KR19990064322A - Stable pharmaceutical dosage preparations containing parathyroid hormone - Google Patents

Abstract

The present invention relates to pharmaceutical compositions which are stable upon storage and which contain basic amino acids and optionally physiologically resistant organic or inorganic acids.

Description

Stable pharmaceutical dosage preparations containing parathyroid hormone

Parathyroid hormone (PTH) is a protein consisting of 84 amino acids involved in regulating calcium and phosphate content in blood and tissues. WO 90/10067; WO 91/06564; EP 0 301 484; From WO 93/15109 it is known that peptides with the appropriate modification of the N-terminal fragment and amino acid sequence of this hormone also have a biological activity similar to PTH (1-84).

However, due to oxidation in the free methionine group in the molecule, it is difficult to stabilize PTH with pharmaceutical dosage formulations. The addition of antioxidants, such as methionine or ascorbic acid, does not provide a dosage form with adequate stability for pharmaceutical purposes. It can be seen from EP 0 619 119 that stabilization can be achieved by freeze-drying the combination of sugar and sodium chloride. However, it can be seen that this form of stabilization tends to form dimers. However, dimers are a problematic pharmaceutical dosage form because of their unintended side effects due to immune responses when administered to a patient. Dimers also reduce protein activity in the dosage form, particularly when the dosage form is stored at inappropriate temperatures for a long time. Lyophilization is also a problem when preparing properly dried pharmaceutical dosage formulations.

It has been surprisingly found that the inclusion of one or several basic amino acids, in particular arginine, lysine or ornithine, as a pharmaceutical adjuvant in the dosage formulation yields a stable pharmaceutical dosage formulation of PTH or a fragment thereof. As a result, the addition of antioxidants or tensides can be omitted. In addition, the addition of basic amino acids can produce stable dosage formulations for long periods of storage. In particular, undesirable aggregate formation can be reduced or substantially avoided. It has also been found that lyophilization can be improved by further addition of acidic and / or neutral amino acids. Stable aqueous dosage formulations are those which contain, in particular, inorganic or organic acids and have a pH value of 4-8, in particular 6-8.

Parathyroid hormone fragments contemplated within the scope of the present invention are in particular human N-terminal fragments of natural proteins, such as fragments (1-34), (1-35), (1-36), (1-37) or (1-38). However, it is equally possible to use fragments of parathyroid hormones with shortened N- and C-terminus, such as shortened one or two amino acids at the N-terminus. It is also possible to use suitable variants or variants of this hormone in which one or several amino acids in the amino acid sequence of PTH (1-84) are substituted by other amino acids. This also applies to the corresponding N- and / or C-terminal short fragments. In particular, pharmaceutical preparations containing parathyroid hormone fragment hPTH 1-37 can be prepared within the scope of the present invention. This fragment is very unstable due to its structure and arrangement. Therefore, the active ingredient content is continually reduced due to the aggregation and degradation of the peptide during long storage, which often results in product instability in the preparation of pharmaceutical formulations containing conventional pharmaceutical adjuvants.

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

In the liquid dosage form, the content of the active parathyroid hormone or parathyroid hormone fragment is 10 mg / ml or less, in particular 5 mg / ml or 2 mg / ml or less. The content of the active ingredient is preferably at least 0.01 mg / ml, at least 0.04 mg / ml or at least 0.1 mg / ml. The content of the active ingredient is more preferably 0.01 to 5 mg / ml, in particular 0.04 to 2 mg / ml. The lyophilized dosage formulation contains an active ingredient in an amount such that a corresponding volume of the physiologically resistant reconstituted solution can be obtained to obtain a corresponding infusion or injection of the active ingredient in the above concentration range. Pharmaceutical dosage formulations within the scope of the present invention must be free of tensides and conventional physiologically resistant antioxidants, in particular those containing sulfhydryl groups or ascorbic acid, such as for example methionine or cysteine. In addition, since dimers of chlorine ions are well formed, it is preferable that chlorine ions are not necessarily present. In particular, the dosage formulations according to the invention do not otherwise contain any other pharmaceutical additives or auxiliaries such as, for example, sugars or physiologically resistant polymers. Dosage formulations within the scope of the invention are characterized in that they contain at least one basic amino acid, although consisting primarily of amino acids and organic or inorganic acids only.

