JP2019156805A - Container filling human pth(1-34) liquid pharmaceutical composition, and method for manufacturing the same - Google Patents

Abstract

To provide a pharmaceutical preparation that contains liquid pharmaceutical composition having human PTH(1-34)(teriparatide) as an active ingredient and excellent storage stability, and a method for manufacturing the preparation.SOLUTION: The present invention relates to a pharmaceutical preparation including: a container; and liquid pharmaceutical composition which contains human PTH(1-34), which is filled in the container and consists of amino acid sequence shown in sequence number 1, saccharide and amino acid in water. The present invention also relates to a method for manufacturing the pharmaceutical preparation which includes: a preparation step of preparing liquid pharmaceutical composition which includes adding the human PTH(1-34) into aqueous solution containing the saccharide and the amino acid in water; and a step of charging the liquid pharmaceutical composition into a container.SELECTED DRAWING: Figure 9

Description

  The present invention relates to a pharmaceutical preparation comprising a liquid pharmaceutical composition containing human PTH (1-34) (teriparatide) filled in a container.

  Human PTH (1-34) (Teriparatide) is a polypeptide (amino acid sequence shown in SEQ ID NO: 1) consisting of the N-terminal 1-34 residues of human parathyroid hormone (human PTH), and is a therapeutic agent for osteoporosis Known as. As an injection containing teriparatide acetate as an active ingredient, Asahi Kasei Pharma Co., Ltd. sells it as “56.5 μg for terrib subcutaneous injection”. “Teribbon Subcutaneous Injection 56.5 μg” is a freeze-dried preparation and requires ergonomic dissolution when used, so the work is complicated and there is a possibility of a medical worker's collection volume error. Therefore, liquidation of teriparatide preparations is desired in the medical field.

  Patent document 1 discloses a human patient comprising human PTH (1-34), a polyol stabilizer for use as a medicament, and a buffer for maintaining the pH of the composition within a range of more than 3 to 7. Discloses a pharmaceutical composition in the form of a storage-stable sterile solution ready for parenteral administration. And mannitol is disclosed as a polyol stabilizer.

  In Patent Document 2, a method for storing a human PTH (1-34) aqueous solution injection under exposure, wherein the pH of the aqueous solution injection is 3 to 5, and methionine is used as a stabilizer for humans. 40 to 4,000 parts by weight with respect to 1 part by weight of PTH (1-34), provided that the aqueous solution injection is exposed under conditions of 600,000 Lux · hr. The said preservation | save method that the residual rate of human PTH (1-34) in an injection is 87% or more is disclosed.

Japanese Patent No. 4405666 Japanese Patent No. 4252260

  In Patent Document 1, 100 μg / mL to 500 μg / mL is described as the concentration of human PTH (1-34) in the pharmaceutical composition in the form of the sterile solution. This concentration is the concentration of teriparatide in the existing product ( 56.5 μg / mL), the pharmaceutical composition in the form of the sterile solution described in Patent Document 1 has poor practicality as an injection.

  In Examples 1 and 2 of Patent Document 2, an aqueous solution injection containing about 30 μg of human PTH (1-34) and about 20 mg of DL-methionine and L-methionine in 1 mL is disclosed. It is disclosed that injections have high storage stability under exposure at 40 ° C. On the other hand, the permissible daily dose of methionine to humans is 0.2 mg / kg body weight for subcutaneous injection and 0.1 mg / kg body weight for intravenous injection (Pharmaceutical Additives Dictionary 2016 (Also, published by Yakuji Nippo), the allowable daily dosage for a human weighing 60 kg is 12 mg for subcutaneous injection and 6 mg for intravenous injection. Therefore, when 1 mL of the aqueous solution injection described in Examples 1 and 2 of Patent Document 2 is administered subcutaneously or intravenously to a human body weight of 60 kg, about 20 mg of methionine will be administered. There is a problem that the daily allowance of intravenous injection is exceeded.

  As a result of intensive studies aimed at providing a pharmaceutical preparation containing a liquid pharmaceutical composition having human PTH (1-34) (teriparatide) as an active ingredient and excellent in storage stability, and a method for producing the same, The present invention has been completed.

