DD267910A5 - METHOD FOR PRODUCING A FORMKOERPER FOR PARENTERAL PROCESSING OF A SOMATOTORPINE TO AN ANIMAL ADJUSTED TO ANIMAL - Google Patents

Abstract

The invention relates to a method for producing a shaped body adapted for parenteral administration of a somatotropin to an animal. It is characterized by bonding together a substantially binder-free and substantially matrix-free formulation containing an effective dose of a solid somatotropin, and optionally an effective amount of a lubricant, and optionally coating the molding on a portion of its surface, which substantially inhibits the release of the somatotropin on the coated surfaces of the molding material, but at least one uncoated release surface is provided.

Description

Hierzj 3 pages drawings

Field of application of the invention:

The invention relates to a process for the preparation of a shaped body adapted for parenteral administration of a somatotropin to an animal.

Characteristic of the known state of the art

Somatotropins are known to be effective in improving growth, feed utilization, milk production and fat-to-lean ratio of various animals when administered parenterally to increase the somatotropic level normally produced by the animal itself. While these goals can often be achieved by applying somatotropin injections, prolonged release systems are easier to administer and often produce superior results. because of the constant concentration in the blood.

In the prior art, a variety of methods have been used to achieve prolonged release of somatotropin. Somatotropin Kt has been dispersed in a biocompatible oil, optionally in the presence of an anti-hydrating agent (see European Patent Application 177478, published April 9, 1986). Somatotropin has been complexed with a water-soluble or water-dispersible carbohydrate polymer such as doxtrin, dextran, and various gums. This complex is administered parenterally as a solution, dispersion or paste (see European Patent Application 193917, published September 10, 1986). Somatotropin was formulated with cholesterol and compressed into a matrix implant (see U.S. Patent 4,452,755, Kent, published June 5, 1984). A large number of systems are known for parenteral implantation of shaped bodies to release a variety of bioactive materials. A wide range of binders and matrix materials are known in the art for use with bioactive materials to form molded articles for parenteral application. Likewise, it is known to implant coated moldings in which the release of a bioactive core material is regulated by diffusion through a polymeric coating or by its erosion. These known systems

are usually one variant. In each of these systems, the release of the active substance is regulated by the matrix, binder or coating. The release is accompanied by a penetration or erosion de. Matrixstorfes, the binder or the coating. Prolonged release of somatotropins occurred in a single sample, with the active somatotropin solution being regulated by another substance.

Object of the invention

The invention is intended to enable the paremeral administration of a somatotropin to an animal without the need for another substance to control the release of the somatotropin. Prior to this invention, it has not been known to administer somatotropin in the form of a shaped body that is substantially free of matrix, substantially binderless, and that has at least one uncoated release surface.

Explanation of the essence of the invention

The invention has for its object to provide a method for producing a suitable for the administration of a somatotropin to an animal shaped body. Administration should be parenteral (by implantation).

The process is characterized by bonding together a substantially binder-free and substantially matrix-free formulation containing an effective dose of solid somatotropin and optionally an effective amount of a lubricant, and optionally coating the shaped article with a coating on a portion of its surface which substantially inhibits the release of the somatotropin on the coated surfaces of the molded article, but at least one uncoated release surface is provided.

The drawings serve to explain the problem underlying the invention. Show it:

Fig. 1: graphical representation of the milk yield of treated cattle and control animals according to Example 2; Fig. 2: graphical representation of the serum somatotropin concentration of treated bovine and control animals according to Example 2; 3: graphic representation of the serum somatotropin concentration of MBS-treated, non-dairy cattle according to Example 2.

The following terms have the following meanings throughout the specification and examples: The term "matrix" means a solid phase support, also referred to as _M matrix material, which forms a network within which somatotropin particles are distributed, thereby increasing the resolution of the somatotropin is inhibited. The release of somatotropin is regulated by erosion of the matrix stool or by diffusion of the somatotropin through openings in the matrix matrix network.

The term "binder" refers to polymeric materials, either of synthetic or biological origin, which are used to increase the adhesion between the particles of the somatotropin or between the particles of the somatotropin and those of a matrix material.

The terms "substantially binderless" and "substantially matrix-free" mean that the molding does not contain sufficient amount of the binder or matrix material to exert a substantial effect on the release of the somatotropin.

The term "at least one uncoated release surface" means that at least a portion of the surface of the molded article has no coating that materially inhibits somatotropin release. This uncoated release surface may be a full front surface of the molded article, e.g. B. one end or both ends of a cylindrical shaped body, or it may be an uncoated surface of a larger surface of the shaped body, for. One or more non-aggressive areas on a spherical shaped body, or one or more uncoated areas on a curved surface of a cylindrical shaped body.

By the term "somatotropin" is meant a polypeptide having a biological activity and a chemical structure similar to that of the somatotropin produced in the pituitary gland of an animal Such somatotropins include natural somatotropins produced by pituitary cells, and alternatively These include somatotropins produced by recombinant DNA technology, where somatotropin is expressed by genetically transformed cells, and suitable systems include tissue culture of transformed Gowebe cells and expression by transformed microorganisms such as E. coli, other bacteria, yeast, etc. Because of production conditions Such somatotropin produced via the recombinant DNA may have an amino acid sequence corresponding to that of the naturally occurring somatotropin, but may also be a variant in which either Ami or otherwise absent from the amino acid sequence of the naturally occurring somatotropin, provided that such additions, deletions or changes in the amino acid sequence have no unacceptable side effects on the bioactivity of the somatotropin. Also included are somatotropins which are salts or complexes with anions or cations. These complexes may be co-precipitated or lyophilized in the presence of an inorganic or organic compound, for example as a salt to form an intimate mixture, or may be formed by moistening a dry mixture of the somatotropin and a compound containing the desired ion , Mixtures of somatotropins or multiply coated somatotropins are also included. Examples of somatotropins useful in the present invention are poultry somatotropins for the treatment of chickens, turkeys and the like; Mammalian somatotropins for the treatment of humans, cattle, swine, sheep, goats and the like; as well as aquatic somatotropin for the treatment of fish and similar organisms. Particularly noteworthy are bovine somatotropin and sudgilline somatotropin.

Because of the ability to produce substantial amounts of somatotropin, recombinant DNA techniques for microwell expression of somatotropin are preferred. Moreover, recombinant DNA techniques allow the production of variants that are either similar or different to the somatotropins with the sequences of natural Sc latotropins. The sequences of naturally occurring bovine and porcine somato-opines have been described by Seeburg et al., DNA, Vol. 2, No. 1, pp. 37-45 (1983), with particular reference to this literature becomes. In the variants below, the N-methionine can sometimes be removed during or after expression. Examples of bovine somatotropin variants include, but are not limited to, polypeptides having the following amino acid sequences with unspecified amino acid residues that are similar to the natural somatotropin:

NH ₂ -met-phe (1 | -pro (2) Ieui126) .... phe (190) -COOH NH ₂ -met-phe (1) -pro (2) val (126) .... phe (190) -COOH NH ₂ -alpha-phn (1) -pro (2) val (126) .... phe (190) -COCH NH ₂ -ala-phe (1) -pro (2) leu (126) .... phe (190> -COOH NHj-phe (1) -pro (2) Ieu (126) .... phe (190) -COOH NH ₂ -phe! 1) -pro (2) val (126) .... phe (190hCOOH

NH ₂ -met-asp-glu-phe (1) -pro (2) .... val (126) ... phe (190) -COOH NH ₂ -met-asp-glu-phe (1) -pro (2) .... leu (126) ... phe (190) -COOH

NH ₂ -rr.ot (4) ser (5) leu (126) .... phe (190) -COOH NH ₂ -met (4) -serine (5j val (126) .... phe (190) -COOH

As can be seen from the above information, these variants may optionally have a methionyl residue at the N-terminus. The first variant in the above list, which has a methionyl residue at the N-terminus and a leucyl residue at position 126, is referred to herein as methinnyl bovine somatotropin or "MBS", and the third variant in the above list, with an alanyl residue at N-terminus and a valyl residue at position 126 is referred to herein as alanyl-valyl-bovine somatotropin or "ala-val BST".

Examples of porcine somatotropin variants include (but are not limited to) polypeptides having the following amino acid sequences that have non-specific amino acid residues that are similar to the natural somatotropin:

NH ₂ -8la-phe (1) .... pne (190) -COOH NH ₂ -met-phe (1) .... phe (190) -COOH NH ₂ -met (4) -pro (5) phe (19O) -COOH NH ₂ -met (6) -ser (7) phe (190) -COOH The first variant in the list above, with an alanyl residue at the N-terminus, is referred to as. Alanyl-pig somatotrop'n or

The second variant in the above list is referred to herein as methionyl-swine somatotropin or "MPS".