All physiologically resistant amino acids having at least one basic side group are considered basic amino acids in the present invention. Basic side groups are, in particular, amino groups which may be optionally substituted with other moieties such as, for example, C ₁ -C ₆ alkyl groups. Especially histidine, lysine, arginine or ornithine are preferably considered as basic amino acids.

As such physiologically resistant amino acids having side groups that do not have sulfhydryl groups (cysteine, methionine), neutral amino acids such as phenylalanine, glycine or isoleucine are contemplated.

The amino acids can in principle be used in racemic form or in optically active form (D or L amino acids). The concentration of amino acid in the liquid dosage form is 100 mg / ml or less. Concentrations of up to 80 mg / ml, in particular up to 60 mg / ml and up to 50 mg / ml or up to 40 mg / ml are particularly advantageous. Amino acid concentrations are advantageously at least 1 mg / ml, in particular at least 5 mg / ml.

In the scope of the present invention, physiologically resistant carboxylic acids, hydroxy carboxylic acids or amino acids, as well as salts thereof, in particular alkali salts, are considered organic acids. Lactic acid, acetic acid, citric acid or aspartic acid are advantageous in the scope of the present invention. When the acids have chiral centers, racemates and also optically active derivatives can be used.

Suitable inorganic salts are physiologically resistant acids such as phosphoric acid, sulfuric acid or salts thereof, which can be used as buffers in aqueous solutions, such as sodium dihydrogen phosphate, potassium dihydrogen phosphate, sodium dihydrogen phosphate, sodium hydrogen sulfate and the like. Organic or inorganic acids may also be used in combination with each other. The amount of acid is selected so that the pH value of the aqueous solution is 4-8, preferably 6-8. In general, the concentration of acid in solution is 100 mg / ml or less, in particular 50 mg / ml or 40 mg / ml or less. The concentration of acid is at least 1 mg / ml, preferably at least 5 mg / ml or 10 mg / ml.

Physiologically resistant adjuvants which are preferably used in the scope of the present invention are in particular combinations of the following additives: a) arginine and phosphoric acid (arginine phosphate), b) arginine, phosphoric acid and aspartic acid or c) arginine, phosphoric acid, aspartic acid and Isoleucine.

Pharmaceutical dosage forms can be provided as injectable injectable solutions or infusions which are contained in suitable ampoules. Alternatively, just prior to administration to a patient, it may also be provided in a suitable lyophilisate which can be converted to an aqueous form by addition of an isotonic solution.

It is convenient to prepare dosage formulations by preparing aqueous solutions of all components and filling them in appropriate ampoules or glass vials. In the case of lyophilized products, it is preferred to dry directly in a glass container filled with the solution.

The invention is illustrated in more detail by the following examples.

Example 1

15 mg of PTH (1-37) and 2.5 g of L-arginine are dissolved in 100 ml of water for injection and the pH is adjusted to pH 6.5 with 99% acetic acid. The solution is filtered and sterilized and 1 ml aliquot of this solution is dispensed into glass vials under nitrogen gas.

Example 2

15 mg of PTH (1-37) and 5.5 g of arginine are dissolved in 100 ml of water for injection and the pH is adjusted to pH 7.4 with 85% phosphoric acid. The solution is filtered and sterilized and 1 ml aliquot of this solution is dispensed into glass vials under nitrogen gas. The vial is lyophilized, sealed and inscribed with crimps.

Example 3

150 mg of PTH (1-37) and 5.5 g of arginine are dissolved in 100 ml of water for injection and the pH is adjusted to pH 7.4 with 85% phosphoric acid. The solution is filtered and sterilized and 1 ml aliquot of this solution is dispensed into glass vials under nitrogen gas. The vial is lyophilized, sealed and nicked.

Example 4

15 mg of PTH (1-37), 3 g of L-arginine and 2 g of aspartic acid are dissolved in 100 ml of water for injection and the pH is adjusted to pH 7.4 with 85% phosphoric acid. The solution is filtered and sterilized and 1 ml aliquot of this solution is dispensed into glass vials under nitrogen gas. The vial is lyophilized, sealed and nicked.

