JP6467102B1 - Formulation with reduced N-formylpiperidine content and / or reduced collapse or shrinkage of lyophilized cake - Google Patents

Abstract

The present invention provides a preparation in which collapse or shrinkage of a freeze-dried cake is suppressed.
A freeze-dried preparation containing teriparatide acetate, which is sealed with a rubber stopper and filled in a medical container, comprising at least one excipient of (1) to (4) Freeze-dried formulation with and without additives:
(1) Glycine;
(2) sodium glutamate;
(3) trehalose;
(4) Purified white sugar.
[Selection figure] None

Description

  The present invention has a reduced N-formylpiperidine content and / or suppressed collapse or shrinkage of a lyophilized cake (hereinafter sometimes simply referred to as cake or cake), Relates to the formulation.

N-Formylpiperidine (hereinafter also referred to as NFP) is a substance sometimes referred to as 1-formylpiperidine, and its CAS number is 2591-86-8. . The structural formula is shown below.

  Rubber stoppers are often used for the purpose of enhancing the sealing performance of drugs filled in injection containers such as vials and prefilled syringes (Patent Document 1).

  Drugs filled in injectable containers may be lyophilized at the time of manufacture for the purpose of improving stability, etc., and are used after being dissolved in glucose injection solution or physiological saline at the time of use. (Non-patent documents 2 to 3).

  The drug may contain a nonionic surfactant (Patent Documents 2 to 4). Preparations containing thrombomodulin or teriparatide are also known (Patent Documents 5 to 6, Non-Patent Documents 2 to 3).

  Moreover, the elcatonin formulation in which the shrinkage | contraction of a lyophilized cake is suppressed is known (patent document 7).

JP 2001-87351 A International Publication No. 95/16460 Japanese Patent Laying-Open No. 2015-168654 JP 2006-222663 A International Publication No. 2013/073545 International Publication No. 2012/169435 JP 05-345729 A JP-T-2006-505543

Journal of the Japan Rubber Association Vol. 53, No. 12 (1980), pages 719-727 Terribbon (registered trademark) 56.5 μg package insert for subcutaneous injection (revised in May 2017 (4th edition) 12800 package insert for intravenous infusion of Lycomodulin (registered trademark) (revised April 2016 (8th edition))

  An object of the present invention is a freeze-dried preparation containing teriparatide acetate as an active ingredient, which is sealed with a rubber stopper and filled in a medical container, and the NFP in the preparation is stored when the preparation is stored. It is to provide a freeze-dried preparation in which an increase with time is suppressed, and a method for suppressing an increase in NFP with time.

  Another object of the present invention is a freeze-dried preparation containing thrombomodulin as an active ingredient, which is sealed with a rubber stopper and filled in a medical container, and the NFP in the preparation is stored when the preparation is stored. It is to provide a freeze-dried preparation in which an increase with time is suppressed, and a method for suppressing an increase in NFP with time.

  Another object of the present invention is to provide a method for suppressing an increase in NFP over time in a lyophilized preparation sealed with a rubber stopper and filled in a medical container. is there.

  Another object of the present invention is a method for producing a lyophilized preparation containing 67.9 μg of teriparatide acetate as an active ingredient, which is sealed with a rubber stopper and filled in a medical container. It is intended to provide a method for producing a lyophilized preparation in which an increase in NFP over time in the preparation is suppressed when stored.

  Another object of the present invention is a freeze-dried preparation containing teriparatide acetate as an active ingredient, which is sealed with a rubber stopper and filled in a medical container, and the freeze-dried cake when the preparation is stored It is to provide a freeze-dried preparation in which collapse or shrinkage of (lyo cake) is suppressed, and a production method thereof.

