SK47298A3 - Stable pharmaceutical forms of administration containing parathormone - Google Patents

Abstract

The invention concerns pharmaceutical preparations of parathormone. The preparations are stable during storage and contain a basic amino acid and optionally an organic or inorganic physiologically tolerated acid.

Description

Stable pharmaceutical dosage forms containing parathyroid hormone

Technical field

The invention relates to pharmaceutical preparations containing as active ingredient parathyroid hormone or fragments thereof, as well as to the corresponding pharmaceutical dosage forms in the form of lyophilisates or injectable solutions.

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BACKGROUND OF THE INVENTION

Parathyroid hormone (PTH) is a protein composed of eighty-four amino acids that is involved in the regulation of calcium and phosphate levels in blood and tissues. It is known from the literature (cf. also WO 90/10067; WO 91/06564; EP 0 301 484; WO 93/15109) that the N-terminal fragments of this hormone, as well as peptides with corresponding modifications in the amino acid sequence, have analogous biological activity as PTH (1-84).

However, due to the oxidation processes at the free methionine groups in the molecule, stabilization of PTH in the pharmaceutical dosage form is problematic. The addition of antioxidants such as methionine or ascorbic acid does not result in dosage forms with sufficient stability for pharmaceutical purposes. It is known from EP 0 619 119 that stabilization can be achieved by freeze drying a combination of carbohydrates and sodium chloride. However, this stabilization method has been shown to promote dimer formation. Dimers in pharmaceutical dosage forms, however, are problematic because, due to immunological reactions, they may lead to undesirable side effects when administered to a patient. In addition, dimers may lead to loss of protein activity in the dosage form, especially when taken

- the dosage forms are stored for a longer period of time or at sub-optimal temperatures. In addition, lyophilization is problematic in the manufacture of the dried pharmaceutical dosage forms. It has surprisingly been found that pharmaceutically stable dosage forms of PTH or fragments thereof are obtained when one or more basic amino acids, in particular arginine, lysine or ornithine, are present as the pharmaceutical excipient in the dosage form. This makes it possible to dispense with the addition of antioxidants or surfactants. In addition, the addition of basic amino acids results in dosage forms that are stable for long periods of storage. In particular, unwanted aggregate formation can be reduced or prevented. It has further been found that by further addition of an acidic amino acid and / or a neutral amino acid, freeze-drying can be improved. Stable aqueous dosage forms are especially those which contain an inorganic or organic acid and whose pH is 4 to 8, in particular 6 to 8.

Particularly suitable parathyroid hormone fragments within the meaning of the invention are human N-terminal fragments of intact protein, for example fragments (1-34), (1-35), (136), (1-37) or (1-38). However, parathyroid hormone fragments that are N- and C-terminally truncated, for example, fragments that are N-terminally truncated by one or two amino acids, may also be used. In addition, suitable variants or modifications of this hormone in which one or more amino acids in the PTH (1-84) amino acid sequence are replaced with other amino acids may also be used. This also applies to appropriately N- and / or C-terminally truncated fragments. In particular, pharmaceutical compositions containing the parathyroid hormone fragment hPTH 1-37 can be produced according to the invention. This fragment is very labile due to its structure and conformation. Therefore, the production of pharmaceutical preparations with conventional pharmaceutical excipients has often led to product instabilities, since the active ingredient content has been constantly reduced by prolonged storage by agglomeration or decomposition of the peptide.

The content of the active ingredient parathyroid hormone or parathyroid hormone fragments in liquid dosage forms is up to 10 mg / ml, in particular up to 5 mg / ml or up to 2 mg / ml. In particular, the active substance content is at least 0.01 mg / ml, 0.04 mg / ml or 0.1 mg / ml. Preferably, the active ingredient content is, for example, about 0.01-5 mg / ml, in particular 0.04-2 mg / ml. The lyophilized dosage forms contain the active ingredient in amounts such that appropriate infusion or injection solutions of the active ingredient with the indicated concentration ranges are obtained by adding a certain volume of physiologically tolerated reconstitution solution. Pharmaceutical dosage forms within the meaning of the present invention are substantially free of surfactants and conventional physiologically tolerated antioxidants, in particular mercapto-containing reagents such as methionine or cysteine, or free of ascorbic acid. In addition, they are essentially substantially free of chloride ions, since chloride ions promote dimer formation. In particular, the dosage forms of the invention do not contain any other pharmaceutical additives or excipients, such as carbohydrates or physiologically compatible polymers. The dosage forms according to the invention are distinguished in particular by essentially consisting of amino acids and organic or inorganic acids, but containing at least one basic amino acid.