While the use of somatotropin variants may be useful and beneficial in some circumstances, one should

to the extent that care should be taken that the variants should not be so far removed from the naturally occurring somatotropin, that no more intolerable side effects occur, or that the variants are not one more; induce tolerable production of unwanted antibodies.

To be biologically active, the somatotropin must be natural, i. H. have a folded structure, the biological Activity guaranteed. While it is preferable to have the presence of inactive polypeptides and inactive polypeptide aggregates

To keep to a minimum, certain amounts may be present, provided that the activity of the bioactive

Somatotropins not inadmissibly inhibieron. In the current state of the art, the release of somatotropin from a prolonged-release system is a result Mechanism in which other components of the system were involved, e.g. the erosion of the matrix, etc. At

According to the present invention, the release of the somatotopin is self-regulated by the properties of the somatotropin and may also to some extent be governed by such factors as the size and shape of the molding and by the nature of the somatotropin

Process for its preparation can be effected. The properties of somatotrcpine that most affect the release characteristics are its solubility and toxicity

its rate of dissolution. This combination of properties is called intrinsic dissolution. The

Self-dissolution and some other properties of the somatotropin can be influenced in many ways. Different somatotropin variants often have quite different solubilities and rates (rates)

resolution, and as a result, self-resolution can sometimes be influenced to some extent by the choice of a variant. The ancestor of the isolation of the solid somatotropin, e.g. B. the pH of the precipitation of a precipitated

Somatotropins or the lyophilization conditions in solution of lyophilized somatotropins can also be used Pretty much influence your own resolution. The intrinsic dissolution can also be influenced by the use of 3yr.es somatotropin, which with another ion

associated with or with another connection, "viz a Saiz. The actual structure of this type of association is not clear, presumably they are salts, complexes, intimate mixtures formed by Co-FSII, or some combinations of these drsi forms.

In this description and in the claims, if necessary, with reference to a somatotropin, which with a

for example, it is referred to as zinc ossozienes bovine sormiotropin, copper-associated porcine somatotropin or bovine somatofropin associated with sodium bicarbonate. Generally referred to, it will be readily referred to as an associated somatotropin.

Associated somatotropins can be formed with nichucxisone amounts of various cations, including

monovalent Matalle and polyvalent metals. Examples of suitable monovalent metals are sodium, potassium and the like.

Examples of suitable polyvalent metals s'nd zinc, iron, calcium .. bismuth, barium. Magnesium, manganese, aluminum, copper, Cobalt, nickel, cadmium and the like. However, one should especially with multivalued Motallen, which under certain

May be toxic, use caution and use non-toxic Associated somatotropins can also be formed with anions such as, for example, hydrogen citrate, aceiate, glycine, borate and the like. Combinations of anions, cations and other compounds can also be used. The ratio of cation, anion or other compound to somatotropin may vary depending on the conditions and the somatotropin used. Some or all of the ions may be associated in a salt of one of the amino acids in the somatotropin, may be included in the folds or crystals of the somatotropin, may be associated as an ion bridge between at least two somatotropin molecules, may be associated as a complex with the somatotropin be associated in another way.

Examples of other üssoziierter somatotropins are a) acid addition salts formed with inorganic acids such as hydrochloric, hydrobromic, sulfuric, phosphoric or nitric acid, or with organic acids such as acetic, oxalic, tartaric, succinic, maleic, ascorbic Benzyl, tannin, pamoe, alginine, polyglutamine, naphthalene, naphthalenedisulfonic or polygalacturonic acid; b / salts with polyfunctional organic compounds such as Ν, Ν'-dibenzylethylenediamine or ethylenediamine or procaine; or c) combinations of two or more of the above types of salts, such as zinc mannate.

Many of the benefits of the associated somatotropins can be achieved by mixing the powdered! Somatotropins can be achieved with a salt of the desired cation or anion or the desired compound before shaping the shaped body. It is believed that the associated somatotropin forms in situ during formation of the molding or after administration of the molding to the animal.

If the associated somatotropin is relatively sparingly soluble, it may be prepared by adding the desired water-soluble salt or water-soluble compound of the desired ion to a solution of the somatotropin at the desired pH to precipitate the desired associated somatotropin. For example, Ζίι, κ-associated bovine or porcine somatotropin can be prepared by adding a water-soluble zinc salt to an aqueous solution of the somatotropin buffered with a suitable buffer. The pH is preferably between about 9 and about 9.5. If the associated somatotropin is relatively more soluble, then it may be produced by lyophilizing a solution of the somatotropin with the desired salt or ions. This solution can be prepared by dialysis or diafiltration of, for example, a solution of urea, followed by dialysis or diafiltration with water to reduce the salt concentration to the desired level. For example, sodium hydrogen carbonate-associated bovine somatotropin may be prepared by diafiltration or dialysis of a urea solution of somatotropin to achieve complete replacement of urea with a sodium hydrogencarbonate solution, followed by further diafiltration with water to remove excess sodium bicarbonate, followed by lyophilization. to produce the sodium bicarbonate-associated somatotropin. Somatotropins have often been prepared and purified in a buffer solution such as sodium bicarbonate. It is preferred to adjust the concentration of the sodium bicarbonate to the desired concentration by dialysis or diafiltration. The pH can be kept within a range in which the somatotropin by addition of a base such. B. NaOH to the dialysis water or somatotropin solution is soluble. The somatotropin can then be obtained by a variety of means such. B. Adjust pH with a suitable acid such as HCl to precipitate the somatotropin followed by diafiltration and lyophilization or by direct lyophilization of the somatotropin solution. It is preferred that the somatotropin prepared in this manner has a concentration of the buffering salt, e.g. Sodium bicarbonate, from about 0% to about 20%, preferably from about 0% to about 5%, and more preferably about 1% or less. A higher concentration of sodium bicarbonate increases the intrinsic dissolution of the somatotropin and results in a product suitable for release over a shorter period of time. Conversely, a lower concentration of sodium bicarbonate will result in a product suitable for long-term release. In this way, the product can be selected for the desired period. Another method of preparing the somatotropin is to isolate and purify the somatotropin, which normally occurs under alkaline conditions, followed by dialysis to remove the buffer, and from the addition of an acid, such as phosphoric acid, to a solution to arrive in acidic form of somatotropin. Subsequently, the somatotropin can be separated from this acidic solution, for example by adding a base until the pH of the solution is raised to a point where the somatotropin precipitates. Care should be taken with this procedure to ensure that the pH is not reduced so much that the somatotropin is denatured. Porcine somatotropin has a self-dissolution that is much higher than that of bovine somatotropin. For this reason, it is preferred to use a copper-associated porcine somatotropin. This copper-assorted porcine somatotropin may contain from about 0.1% to about 3% copper, preferably from about 0.1% to about 2% copper, and more preferably from about 0.1% copper to about 1% copper. A particularly preferred copper-associated porcine somatotropin has a molar ratio of copper to somatotropin of about 1: 1, which is about 0.3% copper. Copper-associated porcine somatotropin results in increased prolonged release and improved yielding efficiency of porcine somatotropin. In addition, there is evidence that copper-associated porcine somatotropin has a reduced tendency to form dimerc and other aggregates, particularly after administration to an animal. The preferred variant is APS. Many advantages of a precipitated copper species of porcine somatotropin can be achieved by physically mixing the porcine somatotropin with the appropriate amount of a copper salt. When dry mixtures are used, additional copper salt should be added to the composition beyond the amount required to yield a copper-associated somatotropin having the desired properties. For example, a dry mixture of a copper salt with porcine somatotropin having a mole ratio of 2: 1 copper to somatotropin has digestibility characteristics similar to a precipitated copper-associated porcine somatotropin with a 1: 1 molar ratio. It is believed that the copper-associated Schwoine somatotropin forms in situ during compaction or, more likely, after administration.

The release of the somatotropin can also be influenced by controlling the release geometry of the shaped body. This can be done for example by selecting the size and shape of the molding or by covering a part of the total surface of the molding with a coating. Both of these procedures are discussed below.

The somatot'opin moldings of the present invention can be prepared by conventional methods such as densification, spray drying, prilling methods, wet paste molding and drying of the molded body, and the like. Compaction is the most preferred procedure for shaping parenteral implants. For simplicity, the present invention will be discussed below with regard to compaction molding. The molded articles of the present invention can be produced on a conventional tableting machine using punches of appropriate size and shape and at conventional pressures. Conventional working and tabletting methods can be used; For example, the somatotropin can be precompressed and comminuted to better handle it and improve flowability.