(1) A pharmaceutical preparation comprising a container and a liquid pharmaceutical composition containing human PTH (1-34) consisting of the amino acid sequence shown in SEQ ID NO: 1, filled with the sugar, and sugar and amino acid in water.
(2) The pharmaceutical preparation according to (1), wherein the sugar is at least one selected from mannitol, sorbitol, and purified sucrose.
(3) The pharmaceutical preparation according to (1) or (2), wherein the amino acid is methionine.
(4) The pharmaceutical preparation according to (3), wherein the liquid pharmaceutical composition contains the methionine at a concentration of 1 to 10 mg / mL.
(5) The pharmaceutical preparation according to any one of (1) to (4), wherein the water is a buffered aqueous solution having a pH of 5.0 or less.
(6) A pharmaceutical preparation comprising a container and a liquid pharmaceutical composition containing human PTH (1-34) consisting of the amino acid sequence shown in SEQ ID NO: 1 filled in the container, sugar and amino acid in water A way to
A preparation step of preparing a liquid pharmaceutical composition comprising adding the human PTH (1-34) to an aqueous solution containing the sugar and the amino acid in water;
Filling the container with the liquid pharmaceutical composition.
(7) The method according to (6), wherein the sugar is one or more selected from mannitol, sorbitol, and purified sucrose.
(8) The method according to (6) or (7), wherein the amino acid is methionine.
(9) The method according to (8), wherein the aqueous solution contains the methionine at a concentration of 1 to 10 mg / mL.
(10) The method according to any one of (6) to (9), wherein water in the aqueous solution is a buffered aqueous solution having a pH of 5.0 or less.

  The pharmaceutical preparation of the present invention can stably store a liquid pharmaceutical composition containing human PTH (1-34) (teriparatide) as an active ingredient.

  According to the method for producing a pharmaceutical preparation of the present invention, degradation of human PTH (1-34) (teriparatide) during production and preservation of the produced product can be suppressed.

The measurement result of the total affinity content in Experiment 1 (comparison of a glass vial) is shown. The measurement result of the total affinity content in Experiment 2 (comparison of rubber plug materials) is shown. The measurement result of the total affinity content in Experiment 3 (comparison of pH of aqueous solution formulation) is shown. The measurement result of the total affinity content in Experiment 4 (comparison of buffer solution) is shown. The measurement result of the total affinity content in Experiment 5 (comparison of isotonic agents) is shown. The measurement result of the total affinity content in Experiment 6 (comparison of L-methionine concentration) is shown. The measurement result of the total relative content rate in Experiment 7 (comparison of the presence or absence of methionine under different ozone concentrations) is shown. The measurement result of the content rate of the related substance of relative retention time 0.76 in Experiment 7 (comparison of the presence or absence of methionine under different ozone concentrations) is shown. The measurement result of the content rate of the related substance of relative retention time 0.98 in Experiment 7 (comparison of the presence or absence of methionine under different ozone concentrations) is shown. The measurement result of the total affinity content in Experiment 8 (comparison of inert gas) is shown.

<Active ingredient>
Human PTH (1-34) is a polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 1. The polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 1 includes those in the form of pharmaceutically acceptable salts. The pharmaceutically acceptable salt is preferably acetate. The human PTH in the form of acetate (1-34), CAS No. teriparatide acetate 99294-7 (molecular formula _{C 181 H 291 N 55 O 51} S 2 · 5CH 3 COOH, molecular weight 4417.97) is known Yes.

<Preferred embodiment of pharmaceutical preparation>
The pharmaceutical preparation of the present invention is excellent in storage stability since the denaturation of human PTH (1-34) into a related substance is suppressed during storage.

The container that can be used for the pharmaceutical preparation of the present invention may be any container that can accommodate a liquid pharmaceutical composition. For example, various types of containers such as vials, ampoules, and syringes can be used. The material of the container is not particularly limited, and examples thereof include glass, polyethylene, polypropylene, silicon, and stainless steel. The container is more preferably a container containing glass, polyethylene, polypropylene or silicon at least on the surface in contact with the liquid pharmaceutical composition. The container is preferably a light-shielded container. When the container is a glass container, at least the surface that comes into contact with the liquid pharmaceutical composition is a surface coated with a SiO ₂ coating (silicoat processing), so that the related substance of human PTH (1-34) during storage Since the modification | denaturation to is suppressed more effectively, it is preferable.

  In the pharmaceutical preparation of the present invention, a liquid pharmaceutical composition containing human PTH (1-34) consisting of the amino acid sequence shown in SEQ ID NO: 1, sugar and amino acid in water is filled and enclosed in the container. The container is sealed, and the oxygen concentration in the gas phase portion in the container is preferably 5% or less, more preferably 4% or less, more preferably 2% or less, even more preferably 1% or less, particularly preferably. 0.5% or less. Such a low oxygen concentration can be achieved by filling the liquid pharmaceutical composition in a container, replacing the gas phase portion with an inert gas such as nitrogen or argon, and then sealing the container.

  Below, preferable embodiment of the said liquid pharmaceutical composition is described concretely.

