poloxamer, for example, polysorbate 20, poloxamer 188, or poloxamer 407 (eg, Pluronic® F68, Lutrol 127, Tween 20), more preferred poloxamer 188. In another aspect, the invention relates to a pharmaceutical formulation of growth hormone, or a growth hormone derivative, comprising an amino acid selected from the group of Asp, He, Val, Leu or His, or a histidine derivative, or a peptide comprising at least one residue of a basic amino acid and at least one residue of an acidic amino acid, and a nonionic detergent, wherein the amount of the nonionic detergent is from about 0.01 to about 10 mg per mg of GH. In a preferred aspect, the invention relates to a pharmaceutical formulation comprising growth hormone (GH) or a growth hormone derivative, and an amino acid selected from the group of Asp, He, Val or Leu, and a detergent not ion, wherein the amount of the non-ionic detergent is from about 0.01 to about 10 mg per mg of the GH. In another preferred aspect, the invention relates to a pharmaceutical formulation, comprising growth hormone (GH), or a growth hormone derivative, and histidine, or a histidine derivative, or a peptide comprising at least one residue of a basic amino acid and at least one residue of an acidic amino acid, and a nonionic detergent, wherein the amount of the non-ionic detergent is from about 0.01 to about 10 mg per mg of the GH. In a preferred embodiment of the above aspect, the amount of the nonionic detergent is from about 0.01 to about 10 mg per mg of the GH, more preferred from about 0.05 to about 5 mg per mg of the GH, even more preferred from about 0.1 to about 3 mg per mg of GH, even more preferred from about 0.1 to about 2 mg per mg of GH. In a preferred embodiment of the above aspect, the non-ionic detergent is selected from a polysorbate or a poloxamer, for example, polysorbate 20, poloxamer 188, or poloxamer 407 (eg, Pluronic® F68, Lutrol 127, Tween 20), more preferred poloxamer 188. Another aspect of the invention is a pharmaceutical formulation comprising growth hormone, or a growth hormone derivative, and an amino acid selected from the group of Asp, He, Val, Leu or His or a histidine derivative, or a peptide comprising at least one basic amino acid and at least one acidic amino acid, and a non-ionic detergent, wherein the of the peptide, or Asp, He, Val, Leu or His or a histidine derivative, is from about 0.01 to about 10 mg per mg of GH, and the amount of the non-ionic detergent is from about 0.1 to about 2 mg per mg of the GH. In a preferred aspect, the invention relates to a pharmaceutical formulation comprising growth hormone or a growth hormone derivative, and an amino acid selected from the group of Asp, Lie, Val or Lue, and a non-ionic detergent, in wherein the amount of Asp, Lie, Val or Leu is from about 0.01 to about 10 mg per mg of the GH, and the amount of the non-ionic detergent is from about 0.1 to about 2 mg per mg of the GH. In another preferred aspect, the invention relates to a pharmaceutical formulation comprising the growth hormone, or a growth hormone derivative and a histidine, or a histidine derivative, or a peptide comprising at least one residue of an amino acid. basic and at least one residue of an acidic amino acid, and a non-ionic detergent, wherein the amount of the peptide or histidine or the histidine derivative is from about 0.01 to about 10 mg per mg of the GH, and the amount of the detergent does not ion is from about 0.1 to about 2 mg per mg of GH. In a preferred embodiment of the above aspect, the amount of the peptide, or Asp, He, Val, Leu or His, or the histidine derivative, is from about 0.05 to about 5 mg per mg of the GH, more preferred from about 0.05 to about 1 mg per mg of GH, even more preferred from about 0.05 to about 0.7 mg per mg of GH, and even more preferred from about 0.05 to about 0.5 mg per mg of GH. In a preferred embodiment of the above aspect, the nonionic detergent is selected from a polysorbate or poloxamer, for example polysorbate 20, poloxamer 188, or poloxamer 407 (for example, Pluronic® F68, Lutrol 127, Tween 20), poloxamer 188 is more preferred.