Performance can be improved by careful selection of size and shape of the molded article. The shaped body can be cylindrical, spherical or egg-shaped or have any other conventional shape. In order to maximize the period of time during which release occurs, it is advantageous to use a mold that has a minimum surface area for a given dose. In the case of the insert of a single shaped body, this can be cemented by producing a spherical implant or by producing a cylindrical implant in which the height of the cylinder is approximately equal to the diameter. However, other shapes and sizes may be provided if shorter periods are desired or other reasons exist.

For example, it is also advantageous to prepare a shaped article using conventional devices such as those used for the more gentle administration of pellets or tablets. When these "implant guns" are used, it is advantageous that the molding of the invention is a cylinder and has a diameter between about 1 mm and about 5 mm and a length between about 1 mm and about 25 mm in diameter from about 0.5 mm to about 12 mm and lengths of about 1 mm and up to about 50 mm, as well as sizes outside these ranges may also be used if desired, and if the size or shape of the molded article necessitates or even the site at which it is to be implanted, then a surgical impression may also be made. Multiple moldings can also be implanted if the desired dose is too large for administration in a molded article, or if it is desirable for other reasons, such as greater flexibility of the dose.

It is preferred that the self-dissolution and the area of the release surface be adjusted so that an average daily release rate occurs over the effective period of the molding in the desired range. For applications related to milk production, the average daily release rate is between about 1 mg to about 75 mg or even 100 mg per day, preferably between about 3 mg and about 50 mg per day, and more preferably between about 6 mg and about 40 mg per day. For administrations to pigs, the average daily release rate may be between about 0.5 mg and about 25 or even 30 mg per day, preferably about 2 mg b's about 15 mg per day. Average release rates outside these ranges can sometimes be useful, depending on the desired outcome. The total dose of somatotropin should be sufficient to maintain the desired release rate for the desired period of time, as discussed below.

The total dose of somatotropin depends on a grin number of factors, such as the size of the animal; Effect for which the somatotropin is administered; Period practiced by the dosing is to take place; and more. The term "effective dose" as used in this specification and in the claims means a dose of bioactive somatotropin contained either in one or more moldings or tablets intended for co-administration and which is effective for prolonged effect Period, sometimes only in a short time such as 1 or 2 days, more often over a period of more than about 7 days, in some cases up to about 28 days or even up to about 48 days or more.

For administration of somatotropin to cows to increase milk yield, it is preferred that the dose be from about 5 mg to about 1000 mg or 1500 mg or more, preferably from about 100 mg to about 800 mg, with administration at about one day intervals until about 28 days or longer, or preferably from about 7 days to about 28 days. For the administration of swine somatotropin to pregnant sows and dairy sows, it is preferred that the dose be from about 5 mg to about 225 mg or more, more preferably from about 75 mg to about 150 mg, with administration at about one hour intervals Day to about 28 days, preferably from about 7 days to about 21 days. For administration to slaughter pigs to increase weight gain and feed utilization or to improve the lean to lean ratio, it is preferred that the doses be from about 10 mg to about 200 mg or more, more preferably from about 10 mg to about 90 mg, and that they are administered at intervals of about 7 days to about 60 days, especially from about 14 days to about 42 days. It may be desirable for slaughtered pigs to administer a dose from about 7 days to about 60 days before slaughter. Administration at shorter intervals in slaughtered pigs may be advantageous, especially towards the end of this period before slaughter. This allows greater flexibility in the administration schedule to better match the animal slaughter date. However, more frequent or less frequent administration may be effectively used in some cases and, under appropriate circumstances, larger or smaller doses may also be used effectively in some cases.

The amount of pressure in forming a shaped somatotropin body is not critical, but performance may be somewhat affected by careful selection of the pressure. Sufficient pressures must be used to form a molded article having sufficient strength and dimensional stability to remain dimensionally stable during handling and implantation.

Another method of controlling the rate of release is to cover a portion of the total surface area of the molded article (with a coating) to reduce the surface area from which dissolution proceeds.

A coating may be used to substantially inhibit or substantially stop the release of the somatotropin from the surface of the molded article, thereby controlling the geometry of the release surface, for example by maintaining a constant area for release. This may help to achieve prolonged zero release of the somatotropin from the uncoated release area, which is the primary release area. For example, the somatotropin molded article of the present invention may be a cylinder having a coating on the curved surface of the cylinder and without coating on one or both ends. In this way, as the somatotropin is released and the shaped body is removed, the area of the release surface remains relatively constant.

otherwise, the A ^f -bau would emanate from all surfaces of the cylinder and thus be released. In addition, the coating may cover the bulk of the molded article except for one or more windows which maintain a constant release area over time.

In order to increase this kind of effect to a maximum, for example, for a zylinririsc ^Ke n article which is coated on the entire curved surface, it is preferred that the coating substantially inhibit release of the somatotropin to the coated surface, and d ß the coating as long as at ^H, he remaining curved. Area dss cylinder remains intact, as the release of the unstressed end (s) takes place. The oar tract may be ablated from one or both ends as the somatotropin is ablated. It is preferred that the coating be effective over the majority of the life of the molded article after V6. Submit ben JIt. It is possible to obtain useful results with a coating which substantially inhibits at least the release of the somatotropin, provided that the primary release surface is uncoated. It may also be preferred, especially when the animal needs to receive repeated administration of somatotropin, that the coating may be absorbed by the animal, which coating must of course be biocompatible. Examples of substances that would be useful for such partial coating on a somatofopin molded article include, but are not limited to, shellac, beeswax, cellulose acetate butyrate, polylactic acid, ethylcellulose, silicones, ethylene-vinyl acetate copolymer, hydroxypropylcellulose, polycarbonate, polycaprolactone , Cellulose acetate, polymethyl methacrylate and a variety of other polymers known for use as a protective overcoat.

A similar result can mar. mistaken by producing a molded article having an outer layer, for example on the curved surface of a cylindrical body, of relatively less soluble somatotropin such as β? represents a zinc-associated somatotropin.

Even a temporary protective coating can be achieved with the present invention. It may be advantageous to provide the somatotropin body with light protection to protect the body during storage and handling, and possibly to assist administration of the body to the animal. For such a case of employing the present invention as protection, the protection may either be removed prior to administration to the animal, for example, a brief soak or wash procedure, or it may be delivered from the body very rapidly prior to administration the animal will be removed. This temporary protective cover would have very little effect on somatotropin release control and, after removal of the coating, would render the somatotropin at least released from an uncoated refractory surface. Since this type of cover has little or no effect on release, it is not considered to be a "coating" for the purposes of this specification and claims This temporary protective cover could simply be a much thinner form of the cover material discussed above Provided that this vially thinner coating must be quickly removed from the molding, either before or immediately after implantation, or it could be a cover that can be quickly removed from the molding, for example because of its solubility, because of melting at body temperature, due to a combination of solubility and melting temperature, or other grindings, suitable materials for this type of cover include, but are not limited to, polyvinyl alcohol, various sugars, polyethylene glycol (such as PEG 8000), and other materials.

In addition to the somatotropin and the possible coating and a possible protective cover, as stated above, it may be advantageous to include other substances in the composition of the shaped body according to the invention. Examples thereof are lubricants, bacteriostats, antioxidants, anti-inflammatory agents, antibiotics and the like.

Conventional lubricants can also be used to assist in the pressing process werdon. Examples of such lubricants are stearates such as magnesium stearate, palmitates such as sodium palmitate and other common lubricants. The amount of lubricant present should be sufficient to allow the somatotropin to flow freely and easily into the mold so that it can be easily removed from the mold without damaging it. However, the lubricant should not be present in such amounts that it acts as a matrix agent or in any way exerts intolerable side effects on the release of the somatotropin. For example, Magnesiumst ⁿ from no or little effect on release of the somatotropin up to a concentration of about 10% stearate. When used in concentrations exceeding 30-50%, magnesium stearate significantly inhibits the release of somatotropin from the molded body. Similarly, sodium palmitate has no or little effect on the release of somatotropin up to a concentration of 3%. When used in concentrations exceeding about 10%, sodium palmitate significantly or completely inhibits release. The effect of concentrations between these specific concentrations can be determined by routine experimentation. Similarly, effective concentration ranges for other conventional lubricants can be determined by simple methods.