  The concentration of human PTH (1-34) in the liquid pharmaceutical composition is not particularly limited, but is preferably a concentration that does not require dilution and can be directly administered to humans as an injection, and is free human PTH (1- 34) The concentration in terms of (teriparatide) is, for example, 1 to 500 μg / mL, preferably 1 to 150 μg / mL, for example 56.5 μg / mL.

  The sugar is not particularly limited as long as it is a pharmaceutically acceptable sugar, and can be a monosaccharide, a disaccharide, or a trisaccharide or higher polysaccharide. The sugar may be composed of one kind of sugar or a mixture of two or more kinds of sugars. In addition to the function as an isotonic agent that regulates the liquid pharmaceutical composition, the sugar has a function of suppressing denaturation of human PTH (1-34) into an analog. Specific examples of the sugar include mannitol, sorbitol, purified sucrose, maltose, glucose and the like, preferably at least one selected from mannitol, sorbitol and purified sucrose, and most preferably mannitol. One or more sugars selected from mannitol, sorbitol, and purified sucrose are preferable because they have a particularly high function of suppressing the denaturation of human PTH (1-34) into a related substance. The sugar may be any of D-form, L-form, and a mixture of D-form and L-form. The concentration of sugar in the liquid pharmaceutical composition is not particularly limited, but is preferably a concentration that does not require dilution and can be directly administered to humans as an injection, for example, 0.5 to 500 mg / mL, preferably 5 to 5 mg / mL. 100 mg / mL. When the sugar is mannitol, the concentration can be exemplified by 52.1 mg / mL.

  The amino acid is not particularly limited as long as it is a pharmaceutically acceptable amino acid, and examples thereof include methionine, lysine, arginine, histidine, tryptophan, ornithine, and methionine is particularly preferable. The amino acid may consist of one amino acid or a mixture of two or more amino acids. The amino acid has a function of suppressing the denaturation of human PTH (1-34) into a related substance. In particular, by coexisting an amino acid with a sugar, the function of suppressing the denaturation of human PTH (1-34), which both substances have alone, into a similar substance is exerted synergistically. Compared with the case where it is used as a preservative for human PTH (1-34), the amount of amino acids used can be greatly reduced. This effect is particularly remarkable when the amino acid is methionine. The amino acid may be any of L-form, D-form, and a mixture of L-form and D-form. The amino acid concentration in the liquid pharmaceutical composition is not particularly limited, but it is preferably a concentration that does not require dilution and can be directly administered to humans as an injection. In an embodiment where the amino acid is methionine, the concentration of methionine in the liquid pharmaceutical composition is preferably 1-10 mg / mL, more preferably 1-5 mg / mL, for example 2 mg / mL. . Even when such an effective amount of human PTH (1-34) is administered to a human, such a liquid pharmaceutical composition having such a low methionine concentration can be used as an acceptable daily dose for humans. Since it is easy to fall below (0.2 mg / kg body weight in the case of subcutaneous injection, 0.1 mg / kg body weight in the case of intravenous injection), it is preferable.

  The water in the liquid pharmaceutical composition may be water that is pharmaceutically acceptable. The water in the liquid pharmaceutical composition may be in the form of a pharmaceutically acceptable buffered aqueous solution having a pH of 5.0 or less. Examples of the buffer aqueous solution having a pH of 5.0 or less include an acetate buffer solution, a phosphate buffer solution, and a citrate buffer solution, and an acetate buffer solution or a phosphate buffer solution is preferable. Examples of the concentration of the acetate buffer, phosphate buffer or citrate buffer include 6.0 to 21.0 mM. In a buffered aqueous solution having a pH of 5.0 or less, human PTH (1-34) is easily retained stably. The buffered aqueous solution having a pH of 5.0 or less is more preferably a buffered aqueous solution having a pH of 3.0 to 5.0, more preferably a pH of 3.5 to 5.0, and particularly preferably a pH of 4.0 to 4.5.

  A particularly preferred embodiment of the liquid pharmaceutical composition is a liquid pharmaceutical composition containing human PTH (1-34), D-mannitol and L-methionine in an acetate buffer having a pH of 5.0 or lower. In said particularly preferred embodiment, human PTH (1-34) is more preferably teriparatide acetate. In the particularly preferred embodiment, the concentration of human PTH (1-34) is more preferably 56.5 μg / mL in terms of free form. In the particularly preferred embodiment, it is further preferred that the concentration of D-mannitol is 52.1 mg / mL. In the particularly preferred embodiment, the concentration of L-methionine is more preferably 2 mg / mL.

  In the pharmaceutical preparation of the present invention, the liquid pharmaceutical composition is excellent in storage stability. For example, the total amount of related substances derived from human PTH (1-34) when stored at 25 ° C. for 1 month is the raw material. It can be 5% or less with respect to human PTH (1-34) used as.