Another aspect of the invention relates to a pharmaceutical formulation comprising the growth hormone or an amino acid selected from the group of Asp, He, Val, Leu or His, or a histidine derivative, or a peptide comprising at least one residue of a basic amino acid and at least one residue of an acidic amino acid, and a poloxamer. In a preferred aspect, the invention relates to a pharmaceutical formulation comprising growth hormone (GH) or a growth hormone derivative, and an amino acid selected from the group of Asp, He, Val or Leu, and a poloxamer. In another preferred aspect, the invention relates to a pharmaceutical formulation comprising the growth hormone, or a growth hormone derivative, and histidine, or a histidine derivative, or a peptide comprising at least one residue of an amino acid basic and at least one residue of an acidic amino acid, and a poloxamer. In a preferred embodiment of the above aspect, the poloxamer is selected from poloxamer 188 or poloxamer 407, more preferred is poloxamer 188 (for example, Pluronic® F68). In another preferred embodiment, the formulation comprises histidine, or a histidine derivative. In a preferred embodiment, the amount of the peptide, or Asp, He, Val, Leu or His, or a histidine derivative is from about 0.05 to about 5 mg per mg of GH, more preferred from about 0.05 to about 1 mg per mg of GH, even more preferred from about 0.05 to about 0. 7 mg per mg of GH, and even more preferred from approximately 0.05 to approximately 0.5 mg per mg of GH. In a preferred embodiment, the amount of the nonionic detergent is from about 0.05 to about 5 mg per mg of the GH, more preferably from about 0.1 mg to about 3 mg per mg of the GH, more preferably from about 0.1 to about 2 mg per mg of GH. The formulation of the invention may be in the form of a lyophilized powder to be reconstituted later using conventional excipients such as distilled water, or water for injection, or it may be in the form of an aqueous solution comprising growth hormone. Such excipients may comprise conventional preservatives such as m-cresol, phenol, and benzyl alcohol. As the augmentation agent (s) for lyophilization, one or more of the members of the group consisting of sugar alcohols, for example, mannitol, and disaccharides, eg, sucrose, may be selected. Lyophilized preparations comprising sucrose due to very high stability are preferred, and preparations comprising sucrose and mannitol which combine very high stability with very good processability to give freeze-dried, firm products which are readily soluble and very stable are especially preferred. in the solution for an extended period of time after dissolution. The further preferred preparations according to the invention are preparations comprising mannitol and sucrose or trehalose as an augmentation agent for lyophilization. The preparations according to the invention comprising mannitol and a disaccharide usually comprise an approximately equal amount of the two constituents in a base by weight. The amount of sucrose present in the preparations of the invention may vary within extended limits. The ratio of growth hormone to sucrose can vary from 0.005 to 1.5 on a weight basis. In this way, the amount of sucrose can be from 0.67 to 200 mg per mg of growth hormone, an amount of 1.1 to 50 mg per mg of growth hormone is preferred. In a preferred embodiment of the invention, the growth hormone is human growth hormone (hGH) and the pharmaceutical composition is further comprised of a carrier in the form of an aqueous buffer. Such an aqueous preparation is in a ready-to-use form and can be stored and shipped as such without any considerable degradation. A buffer to be used in a growth hormone solution can be, for example, a histidine, citrate, tartrate, or phosphate buffer. The L-histidine has a pKA of 6.0 and is therefore suitable as a buffer itself in a pH range of about 5 to about 7. In a further aspect, the invention relates to a method for the preparation of a pharmaceutical formulation comprising a growth hormone or a growth hormone derivative, and an amino acid selected from the group of Asp, He, Val, Leu or His, or a histidine derivative or a peptide which comprises at least one residue of a basic amino acid and at least one residue of an acidic amino acid, and a nonionic detergent, wherein the appropriate amount of the growth hormone, or the growth hormone derivative, is dissolved in a solution comprising an amino acid selected from the group of Asp, He, Val, Leu or His, or a derivative of histidine, or a peptide comprising at least one residue of a basic amino acid and at least one residue of an acidic amino acid in water of sionized, and a non-ionic detergent, for example, a polysorbate or a poloxamer, optionally containing a preservative, and optionally containing an agent for adjusting the tonicity, and optionally adjusting the pH to a value of about 6.0 to about 8.8.; the appropriate amount. of GH which is what causes the amount of the peptide, Asp, He, Val, Leu or His, or the histidine derivative, to vary from about 0.01 to about 10 mg per mg of GH. In a further aspect, the invention relates to a method for preparing a pharmaceutical formulation comprising a growth hormone, or a growth hormone derivative, and an amino acid selected from the group of Asp, He, Val, Leu, or His , or a histidine derivative or a peptide comprising at least one residue of a basic amino acid and at least one residue of an acidic amino acid, and a nonionic detergent, wherein the growth hormone, or the growth hormone derivative , is dissolved in a solution comprising an amino acid selected from the group of Asp, He, Val, Leu, or His, or a histidine derivative or a peptide comprising at least one residue of a basic amino acid and at least one residue of an acidic amino acid in deionized water, and an appropriate amount of a non-ionic detergent, for example, a polysorbate or a poloxamer, optionally containing a preservative, and optionally containing an agent for using the tonicity, and optionally adjusting the pH to a value of about 6.0 to about 8.8; the appropriate amount of the non-ionic detergent which is what causes the amount of the non-ionic detergent to vary from about 0.01 to about 10 mg per mg of the GH. In a further aspect, the invention relates to a method for the preparation of a pharmaceutical formulation comprising a growth hormone, or a growth hormone derivative, and an amino acid selected from the group of Asp, He, Val, Leu, or His, or a histidine derivative, or a peptide comprising at least one residue of a basic amino acid and at least one residue of an acidic amino acid, and a nonionic detergent, comprising the dissolution of such a quantity of the hormone of the growth, or a growth hormone derivative, which causes the amount of the peptide, or Asp, He, Val, Leu or His, or a histidine derivative to vary from about 0.05 to about 0.5 mg per mg of GH, in a solution comprising an amino acid selected from the group of Asp, He, Val, Leu or His, or a histidine derivative, or a peptide comprising at least one residue of a basic amino acid and at least one residue of an acidic amino acid, and a Such a quantity of a non-ionic detergent, for example a polysorbate or a poloxamer, wherein the amount of the non-ionic detergent is from about 0.1 to about 2 mg per mg of the GH, in deionized water, optionally containing a preservative, and optionally contains an agent for adjusting the tonicity, and optionally adjusting the pH to a value of about 6.0 to about 8.8. In a further aspect, the invention relates to a method for the preparation of a pharmaceutical formulation comprising a growth hormone, or a growth hormone derivative, and an amino acid selected from the group of Asp, Lie, Val, Leu, or His, or a histidine derivative, or a peptide comprising at least one residue of a basic amino acid and at least one residue of an acidic amino acid, and a poloxamer, wherein the growth hormone, or a derivative of the hormone of the growth, is dissolved in a solution comprising an amino acid selected from the group of Asp, He, Val, Leu, or His, or a histidine derivative or a peptide comprising at least one residue of a basic amino acid and at least one residue of an acidic amino acid in deionized water, and a poloxamer, optionally containing a preservative, and optionally containing an agent for adjusting the tonicity, and optionally adjusting the pH to a value of approximately 6.0 to approximately 8.8. The concentration of the amino acid selected from the group of Asp, He, Val, Leu, or His, or a derivative of histidine, or a peptide which comprises at least one basic amino acid and at least one acidic amino acid, is preferably from about 1 mM to about 100 mM. More preferred, the concentration of Asp, He, Leu, or His, or a histidine derivative, or a peptide comprising at least one basic amino acid and at least one acidic amino acid, is from about 2 mM to about 20 mM, more preferred from about 5 mM to about 15 mM. The pharmaceutical formulations of the invention can be formulated for administration in any suitable manner, for example, by parenteral or oral administration or administration to a mucosal membrane, for example, nasal administration. The pharmaceutical formulation can be presented in the form of a dose contained in a vial or cartridge or any other suitable container such as a prefilled syringe or a storage device. The pharmaceutical preparation of the invention may further comprise salts conventionally used in order to facilitate processing thereof, for example, lyophilization or reconstitution.