Other ingredients such as antioxidants, anti-inflammatory agents, bacteriostats, antibiotics, agents for inhibiting the aggregation of somatotropin and the like can be incorporated by known methods. However, these ingredients should not be present in an amount such that they act as matrix material or otherwise exert intolerable side effects on the somatotropin releasing agent. The effective concentrations can be determined in a similar manner as previously discussed for the lubricants. In some cases, sterilization can be by irradiation; however, it should be noted that the radiation can reduce the biocompatibility of the somatotropin under regressions.

It may be advantageous for some somatotropins to adjust the amount of water present in the somatotropin, either by careful control of the drying process such as lyophilization, or by exposing the lyophilized somatotropin to a water vapor-containing atmosphere until the desired amount of water is present in the composition. However, these settings should not be performed under conditions that accelerate the denaturation of somatotropin, aggregate formation, or otherwise somatotropin degradation or efficacy. This can be done either before or after the molding of the molding and completed by other techniques.

The shaped article according to the invention can be administered in the usual way, either subcutaneously in the adipose tissue or intramuscularly. It can then be administered either surgically or using conventional implantation devices.

The following examples illustrate the invention and are not intended to be limiting. All parts and proactive details are lied to the fair, unless otherwise stated.

embodiments example 1

Microbially expressed MBS was produced by expression of recombinant DNA essentially as described in Example A of EP Application 177478, published April 9, 1936. Fragments were isolated by cell disruption followed by centrifugation. The fragments were then solubilized, folded and oxidized in a solution of urea and tris-hydroxymethyl-aminomethane (TRIS) buffer. Never buffered MBS solution was passed through a chromatographic column containing DE-52 ion exchangers. The concentration of this solution was adjusted to 22 mg / ml MBS in a solution of 4.5M urea and 50 mM Tris-hydroxymethylaminomethane (TRIS) buffer at pH 9.3-9.5. This solution was sterile filtered through a 0.22 micron filter cartridge. The filtered solution was dialysed against a seven-fold volume of 25 mM sodium bicarbonate buffer (at pH 9.5), replacing the buffer solution 8 times every 8-12 hours. Thereafter, the buffer solution was replaced with WFI grade water, which was changed every 8-12 hours. When the pH dropped to 8 to 8.5, the MBS began to precipitate out of the solution. Dialysis was continued for a further 4-6 changes with water to further reduce the concentration of residual buffer components until the pH ranged from 6.5 to 7.5. The precipitated MBS was then lyophilized and recovered as a dry white powder. MBS prepared in this manner was used to make pressed molded capsules for the parenterr's implantation in dairy cows (described below).

Belsple'2

Pressed somatotropin moldings were made from MBS as prepared in Example 1. 50mg of this MBS was added to a 6.4mm diameter tablet form and placed in a 12 ton Carvar Mooell C tablet press with a 21.25 cm ² (3.294 square inch) hydraulic punch at a loading of 17790 Newton (4000 pound), read on the pressure gauge, pressed. Ten of these 50 mg molded tablets were surgically subcutaneously implanted in the postal scapular area of boi eight lactating Holstein cows on day zero, day 14, and day 28. In each of the treated cows, a total of 30 molded articles were implanted. The milk production of the treated cows was compared over a period of 56 days to a group of 9 control animals that had not received somatotropin. The increase in production of milk and 3.5% fat-corrected (3.5 cc) milk of the treated cows compared to the untreated cows is summarized in Table 1 and shown graphically in FIG.

Table I

days Increase in milk Increase in % Increase in (Kg / day) 3.5 FC milk (kg / day) FC milk against about control Ibis 14 3.7 4.1 14.7 15bis28 11.6 11.9 '3' 29 to 42 9.6 9.6 ! "4.9 43bis49 0.5 10.6 39.1 50 to 56 6.3 5.1 19.2

Blood samples were taken daily over the entire time period of the study and blood serum was analyzed for somatotropin by padioimmunoassay (RIA). The detected serum levels are shown graphically in FIG. Milk production was consistent with the observed Servm somatotropin profile.

Example 3

A group of 14 non-lactating Holstein cows was divided into two groups of 7 cows each. One group received the surgical implantation of i0 of the 50 mg MBS shaped body described in Example 2, prepared as described in Example 2, and the other group were subjected to the chiral measures as a blank test. Ten blood samples were taken on day 1, 6 samples on days 2 and 3, three on days 4 to 14, and 2 samples on days 15 to 21. Blood serum somatotropin levels were determined by RIA and the results are in Fig. 3 can be seen. The serum levels of this study were slightly lower than in the milk dairy cows study of Example 2. The reason for this lower level was not clear. Presumably these were normal fluctuations in natural somatotroph production. However, a steady increase was observed over 17 days or 18 days.

Example 4

Five of the 50 mg MBS moldings prepared as in Example 2 were surgically implanted on 5 non-lactating Holstein stains in either the right or left anterior or posterior postscapular region on days 0.3, 7 and 14 of the study. On day 14, the animals were sacrificed and tissue analysis was performed. A non-normal tissue reaction as a side effect was not observed, which speaks for the good biocompatibility.

Bolsplel5

Microbial expressions APS was prepared by expressing recombinant DNA essentially as described in Example 3B of European Patent Application 193,515 (previously published on Sep. 3, 1986). Reference is made to this EP application. The APS fragments were isolated, annealed, folded and purified as described in Example 1 by means of the ion exchange step. The APS-buffered solution was dialyzed against a sodium bicarbonate buffer solution and the resulting bicarbonate-buffered solution was sterile filtered and lyophilized. This materia! 5.4 g of this APS was dissolved in 250 ml of deionized water and dialyzed against four changes of 16 volumes of water for 24 hours at 4 ° C. The dialyzed APS became pH 6 to 8 precipitated with 30 micromoles of a 10OmM copper sulfate solution. Dar by addition of copper sulfate formed precipitate was isolated by centrifugation and Ivophilisiert. Dm product obtained had a copper content of 0/3%.

Examples

Shaped bodies were made of APS bulk powder and of copper-associated APS. Both components. (s were prepared as described in Example 5. The copper-associated APS was prepared by maintaining a copper to APS molar ratio of 1.2 to 1.4 to 1. The compression process was generally as in Example 2 except that 1% sodium palmitate (NaP) or 5% magnesium stearate (MgS) was used as the lubricant, exceptions are indicated. Sufficient amount of somatotropin formulation to give 60mg and 90mg doses of somatotropin. was placed in a 2.4 mm or 3.2 mm mold and compressed using an applied load of 4448 Newton (1000 pounds) .These molded articles were parenterally implanted in commercial, cross-bred pigs at the end of the fattening period The average daily weight gain in kg / day (ADG) and feed conversion (FE), divided as daily feed intake by the ADG (a lower FE is a better FE) and the percentage verb FR (% Imp. FE) in the implanted pigs was compared to negative control pigs that did not contain somatotropin. These results and the results for a positive control group of pigs who had recovered daily injections of 2 mg APS in solution are summarized in Table II. The first part of the trial period was 14 Ta'je, followed by a 7-day Intenms period, and then the second part of the trial period followed, at 14 days. The results are summarized in Table II.

Table II

Part 1 (-!Control (+! Control 1.11 1.02 Type: Cu Cu Cu APS APS (no treatment) (2 mg / day) 3.24 3.78 Dia .: 3mm 3mm 3mm 3mm 3mm - - Dose: 60mg 60mg 60 mg 2 χ 30 mg 2 χ 30 mg 0.98 Gleitm .: NaP NaP N / A' NaP NaP ' 4.10 Location: ear ear Fet ear ear - 1.07 1.03 1, W 0.95 0.87 ADG (kg) (+) Control 3.63 3.43 3.61 3.73 4.29 FE (feed / increase) (no treatment) (2 mg / day) 24.5% 16.3% 12.0% 9.0% - % Imp. FE Section 2 (-)Control 0.90 Type: Cu Cu Cu Cu 4.81 dia .; 3mm 2mm 2mm 3mm - Dose: 60mg 60 mg 60 mg 90 mg Gleitm .: NaP NaP ² MgS NaP Location: fat ear ear ear 1.05 1.02 1.08 1.12 ADu (kg) 3.74 3.61 3C · " 3.19 FE (feed / increase) 22.2 25.0 : O, 6 33.7 % Imp. FE

1 10% sodium palmitate

2% sodium palmitate

The implanted tablets of the present invention performed favorably against the daily injection and resulted in significantly improved weight gain and food wastage compared to the untreated pigs. The sodium palmitate level of 10% resulted in a decrease in somatotropin release, so that 10% Nstriumpalmitat is higher than the actual amount of lubricant.