  Another object of the present invention is to provide a quality inspection method for a freeze-dried preparation sealed with a rubber stopper and filled in a medical container. Another object of the present invention is to provide a method for preparing a high-quality chemical solution using a freeze-dried preparation sealed with a rubber stopper and filled in a medical container. Another subject of this invention is providing the method of suppressing NFP in the chemical | medical solution obtained by melt | dissolving the freeze-dried formulation sealed with the rubber stopper and filled in the medical container with a solvent.

  One aspect of the freeze-dried preparation of the present invention is a freeze-dried preparation containing teriparatide acetate as an active ingredient, which is sealed with a rubber stopper and filled in a medical container, and comprises a specific excipient and / or Alternatively, it is a freeze-dried preparation containing additives. One aspect of the production method of the present invention is a method for producing a freeze-dried preparation containing teriparatide acetate as an active ingredient, which is sealed with a rubber stopper and filled in a medical container, It includes a step of incorporating specific excipients and / or additives into the preparation before lyophilization in the production process. In the freeze-dried preparation of this embodiment and the freeze-dried preparation obtained by the production method of this embodiment, an increase in the amount of NFP in the preparation during storage is suppressed over time.

  Another aspect of the freeze-dried preparation of the present invention is a freeze-dried preparation containing thrombomodulin as an active ingredient, which is sealed with a rubber stopper and filled in a medical container, and comprises a specific excipient and / or It is a lyophilized formulation containing an additive. In addition, one aspect of the suppression method of the present invention is to increase N-formylpiperidine over time in a freeze-dried preparation containing thrombomodulin as an active ingredient, which is sealed with a rubber stopper and filled in a medical container. A method of inhibiting, comprising a step of incorporating a specific nonionic surfactant into the preparation before lyophilization treatment in the production process of the preparation. In the freeze-dried preparation of this embodiment and the freeze-dried preparation obtained through the suppression method of this embodiment, the increase in the amount of NFP in the preparation during storage is suppressed over time.

  Another aspect of the lyophilized formulation of the present invention is a lyophilized formulation that is sealed with a rubber stopper that uses a specific material and / or exhibits specific physical properties, and is filled in a medical container. In the freeze-dried preparation of this embodiment, the increase in the amount of NFP in the preparation during storage is suppressed over time.

  Another aspect of the production method of the present invention is a method for producing a freeze-dried preparation containing 67.9 μg of teriparatide acetate as an active ingredient, which is sealed with a rubber stopper and filled in a medical container. And a step of adding sodium chloride to the preparation before lyophilization treatment in the production process of the preparation. In the freeze-dried preparation obtained by the production method of this aspect, the increase in the amount of NFP in the preparation during storage is suppressed over time.

  Another aspect of the freeze-dried preparation of the present invention is a freeze-dried preparation containing teriparatide acetate as an active ingredient, which is sealed with a rubber stopper and filled in a medical container, and includes 1) teriparatide acetate and A lyophilized formulation containing specific excipients and no additives, or 2) teriparatide acetate and specific excipients and specific additives. Another aspect of the production method of the present invention is a method for producing a freeze-dried preparation containing teriparatide acetate as an active ingredient, which is sealed with a rubber stopper and filled in a medical container. Including a step of freeze-drying a liquid medicine containing teriparatide acetate and a specific excipient and not containing an additive, or 2) a liquid medicine containing teriparatide acetate and a specific excipient and a specific additive . In the freeze-dried preparation of this embodiment and the freeze-dried preparation obtained by the production method of this embodiment, collapse or shrinkage of the freeze-dried cake (lyo cake) in the preparation at the time of storage is suppressed.