  In the pharmaceutical preparation of the present invention, the liquid pharmaceutical composition is excellent in storage stability. For example, in the Examples among related substances derived from human PTH (1-34) when stored at 25 ° C. for 1 month. The amount of a related substance having a relative retention time of 0.76 to be defined can be 0.5% or less based on human PTH (1-34) used as a raw material.

<Preferred embodiment of a method for producing a pharmaceutical preparation>
The method for producing the pharmaceutical preparation of the present invention comprises:
A preparation step of preparing a liquid pharmaceutical composition comprising adding human PTH (1-34) in an aqueous solution comprising a sugar and an amino acid in water;
And a filling step of filling the liquid pharmaceutical composition into a container.

  Here, preferred embodiments of human PTH (1-34), sugar, amino acid, water, container, pharmaceutical preparation and liquid pharmaceutical composition are as described above for the pharmaceutical preparation. The preferred concentration of sugar or amino acid in the aqueous solution is the same as the preferred concentration of sugar or amino acid in the liquid pharmaceutical composition.

  In the preparation step, an aqueous solution containing a sugar and an amino acid in water is prepared in advance, and human PTH (1-34) is added to the aqueous solution, thereby suppressing deterioration of human PTH (1-34) to a minimum. it can.

  Although it is not necessary to perform the said preparation process in the nitrogen atmosphere which hardly contains ozone, degradation of human PTH (1-34) in the said preparation process is suppressed by performing in the atmosphere with low ozone concentration. Therefore, it is preferable.

  In the preparation step and the filling step, the aqueous solution or the prepared liquid pharmaceutical composition is preferably handled while being bubbled with an inert gas.

  In the filling step, when filling the liquid pharmaceutical composition into a container, it is preferable to replace the gas phase portion in the container with an inert gas and enclose it.

<Procedure for preservation test>
Fill the vial with 1.2 mL of an aqueous solution prepared by mixing 60.6 μg teriparatide acetate (56.5 μg as teriparatide) and other components described in each experiment, and inactivate the gas phase. A sample for storage test was substituted with a gas and sealed with a rubber stopper having a Teflon laminate in the wetted part.

  When preparing the aqueous solution preparation, preparation of the raw material and filling into the vial are performed in an air atmosphere at room temperature (22 ° C.) unless otherwise specified, and it takes about 5 hours from preparation of the raw material to completion of filling. went. When preparing the raw materials, components other than teriparatide acetate were first prepared, and then teriparatide acetate was prepared. The raw materials were prepared under non-light-shielding conditions while bubbling with nitrogen in water or an aqueous solution. Nitrogen was used as an inert gas for bubbling and replacing the gas phase in the vial unless otherwise specified.

  Samples for storage tests were stored at the temperature and time conditions described in each experiment. The inside of the stability tester where the storage test was performed is shielded from light.

  The content of the related substance of teriparatide in the sample for storage test stored under each condition was measured by the procedure described later.

  The percentage of the total weight of related substances in the sample for storage test measured after storage under each condition with respect to the teriparatide converted weight (56.5 μg) of teriparatide acetate added as a raw material, expressed as a percentage It was set as content rate (%).

  For the aqueous solution preparation immediately before encapsulation, the total related content (%) was determined in the same procedure.

The total related content (%) in the aqueous solution preparation immediately before encapsulation is A _IN (%), and the total related content (%) in the sample for storage test after storage under each condition is A _{storage condition} (%) It was.

A _IN (%) is the ratio of related substances generated at the stage of filling the formulation of the aqueous solution formulation.

The range of change from A _IN (%) to A _{storage conditions} (%) is an index of the increase in related substances during storage under each condition, that is, the degree of deterioration.

Furthermore, the teriparatide residual rate (%) after storage under each condition was calculated by the following formula.
Teriparatide remaining rate (%) = 100 × (100-A _{storage condition} ) / (100-A _IN )
The teriparatide residual rate is an indicator of the stability of teriparatide during storage in a vial.

<Measurement of total amount of related substances>
Related substances were measured under the following conditions.
Sample solution: An amount corresponding to about 56.5 μg of teriparatide of the sample for preservation test stored under each condition (equivalent to 1 mL of this product) is taken, and a solution (3 g of benzalkonium chloride is dissolved in a sulfate buffer solution, and the total amount is 2000 mL) 80 μL and 120 μL of acetonitrile for liquid chromatography are added to obtain a sample solution.
Detector: UV absorptiometer (measurement wavelength: 214 nm)
Column: A stainless tube having an inner diameter of 4.6 mm and a length of 15 cm packed with octadecylsilylated silica gel for liquid chromatography having a thickness of 3.5 μm.
Column temperature: constant temperature around 40 ° C. Mobile phase A: 0.05 mol / L sulfate buffer / acetonitrile mixture for liquid chromatography (9: 1)
Mobile phase B: 0.05 mol / L sulfate buffer / acetonitrile mixture for liquid chromatography (3: 7)
Mobile phase liquid feeding: The concentration gradient is controlled by changing the mixing ratio of mobile phase A and mobile phase B as follows.