Example 7

Pressed molded articles were produced from microbially expressed APS bulk powder, which was isolated and purified as in Example 5. The process was carried out as generally described in Example 2 except that 8.4mm diameter 30mg molded bodies were made using a force of 17790 Newtons (4000 pounds) and 60mg, 6 diameter shaped bodies. 4mm when using a force of 17 790 Newton (4000 pounds). These moldings were surgically implanted in the fat of the neck back on the first day of the study. A second set of 3.2 mm diameter moldings was made at 4448 Newtons (1000 pounds) with 30 mg each and 60 mg somatotropin. These shaped bodies were implanted in the fat of the neck back with an implant gun. This second set of 3.2 mm diameter moldings was implanted on day 15 of the stuaia. The pigs used were commercial, by crossing

bred pigs at the end of the fattening period, with 10 pigs receiving the 30mg tablets, 10 pigs receiving the 60mg tablets, and 10 pigs controlling which, similar to the treated pigs, the surgical procedures were performed as a blank. ACI, FE and% improvement in FE were determined for the three pigs groups and presented in Table III.

Table III

control 30mg 60 mg week 1 ADG (kg / 0.87 0.94 1.02 FE (feed / increase) 3.84 2.78 2.53 Week 2 ADG (kg) 0.82 0.77 0.67 FE (Futtar / increase) 4.27 4.62 4.58 Week 1-2 ADG (kg) 0.85 0.85 0.84 FE (feed / increase) 3.94 3.49 3.30 % Imp. PE - 11.4% 16.2% Week 3 ADG (kg) 0.84 1.15 1.00 FE (feed / increase) 4.41 2.97 3.17 Week 4 ADG (kg) 0.95 0.71 0.62 FE (feed / increase) 3.95 6.70 6.31 Week 3-4 ADG (kg) 0.89 0.93 0.81 FE (feed / increase) 4.01 3.75 4.29 % Imp. FE - 6.5% - Weeks 1-4 ADG (kg) 0.87 0.89 0.81 FE (feed / increase) 3.91 3.60 3.66 % Imp. FE - ?, 9% 6.4%

These results show that this pressed. Shaped bodies have an effectively extended freezing period of about one week. During the second week of each implant cycle, the treated pigs showed the expected reduction in feed conversion, which is usually observed when somatotropin is discontinued in swine. Even with these reduced feed conversion rates, both regimens showed a 7.9% and 6.4% increase in feed conversion through the 4-week study.

Example 8

Pressed somatotropin moldings were prepared from MBS prepared in the manner described in Example 1. The pressing procedure was the same as in Example 2, except that the tablet form had a diameter of 3.2 mm and the load applied was 8895 Newton (2000 pound). The 50mg moldings were about 5mm long. Fine group of 9 lactating Holstein cows received two subcutaneous implants in the postscapular area of 5 moldings, for a total of 500mg, and a second group of 9 cows received three similar implants with a total of 750mg. The increase in actual milk production and 3.5% fat corrected milk production (FC milk) of these treated cows versus a group of 10 control cows that had not received somatotropin over a 28 day test period and the percentage increase in the 3.5% fat-corrected milk of the treated cows compared to the control cows is summarized in Table IV.

Table IV

Increase in Increase in % Rise Actuals. FC milk bsi FC milk Day / dose Milk (kg / day) (Kg / day) Days Ibis 14 500mg 4.7 5.2 1S, 6 750mg 6.4 6.8 2E, 7 Days 15 to 28 öOOmg 0.8 2.0 8.2 750mg 1.1 1.0 4.1

Example 9

Pressed somatotropin moldings were prepared from MBS, the preparation of which was described in Example 1. For this 1% magnesium stearate was used as lubricant. The press run was similar to that of Example 2, except that the tablet form had a diameter of 4.0 mm and the load applied was 8895 Newton (2000 pound). The 50mg moldings were about 3mm long. A group of 10 moldings was found in a group of 8

lactating cows (Holstein) in the postscapular area on day zero and implanted on day 21. The 10 control cows did not receive somatotropin. During days 1 to 21, the 3.5% FeV corrected milk intake was 8.5% over control and 14.3% on days 22-42. The results of this example were influenced by the fact that in one number of cows in the study there was a marked decrease in milk production as they neared the end of the lactation cycle.

Example 10

Microbially expressed MBS was prepared as described in Example 1 and goreinigt with the following exceptions. The pH of the concentrated MBS solution in urea and in TSIS was adjusted to about 10 by the addition of NaOH. This solution was diafiltrated against 10 volumes of WFI-Wasner, with additional addition of NsOH periodically to maintain the pH at about 10. After filtration, the MBS solution was lyophilized to give a dry powder. This product has been referred to as the "Solution Lyophilizer" MBS ".

Example 11

Microbially expressed MBS was prepared and purified, as described in Example 10, except that the diafiltriv.; A? 'SS solution at pH 10 with HCl neutralizes λ jrde to precipitate the MBS. The precipitated MBS was lyophilized. to get to or dry powder. This product has been named "HCI-precipitated MBS".

Example 12

Microbially expressed MBS was prepared and purified essentially as described in Example 1, except that the pH of the de & MBS concentrated solution in urea and Tris was adjusted to about 4 by the addition of H ₃ PO *. This solution was diafiltered against 10 volumes of WFI water. This diafiltered solution was neutralized with KOH to precipitate the MBS which was lyophilized. A dry powder was obtained. This product was designated as "KOH-precipitated MBS".

Example 13

Pressed somatotropin molded articles were prepared from solution-lyophilized MBS according to Example 10 and HCI-precipitated MBS according to Example 11. The pressing procedure was the same as in Example 2, except that 1% magnesium stearate was added as a lubricant, the diameter of the mold was 4 mm, and the load applied was 8895 Newtons (2000 pound). The 50mg moldings were about 3mm long. One group of 7 Holstein cows received implants of 10 of the 50mg (500mg total dose) molds from solution-lyophilized MBS, and another group of 7 Holstein cows received implants from 10 of the 50mg HCI-precipitated MBS moldings. Implants were implanted subcutaneously in the postscapular area on day 0 and day 21. A group of 7 control cows received blind implants. The actual milk production and the 3.5% fat corrected milk production of the treated cows compared to the control cows which did not receive somatotropin are listed in Table V.

Table V

rise Increase in % Increase in with milk 3,5FCMilch 3,5 FC milk Weeks / MBS Type (Kg / day) (Kg / day) Weeks 1-3 Solution Iyo. 5.3 4.9 20.6 HCI-like 2.6 2.8 11.3 Weeks 4-6 Solution Iyo. 5.9 5.9 27.1 HCI-like 2.8 3.5 16.1

Milk production during week 7 showed a slight increase in the number of cows treated with solution-lyophilized MBS compared to control cows. However, there was no increase with HCI-precipitated MBS.

Example 14

A study similar to that of Example 13 was carried out except that 8 Holstein cows received HCI-precipitated MBS, 8 Holstein cows received KOH-precipitated MBS prepared according to Example 12, and 8 Holstein cows received blind implants. Over a period of 42 days of the study, the cows that received the solution-lyophilized MBS received the control cows in the milk production around 4.5kg / day. The cows that received the HCI-precipitated MBS exceeded the control cows by 4.6 kg / day, and the cows that received the KOH-precipitated MBS exceeded the control cows by 4.0 kg / day.

Example 15

Pressed moldings were prepared from solution lyophilized MBS according to Example 13. A group of 8 Holstein cows received postscapular subcutaneous implantation of 6 of the 50 mg (300 mg dose) molds every 21 days. Over a 42-day study, the group that received 300 mg every 14 days showed an increase in milk production of 5.4 kg / day versus milk production from the same cows before treatment. Similarly, the group that received 500mg every 21 days during the 42-day study showed an increase in milk production of 5.4kg / day over the miich production that these cows had before treatment.

Example 16 Pressed somatctropin moldings were prepared from 25 mg solution-lyophilised MBS prepared according to Oeispel 13

had been made. The pressing procedure was similar to that in the example , with the exception that the tablet form had a

Diameter of 2mrr. the load applied was 1112 Newton (220pound) and 1% magnesium di-ene Lubricant was added. Five groups of 9 Holstein cows each received subcutaneous implants in the Postscapular area with different doses and at different distances. A group of 9 control cows are all 14 Days blind implants. During the study period of 84 days, milk production exceeded:

cows having received 100mg cllo for 14 days, control cows at 5.6kg / day, cows receiving 200mg every 14 days, control cows by θ, 4kg / day, cows, who received Ί0Ο mg every 14 days, the control cows 7.7 kg / day, the cows 600 mg every 21 days, the control cows 7.9 kg / day;

Example 17 Two groups of pressed moldings from solution-lyophilised MBS prepared as in Example 10, to which 1%

\ jpinderstftarat was added as a lubricant, were prepared. The pressing process was similar to Example 2, with the following exceptions. One group was 25mg molded bodies of 2mm diameter pressed with a load of 1112 Newton (250 pound). The second group were 50 mg 4mm diameter moldings pressed at a load of 4448 Newton (1000 pound).