For reasons of stability, the pH of the solution is preferably adjusted to a value in the range of from about 6.0 to about 8.8, preferably from about 6.0 to about 7.0, more preferred from about 6.0 to about 6.8, even more preferred. from about 6.0 to 6.3 and much more preferred from about 6.0 to 6.2 (for example, about 6.1). The pH can be adjusted by adding an acid which does not have an adverse effect on the growth hormone, preferably a physiologically acceptable acid, for example, a mineral acid such as hydrochloric acid, sulfuric acid or nitric acid or an organic acid such as acetic acid or citric acid. The pharmaceutical formulation of the invention may further comprise salts for adjusting the tonicity and optionally an excipient in order to facilitate processing thereof, for example, lyophilization and rapid and complete dissolution of a lyophilized formulation when the formulation is reconstituted prior to use. Such salts may be selected from conventional additives such as alkali metal, alkaline earth metal or ammonium salts of organic acids such as citric acid, tartaric acid or acetic acid, for example, sodium citrate, sodium tartrate or sodium acetate, or mineral acids such as hydrochloric acid, for example, sodium chloride. An excipient of the disaccharides such as lactose, trehalose, and sucrose, sugar alcohols such as mannitol, xylitol, erythritol, treitol, sorbitol or glycerol, polysaccharides such as the polymers marketed as Dextran® products such as Dextran® 40, can be selected. Dextran® 70 or Dextran® 75, and Ficoll® and polyvalent alcohols such as polyethylene glycol or polyvinyl alcohol or a combination of two or more of these. In the present context, "growth hormone" or "GH" can be a growth hormone of any origin such as growth hormone of poultry, cattle, eguines, humans, sheep, swine, salmon, trout or tuna, preferably growth hormone of bovine, human or porcine, the growth hormone of human gue is the most preferred. The growth hormone used according to the invention can be a natural growth hormone isolated from a natural source, for example, by extracting the pituitary glands in a conventional manner, or a growth hormone produced by recombinant techniques, for example, as is described in EB Jensen and S. Carlsen in Biotech and Bioeng. 36, 1-11 (1990). The "growth hormone derivative" can be a truncated form of a growth hormone in which one or more amino acid residues has been deleted (s); an analogous part thereof wherein one or more amino acid residues in the natural molecule has been (are) substituted by another residue of an amino acid, preferably the residue of an amino acid which occurs naturally, since the substitution does not have any adverse effects such as antigenicity or reduced action; or a derivative thereof, for example, deamidated or sulfoxidated forms of growth hormone or forms having an N- or C-terminal extension such as Met-hGH, Met-Glu-Ala-Glu-hGH or Ala- Glu-hGH. The preferred growth hormone is hGH. The growth hormone can be a concentrate obtained, directly from fermentation in broth or a freeze-dried, conventional preparation that is dissolved in an appropriate solvent. The term "histidine derivatives" is used, for the present purpose, to designate amides and esters of histidine such as methyl or ethyl ester dipeptides such as His-Gly, His-Ala, His-Leu, His-Lys , His-Ser, His-Phe, and dipeptides such as Ala-His, Gly-His, Val-His, Glu-His, Met-His, Arg-His, Asp-His, Leu-His, Ser-His, and analogs or derivatives of His such as imidazole, des-amino-His or poly-His. In the present context "non-ionic detergent" or "non-ionic surfactant" may be polysorbates, such as polysorbate 20 or 80, etc., and poloxamers, such as poloxamer 188 or 407, Pluronic® polyols, and other ethylene / polypropylene polymers, etc. The amounts effective to provide an aqueous, stable formulation will be used, usually in the range of about 0.01 to about 10 mg of detergent per mg of growth hormone, preferably from about 0.05 to about 5 mg of detergent per mg of the Growth hormone, more preferred from about 0.1 to about 2 mg of detergent per mg of growth hormone. The use of nonionic detergents allows the formulation to be exposed to shear stress and surface tension without causing denaturation of the protein. The amount of the nonionic detergent in the formulations may be in the range of 0.01 to about 10 mg of detergent per mg of growth hormone, preferably 0.05 to about 5 mg of detergent per mg of growth hormone, more preferred from 0.1 to about 2 mg of detergent per mg of growth hormone. In the present context. "disaccharide" is used to designate naturally occurring disaccharides such as sucrose, trehalose, maltose, lactose, sepharose, turanosa, laminaribiose, isomaltose, gentiobiose or melibiose. A peptide comprising at least one residue of a basic amino acid and at least one residue of an acidic amino acid to be used in accordance with the present invention is preferably comprised of naturally occurring a-amino acid residues. The amino acid (s) can be the amino acid (s) 1 or d or a mixture of the same (s). A preferred peptide comprising at least one residue of a basic amino acid and at least one residue of an acidic amino acid is Lys-Gly-Asp-Ser. In the present context "acidic amino acid residues" are, for example, Glu or Asp, and "basic amino acid residues" are, for example, Lys or Arg. The solvent used in the method of the invention may be water, alcohols such co or ethyl, n-propyl or isopropyl, butyl alcohol or mixtures thereof. To retard microbial growth, the solvent may comprise a preservative such as m-cresol, phenol, or benzyl alcohol. The term "dose" of growth hormone refers to an amount that provides a therapeutic effect in a regimen of administration. The formulations thereof are prepared containing amounts of the hGH of at least about 0.1 mg / ml, preferably above about 1 mg / ml, preferably from about 1 mg / ml to about 40 mg / ml, of more preferably from about 1 mg / ml to about 25 mg / ml, more preferably from about 1 mg / ml to about 15 mg / ml, for example, from 1 mg / ml to about 10 mg / ml, calculated in the formulation ready for use. For the use of those compositions in administration to humans suffering from hypopituitary dwarfism, for example, those formulations contain from about 0.1 mg / ml to about 10 mg / ml, corresponding to the dosage regimen currently contemplated for the proposed treatment. The concentration range is not critical to the invention and can be varied by the physician supervising the administration. Another aspect of the invention relates to a method for the treatment of a patient who may be affected by growth hormone which comprises treating the patient with an amount of the pharmaceutical formulation according to the invention effective to treat said disorder. The invention is explained in more detail in the following Examples, which illustrate the invention. These are not considered as limiting the scope of the invention, being defined by the appended claims.