Example 5

15 mg of PTH (1-37), 3.0 g of L-arginine and 2.0 g of isoleucine are dissolved in 100 ml of water for injection and the pH is adjusted to pH 7.4 with 85% phosphoric acid. The solution is filtered and sterilized and 1 ml aliquot of this solution is dispensed into glass vials under nitrogen gas. The vial is lyophilized, sealed and nicked.

Example 6

15 mg of PTH (1-37), 3.0 g of arginine, 1.0 g of aspartic acid and 1.0 g of isoleucine are dissolved in 100 ml of water for injection and the pH is adjusted to pH 7.4 with 85% phosphoric acid. The solution is filtered and sterilized and 1 ml aliquot of this solution is dispensed into glass vials under nitrogen gas. The vial is lyophilized, sealed and nicked.

Example 7

15 mg of PTH (1-37), 2.0 g of sucrose and 100 mg of sodium chloride are dissolved in 100 ml of water for injection. The solution is filtered and sterilized and 1 ml aliquot of this solution is dispensed into glass vials under nitrogen gas. The vial is lyophilized, sealed and nicked.

Example 8

In this example, the formulation of Example 2 is prepared at various pH values. The amount of arginine also changes at the same time.

Formulation Arginine pH value 8a 5.8 g 8.0 8b 5.0 g 6.5 8c 3.2 g 5.0 8d 3.2 g 3.0

PTH and arginine are dissolved in 100 ml of water for injection and the pH value is adjusted to each pH value with 85% phosphoric acid. The solution is filtered sterilized and 1 ml aliquot of this solution is dispensed into vials under nitrogen gas. The vial is lyophilized, sealed and nicked.

Example 9

The formulation of Example 9 is prepared similarly to the formulation of Example 2. In formulation 9a) 10 mg of methionine is used. In formulation 9b) 10 mg of ascorbic acid is added. Adjust the pH value of both solutions to 7.4. The solution is filtered sterilized and 1 ml aliquots of this solution are dispensed into vials under nitrogen gas, the vials lyophilized, sealed and nicked.

Example 10

In Example 10 the formulation of Example 2 is used with the addition of 10 mg of Tween 20. In this case, PTH (1-84), arginine and Tween 20 are dissolved in water for injection and the pH value is adjusted to pH 7.4 with 85% phosphoric acid. The solution is filtered sterilized and 1 ml aliquot of this solution is dispensed into vials under nitrogen gas. The vial is lyophilized, sealed and nicked.

Example 11

15 mg of PTH (1-37) and 3.5 g of histidine are dissolved in 100 ml of water for injection and the pH is adjusted to pH 7.4 with 85% phosphoric acid. The solution is filtered sterilized and 1 ml aliquot of this solution is dispensed into vials under nitrogen gas. The vial is lyophilized, sealed and nicked.

Example 12

15 mg of PTH (1-84) (manufactured by Sigma Corporation) and 5.5 g of L-arginine are dissolved in 100 ml of water for injection, and the pH is adjusted to pH 7.4 with 85% phosphoric acid. The solution is filtered sterilized and 1 ml aliquot of this solution is dispensed into vials under nitrogen gas. The vial is lyophilized, sealed and nicked.

In the table:

Table 1 summarizes the results of the stability test for one or three months at refrigeration temperature and 50 ° C. for this example of the present application.

No dimer was found in the dosage form according to the invention as measured by the SDS-PAGE method. It has also been found that the PTH content is greater than 98% after 1 or 3 months storage at about 4 ° C. Even above temperature (50 ° C.) no significant loss of PTH content was seen.

On the other hand, the dosage formulations described in Examples 7, 9 and 10 can be seen to form dimers and lower the PTH content after storage for 1 or 3 months, thus making them less suitable pharmaceutical dosage formulations with regard to storage stability. It can be thought of. In principle the addition of chlorine ions adversely affects the storage stability, as a result of which it has been found that the chlorine-free form is preferred for the dosage formulations according to the invention.