A group of 10 Holstein cows received 12 of the 25mg moldings (300mg) every 14 days. Another group of 10 Holstein cows received 6 of the 50 mg moldings (300mg) every 14 days. A third group of 10 Holstein cows received every 14 days Blind implants. The implants were inserted subcutaneously in the postscapular area. In the period of the 42-day study

exceeded the milk production of the cows which had received the 25 mg tablets, that of the control cows by 4.2 kg / day; and the

Much production of the cows that had received the 50mg moldings exceeded that of the control cows by 4,3kg / day. If the

When the first implantation cycle is considered to be the transitional period between an untreated and a treated condition, the increases at days 15-42 of the study in terms of milk production were 5.0 and 5.4 kg / day versus the control cows, based on the 25 mg tablets and the 50 mg shaped bodies.

Example 18

Microbially expressed Ala-VaI-BST was produced by expression of recombinant DNA essentially as described in Example 3a of European Patent Application 193515, published September 3, 1986. This application is hereby incorporated by reference. The Ala-Val-BST fragments were isolated, solubilized, folded and purified as described in Examples 10 and 1 to give a solution designated as "lyophilized Ala-VaI-BST".

Example 19

Pressed moldings were prepared from solution Iyophilized MBS prepared as in Example 10 and solution-lyophilized Ala-VaI-BST prepared as in Example 18. As described in Example 1 β Zwel-Mllümeter molded bodies were prepared with 1% magnesium stearate. Four groups of 9 Holstein cows each received subcutaneous implants of solution-lyophilized MBS in the postscapular area with the dose levels and states described below. A group of 9 Holstein cows received subcutaneous implants of solution-lyophilized AIa-VaI-BST in the post-scapular area with a 250 mg dose, implanted at 14-day intervals. A group of 9 cows received subcutaneous implants between the hind legs, midway between the udder and vulva. A group of 9 control cows received a blind implant in the postscapular period at 14-day intervals. After 42 days, the treated cows showed the following increases in milk production over the control cows: 125 mg MBS every 14 days, increase of 3.1 kg / day; 250 mg M3S every 14 days, increase of 4.0 kg / day; 250 mg AIa-VnI-BST every 14 days, increase of 6.6 kg / day; 250mg MBS implanted above the udder every 14 days, increase of 4.4kg / day; 375mg MBS every 14 days, increase of 5.0kg / tap; and 600mg MB5 every 2Ί days, increase of 5.8k pounds / day.

Considering the first implantation cycle as a transitional period between an untreated condition and a treated condition, the following increases were seen in the production of treated cows versus control cows, excluding the first treatment cycle: 125 mg MBS every 14 days, increase of 3.7 kg / day; 250 mg MBS every 14 days, increasing to 4.7 kg / day ; 250mg AIa-VaI-BST every 14 days, increase of 7.4kg / day;

250mg MBS implanted above the udder, every 14 days, increase of 5.1 kg / day; 375mg MBS every 14th tape, increase of 5.5kg / day; and 500mg MBS every 21 days, increase of 6.4kg / day.

Example 20 Pressed moldings were made of APS bulk powder and copper-associated APS prepared as in Example 5. The copper-associated APS was prepared at a copper to APS mole ratio of about 2: 1. The pressing process

was generally similar to that of Example 2, except that 5% magnesium stearate was wound as a lubricant.

Eo was given a sufficient amount of the somatotropin formulation to give 60mg and 90mg doses in 2.4mm or

3.2 mm molds and compressed at a loaded load of 4448 Newton (1000 pounds). These shaped bodies were implanted in the fat of the neck back in groups of 10 commercial, crossbred pigs at 14 and 21 day intervals. The study ran for 42 days.

Over the period of 42 days, ADG, FE and% were improved over the negative feed conversion Control animals determined. The results are listed in Table VI, compared to a negative control where no

Somatotropin was given and a positive control where a daily injection of 2 mg APS in solution was given. The results show an effective prolonged release of about one week with the expected reduction in feed conversion after the release period was over. All implantation regimens over the entire 6-week study showed improved feed conversion versus negative control, and most showed feed conversion approximately equivalent to that seen in the daily positive control injection.

Table VI

(-) control (+ IKontr. Dosk. 1.33 90 mg 60 mg APS 60mg Cu APS 90 mg 60 mg no loading (2 mg / day) Interv: 2.73 3Woch. 2Woch. 90 mg 2Woch. 3Woch. 2Woch. plot By n .: 3mm 3mm 2.Woch. 2mm 3mm 2 mm 0.99 3mm week 1 0.99 3.72 1.03 1.05 1.14 1.12 1.16 ADG (kg / day) 3.40 2.99 2.81 1.17 2.71 2.21 2.34 FE (feed / ZI) 1.13 2.69 Week 2 0.98 3.16 0.78 0.80 0.70 0.62 0.65 ADG (kg / day) 3, R4 4.57 4.46 ü, 75 5.25 5.21 4.96 FE (feed / to.) 1.01 4.50 Week 3 0.99 3.74 0.82 1.01 1.19 0.92 1.26 ADG {kg / day) 3.83 4.00 2.84 1.16 3.05 3.90 2.58 FE (feed / to.) 1.03 2.76 Week 4 0.84 3.78 1.28 0.95 0.82 1.23 0.71 ADG (kg / day) 4.62 3.20 4.16 0.70 4.72 2.60 5.21 FE (Filtt./Zun.) 1.19 5.27 Week 5 0.79 3.43 0.97 0.93 1.05 0.69 1.17 ADG (kg / day) 4.34 3.94 3.70 1.04 3.64 6.08 3.16 FE (Futt./Zunah.) 1.11 3.39 Week β 1.00 3.34 0.78 0.99 1.03 0.87 0.82 ADG (kg / day 3.88 - 5.87 3.75 1.06 3.84 4.75 4.84 FE (Futt./Zun.) 3.83 Weeks Ibis β 0.93 0.95 0.96 0.99 0.91 0.96 ADG (kg / day) 3.98 3.82 3.43 0.98 3.58 3.54 3.45 FE (Futt./Zun.) - 4.0 13.8 3.45 10.1 11.1 13.3 % Imp. FE 13.3

Example 21

Pressed moldings were made from APS bulk powder and copper-associated APS (2: 1 molar ratio copper to APS) with 5% magnesium stearate lubricant as in Example 2, with the following exceptions. The moldings were made in 60 mg and 30 mg sizes of 3.2 mm diameter, spread at 4448 Newton (1000 pound) load and 4 mm diameter pressed at 8895 Newton (2000 pound) load. The 4 mm molds had a height approximately equal to the diameter, so that the surface of the cylinder was minimized. A set of 3.2 mm diameter 60 mg moldings and a set of 3.2 mm diameter 30 mg moldings were placed in 2 mm stretchable silaatik tubes which were not over-coated at the ends. In commercial cross-bred pigs, 60 mg dose levels were implanted in the fat on the back of the neck at 4-week intervals. The results of a 5-week period are shown in Table VII and compared to control pigs that had not received somatotropin. The results of the two groups of pigs that had received the implants in Silas'.ikröhrchon show the ability to influence the performance by influencing the size of the release surface. The pigs receiving 2 of the 30 mg molded bodies had improved performance over the pigs which had received an oOmg shaped body. Since the implants had open ends, the implants of the 30 mg molded bodies had twice the release surface area compared to the 80 mg molded body, whereupon their improved performance was due.