Flow rate: 1.0 mL per minute
Area measurement range: 50 minutes after sample solution injection The relative retention time (RRT) of the related substance indicates the relative retention time when the relative retention time of the main peak of human PTH (1-34) is 1.0.

<Experiment 1>
As an aqueous solution preparation, an aqueous solution preparation containing 60.6 μg / mL teriparatide acetate (56.5 μg / mL as teriparatide) in water and adjusted to pH 4.5 by adding glacial acetic acid was used.

As a vial, a 5 mL capacity borosilicate glass vial coated with a SiO ₂ coating (silico-coated) (CS-5 silicate coating), a 2 mL capacity borosilicate glass vial (CS-2 silicate coated) ) Three types of brown borosilicate glass vials (CS-5) whose inner surfaces with a volume of 5 mL were not subjected to silico coating were used.

  The storage conditions of the sample for storage test were 3 months at 5 ° C, 2 weeks at 10 ° C, 1 month and 3 months, 2 weeks and 1 month at 25 ° C, 1 week and 2 weeks at 40 ° C. .

  The measurement results of the total affinity content are shown in FIG.

  Table 2 shows the residual ratio of teriparatide.

Coating the inner surface of SiO ₂ film (Shirikoto processing) the borosilicate glass vial (CS-5 Shirikoto processing, CS-2 Shirikoto processing) by using, stability is increased analogues teriparatide in the vial during storage It was confirmed that the production of substances was suppressed. In addition, it was recognized that the use of 2 mL vials tended to increase the stability of teriparatide and suppress the generation of related substances, compared to the use of 5 mL vials.

  In the following experiments, borosilicate glass vials (CS-5 silicoat processing) whose inner surfaces were 5 mL in volume were coated unless otherwise specified.

<Experiment 2>
As an aqueous solution preparation, an aqueous solution preparation containing 60.6 μg / mL teriparatide acetate (56.5 μg / mL as teriparatide) in water and adjusted to pH 4.5 by adding glacial acetic acid was used.

  Two types of rubber plugs were used: a rubber plug made of chlorinated butyl rubber and a rubber plug made of n-butyl rubber.

  The storage conditions of the sample for storage test were 3 months at 5 ° C, 2 weeks at 10 ° C, 1 month and 3 months, 2 weeks and 1 month at 25 ° C, 1 week and 2 weeks at 40 ° C. .

  The measurement results of the total affinity content are shown in FIG.

  Table 3 shows the residual ratio of teriparatide.

  With either rubber stopper, no difference was observed regarding the total content of teriparatide and the residual ratio of teriparatide.

  In the following experiments, unless otherwise specified, a rubber plug made of chlorinated butyl rubber was used as the rubber plug.

<Experiment 3>
As an aqueous solution preparation, it contains 60.6 μg / mL teriparatide acetate (56.5 μg / mL as teriparatide) and 0.17 mM glacial acetic acid in water, and an appropriate amount of hydrochloric acid or sodium hydroxide is added to adjust the pH. The aqueous solution formulation adjusted to 3.5, 4.0, 4.5, 5.0, or 5.5 was used.

  The storage conditions of the sample for storage test were 3 months at 5 ° C, 2 weeks at 10 ° C, 1 month and 3 months, 2 weeks and 1 month at 25 ° C, 1 week and 2 weeks at 40 ° C. .

  The measurement result of the total affinity content is shown in FIG.

  Table 4 shows the residual ratio of teriparatide.

  It was confirmed that when the pH was 4.0 or 4.5, the increase in the total related content of teriparatide during the storage period and the decrease in the teriparatide residual rate were suppressed, and the teriparatide was stabilized. In particular, when pH is 4.5, the effect of stabilizing teriparatide is high.

  There is no difference between pH with respect to the total relative content at the beginning (IN in FIG. 3) reflecting the degradation of teriparatide until filling the vial with the aqueous formulation.

<Experiment 4>
Five aqueous solution formulations having different compositions were prepared.