EXPERIMENTAL PART Example 1: Inhibition of aggregation The liquid formulations of hGH were prepared by dissolving hGH in a solution of excipients and adjusting the pH with HCl / NaOH. The composition of the formulations was: hGH 5 mg / ml Mannitol 45 mg / ml L-histidine 0.62 mg / ml Phenol 2.5 mg / ml Surfactant x mg / ml pH = y (from 6.1 to 6.8) The appearance of the solution was examined visually after the solutions had been stored 1 day at 8 ° C and rotated for 19 hours (20 rpm) at 25 ° C, respectively: Example 2: Deamidation rate The eight formulations tabulated below were prepared by adding hGH to a solution containing the excipients. The pH was adjusted to 6.1. The preparations were then placed for 28 days at 25 ° C and for 14 days at 37 ° C and analyzed for the formation of deaminated forms of hGH. The evaluation constants for the deamidation were calculated based on the stability data. In the following table the contents of hGH and the excipients are given in mg / ml and the evaluation constants k25 = c and 37 ° c are given in days-1 Example 3 Formation of deamidated forms The formulations tabulated below were prepared by adding the hGH to a solution containing the excipients and adjusting the pH. The preparations were then placed for 3 months at 8 ° C and 25 ° C and analyzed for the formation of the deamidated forms of hGH. 1): The type of surfactant is given in parentheses F68 = Pluronic F68 = poloxamer 188 F127 = Lutrol 127 = poloxamer 407 T20 = Tween 20 = polysorbate 20 Example 4: The 24 formulations were prepared from the following formula: hGH 3.33 at 13.1 mg / ml L-histidine 0.78 mg / ml Mannitol 22 mg / ml Sucrose 21 mg / ml Poloxamer 188 1.33 - 5.33 mg / ml Preservative: Benzyl alcohol (0 20 mg / ml) or Phenol (0 - 5 mg / ml) The formulations were prepared by adding hGH to a solution of the excipients. The pH was adjusted to 6.8. The formulations were maintained at 25 ° C for 4 weeks and then the clarity of the solutions was measured as the absorbance at 340 nm. The absorbance at 340 nm indicates the degree of aggregation in the solutions. the dissolution of hGH was not possible.

REFERENCES 1) Y. -C-J. Wang and M.A. Hanson. Parenteral Formulations of Proteins and Peptides: Stability and Stabilizers. J. Parenteral Science and Technology 42 (Suppl.) (1988) 53-525. 2) M.C. Manning, K. Patel, R.T. Borchardt. Stability of Protein Pharmaceuticals. Pharmaceutical Research 6 (11) (1989) 903-918. 3) B.A. Johnson, J.M. Shirokawa, W.S. Hancock, M.W.

Spellman, L.J. Basa and D.W. Asward.

J.Biol. Chem.264, 1462-71 (1989). 4) L.C. Teh et al., J. Biol. Chem., 262, 785-794 (1987). 5) G.W. Becker et al., Biotech Appl. Biochem. , 10, 326-337 (1988). 6) R.A. Houghten and. collaborators, Arch. Biochem. Biophys. , 178, 350-355 (1977). 7) R.M. Riggin et al., Anal. Biochem. , 167, 199-209 (1987). 8) P-Gellerfors et al. Acta Paediatr. Scand (suppl), 370, 93-100 (1990). 9) M.J. Kaufman, Pharm. Res., 7 (3) 289-292 (1990).

It is noted that in relation to this date, the best method known to the applicant to carry out the aforementioned invention, is that which is clear from the present description of the invention.

Having described the invention as above, the content of the following claims is claimed as property.

- ^ YES_? 3sáSl

Claims (75)

1. A pharmaceutical formulation characterized in that it comprises a) a growth hormone (GH) or a growth hormone derivative, b) an amino acid selected from the group of Asp, He, Val, Leu, or His, or a histidine derivative, or a peptide comprising at least a residue of a basic amino acid and at least one residue of an acidic amino acid, c) and a nonionic detergent, wherein the amount of the peptide, or Asp, He, Val, Leu or His, or the histidine derivative is about 0.01 to about 10 mg per mg of GH.