Table VII

control 0.72 Type: APS APS CU Cu » Cu · 5.35 Dose: 60 mg 60mg 6bfi-.y 60mg 2-30 mg 0: 3 mm 4mm 4mm 3mm 3mm week 1 ADG (kg / day) 0.84 1.PS 0.94 0.82 0.93 0.83 FE (feed / increase) 4,? 3 a, 53 3.14 3.58 3.33 3.53 Week 2 ADG (kg / day) 3.95 0.63 0.69 0.73 0.65 0.64 FE (feed / increase) 3.69 4:44 4.64 4.08 4.92 9..7S Week 3 ADG (kg / day) 1.02 1.05 0.76 0.92 0.81 0.75 FE (feed / increase) 3.66 3.50 4.09 3.44 4.19 3.97 Week 4 (pigs reimplanted) ADG (kg / day) 0.87 0.89 0.74 0.74 0.84 FE (feed / increase) 3.92 3.84 4.42 4.29 3.90

Control Type: APS APS Cu Cu · Cu » Dose: 60 mg 60 mg 60 mg 60 mg 2-30 mg 60 mg

0: 3mm 4mm 4mm 3mm 3mm

Week 5 0.83 0.82 0.78 0.99 0.78 0.99 ADG (kg / day) 5.58 4.15 3.89 3.15 4.16 3.30 FE (feed / increase) Weeks 1-5 0.87 0.88 0.81 0.84 0.78 ο, αι ADG 3.90 3.53 3.65 3.55 3.98 3.65 FE (feed / increase) - 9.5% 6.4% 9.0% - 6.4% % Imp. FE

• in silastic tubes

Example 22

Coupon-associated APS was prepared by dissolving APS bulk powder prepared according to Example 5 in deionized water at 60 mg / ml. Sufficient 10OmM CuSO 4 solution was added to achieve the desired Cu 2 HP ratio. The pH was adjusted to 7 by the addition of sulfuric acid to precipitate the somatotropin. The precipitate was isolated and lyophilized. From these copper-associated somatotropins pressed moldings were produced which had the said Cu: APS ratio. The press method used was generally that of Example 2, except that the diameter was 3.2 mm, 5% magnesium stearate was added as a lubricant, the load applied was 4448 Newtons (1000 pounds), and 60 mg and 30 mg somatotropin quantities were used. as listed. These moldings were implanted every 14 days in the fat of the neck back of groups of 10 pigs. The pigs were observed over the study period of 6 weeks. The results are listed in Table VIII and compared to a negative control that did not receive somatotropin and a positive control that received a daily injection of APS in solution. The effective duration of each of the treatments varied, but treatments with lower ratios of copper had a longer effective duration than the treatments that had higher ratios.

Table VIII

'-{Control (+) Control 1.0Ü 60 mg 60 mg 2 x 30mg 3 x 30mg 2 x 30mg (no Be (2 mg / day) Dose: 2.73 1: 1 2: 1 1: 1 2: 1 1.5: 1 plot) Cu: APS: week 1 0.98 1.05 1.01 0.99 0.89 0.92 ADG (kg / day) 1.00 2.97 2.57 3.08 2.97 2.90 2.95 FE (feed / increase) ?, 83 Week 2 1.02 0.86 0.69 0.81 0.87 0.92 ADG (kg / day) 0.79 2.76 3.73 4.64 3.72 3.33 3.19 FE (feed / increase) 4.24 Week 3 1.13 1.05 1.21 0.91 1.00 1.07 ADG (kg / day) 0.95 2.75 , 2.35 2.41 2.87 2.84 2.66 FE (feed / increase) 3.36 'Voche * 0.97 0.64 0.70 0.93 1.01 0.81 ADG (lg / day) 0.99 3.15 4.69 6.71 3.32 3.24 3.90 FE (feed / increase) 3.28 Week δ 0.68 1.13 1.11 1.05 0.99 1.00 ADG (kg / day) 0.81 4.16 2.40 2.95 2.58 3.16 2.94 FE (feed / increase) 3.90 Week 6 0.97 0.68 0.69 0.29 0.63 0.38 ADG (kg / day) 0.75 2.85 5.86 5.03 7.05 5.69 4.72 FE (feed / increase) 4.23 - Weeks 1-β 0.91 0.90 0.83 0.90 0.85 ADG (kg / day) 0.88 288 3.31 3.17 3.23 3.38 FE (feed / increase) 3.43 16.0% 3.5% 7.6% 5.8% 1.5% % Imp. FE -

Example 23

Pressed molded articles were dry blended from APS bulk powder prepared as described in Example 5 with a sufficient amount of CuSO ₄ to give the reported Cu to APS molar ratio and from the precipitated copper-associated APS with the indicated Cu-to-APS ratio. APS molar ratio made. The pressing procedure was generally as described in Example 2, except that 5% magnesium stearate was added as a lubricant and the load applied was either 1000 Newtons (225 pounds) or 2224 Newtons (500 pounds) as indicated. The moldings were either weekly or every 2 weeks, as indicated, in groups of 10 pigs bred commercially by crossbreeding in the fat of the back of the neck. The results of the study are listed in Table IX below.

Table IX

Treatment* 1 2 3 1.00 4 1.24 5 6 1.19 7 week 1 2.71 2.42 2.34 ADG (kg / day) 0.93 1.24 1.13 1.19 FE (F / Z) 3.05 2.31 0.88 1.05 2.44 0.95 2.34 Week 2 3.28 3.17 3.17 ADG (kg / day) 0.89 1.11 0.91 0.84 FE (F / Z) 4.19 2.76 0.97 1.09 3.60 1.20 3.62 Week 3 3.32 3.19 2.70 ADG (kg / day) 0.93 1.13 1.14 1.04 Feif / Z) 3.99 2.96 1.09 1.11 3.06 1.02 3.36 Week 4 3.08 3.40 3.62 ADG (kg / day) 1.00 1.21 1.08 0.94 Fe (F / Z) 3.31 2.83 1.12 1.21 3.27 1.08 3.83 Week 5 3.08 3.08 3.12 ADG (kg / day) 1.02 1.18 1.02 1.30 Feif / Z) 3.54 3.11 0.75 1.01 3.37 1:01 2.68 Week 6 4.47 4.18 4.43 ADG (kg / day) 0.69 0.98 0.73 0.63 Feif / Z) 5.29 4.89 0.97 1.12 6.00 1.03 7:06 Weeks 1 to 6 3.23 3.15 3.07 ADG {kg / day) 0.91 1.14 11.0 13.2 1.00 15.4 0.99 FE (F / Z) 3.63 2.93 3.35 3.32 % Imp. FE - - 7.7 8.5

FZ = feed / increase

'Treatment Ί = negative control, did not receive somatotropin treatment 2 = positive control, received 2mg / day APS in solution

Treatment 3 = 21 mg APS administered weekly, no Cu, 2.4mm diameter, 10Oonewtons (22S pound) treatment 4 = 21 mg APS administered weekly, mixed dry. Ratio 2: 1 Cu: APS, 2.4mm diameter, 1000 Newtons, (225 pound)

Treatment 5 = 42mg APS given every 14 days, mixed dry. Ratio 4: 1 Cu: APS, 3.2 mm diameter, 2224 Newton's (500 pound) treatment β = 60 mg APS administered biweekly. Precipitate, 1: 1 ratio Cu: APS, 3.2mm diameter, 2224 Newton

(500 pounds) treatment? = 42 mg APS administered biweekly, precipitate, ratio Cu: APS such as 1: 1.3mm diameter, 2224 Newton (550 pound).

Example 24

Pressed moldings were prepared from precipitated copper-associated APS having a Cu: APS molar ratio of 1: 1, prepared as in Example 5, and a dry mixture of CuSO ₄ and APS bulk powder having a Cu.APS molar ratio of 4: 1 , The pressing procedure was generally similar to that of Example 2, except that the pressed molded articles contained 14 mg and 21 mg of somatotropin, had 2.4 mm diameter, contained 5% magnesium stearate as a lubricant and pressed at a load of Ί 000 Newtons (225 pounds) were. The implantation was carried out weekly in groups of 10 pigs bred commercially by crossing in the fat of the nipple hernia. The results are included in Table X and compared to a negative control which had not received somatotropin and a positive control which had received 2 mg / day of APS administered as a solution.