  The “acetate buffer (6.8 mM)” test group contained 60.6 μg / mL teriparatide acetate (56.5 μg / mL as teriparatide) in a 6.8 mM acetate buffer aqueous solution, and an appropriate amount of hydrochloric acid or water. It is a test section using an aqueous solution formulation adjusted to pH 4.5 by adding sodium oxide.

  The “glacial acetic acid + NaOH” test group contained 60.6 μg / mL teriparatide acetate (56.5 μg / mL as teriparatide) and 6.8 mM glacial acetic acid in water, and an appropriate amount of hydrochloric acid or sodium hydroxide was added. This is a test section using an aqueous solution preparation having a pH adjusted to 4.5.

  The “acetate buffer (20.4 mM)” test group contained 60.6 μg / mL teriparatide acetate (56.5 μg / mL as teriparatide) in a 20.4 mM acetate buffer aqueous solution, and contained an appropriate amount of hydrochloric acid or water. It is a test section using an aqueous solution formulation adjusted to pH 4.5 by adding sodium oxide.

  The “citrate buffer solution (6.8 mM)” test group contained 60.6 μg / mL teriparatide acetate (56.5 μg / mL as teriparatide) in a 6.8 mM citrate buffer aqueous solution, and contained an appropriate amount of hydrochloric acid. Or it is a test section using the aqueous solution formulation which added sodium hydroxide and adjusted pH to 4.5.

  The “phosphate buffer (6.8 mM)” test group contained 60.6 μg / mL teriparatide acetate (56.5 μg / mL as teriparatide) in a 6.8 mM phosphate buffer aqueous solution, and contained an appropriate amount of hydrochloric acid. Or it is a test section using the aqueous solution formulation which added sodium hydroxide and adjusted pH to 4.5.

  The storage conditions of the sample for storage test were 3 months at 5 ° C, 2 weeks at 10 ° C, 1 month and 3 months, 2 weeks and 1 month at 25 ° C, 1 week and 2 weeks at 40 ° C. .

  The measurement result of the total affinity content is shown in FIG.

  Table 5 shows the residual ratio of teriparatide.

  In the “citrate buffer solution (6.8 mM)” test group, an increase in the total content and a decrease in the teriparatide residual rate during the storage period were observed at 40 ° C., and there was no difference in the other test sections.

  There was no difference between the test sections with respect to the total content ratio at the beginning (IN in FIG. 4) reflecting the deterioration of teriparatide until the vial was filled with the aqueous preparation.

<Experiment 5>
Five aqueous solution formulations having different compositions were prepared.

  The “D-mannitol” test group contained 60.6 μg / mL teriparatide acetate (56.5 μg / mL as teriparatide) and 52.1 mg / mL D-mannitol in a 6.8 mM acetate buffer aqueous solution. This is a test section using an aqueous solution preparation adjusted to pH 4.5 by adding an appropriate amount of hydrochloric acid or sodium hydroxide.

  The “purified sucrose” test group contains 60.6 μg / mL teriparatide acetate (56.5 μg / mL as teriparatide) and 97.9 mg / mL purified sucrose in a 6.8 mM acetate buffer aqueous solution. It is a test group using an aqueous solution preparation adjusted to pH 4.5 by adding hydrochloric acid or sodium hydroxide.

  The “D-sorbitol” test group contained 60.6 μg / mL teriparatide acetate (56.5 μg / mL as teriparatide) and 52.1 mg / mL D-sorbitol in a 6.8 mM acetate buffer aqueous solution. This is a test section using an aqueous solution preparation adjusted to pH 4.5 by adding an appropriate amount of hydrochloric acid or sodium hydroxide.

  The “maltose monohydrate” test group was prepared by adding 60.6 μg / mL teriparatide acetate (56.5 μg / mL as teriparatide) and 51.0 mg / mL maltose monohydrate in a 6.8 mM acetate buffer aqueous solution. This is a test section using an aqueous solution preparation containing a Japanese product and adjusted to pH 4.5 by adding an appropriate amount of hydrochloric acid or sodium hydroxide.

  The “D-glucose” test group contained 60.6 μg / mL teriparatide acetate (56.5 μg / mL as teriparatide) and 51.0 mg / mL D-glucose in a 6.8 mM acetate buffer aqueous solution. This is a test section using an aqueous solution preparation adjusted to pH 4.5 by adding an appropriate amount of hydrochloric acid or sodium hydroxide.

  The storage conditions of the sample for storage test were 3 months at 5 ° C, 2 weeks at 10 ° C, 1 month and 3 months, 2 weeks and 1 month at 25 ° C, 1 week and 2 weeks at 40 ° C. .

  The measurement results of the total affinity content are shown in FIG.

  Table 6 shows the residual ratio of teriparatide.