2. A pharmaceutical formulation according to claim 1, characterized in that it comprises a) a growth hormone (GH) or a growth hormone derivative, b) an amino acid selected from the group of Asp, He, Val, Leu, c) and a non-ionic detergent, wherein the amount of the amino acid selected from the group of Asp, He, Val or Leu is from about 0.01 to about 10 mg per mg of the GH.

3. A pharmaceutical formulation according to claim 1, characterized in that it comprises a) a growth hormone (GH) or a growth hormone derivative, b) histidine or a histidine derivative or a peptide comprising at least one residue of a basic amino acid and at least one residue of an acidic amino acid, c) and a nonionic detergent, wherein the amount of the peptide or histidine or the histidine derivative is from about 0.01 to about 10 mg per mg of the GH.

4. A pharmaceutical formulation according to claim 1 or 3, characterized by comprising a) a growth hormone (GH) or a growth hormone derivative, b) histidine or a histidine derivative, c) and a nonionic detergent.

5. A pharmaceutical formulation according to any one of the preceding claims, characterized in that the amount of the peptide, or Asp, He, Val, Leu or His, or a histidine derivative is from about 0.05 to about 5 mg per mg of the GH .

6. A pharmaceutical formulation according to any of the preceding claims, characterized in that the amount of the peptide, or Asp, He, Val, Leu or His, or a histidine derivative is from about 0.05 to about 1 mg per mg of the GH.

7. A pharmaceutical formulation according to any of the preceding claims, characterized in that the amount of the peptide, or Asp, He, Val, Leu or His, or a histidine derivative is from about 0.05 to about 0.5 mg per mg of the GH.

8. A formulation according to any of the preceding claims, characterized in that the detergent is a polysorbate, for example Tween 20.

9. A formulation according to any of the preceding claims 1-7, characterized in that the detergent is a poloxamer, for example poloxamer 188 or poloxamer 407.

10. A formulation according to any of the preceding claims, characterized in that it also comprises one or more agents that give isotonicity, for example, sugar alcohols and / or salts.

11. A formulation according to claim 10, characterized in that the agents that give the isotonicity are selected from mannitol, sorbitol and sodium chloride

12. A formulation according to any of the preceding claims, characterized in that it is an aqueous formulation.

13. A formulation according to any of the preceding claims, characterized in that it also comprises one or more preservatives, for example, phenol, benzyl alcohol.

14. A formulation according to any of the preceding claims, characterized in that the pH value is adjusted to be between 6.0 and 8.8.

15. A pharmaceutical formulation, stable according to any of the preceding claims, characterized in that the growth hormone is a human growth hormone.

16. A pharmaceutical formulation characterized in that it comprises a) a growth hormone (GH) or a growth hormone derivative, b) an amino acid selected from the group of Asp, He, Val, Leu or His, or a derivative of histidine or a peptide comprising at least one residue of a basic amino acid and at least one residue of an acidic amino acid, c) and a nonionic detergent, wherein the amount of the nonionic detergent is from about 0.01 to about 10 mg per mg of the GH.

17. A pharmaceutical formulation according to claim 16, characterized by comprising a) a growth hormone (GH) or a growth hormone derivative, b) an amino acid selected from the group of Asp, He, Val, or Leu, c) and a non-ionic detergent, wherein the amount of the non-ionic detergent is from about 0.01 to about 10 mg per mg of the GH.

18. A pharmaceutical formulation according to claim 16, characterized in that it comprises a) a growth hormone (GH) or a growth hormone derivative, b) histidine, or a histidine derivative or a peptide comprising at least one residue of a basic amino acid and at least one residue of an acidic amino acid, c) and a nonionic detergent, wherein the amount of the non-ionic detergent is from about 0.01 to about 10 mg per mg of the GH.

19. A pharmaceutical formulation according to claim 16 or 18, characterized in that it comprises a) a growth hormone (GH) or a growth hormone derivative, b) histidine, or a histidine derivative, c) and a detergent not ionic

20. A pharmaceutical formulation according to any of the preceding claims 16-19, characterized in that the amount of the non-ionic detergent is from about 0.05 to about 5 mg per mg of the GH.

21. A pharmaceutical formulation according to any of the preceding claims 16-20, characterized in that the amount of the non-ionic detergent is from about 0.1 to about 2 mg per mg of the GH.

22. A formulation according to any of the preceding claims 16-21, characterized in that the detergent is a polysorbate, for example, Tween 20.

23. A formulation according to any of the preceding claims 16-21, characterized in that the detergent is a poloxamer, for example, poloxamer 188 or poloxamer 407.

24. A formulation according to any of the preceding claims 16-23, characterized in that it further comprises one or more agents that give isotonicity, for example, sugar alcohols and / or salts.

25. A formulation according to claim 24, characterized in that the agents which give isotonicity are selected from mannitol, sorbitol, and natrium chloride.

26. A formulation according to any of the preceding claims 16-25, characterized in that it is an aqueous formulation.

27. A formulation according to any of the preceding claims 16-26, characterized in that it also comprises one or more preservatives, for example, phenol, benzyl alcohol.

28. A formulation according to any of the preceding claims 16-27, characterized in that the pH value is adjusted to be between 6.0 and 8.8.

29. A pharmaceutical formulation, stable according to any of the preceding claims 16-28, characterized in that the growth hormone is a human growth hormone.