Table X (-) control (+! Control Type: precipitation dry mixture 4: 1 no loading 2 mg / day Cu: APS Ratio .: 1: 1 4: 1 14mg plot Dose: 14mg 21mg 1.21 week 1 1.23 1.5 1.03 1.06 2.32 ADG (kg / day) 2.44 2.32 2.74 2.49 FE (feed / increase) 0.93 Week 2 1.08 1.00 0.31 0.83 3.44 ADG (kg / day) 3.30 3.16 3.15 3.52 FE (feed / increase) 1.00 Week 3 1.02 1.05 0.96 1.01 3.22 ADG (kg / day) 3.47 2.64 3.11 2.54 FE (feed / increase) 0.82 Week 4 1.05 0.99 0.84 0.79 4.34 ADG (kg / day) 3.62 3.15 4.02 3.89 FE (feed / increase) 0.84 Week 5 0.78 0.86 0.99 0.75 4.01 ADG (kg / day) 5.22 3.49 3.62 4.10 FE (feed / increase)

(-) control (+ {Control Type: precipitation -15- 267 910 4: 1 no loading 2 mg / day Cu: APS Ratio .: 1: 1 tiockenes mixture 14mg plot Dose: 14 mg 4: 1 21mg 1.02 Week 6 0.79 1.27 0.88 3.95 ADG (kg / day) 6,4b 3.32 4.25 0.99 FE (feed / increase) 4.95 0.97 Weeks 1 bite 0.99 1.05 0.93 3.28 ADG 3.56 2.73 3.30 0.90 7.9 FE (feed / increase) - - 7.3 3.17 % Imp. FE 11.0

Example 25

Extruded moldings were prepared from a dry mixture of APS bulk powder and CuSO ₄ (2: 1 Cu: APS molar ratio) and copper-associated APS prepared by lyophilizing a solution of APS and CuSO ₄ at a CutAPS molar ratio of 1: 1 , The pressing procedure was generally that of Example 2, except that the diameter was 2.4 mm, 5% magnesium stearate was added as a lubricant, the load applied was 1000 Newtons (225 pounds), and 21 mg somatotropin per dose was used. Implants were performed weekly for 4 weeks on groups of 10 commercial crossbred pigs in the fat of the neck back. The results are listed in Table XI and compared to a group of negative control pigs that had not received samototropin and to a group of positive control pigs that had received 2 mg of APS in solution per day. The solution lyophilized APS in the ratio 1: 1 with copper-associated APS and the dry mixture with 2: 1 molar ratio proved to be substantially equivalent.

Table Xl

(-) control (+ (Control Lsg.lyophilisiert trock. (no (2 mg / day) mixture plot) week 1 ADG (kg / day) 1.04 1.32 0.91 1.2 FE (feed / increase) 2.88 1.99 2.70 2.10 Week 2 ADG (kg / day) 0.79 1.04 0.90 0.79 FE (feed / increase) 4.09 2.84 3.22 3.59 Week 3 ADG (kg / day) 1.09 1.26 0.99 0.96 FE (feed / increase) 3.18 2.47 3.16 3.26 Week 4 ADG (kg / day) 0.76 0.59 0.86 0.77 FE (feed / increase) 4.86 3.50 3.73 3.84 Weeks IbIs 4 ADG (kg / day) 0.92 1.05 0.91 0.94 FE (food / increase) 3.43 2.77 3.05 3.05 % Imp. FE - - 11.1% 11.1%

Example 26

Pressed moldings were prepared using a dry mixture of CuSO ₄ and APS Polkpulver with the indicated molar ratios and dosages. The pressing procedure was generally similar to that of Example 2, except that 5% magnesium stearate was added as a lubricant, a 2.4 mm mold was used, and the load applied was 1000 Newtons (225 pounds). These pressed bodies were implanted weekly over a six-week period in groups of 10 commercial cross-bred pigs on the back of the neck behind the ear. The results of the study are shown in the following Table XII, comparing with a negative control group that had not received somatotropin and with a positive control group that had received 2 mg / day of APS-administered solution.

Table XII

(-(Control (+ "Control verh .: 1: 1 2: 1 3: 1 2: 1 2: 1 2: 1 (no handicap) (2 mg / day) Dose: 21mg 21mg 21mg 17mg 25mg 21mg week 1 ADG (kg / day) 0, S3 1.03 1.02 1.08 1.01 0.98 1.09 1.14 Fe (forage / Zun.) 3.79 3.58 3.29 3.11 3.51 3.30 3.68 3.23 Week 2 ADG (kg / day) 1.00 1.20 0.98 1.09 1.10 0.92 1.13 1.06 Fe (forage / Zun.) 3.88 3.25 3.49 3.30 3.19 3.82 3.11 3.Ί6

(-) control {+ »control ratio: (no handling) (2mg / day) dose:

1: 1 2: 1 3: 1 2: 1 2: 1 2: 1

21mg 21mg 21mg 17mg 25mg 21mg

Week 3 0.81 0.94 ADG (kg / day) 4.71 3.97 Fe (forage / Zun.) Week 4 0.86 1.01 ADG (kg / day) 4.56 3.72 Fe (forage / Zun.) Week 5 0.79 1:02 ADG (kg / day 5.11 3.56 Fe (forage / Zun.) Week β 0.87 1.06 ADG (kg / day) 4.55 3.73 Fe (forage / Zun.) Weeks 1 bit β 0.88 1.04 ADG (kg / day) 4.29 3.56 FE (feed / increase) - % Imp. FE

0.94 3.54 0.79 4.02 0.93 3.75 0.92 3.76 0.97 3.63 1.01 3.72 0.95 3.73 0.90 3.96 1, 01 3.43 0.97 3.82 0.93 3.87 0.89 7.06 1.05 3.61 1.05 3.75 0.94 3, Γ8 0.84 4.63 0.32 4.23 1.00 4.16 0.99 3.94 1.03 3.56 1.17 3.51 0.97 3.95 1.23 3.46 1.09 3.73 0.99 3.52 17.9 0.99 3.45 19.6 1.03 3.46 19.3 0.93 3.76 12.4 1.05 3.53 17.7 1.03 3.71 13.5

Example 27

Pressed moldings were prepared from a dry mixture of CuSO ₄ and APS bulk powder at a 2: 1 molar ratio of CutAPS. The pressing procedure was generally similar to that of Example 2, except that 5% magnesium stearate or 1% magnesium stearate was added as lubricant as indicated, the load applied was 1000 Newtons (225 pounds) and the diameter was 2.4 mm. Implants of 21 mg per week are performed for 6 weeks in a group of 10 commercial cross bred pigs in the nape of the neck just behind the ear. The results are shown in Table XIII, which was preceded by a negative control group of pigs that had not received somatotropin, and a positive control group of pigs that had received 2 mg of APS per day as a solution. Over the six-week period of the study, 1% magnesium stearate and 5% magnesium stearate treatments had similar results.

Table XIII

(-) Contr. (+ JKontr. • 5% MgSt 1% MgSt no loading (Mg / day) plot week 1 ADG (kg / day) 1.19 1.41 1.26 1.20 FE (feed / increase) 2.48 2.10 2.31 2.36 Week 2 ADG (kg / day) 0.92 0.97 0.80 0.92 FE (feed / increase) 3.87 3.27 3.91 4.61 Week 3 ADG (kg / day) 0.92 1.03 0.99 0.99 FE (feed / increase) 4.28 2.93 3.23 6.02 Week 4 ADG (kg / day) 0.72 1.13 0.85 0.96 FE (feed / increase) 3.96 3.07 3.88 3.51 Week 5 ADG (kg / day) 0.81 1.05 1.03 0.80 FE (Futtor / Increase) 5.05 3.52 3.46 4.31 Week β ADG (kg / day / 0.83 0.96 0.86 1.03 FE (feed / increase) 4.52 3.83 4.28 3.66 Weeks 1 to β ADG (kg / day) 0.90 1.09 0.97 0.94 FE (feed / increase) 3.80 2.99 3.27 3.28 % Imp. FE - - 13.9 13.9

The above examples are illustrative only and not restrictive.

Claims (11)

A process for preparing a shaped body adapted for parenteral administration of a somatotropin to an animal, which comprises combining a substantially binder-free and substantially matrix-free formulation containing an effective dose of solid somatotropin and optionally an effective amount of a lubricant; and coating the shaped article optionally with a coating on a portion of its surface which substantially inhibits the release of the somatotropin on the coated surfaces of the shaped article, but at least one uncoated release surface is provided. 2. The method according to claim 1, characterized in that the somatotropin is a microbially expressed somatotropin. A method according to claim 1, characterized in that the somatotropin is selected from the group consisting of bovine somatotropin and porcine somatotropin. 4. The method according to claim 3, characterized in that the somatotropin is a microbially expressed bovine somatotropin. 5. The method according to claim 4, characterized in that the microbially expressed bovine somatotropin is MBS. 6. The method according to claim 4, characterized in that the microbially expressed bovine somatotropin is Ala-VaI-BST. 7. Method according to claim 3, characterized in that the somatotropin is a microbially expressed porcine somatotropin. 8. The method according to claim 7, characterized in that the microbially expressed pig somatotropin is APS. A method according to claim 7, characterized in that the microbially expressed porcine somatotropin is copper-associated porcine somatotropin or a dry mixture of a copper salt and a porcine somatotropin. 10. The method according to claim 9, characterized in that the shaped body has a copper content of about 0.1% to about 3%. 11. The method according to claim 1, characterized in that the lubricant is selected from the group consisting of stearates and palmitates.