  In the test section using D-mannitol, purified sucrose, or D-sorbitol as an isotonic agent, an increase in the total content and a decrease in the teriparatide residual rate during storage at 40 ° C. It was confirmed to be small and high in storage stability compared to the test group using hydrate or D-glucose.

  There was no difference between the test sections with respect to the total content ratio at the beginning (IN in FIG. 5) reflecting the deterioration of teriparatide until filling the vial with the aqueous preparation.

<Experiment 6>
Three aqueous solution formulations having different compositions were prepared.

  The “L-methionine (1 mg / mL)” test group was prepared by adding 60.6 μg / mL teriparatide acetate (56.5 μg / mL as teriparatide) and 52.1 mg / mL in a 6.8 mM acetate buffer aqueous solution. This is a test section using an aqueous solution preparation containing D-mannitol and 1 mg / mL L-methionine and adjusted to pH 4.5 by adding an appropriate amount of hydrochloric acid or sodium hydroxide.

  The “L-methionine (2 mg / mL)” test group was prepared by adding 60.6 μg / mL teriparatide acetate (56.5 μg / mL as teriparatide) and 52.1 mg / mL in a 6.8 mM acetate buffer aqueous solution. This is a test section using an aqueous solution preparation containing D-mannitol and 2 mg / mL L-methionine and adjusted to pH 4.5 by adding an appropriate amount of hydrochloric acid or sodium hydroxide.

  The “L-methionine (10 mg / mL)” test group was prepared by adding 60.6 μg / mL teriparatide acetate (56.5 μg / mL as teriparatide) and 52.1 mg / mL in a 6.8 mM acetate buffer aqueous solution. This is a test section using an aqueous solution preparation containing D-mannitol and 10 mg / mL L-methionine and adjusted to pH 4.5 by adding an appropriate amount of hydrochloric acid or sodium hydroxide.

  The storage conditions of the sample for storage test were 3 months at 5 ° C, 2 weeks at 10 ° C, 1 month and 3 months, 2 weeks and 1 month at 25 ° C, 1 week and 2 weeks at 40 ° C. .

  The measurement results of the total affinity content are shown in FIG.

  Table 7 shows the residual ratio of teriparatide.

  When D-mannitol and L-methionine are used in combination, the same effect is obtained with respect to the suppression of teriparatide degradation and the generation of related substances regardless of whether the concentration of L-methionine is 1 mg / mL, 2 mg / mL, or 10 mg / mL. Was recognized.

<Experiment 7>
The following tests were conducted to examine the influence of the ozone concentration in the atmosphere at the time of preparation of the aqueous solution preparation and the presence or absence of addition of methionine on the production of related substances of teriparatide.

  The “0 ppm (with methionine)” test zone and the “0 ppm (without methionine)” test zone are the test zones in which the process of about 5 hours from preparation of the raw material of the aqueous solution preparation to filling was performed in a nitrogen atmosphere not containing ozone. It is.

  The “0.01 ppm (with methionine)” test section and the “0.01 ppm (without methionine)” test section are the steps of about 5 hours from preparation of the raw material of the aqueous solution preparation to filling, and air with an ozone concentration of 0.01 ppm. This is a test section conducted in an atmosphere.

  The "0.1-0.5 ppm (with methionine)" test section and the "0.1-0.5 ppm (without methionine)" test section are steps of about 5 hours from preparation of the raw material of the aqueous solution preparation to filling. This is a test section conducted in an air atmosphere with an ozone concentration of 0.1 to 0.5 ppm.

  The “0 ppm (with methionine)” test section, the “0.01 ppm (with methionine)” test section, and the “0.1 to 0.5 ppm (with methionine)” test section are all 6.8 mM as an aqueous solution formulation. In an acetate buffer aqueous solution, containing 60.6 μg / mL teriparatide acetate (56.5 μg / mL as teriparatide), 52.1 mg / mL D-mannitol, and 2 mg / mL L-methionine, An aqueous solution formulation adjusted to pH 4.5 by adding hydrochloric acid or sodium hydroxide was used.

  The “0 ppm (without methionine)” test section, the “0.01 ppm (without methionine)” test section, and the “0.1 to 0.5 ppm (without methionine)” test section are all 6.8 mM as an aqueous solution formulation. In an acetate buffer aqueous solution, 60.6 μg / mL teriparatide acetate (56.5 μg / mL as teriparatide) and 52.1 mg / mL D-mannitol were added, and an appropriate amount of hydrochloric acid or sodium hydroxide was added. An aqueous solution formulation having a pH adjusted to 4.5 was used.

  The storage conditions of the sample for storage test were 3 months at 5 ° C, 2 weeks at 10 ° C, 1 month and 3 months, 2 weeks and 1 month at 25 ° C, 1 week and 2 weeks at 40 ° C. .