30. A pharmaceutical formulation characterized in that it comprises a) a growth hormone (GH) or a growth hormone derivative, b) an amino acid selected from the group of Asp, He, Val, Leu or His, or a derivative of histidine, or a peptide comprising at least one residue of a basic amino acid and at least one residue of an acidic amino acid, c) and a nonionic detergent, wherein the amount the amount of the peptide or Asp, He, Val, Leu, or His, or a histidine derivative is from about 0.01 to about 10 mg per mg of the GH, and the amount of the non-ionic detergent is from about 0.01 to about 2 mg per mg of the GH.

31. A pharmaceutical formulation according to claim 30, characterized in that it comprises a) a growth hormone (GH) or a growth hormone derivative, b) an amino acid selected from the group of Asp, He, Val, or Leu, c) and a non-ionic detergent, wherein the amount of Asp, He, Val, or Leu is from about 0.01 to about 10 mg per mg of the GH, and the amount of the non-ionic detergent is from about 0.1 to about 2 mg per mg. of the GH.

32. A pharmaceutical formulation according to claim 30 characterized by comprising a) a growth hormone (GH) or a growth hormone derivative, b) histidine, or a histidine derivative or a peptide comprising at least one residue of a basic amino acid and at least one residue of an acidic amino acid, c) and a nonionic detergent, wherein the amount of the peptide or histidine or a histidine derivative is from about 0.01 to about 10 mg per mg of the GH, and the amount of the non-ionic detergent is from about 0.1 to about 2 mg per mg of the GH.

33. A pharmaceutical formulation according to claim 30 or 32, characterized in that it comprises a) a growth hormone (GH) or a growth hormone derivative, b) histidine, or a histidine derivative, c) and a non-ionic detergent .

34. A pharmaceutical formulation according to any of the preceding claims 30-33, characterized in that the amount of the peptide, or Asp, He, Val, Leu or His, or a histidine derivative is from about 0.05 to about 5 mg per mg of the hGH.

35. A pharmaceutical formulation according to any of the preceding claims 30-34, characterized in that the amount of the peptide, or Asp, He, Val, Leu or His, or a histidine derivative is from about 0.05 to about 1 mg per mg of the hGH.

36. A pharmaceutical formulation according to any of the preceding claims 30-35, characterized in that the amount of the peptide, Asp, He, Val, Leu or His or a histidine derivative is from about 0.05 to about 0.5 mg per mg of the hGH.

37. A formulation according to any of the preceding claims 30-36, characterized in that the detergent is a polysorbate, for example Tween 20.

38. A formulation according to any one of the preceding claims 30-36, characterized in that the detergent is a poloxamer, poloxamer 188 or poloxamer 407.

39. A formulation according to any of the preceding claims 30-38, characterized in that it further comprises one or more agents that give isotonicity, for example, sugar alcohols and / or salts.

40. A formulation according to claim 39, characterized in that the isotonicity-giving agents are selected from mannitol, sorbitol, and natrium chloride.

41. A formulation according to any of the preceding claims 30-40, characterized in that it is an aqueous formulation.

42. A formulation according to any of the preceding claims 30-41, further characterized in that it comprises one or more preservatives, for example, phenol, benzyl alcohol.

43. A formulation according to any of the preceding claims 30-42, characterized in that the pH value is adjusted to be between 6.0 and 8.8.

44. A pharmaceutical formulation, stable according to any of the preceding claims 30-43, characterized in that the growth hormone is a human growth hormone.

45. A pharmaceutical formulation characterized in that it comprises a) a growth hormone (GH) or a growth hormone derivative, b) an amino acid selected from the group of Asp, He, -Val, Leu, or His, or a derivative of histidine, or a peptide comprising at least one residue of a basic amino acid and at least one residue of an acidic amino acid, c) and a poloxamer.

46. A pharmaceutical formulation according to claim 45, characterized in that it comprises a) a growth hormone (GH) or a growth hormone derivative, b) an amino acid selected from the group of Asp, He, Val or Leu, c) and a poloxamer.

47. A pharmaceutical formulation according to claim 45, characterized by comprising a) a growth hormone (GH) or a growth hormone derivative, b) histidine, or a histidine derivative or a peptide comprising at least one residue of a basic amino acid and at least one residue of an acidic amino acid, c) and a poloxamer.

48. A pharmaceutical formulation according to claim 45 or 47, characterized in that it comprises a) a growth hormone (GH) or a growth hormone derivative, b) histidine or a histidine derivative, c) and a poloxamer.

49. A pharmaceutical formulation according to any of the preceding claims 45-48, characterized in that the amount of the peptide, or Asp, He, Val, Leu or His, or a histidine derivative is from about 0.01 to about 10 mg per mg of the GH

50. A pharmaceutical formulation according to any of the preceding claims 45-49, characterized in that the amount of the peptide, or Asp, He, Val, Leu or His, or a histidine derivative is from about 0.05 to about 5 mg per mg of the GH.

51. A pharmaceutical formulation according to any of the preceding claims 45-50, characterized in that the amount of the peptide, or Asp, He, Val, Leu or His, or a histidine derivative is from about 0.05 to about 1 mg per mg of the GH

52. A pharmaceutical formulation according to any of the preceding claims 45-51, characterized in that the amount of the peptide, or Asp, He, Val, Leu or His, or a histidine derivative is from about 0.05 to about 0.5 mg per mg of the GH

53. A formulation according to any of the preceding claims 45-52, characterized in that the poloxamer is poloxamer 188 or poloxamer 407.

54. A formulation according to any of the preceding claims 45-53, characterized in that the amount of the poloxamer is from about 0.01 to about 10 mg per mg of GH.

55. A formulation according to any of the preceding claims 45-54, characterized in that the amount of the poloxamer is from about 0.05 to about 5 mg per mg of the GH.