  One aspect of the quality inspection method of the present invention is a quality inspection method of a freeze-dried preparation sealed with a rubber stopper and filled in a medical container, wherein the presence of NFP in the preparation is confirmed, and / or Quantifying the abundance. Moreover, one aspect of the method for preparing a chemical solution of the present invention includes a step of adding a solvent to a freeze-dried preparation sealed with a rubber stopper and filled in a medical container, and a step of twisting the medical container including. Furthermore, another aspect of the lyophilized preparation of the present invention is a lyophilized preparation sealed with a rubber stopper and containing teriparatide acetate as an active ingredient, comprising a specific excipient, and containing an additive. A lyophilized formulation that does not contain, or a lyophilized formulation that contains certain excipients and certain additives. The chemical solution obtained through the quality inspection method of this aspect, the chemical solution obtained by the chemical preparation method of this aspect, and the chemical liquid obtained from the freeze-dried preparation of this aspect have the NFP content in the chemical solution suppressed. And high quality.

That is, the present invention relates to the following inventions and the like.
[1]
A freeze-dried preparation sealed with a rubber stopper and filled in a medical container, containing teriparatide acetate as an active ingredient, and further containing at least any one of (1) to (4) Lyophilized preparation:
(1) sodium glutamate or a hydrate thereof;
(2) succinic acid;
(3) sorbitol;
(4) Sodium acetate or a hydrate thereof.
[2]
The freeze-dried preparation according to [1] above, comprising 67.9 μg of teriparatide acetate.
[3]
The freeze-dried preparation according to [1] or [2] above, wherein the rubber plug material is butyl rubber.
[4]
A freeze-dried preparation containing thrombomodulin as an active ingredient, which is sealed with a rubber stopper and filled in a medical container, and further containing the ingredient (1):
(1) A nonionic surfactant having an HLB (Hydrophilic-Lipophilic Balance) value of 10.5 or more.
[5]
The freeze-dried preparation according to [4], wherein the component (1) is polyoxyethylene (POE) sorbitan fatty acid ester or POE hydrogenated castor oil.
[6]
The freeze-dried preparation according to [4] or [5] above, wherein the content of the component (1) is 0.1 mg or more.
[7]
The lyophilized preparation according to any one of [4] to [6] above, wherein the thrombomodulin is a peptide represented by any of the following amino acid sequences and has thrombomodulin activity:
(1) the amino acid sequence set forth in SEQ ID NO: 2;
(2) an amino acid sequence in which one or more amino acids of the amino acid sequence of SEQ ID NO: 2 are substituted, deleted, or added;
(3) An amino acid sequence having 80% or more homology with the amino acid sequence set forth in SEQ ID NO: 2.
[8]
The freeze-dried preparation according to any one of [4] to [7], wherein the rubber plug material is butyl rubber.
[9]
The freeze-dried preparation according to any one of [1] to [8], wherein the medical container is a vial.
[10]
A freeze-dried preparation containing teriparatide acetate, sealed with a rubber stopper and filled in a medical container, containing at least one excipient of (1) to (4) and added Freeze-dried preparation without food:
(1) Glycine;
(2) sodium glutamate;
(3) trehalose;
(4) Purified white sugar.
[11]
A freeze-dried preparation containing teriparatide acetate, which is sealed with a rubber stopper and filled in a medical container, containing purified white sugar, and an additive of at least one of (1) to (6) Freeze-dried formulations containing:
(1) histidine, a salt thereof, or a hydrate thereof;
(2) ascorbic acid;
(3) sodium acetate or a hydrate thereof;
(4) sodium bisulfite;
(5) benzalkonium chloride;
(6) Methionine.
[12]
The freeze-dried preparation according to [10] or [11] above, wherein the amount of teriparatide acetate in the freeze-dried preparation is 67.9 μg.
[13]
A method for suppressing an increase in N-formylpiperidine over time in a freeze-dried preparation containing teriparatide acetate as an active ingredient, which is sealed with a rubber stopper and filled in a medical container, (1) A control method comprising a step of incorporating at least any one of the components in (4) into the preparation before lyophilization treatment in the production process of the preparation:
(1) sodium glutamate or a hydrate thereof;
(2) succinic acid;
(3) sorbitol;
(4) Sodium acetate or a hydrate thereof.
[14]
The method for inhibiting according to the above [13], wherein the amount of teriparatide acetate in the freeze-dried preparation is 67.9 μg.
[15]
The suppression method according to [13] or [14], wherein the material of the rubber plug is butyl rubber.
[16]
A method for suppressing an increase in N-formylpiperidine over time in a lyophilized preparation containing 67.9 μg teriparatide acetate as an active ingredient, sealed with a rubber stopper and filled in a medical container, And a method of inhibiting, comprising a step of adding sodium chloride to the preparation before freeze-drying treatment in the preparation process of the preparation.
[17]
The suppression method according to [16], wherein the rubber plug material is butyl rubber.
[18]
A method for suppressing an increase in N-formylpiperidine over time in a freeze-dried preparation containing thrombomodulin as an active ingredient, which is sealed with a rubber stopper and filled in a medical container, the ingredient of (1) Inhibiting method characterized by comprising adding to the preparation before lyophilization treatment in the preparation process of the preparation:
(1) A nonionic surfactant having an HLB (Hydrophilic-Lipophilic Balance) value of 10.5 or more.
[19]
The suppression method according to [18], wherein the component (1) is polyoxyethylene (POE) sorbitan fatty acid ester or POE hydrogenated castor oil.
[20]
The suppression method according to [18] or [19] above, wherein the content of the component (1) is 0.1 mg or more.
[21]
The suppression method according to any one of [18] to [20], wherein the material of the rubber plug is butyl rubber.
[22]
A method for suppressing a time-dependent increase of N-formylpiperidine in a preparation sealed with a rubber stopper and filled in a medical container, wherein the preparation process includes the following steps 1-2. :
Step 1) Seal the preparation with a rubber stopper made of halogenated butyl rubber;
Step 2) The preparation is wound with an aluminum cap.
[23]
A method for suppressing a time-dependent increase of N-formylpiperidine in a preparation sealed with a rubber stopper and filled in a medical container, wherein the preparation process includes the following steps 1-2. :
Step 1) The preparation is sealed with a rubber plug that satisfies the following conditions 1 to 3;
Condition 1) The material of the rubber plug is butyl rubber;
Condition 2) Rubber stopper is laminated;
Condition 3) The water vapor permeability of the rubber plug is less than 0.57 (g / m ² · 24 h);
Step 2) The preparation is wound with an aluminum cap.
[24]
A method for suppressing a time-dependent increase of N-formylpiperidine in a preparation sealed with a rubber stopper and filled in a medical container, wherein the preparation process includes the following steps 1-2. :