  The measurement results of the total affinity content are shown in FIG.

  Table 8 shows the residual ratio of teriparatide.

  Table 9 also shows the total related content (upper) and teriparatide residual rate (lower) after storage test at 5 ° C for 3 months, 10 ° C for 3 months, and 25 ° C for 1 month. The numerical value of is shown.

  Under the conditions in which the process of preparing the teriparatide aqueous solution preparation for about 5 hours from the preparation of the raw material to filling is performed in an atmosphere of the same ozone concentration, the total related content at the start of the storage test (FIG. 7, Table 9). In) was significantly lower when methionine was added (with methionine) than when methionine was not added (without methionine).

  The total relative content at the start of the storage test reflects the degradation of teriparatide until the vial is filled with the aqueous formulation. It was shown that methionine in the aqueous solution preparation suppresses the degradation of teriparatide by ozone in the atmosphere during the preparation of the aqueous solution preparation and suppresses the formation of related substances.

  Among the related substances of teriparatide in the aqueous solution preparation of each test section during the storage period, the related substance (RRT 0.76) with a relative retention time of 0.76 in HPLC, and the relative retention time in HPLC of 0.98 8 and FIG. 9 show the content ratio of the related substance (RRT0.98) to the raw material teriparatide, respectively.

  From FIG. 8, under the condition that the preparation of teriparatide aqueous solution is prepared in an atmosphere of the same ozone concentration, the content of the related substance RRT0.76 at the start of the storage test (IN in FIG. 8) is the case where methionine is not added (methionine) It was confirmed that it was significantly lower when methionine was blended (with methionine) compared to (no). The related substance RRT0.76 is presumed to be a related substance in which the methionine residue of teriparatide is denatured, and the addition of methionine is presumed to suppress denaturation by ozone of the methionine residue of teriparatide.

<Experiment 8>
As an aqueous formulation, 60.6 μg / mL teriparatide acetate (56.5 μg / mL as teriparatide), 52.1 mg / mL D-mannitol, 2 mg / mL L in 6.8 mM acetate buffer aqueous solution. -An aqueous solution preparation containing methionine and adjusted to pH 4.5 by adding an appropriate amount of hydrochloric acid or sodium hydroxide was used.

  The “nitrogen” test group is a test group using nitrogen gas as an inert gas for bubbling during preparation of an aqueous solution preparation and replacement of the gas phase in the vial.

  The “Argon” test section is a test section using argon gas as an inert gas for bubbling during preparation of an aqueous solution preparation and replacement of the gas phase in the vial.

  The storage conditions of the sample for storage test were 3 months at 5 ° C, 2 weeks at 10 ° C, 1 month and 3 months, 2 weeks and 1 month at 25 ° C, 1 week and 2 weeks at 40 ° C. .

  The measurement results of the total affinity content are shown in FIG.

  Table 10 shows the teriparatide residual ratio.

  It was confirmed that teriparatide can be held equally when nitrogen is used as an inert gas and when argon is used.

Claims (10)

  A pharmaceutical preparation comprising a container, and a liquid pharmaceutical composition containing human PTH (1-34) consisting of the amino acid sequence shown in SEQ ID NO: 1, filled with sugar, and an amino acid in water.   The pharmaceutical preparation according to claim 1, wherein the sugar is at least one selected from mannitol, sorbitol, and purified sucrose.   The pharmaceutical preparation according to claim 1 or 2, wherein the amino acid is methionine.   The pharmaceutical preparation according to claim 3, wherein the liquid pharmaceutical composition comprises the methionine at a concentration of 1 to 10 mg / mL.   The pharmaceutical preparation according to any one of claims 1 to 4, wherein the water is a buffered aqueous solution having a pH of 5.0 or less. A method for producing a pharmaceutical preparation comprising a container, and a liquid pharmaceutical composition containing human PTH (1-34) consisting of the amino acid sequence shown in SEQ ID NO: 1 filled in the container, a sugar and an amino acid in water There,
A preparation step of preparing a liquid pharmaceutical composition comprising adding the human PTH (1-34) to an aqueous solution containing the sugar and the amino acid in water;
Filling the container with the liquid pharmaceutical composition.   The method according to claim 6, wherein the sugar is one or more selected from mannitol, sorbitol, and purified sucrose.   The method according to claim 6 or 7, wherein the amino acid is methionine.   9. The method of claim 8, wherein the aqueous solution comprises the methionine at a concentration of 1-10 mg / mL.   The method according to any one of claims 6 to 9, wherein the water in the aqueous solution is a buffered aqueous solution having a pH of 5.0 or less.

Images