Suitable basic amino acids for the purposes of the present invention are all physiologically tolerable amino acids with at least one basic side group. In particular, the basic side groups are amino groups which may optionally be substituted by other radicals (such as, for example, C 1 -C 6 alkyl). Suitable basic amino acids are, in particular, histidine _from lysine, arginine or ornithine.

Suitable neutral amino acids include physiologically tolerable side-grouped amino acids having no mercapto groups (cysteine, methionine), for example phenylalanine, glycine or isoleucine.

The amino acids can be used in principle in the form of their racemates or optically active forms (D- or Laminic acids). The concentration of amino acids in the liquid dosage form is in the range up to 100 mg / ml. Concentrations of up to 80 mg / ml, in particular up to 60 mg / ml or up to 50 mg / ml or 40 mg / ml are particularly preferred. The amino acid concentration is preferably at least 1 mg / ml, in particular at least 5 mg / ml.

Suitable organic acids for the purposes of the present invention are physiologically tolerable carboxylic acids, hydroxy acids or amino acids, and salts thereof, in particular alkali metal salts. Lactic acid, acetic acid, citric acid or aspartic acid are preferred for the purposes of the invention. If these acids have a chiral center, racemates or optically active derivatives can be used.

Suitable inorganic acids are physiologically tolerable acids, for example phosphoric acid or sulfuric acid or salts thereof, which can also be used in aqueous solution as buffers, for example sodium dihydrogen phosphate, potassium dihydrogen phosphate, sodium hydrogen phosphate, sodium hydrogen sulphate, etc. The organic or inorganic acids may also be used in combination. The amount of acid is selected such that the aqueous solution has a pH of 4-8, preferably 6-8. As a rule, the acid concentration in the solution is up to 100 mg / ml, in particular up to 50 mg / ml or up to 40 mg / ml. The acid concentration is at least 1 mg / ml, preferably at least 5 mg / ml or 10 mg / ml.

In particular, the following combinations of additives are used as physiologically tolerable excipients for the purposes of the present invention: a) arginine and phosphoric acid (arginine phosphate);

(b) arginine, phosphoric acid and aspartic acid; or

c) arginine, phosphoric acid, aspartic acid and isoleucine.

The pharmaceutical dosage forms may be made available as ready-to-use injection or infusion solutions in suitable ampoules. Alternatively, suitable lyophilisates can similarly be made available, which can be converted to an aqueous form shortly for administration to the patient by the addition of isotonic solvents.

Suitably, the preparation of the dosage forms is effected by producing an aqueous solution of all the components and pouring them into suitable ampoules or glass vials. In the case of lyophilisates, the drying takes place primarily directly from the glass containers into which the solution has been filled.

The invention is illustrated by the following examples.

DETAILED DESCRIPTION OF THE INVENTION

Example 1 mg of PTH (1-37) and 2.5 g of L-arginine were dissolved in 100 ml of water for injection and the pH was adjusted to 6.5 with 99% acetic acid. This solution was sterile filtered and 1 ml of this solution was filled into glass vials with nitrogen gasification.

Example 2 mg of PTH (1-37) and 5.5 g of arginine were dissolved in 100 ml of water for injection and the pH was adjusted to 7.4 with 85% phosphoric acid. This solution was sterile filtered and 1 ml of this solution was filled into glass vials with nitrogen gasification. These vials were lyophilized, capped and flanged.

Example 3

150 mg of PTH (1-37) and 5.5 g of arginine were dissolved in 100 ml of water for injection and the pH was adjusted to 7.4 with 85% phosphoric acid. This solution was sterile filtered and 1 ml of this solution was filled into glass vials with nitrogen gasification. These vials were lyophilized, capped and flanged.

Example 4 mg of PTH (1-37), 3 g of L-arginine and 2 g of aspartic acid were dissolved in 100 ml of water for injection and the pH was adjusted with 85% phosphoric acid to

7.4. This solution was sterile filtered and 1 ml of this solution was filled into glass vials with nitrogen gasification. These vials were lyophilized, capped and flanged.

Example 5 mg of PTH (1-37), 3.0 g of L-arginine and 2.0 g of isoleucine were dissolved in 100 ml of water for injection and the pH was adjusted to 7.4 with 85% phosphoric acid. This solution was sterile filtered and 1 ml of this solution was filled into glass vials with nitrogen gasification. These vials were lyophilized, capped and flanged.