Results for stability survey after 3 months Example PTH content Basic amino acid / sugar Organic acid / inorganic acid Neutral Amino Acids / Other Ingredients pH PTH content% (RP-HPLC) Dimer (SDS-page) start 1 month4 ° 1 month50 ° 3 months ° 3 months50 ° One 0.15 mg Arginine Acetic acid - 6.5 99.2 98.6 92.4 98.2 85.7 - 2 0.15 mg Arginine Phosphoric Acid - 7.4 99.0 98.9 98.5 98.2 98.5 - 3 1.50 mg Arginine Phosphoric Acid - 7.4 99.2 99.1 98.7 98.7 98.5 - 4 0.15 mg Arginine Phosphoric Acid / Aspartic Acid - 7.4 99.9 97.8 98.1 - 5 0.15 mg Arginine Phosphoric Acid Isoleucine 7.4 98.6 98.2 96.4 - 6 0.15 mg Arginine Phosphoric Acid / Aspartic Acid Isoleucine 7.4 99.3 99.1 98.4 98.2 96.6 - 7 0.15 mg Sucrose - NaCl 5.1 98.0 98.9 98.6 98.9 98.6 ++ 8 0.15 mg Arginine Phosphoric Acid - 8.0 97.7 98.5 97.4 - 8 0.15 mg Arginine Phosphoric Acid - 6.5 98.0 98.6 97.1 98.1 96.7 - 8a 0.15 mg Arginine Phosphoric Acid - 5.0 98.6 98.4 96.8 - 8b 0.15 mg Arginine Phosphoric Acid - 3.0 98.2 98.1 96.4 - 9a 0.15 mg Arginine Phosphoric Acid Methionine 7.4 97.5 96.7 96.3 + 9b 0.15 mg Arginine Phosphoric Acid Iscorbic acid 7.4 92.9 89.8 85.9 ++

Claims (17)

Pharmaceutical formulations that are stable upon storage containing parathyroid hormone or parathyroid hormone fragments as active ingredients, as well as one or several basic amino acids. The pharmaceutical formulation of claim 1, wherein adjusting the pH value to 3.0 to 8.0 further comprises a suitable organic or inorganic acid. The pharmaceutical preparation according to claim 1 or 2, which further contains one or several neutral amino acids. The pharmaceutical formulation according to any one of claims 1 to 3, wherein the basic amino acid is histidine, lysine or arginine. The pharmaceutical preparation according to any one of claims 1 to 4, wherein the organic acid is selected from the group of carboxylic acids or amino acids, in particular lactic acid, acetic acid, citric acid or aspartic acid. The pharmaceutical preparation according to any one of claims 1 to 4, wherein the inorganic acid is phosphoric acid or sulfuric acid. The pharmaceutical preparation according to any one of the preceding claims, characterized in that it contains a combination of organic and inorganic acids, in particular a combination of phosphoric acid and aspartic acid or a physiologically resistant salt thereof. The pharmaceutical preparation according to claim 3, wherein the neutral amino acid contains sulfhydryl group-free amino acids, in particular phenylalanine, glycine or isoleucine. The pharmaceutical formulation according to any one of claims 1 to 8, which is a dosage form in the form of a liquid for injection or infusion solution, characterized in that the pH value of the solution is in the range of 4 to 8. A pharmaceutical formulation according to any one of the preceding claims in the form of a lyophilizate for the preparation of infusions or injections with a pH value of 4 to 8, in particular a pH value of 6 to 8. Pharmaceutical preparation according to any one of the preceding claims, characterized in that the active ingredient is contained in a concentration of 0.01 to 10 mg / ml, preferably 0.04 to 2 mg / ml. Pharmaceutical formulation according to any one of the preceding claims, characterized in that it contains a basic amino acid at a concentration of 1 to 100 mg / ml, preferably 5 to 60 mg / ml. 13. A pharmaceutical formulation according to any one of the preceding claims which contains an organic or inorganic acid at a concentration of 1 to 50 mg / ml, preferably 5 to 40 mg / ml. 14. A pharmaceutical formulation according to any one of claims 1 to 13, consisting essentially of amino acids and organic or inorganic acids and containing at least one basic amino acid. A solution or suspension of the active ingredient is prepared in a physiologically tolerant solvent, one or several basic amino acids and one or several organic or inorganic acids are added and then the solution is processed into an injectable or injectable solution which can be directly injected or the solution is A method for preparing a pharmaceutical formulation according to any one of claims 1 to 13, characterized by lyophilization. Use of the pharmaceutical dosage formulation of any one of claims 1 to 13 for the treatment of calcium metabolic diseases, in particular osteoporosis. Use of a basic amino acid to prepare a storage stable pharmaceutical dosage formulation containing parathyroid hormone or parathyroid hormone fragments.