Step 1) The preparation is sealed with a rubber plug having a dehydration rate of 33% / h or less according to the following definition;
Definition) The rate of change in the amount of water contained in a rubber plug obtained after a sterilization treatment at 121 ° C. for 30 minutes as a water-containing treatment for a rubber plug, followed by a drying treatment for 1 hour. Water content of rubber plug after treatment / Water content of rubber plug after water treatment × 100 (% / hour);
Step 2) The preparation is wound with an aluminum cap.
[25]
A method for suppressing a time-dependent increase of N-formylpiperidine in a preparation sealed with a rubber stopper and filled in a medical container, wherein the preparation process includes the following steps 1-2. :
Step 1) The preparation is sealed with a rubber plug that satisfies the following conditions 1 and 2:
Condition 1) The material of the rubber stopper is butyl rubber or polyisobutylene;
Condition 2) The density of the rubber plug is less than 1.3 g / cm ³ ;
Step 2) The preparation is wound with an aluminum cap.
[26]
The suppression method according to any one of [13] to [25], wherein the medical container is a vial.
[27]
A method for inspecting the quality of freeze-dried preparations sealed with rubber stoppers and filled in medical containers, confirming the presence of N-formylpiperidine in the preparation and / or quantifying the amount present Inspection method including the process to do.
[28]
A method for preparing a chemical solution including the following steps:
Step 1) A step of adding a solvent to a freeze-dried preparation sealed with a rubber stopper and filled in a medical container;
Step 2) A step of twisting the medical container, the number of times being 1 to 5 times.
[29]
A method for suppressing the N-formylpiperidine content in a chemical obtained by dissolving a lyophilized preparation with a solvent, comprising the following steps:
Step 1) A step of adding a solvent to the freeze-dried preparation sealed with a rubber stopper and filled in a medical container;
Step 2) A step of twisting the medical container, the number of times being 1 to 5 times.
[30]
A method for producing a freeze-dried preparation sealed with a rubber stopper and filled in a medical container containing 67.9 μg teriparatide acetate as an active ingredient, wherein sodium chloride is freeze-dried in the preparation process A production method comprising a step of adding the preparation to the preparation before treatment.
[31]
The method according to [30], wherein the material of the rubber stopper is butyl rubber.
[32]
A method for producing a freeze-dried preparation containing teriparatide acetate, which is sealed with a rubber stopper and filled in a medical container, wherein teriparatide acetate and at least one of (1) to (4) are added. A freeze-dried formulation comprising a step of freeze-drying a drug solution containing a form and containing no additives, wherein collapse or shrinkage of a freeze-dried cake (lyo cake) in the freeze-dried formulation is suppressed Production method:
(1) Glycine;
(2) sodium glutamate;
(3) trehalose;
(4) Purified white sugar.
[33]
A method for producing a freeze-dried preparation containing teriparatide acetate, which is sealed with a rubber stopper and filled in a medical container, wherein teriparatide acetate, purified white sugar, and at least one of (1) to (6) A method for producing a freeze-dried preparation, comprising the step of freeze-drying a drug solution containing the additive of 1, wherein the collapse or shrinkage of the freeze-dried cake (lyo cake) in the freeze-dried preparation is suppressed:
(1) histidine, a salt thereof, or a hydrate thereof;
(2) ascorbic acid;
(3) sodium acetate or a hydrate thereof;
(4) sodium bisulfite;
(5) benzalkonium chloride;
(6) Methionine.