56. A formulation according to any of the preceding claims 45-55, characterized in that the amount of the poloxamer is from about 0.1 to about 2 mg per mg of the GH.

57. A formulation according to any of the preceding claims 45-56, characterized in that it further comprises one or more agents that give isotonicity, for example sugar alcohols and / or salts.

58. A formulation according to claim 57, characterized in that the isotonicity-giving agents are selected from mannitol, sorbitol and sodium chloride

59. A formulation according to any of the preceding claims 45-58, characterized in that it is an aqueous formulation.

60. A formulation according to any of the preceding claims 45-59, characterized in that it also comprises one or more preservatives, for example, phenol, benzyl alcohol.

61. A formulation according to any of claims 45-60, characterized in that the pH value is adjusted to a value between 6.0 and 8.8.

62. A pharmaceutical formulation, stable according to any of the preceding claims 45-61, characterized in that the growth hormone is human growth hormone.

63. A method for preparing a formulation, comprising a growth hormone, or a growth hormone derivative, and an amino acid selected from the group of Asp, He, Val, Leu or His, or a histidine derivative, or a peptide comprising at least one residue of a basic amino acid and at least one residue of an acidic amino acid, and a detergent nonionic, characterized in that it comprises dissolving such a quantity of growth hormone, or a growth hormone derivative, that it causes the amount of the peptide, or Asp, He, Val, Leu or His, or a derivative of histidine, varies from about 0.01 to about 10 mg per mg of GH in a solution comprising the amino acid selected from the group of Asp, He, Val, Leu or His, or a histidine derivative, or a peptide comprising at least one residue of a basic amino acid and at least one acidic amino acid residue in deionized water, and a nonionic detergent, for example, a polysorbate or a poloxamer, optionally containing a preservative, and optionally containing an agent for tonicity adjustment, optionally adjusting the pH to a value of about 6.0 to about 8.8. 64. A method according to claim 63, for preparing a formulation comprising a growth hormone, or a growth hormone derivative and an amino acid selected from the group of Asp, He, Val or Leu, and a non-ionic detergent. , characterized in that it comprises dissolving such an amount of the growth hormone or a growth hormone derivative, which causes the amount of Asp, He,

Val or Leu, ranging from about 0.01 to about 10 mg per mg of GH in a solution comprising the selected amino acid in deionized water, and a nonionic detergent, for example a polysorbate or poloxamer, which optionally contain a preservative, and optionally containing an agent for adjusting the tonicity, and optionally adjusting the pH to a value of about 6.0 to about 8.8.

65. A method according to claim 63 for preparing a formulation comprising a growth hormone, or a growth hormone derivative, and histidine, or a histidine derivative or a peptide comprising at least one residue of a basic amino acid and at least one residue of an acidic amino acid, and a nonionic detergent, characterized by comprising dissolving such a quantity of the growth hormone, or a growth hormone derivative, which causes the amount of the peptide or histidine or a derivative of histidine, ranging from about 0.01 to about 10 mg per mg of GH in a solution comprising histidine, or a histidine derivative, or a peptide comprising at least one residue of a basic amino acid and at least one residue of an acidic amino acid in deionized water, and a non-ionic detergent, for example a polysorbate or a poloxamer, optionally containing a preservative, and optionally containing An agent for adjusting the tonicity, and optionally adjusting the pH to a value of about 6.0 to about 8.8.

66. A method for preparing a formulation comprising a growth hormone, or a growth hormone derivative, and an amino acid selected from the group of Asp, He, Val, Leu or His, or a histidine derivative, or a peptide comprising at least one residue of a basic amino acid and at least one residue of an acidic amino acid, and a non-ionic detergent, characterized by comprising dissolving the growth hormone, or a growth hormone derivative, in a solution comprising the selected amino acid from the group of Asp, He, Val, Leu, His or a histidine derivative, or a peptide comprising at least one residue of a basic amino acid and at least one residue of an acidic amino acid in deionized water, and such an amount of a nonionic detergent, for example a polysorbate or a poloxamer, wherein the amount of the non-ionic detergent is from about 0.01 to about 10 mg per mg of the GH, optionally containing a preservative or, and optionally containing an agent for adjusting the tonicity, and optionally adjusting the pH to a value of about 6.0 to about 8.8.

67. A method according to claim 66, for preparing a formulation comprising a growth hormone, or a growth hormone derivative, and an amino acid selected from the group of Asp, He, Val, or Leu and a non-ionic detergent, characterized in that it comprises dissolving the growth hormone, or a growth hormone derivative, in a solution comprising an amino acid selected from the group of Asp, He, Val, or Leu in deionized water, and such a quantity of a non-ionic detergent. , for example a polysorbate or a poloxamer, wherein the amount of the non-ionic detergent is from about 0.01 to about 10 mg per mg of the GH, optionally containing a preservative, and optionally containing an agent for adjusting the tonicity, and optionally adjusting the pH to a value of about 6.0 to about 8.8.

68. A method according to claim 66, for preparing a formulation comprising a growth hormone, or a growth hormone derivative, and histidine, or a histidine derivative or a peptide comprising at least one residue of a basic amino acid and at least one residue of an acidic amino acid, and a non-ionic detergent, characterized in that it comprises dissolving the growth hormone, or a growth hormone derivative, in a solution comprising histidine or a histidine derivative, or a peptide comprising at least one residue of a basic amino acid and at least one residue of an acidic amino acid in deionized water, and a such amount of a non-ionic detergent, for example a polysorbate or a poloxamer, that the amount of the non-ionic detergent is from about 0.01 to about 10 mg per mg of the GH, optionally containing a preservative, and optionally containing an agent for adjust the tonicity, and optionally adjust the pH to a value of about 6.0 to about 8.8.