Example 6 mg of PTH (1-37), 3.0 g of arginine, 1.0 g of aspartic acid and 1.0 g of isoleucine were dissolved in 100 ml of water for injection and the pH was adjusted to 7 with 85% phosphoric acid. 4th This solution was sterile filtered and 1 ml of this solution was filled into vials with nitrogen gasification. These vials were lyophilized, capped and flanged.

Example 7 mg of PTH (1-37), 2.0 mg of sucrose and 100 mg of sodium chloride were dissolved in 100 ml of water for injection. This solution was sterile filtered and 1 ml of this solution was filled into vials with nitrogen gasification. These vials were lyophilized, capped and flanged.

Example 8

In this example, the formulation of Example 2 is prepared with different pH values. At the same time, the amounts of arginine are varied.

formula arginine PH value 8 a 5.8 g 8.0 8 b 5.0 g 6.5 8 c 3.2 g 5.0 9 d 3/2 g 3.0

PTH and arginine were dissolved in 100 water for injection and the pH was adjusted to 85 using phosphoric acid. The solutions were sterile filtered and 1 ml of solution was filled into vials with nitrogen gas. These vials were lyophilized, capped and flanged.

Example 9

The formulations in Example 9 were prepared similarly to the formulation in Example 2. In formulation 9 a), 10 mg of methionine was used. 10 mg of ascorbic acid was added to formulation 9b). Both solutions were adjusted to pH 7.4. The solutions were sterile filtered and 1 ml of this solution was filled into vials with nitrogen gas. These vials were lyophilized, capped and flanged.

Example 10

In Example 10, the formulation of Example 2 was added with the addition of 10 mg Tween 20. PTH (1-84), arginine and Tween 20 were dissolved in water for injection purposes and the pH was adjusted to ⁷ · 1 with 85% phosphoric acid to 7.4. This solution was sterile filtered and 1 ml of the solution was filled into vials with nitrogen gasification. These vials were lyophilized, capped and flanged.

Example 11 mg of PTH (1-37), 3.5 g of histidine were dissolved in 100 ml of water for injection and the pH was adjusted to 7.4 with 85% phosphoric acid. This solution was sterile filtered and 1 ml of this solution was filled into vials with nitrogen gasification. These vials were lyophilized, capped and flanged.

Example 12 mg of PTH (1-84) (manufactured by Sigma Corporation) and 5.5 g of Larginine were dissolved in 100 ml of water for injection and the pH was adjusted with 85% phosphoric acid to

7.4. This solution was sterile filtered and 1 ml of this solution was filled into vials with nitrogen gasification. These vials were lyophilized, capped and flanged.

Table notes:

The results of the stability test after a shelf life of one or three months at refrigerator temperature (KS) and at 50 ° C are summarized for the above embodiments in Table 1.

Within the dosage forms of the invention, no dimers could be detected in the SDS-PAGE assay. Moreover, it turned out that the PTH content after a storage time is- 'day, optionally three months at about 4 ^and C is at least 98%. · Even at higher temperatures (50 ° C) could not detect any significant loss of the PTH content.

The dosage forms described in Examples 7, 9 and 10, on the other hand, promote dimer formation, or possibly exhibit less PTH after a shelf life of one or three months, and thus may be classified as less suitable pharmaceutical dosage forms due to storage stability. In principle, it has been shown that the chloride ion content is negative with regard to storage stability, so that the chloride-free forms are preferably used in the application forms according to the invention.

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Stability test results after 3 months