  ADVANTAGE OF THE INVENTION According to this invention, the freeze-dried formulation etc. with which NFP content is reduced and / or the collapse or shrinkage | contraction of a freeze-dried cake are suppressed are provided.

FIG. 1 illustrates the relationship between the polysorbate 80 content (surfactant amount (mg)) contained in a lyophilized formulation and the NFP content (μg / vial) in the formulation after 2 weeks from the production. . FIG. 2 shows the relationship between the dehydration rate (% / hour) of the rubber stopper used for sealing the preparation and the NFP content (μg / vial) in the preparation after storage for 1 week. FIG. 3 illustrates the relationship between the density (g / cm ³ ) of the butyl rubber stopper and polyisobutylene stopper used for sealing the preparation and the NFP content (μg / vial) in the preparation after storage for 1 week. . FIG. 4 illustrates the relationship between the material of the rubber stopper used to seal the formulation and the NFP content (μg / vial) in the formulation after 1 week of storage. FIG. The high performance liquid chromatography result of one sample is shown. The peak observed at an elution time of 4.682 minutes is an NFP-derived peak. FIG. 6 shows the result of photographing a freeze-dried cake of a freeze-dried preparation containing a surfactant with an electron microscope (imaging magnification is 60 times). FIG. 7 shows the result of photographing a freeze-dried cake of a freeze-dried preparation containing no surfactant with an electron microscope (imaging magnification is 60 times). FIG. 8 shows the result of photographing a freeze-dried cake of a freeze-dried preparation containing a surfactant with an electron microscope (imaging magnification is 300 times). The upper row is a photograph of the inside of the cake, and the lower row is a photograph of the cake surface. FIG. 9 is a result of photographing a freeze-dried cake of a freeze-dried preparation containing no surfactant with an electron microscope (imaging magnification is 300 times). The upper row is a photograph of the inside of the cake, and the lower row is a photograph of the cake surface. FIG. 10 is a result of photographing a freeze-dried cake of a freeze-dried preparation containing a surfactant with an electron microscope (imaging magnification is 1000 times). The upper row is a photograph of the inside of the cake, and the lower row is a photograph of the cake surface. FIG. 11 shows the results of photographing a freeze-dried cake of a freeze-dried preparation containing no surfactant with an electron microscope (imaging magnification is 1000 times). The upper row is a photograph of the inside of the cake, and the lower row is a photograph of the cake surface. FIG. 12 shows the relationship between the HLB of a lyophilized preparation containing a non-surfactant and the NFP content (μg / vial) in the preparation after 2 weeks from the production. FIG. 13 shows the results of measuring the NFP content in the preparation after storing the lyophilized preparation (formulations 21 to 31) containing various excipients / additives. The upper number shown in the lower part of the graph is the formulation number, and the lower number is the NFP content (NFP content in one vial; μg). FIG. 14 is a photograph of a freeze-dried preparation taken after storing a freeze-dried preparation (formulations 25 to 28) containing various excipients / additives. Each formulation is sealed with a rubber stopper and filled in a vial container. In the photograph, the preparation marked with “5” is the freeze-dried preparation after storage (prescription 25), and the preparation marked with “6” is the freeze-dried preparation after storage (prescription 26) with “7”. The preparations marked with indicate freeze-dried preparations (prescription 27) after storage, and the preparations marked with “8” indicate freeze-dried preparations (prescription 28) after storage. FIG. 15 shows the results of measuring the NFP content in a preparation after storing a teriparatide lyophilized preparation (formulations 32-46) containing various excipients / additives. The upper number shown in the lower part of the graph is the formulation number, and the lower number is the NFP content (NFP content in one vial; μg). FIG. 16 is a photograph of lyophilized preparations taken after storage of teriparatide lyophilized preparations (formulations 33, 39, 40, 41, 45) containing various excipients and additives. Each formulation is sealed with a rubber stopper and filled in a vial container. In the photograph, the preparation marked with “2” is the freeze-dried preparation after storage (prescription 33), and the preparation marked with “8” is the freeze-dried preparation after storage (prescription 39) with “9”. The preparations marked with are freeze-dried preparations after storage (prescription 40), the preparations marked with “10” are freeze-dried preparations after storage (prescription 41) and the preparations marked with “14” are The freeze-dried preparation (prescription 45) after storage is shown respectively. FIG. 17 is a photograph of a lyophilized preparation taken after storage of teriparatide lyophilized preparation containing various excipients / additives. Each formulation is sealed with a rubber stopper and filled in a vial container. In the photograph, the preparations marked with “1” to “15” indicate freeze-dried preparations (prescriptions 32-46) after storage, respectively.

  Hereinafter, the present invention will be described in detail according to specific embodiments. However, the present invention is not limited to the following embodiments, and can be implemented in any form without departing from the spirit of the present invention.

1. Lyophilized formulation:
(1) Medicinal component (component 1):
The active ingredient according to the present invention is not particularly limited, and can be an ingredient effective for treatment, prevention, diagnosis and the like for mammals including humans. The structure of the medicinal component is not particularly limited, and may be a low molecular compound, peptide, protein, nucleic acid compound, polysaccharide, and the like. The medicinal component is preferably a peptide or protein, more preferably a physiologically active peptide or a physiologically active protein.

  Examples of the bioactive peptide or bioactive protein include thrombomodulin, interleukin, colony stimulating factor, interferon, tumor necrosis factor, insulin, glucagon, parathyroid hormone (PTH), teriparatide or a salt thereof, gastrin, selectin and the like. Among these, thrombomodulin, parathyroid hormone (PTH), and derivatives thereof are preferable as medicinal ingredients.