69. A method for preparing a formulation comprising a growth hormone, or a growth hormone derivative, and an amino acid selected from the group of Asp, He, Val, Leu or His, or a histidine derivative, or a peptide comprising at least one residue of a basic amino acid and at least one residue of an acidic amino acid, and a nonionic detergent, characterized in that it comprises dissolving such an amount of the growth hormone, or a derivative of the growth hormone, which causes the amount of the peptide, or Asp, He, Val, Leu, or His, or a histidine derivative, ranges from about 0.05 to about 0.5 mg per mg of GH, in a solution comprising the amino acid selected from the group of Asp, He , Val, Leu or His, or a histidine derivative, or a peptide comprising at least one residue of a basic amino acid and at least one residue of an acidic amino acid in deionized water, and such a quantity of a non-ionic detergent, per example a polysorbate or. a poloxamer, that the amount of the non-ionic detergent is from about 0.1 to about 2 mg per mg of the GH, optionally containing a preservative, and optionally containing an agent for adjusting the tonicity, and optionally adjusting the pH to a value of approximately '6.0 to approximately 8.8.

70. A method according to claim 69 for preparing a formulation comprising a growth hormone, or a growth hormone derivative, and an amino acid selected from the group of Asp, He, Val, or Leu, and a non-ionic detergent, characterized in that it comprises dissolving such a quantity of growth hormone, or a growth hormone derivative, that causes the amount of Asp, He, Val, or Leu, to vary from about 0.05 to about 0.5 mg per mg of GH in a solution comprising the amino acid selected from the group of Asp, He, Val or Leu in deionized water, and such a quantity of a non-ionic detergent, for example a polysorbate or a poloxamer, that the amount of the non-ionic detergent is about 0.1 to about 2 mg per mg of the GH, optionally containing a preservative, and optionally containing an agent for adjusting the tonicity, and optionally adjusting the pH to a value of about 6.0 to approximately 8.8.

71. A method according to claim 69 for preparing a formulation comprising a growth hormone, or a growth hormone derivative, and histidine, or a histidine derivative, or a peptide comprising at least one residue of a basic amino acid and at least one residue of an acidic amino acid, and a non-ionic detergent, characterized in that it comprises dissolving such a quantity of growth hormone, or a growth hormone derivative, that it causes the amount of the peptide or histidine or a derivative of histidine, ranging from about 0.05 to about 0.5 mg per mg of GH in a solution comprising histidine, or a histidine derivative, or a peptide comprising at least one residue of a basic amino acid and at least one residue of an amino acid acidic in deionized water, and such a quantity of a non-ionic detergent, for example, a polysorbate or a poloxamer, that the amount of the non-ionic detergent is from about 0.1 to about 2 mg per mg of GH, optionally containing a preservative, and optionally containing an agent for adjusting the tonicity, and optionally adjusting the pH to a value of about 6.0 to about 8.8.

72. A method for preparing a formulation comprising a growth hormone, or a growth hormone derivative, and an amino acid selected from the group of Asp, He, Val, Leu or His, or a histidine derivative, or a peptide comprising at least one residue of a basic amino acid or at least one residue of an acidic amino acid, and a non-ionic detergent, characterized in that it comprises dissolving the growth hormone, or a growth hormone derivative, in a solution comprising the amino acid selected from the group of Asp, He, Val, Leu or His, or a histidine derivative, or a peptide comprising at least one residue of a basic amino acid and at least one residue of an acidic amino acid in deionized water, and a poloxamer, for example poloxamer 188 or 407, optionally containing a preservative, and optionally containing an agent for adjusting the tonicity, and optionally adjusting the pH to a value from about 6.0 to about 8.

73. A method according to claim 72 for preparing a formulation comprising a growth hormone, or a growth hormone derivative, and an amino acid selected from the group of Asp, He, Val, or Leu, and a non-ionic detergent, characterized by comprising dissolving the growth hormone, or a growth hormone derivative, in a solution comprising an amino acid selected from the group of Asp, He, Val, or Leu in deionized water, and a poloxamer, for example, poloxamer 188 or 407, optionally containing a preservative, and optionally containing an agent for adjusting the tonicity, and optionally adjusting the pH to a value of about 6.0 to about 8.

74. A method according to claim 72 for preparing a formulation comprising a growth hormone, or a growth hormone derivative, and histidine, or a histidine derivative, or a peptide comprising at least one residue of a basic amino acid and at least one residue of an acidic amino acid, and a non-ionic detergent, characterized in that it comprises dissolving the growth hormone, or a growth hormone derivative, in a solution comprising histidine, or a histidine derivative, or a peptide comprising at least one residue of a basic amino acid and at least one residue of an acidic amino acid in deionized water, and a poloxamer, for example, poloxamer 188 or 407, optionally containing a preservative, and optionally containing an agent for adjusting the Fc 73 tonicity, and optionally adjusting the pH to a value of about 6.0 to about 8.

75. A method for treating a disorder in a patient affected by growth hormone, characterized in that it comprises treating the patient with an amount of the pharmaceutical formulation according to any of claims 1-62, effective to treat such a disorder. *? i * r- * 74 SUMMARY OF THE INVENTION The invention relates to pharmaceutical formulations comprising a growth hormone, an amino acid selected from the group of Asp, He, Val, Leu, or His, or a histidine derivative, or a peptide comprising at least one basic amino acid and less an acidic amino acid, and a non-ionic detergent, for example, a 10 polysorbate or a poloxamer. The formulation is synthesized against deamidation and aggregation. The formulation may be an aqueous formulation. Growth hormone can be a human growth hormone. fifteen