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dimers (SDS Page) 1 1 ' 1 1 1 1 + + and and 1 + + + 0 o O Q V> cd '55 85.7 98.5 98.5 9 * 96 9 * 86 96.7 ’-HPLC) 3m < ABOUT 98.2 98.2 98.7 .98,2 6 * 86 98.1 Gi in"/ ABOUT C 50 1 92.4 98.5 98.7 1 * 86 96.4 98.4 9 * 86 TT Γ " ' σ \ 97.1 8 * 96 96.4 96.3 85.9 Contents P 1 mi 4 ° 9 * 86 98.9 1 * 66 97.8 _i 98.2 -1 99.1 6 * 86 98.5 9 * 86 98.4 98.1 96.7 89,8 C w »CZJ 99.2 0 * 66 99.2 99.9 98.6 99.3 0 86 * 97.7 98.0 9 * 86 98.2 97.5 92.9 PH \ABOUT in · t ' in The IN Γ · 7.4 r- " · * m 8.0 \ABOUT CD tn m * 'd r * ΤΓ r * · * Neutral. AMK / ost. ingredients 1 1 1 1 isoleucine isoleucine: NaCl and 1 1 and methionine Kys. ascorbic org./anorg. acid Kys. acid Kys. phosphoric Kys. phosphoric Kys. Phosphoric Kys. aspartic Kys. phosphoric Kys. phosphoric / Kys. aspartic 1 Kys. phosphoric Kys. phosphoric Kys.'fosforečná Kys. phosphoric Kys. phosphoric Kys. phosphoric Again. AMK / saccharide arginine arginine arginine arginine arginine arginine sucrose arginine arginine arginine arginine arginine arginine PTH content 0.15 mg 0.15 mg 1,50 mg 0.15 mg _1 0.15 mg 60 E «n about 0.15 mg 0.15 mg 00 E • t about 0.15 mg 0.15 mg 0.15 mg 0.15 mg Example - CM m 'J · «Λ about T OO 00 CD 00 X) 00 9 a 9b

amino acid

Claims (17)

PATENT CLAIMS A storage-stable pharmaceutical formulation in the form of a lyophilisate or solution for injection or infusion comprising as active ingredient parathyroid hormone or a parathyroid hormone fragment, characterized in that the composition comprises one or more basic amino acids and is substantially free of physiologically tolerable antioxidants. The pharmaceutical composition of claim 1, further comprising an organic or inorganic acid in an amount suitable to adjust the pH of the solution for injection or infusion to a pH range of 4 to 8. Pharmaceutical preparation according to claim 1 or 2, characterized in that it additionally contains one or more neutral amino acids. Pharmaceutical preparation according to one of claims 1 to 3, characterized in that the basic amino acids are histidine, lysine or arginine. Pharmaceutical preparation according to one of claims 14, characterized in that the organic acid is selected from the group comprising carboxylic acids or amino acids, in particular lactic acid, acetic acid, citric acid or aspartic acid. Pharmaceutical preparation according to one of Claims 1 4, characterized in that the inorganic acid is phosphoric or sulfuric acid. Pharmaceutical preparation according to one of Claims 1 to 6, characterized in that it contains combinations of organic and inorganic acids, in particular phosphoric acid and aspartic acid or their physiologically tolerable salts. Pharmaceutical preparation according to one of Claims 37, characterized in that it contains amino acids without mercapto groups, in particular phenylalanine, glycine or isoleucine, as neutral amino acids. Pharmaceutical preparation according to one of Claims 18 in the form of a liquid dosage form for injection or infusion, characterized in that the pH of the solution is in the range of 4 to 8. Pharmaceutical preparation according to one of the claims 1-8 in the form of a lyophilisate for the manufacture of a solution for infusion or injection with a pH of 4 to 8, in particular a pH of 6 to 8 8 Pharmaceutical preparation according to one of Claims 1 - 10, characterized in that it contains the active ingredient in a concentration of 0.01 to 10 mg / ml, preferably 0.04 to 2 mg / ml. Pharmaceutical preparation according to one of Claims 1 - 11, characterized in that it contains a basic amino acid at a concentration of 1-100 mg / ml, preferably 5-60 mg / ml. Pharmaceutical preparation according to one of the claims 1-12, characterized in that it contains an organic or inorganic acid at a concentration of 1-50 mg / ml, preferably 5-40 mg / ml. Pharmaceutical composition according to one of claims 1 13 consisting essentially of amino acids and one organic or inorganic acid, comprising at least one basic amino acid. Process for the production of pharmaceutical preparations according to one of Claims 1 - 14, characterized in that a solution or suspension of the active substance in a physiologically tolerated solvent is prepared and one or more basic amino acids and one or more organic or inorganic acids are added immediately thereafter. the solution is made up into ready-to-use solutions for injection or infusion or lyophilized. Use of pharmaceutical dosage forms according to one of Claims 1 - 14 for the manufacture of medicaments for the treatment of disorders of calcium metabolism, in particular osteoporosis. Use of a basic amino acid for the manufacture of storage stable pharmaceutical dosage forms comprising a parathyroid hormone or a parathyroid hormone fragment, wherein the dosage forms are substantially free of physiologically tolerable antioxidants.