(1-1) Thrombomodulin:
Thrombomodulin is known as a substance that specifically binds to thrombin and inhibits thrombin's blood clotting activity, and at the same time significantly enhances the protein C activation ability of thrombin, and is known to have a strong blood clotting inhibitory action. (Patent Document 5).

  The thrombomodulin according to the present invention preferably has an action of (1) selectively binding to thrombin, (2) promoting the activation of protein C by thrombin, and (3) prolonging the clotting time by thrombin. More preferably, it has an action and (4) also has an action of suppressing platelet aggregation by thrombin. In addition to the actions (1) to (4), the thrombomodulin according to the present invention more preferably has (5) an anti-inflammatory action. Any one or more actions selected from (1) to (5) may be referred to as thrombomodulin activity.

  The binding action of thrombomodulin with thrombin is described in various known literatures such as Thrombosis and Haemostasis 1993 70 (3): 418-422 and The Journal of Biological Chemistry 1989 264 (9): 4872-4876. It can be confirmed by the method. The action of accelerating the activation of protein C by thrombin is, for example, the action of accelerating the activation of protein C by a test method clearly described in various known literatures including JP-A-64-6219. The amount of activity and its presence can be easily confirmed. In addition, the action of prolonging the coagulation time by thrombin and / or the action of suppressing platelet aggregation by thrombin can be easily confirmed in the same manner. Furthermore, the anti-inflammatory action can also be confirmed by test methods described in various known literatures such as Blood 2008 112: 3361-3670 and The Journal of Clinical Investigation 2005 115 (5): 1267-1274.

  The thrombomodulin in the present embodiment is not particularly limited as long as it has thrombomodulin activity, but is preferably soluble thrombomodulin that is soluble in water in the absence of a surfactant. A preferable example of the solubility of soluble thrombomodulin is 1 mg / mL in water, for example, distilled water for injection (in the absence of a surfactant such as Triton X-100 or polidocanol, usually near neutrality). Or 10 mg / mL or more, preferably 15 mg / mL or more, or 17 mg / mL or more, more preferably 20 mg / mL or more, 25 mg / mL or more, or 30 mg / mL or more. Preferably, 60 mg / mL or more is mentioned, and depending on the case, 80 mg / mL or more, or 100 mg / mL or more is mentioned, respectively. In determining whether or not soluble thrombomodulin could be dissolved, it was clear and clearly recognized when dissolved, for example, directly under a white light source at a brightness of about 1000 lux. It is understood that the absence of such an insoluble material is a simple indicator. Moreover, it can also filter and can confirm the presence or absence of a residue.

  The molecular weight of the thrombomodulin according to the present invention is not limited, but is preferably 100,000 or less, more preferably 90,000 or less, still more preferably 80,000 or less, and particularly preferably 70,000 or less. The molecular weight of thrombomodulin is more preferably 50,000 or more, and particularly preferably 60,000 or more.

  The molecular weight of the soluble thrombomodulin according to the present invention can be easily measured by an ordinary method for measuring the molecular weight of a protein, but is preferably measured by mass spectrometry, and measured by a MALDI-TOF-MS method. It is more preferable. Soluble thrombomodulin can be obtained by culturing transformed cells prepared by transfecting host cells with DNA encoding soluble thrombomodulin, and further subjecting to fractionation treatment by column chromatography or the like. A soluble thrombomodulin having a molecular weight of can be obtained.

  The thrombomodulin according to the present invention includes the amino acid sequence at positions 19 to 132 of SEQ ID NO: 1, which is known as the central site of thrombomodulin activity in human thrombomodulin (hereinafter also referred to as human TRM 19-132). It is preferable. The thrombomodulin according to the present invention is an amino acid sequence in which one or more amino acids are substituted, deleted, or added in human TRM19-132, and has a certain homology (for example, to human TRM19-132). : 50% or more, 70% or more, 80% or more, 90% or more, 95% or more, 98% or more, etc.), and a thrombomodulin variant that exhibits thrombomodulin activity.

The thrombomodulin according to the present invention is a peptide represented by any one of the following amino acid sequences A to C, and may further be a peptide exhibiting thrombomodulin activity.
Sequence A: The amino acid sequence shown by SEQ ID NO: 2.
Sequence B: An amino acid sequence obtained by substituting, deleting, or adding one or more amino acids in the amino acid sequence represented by SEQ ID NO: 2.
Sequence C: an amino acid having a certain homology (eg, 50% or more, 70% or more, 80% or more, 90% or more, 95% or more, 98% or more, etc.) with respect to the amino acid sequence represented by SEQ ID NO: 2 An array.

  The amount of thrombomodulin contained in the lyophilized preparation of the present invention is not particularly limited, but it is preferable to contain 12,800 active units (about 2 mg) of thrombomodulin. The amount of thrombomodulin per administration contained in the freeze-dried preparation of the present invention is not particularly limited. For example, when used for intravenous infusion, the upper limit is preferably 1 mg / kg or less. 5 mg / kg or less, 0.1 mg / kg or less, or 0.08 mg / kg or less is particularly preferable. Further, the lower limit is particularly preferably 0.005 mg / kg or more, 0.01 mg / kg or more, 0.02 mg / kg or more. A preferred example of the amount of thrombomodulin contained in the freeze-dried preparation for intravenous administration according to the present invention is 0.06 mg / kg. When administered to a patient whose body weight exceeds 100 kg, it may be preferable to administer a fixed dose of 6 mg from the viewpoint that the blood volume is not proportional to the body weight and the blood volume relative to the body weight is relatively reduced. .

(1-2) Teriparatide or a salt thereof:
In the present invention, human PTH (1-34) consists of amino acid residues from the 1st to the 34th amino acid sequence as viewed from the N-terminal side in the amino acid sequence of human PTH (1-84), which is a human parathyroid hormone. A peptide represented by a partial amino acid sequence. In the present invention, teriparatide means free human PTH (1-34). Teriparatide can also be in the form of a salt.

  The salt of teriparatide according to the present invention includes any salt formed by teriparatide and one or more volatile organic acids. Examples of the volatile organic acid include trifluoroacetic acid, formic acid, acetic acid and the like. The ratio of the free teriparatide and the volatile organic acid when forming a salt is not particularly limited as long as the salt is formed. Among them, acetic acid is preferable as the volatile organic acid. That is, teriparatide acetate is preferably exemplified as the teriparatide salt according to the present invention.

  The amount of teriparatide or a salt thereof contained in the freeze-dried preparation of the present invention is not particularly limited, but it is preferable to contain 63.3 μg of teriparatide or 67.9 μg of teriparatide acetate. The amount of teriparatide or a salt thereof per administration contained in the freeze-dried preparation of the present invention is not particularly limited, but teriparatide is preferably 56.5 μg.

(2) Other components (component 2):
The lyophilized preparation of the present invention may contain any other components that are pharmacologically or physiologically acceptable in addition to the above-mentioned medicinal components.

  As other components of the present invention, for example, one or two or more kinds of excipients can be mentioned, and if necessary, one or more kinds of solubilizers, stabilizers, antioxidants, Additives such as anti-aggregating agents, tonicity agents, pH adjusting agents, preservatives (preservatives) and the like can be included in the lyophilized preparation of the present invention.

(2-1) Excipients and additives:
The excipient used in the lyophilized preparation of the present invention is not particularly limited, but the increase in NFP over time during the storage process of the lyophilized preparation of the present invention is suppressed by being alone or in combination with an additive. The resulting excipient is preferred.

  The additive used in the freeze-dried preparation of the present invention is not particularly limited, but the increase in NFP over time in the freeze-dried preparation of the present invention can be suppressed by using alone or in combination with excipients and other additives. It is preferred that it be an additive that can be made. Further, the additive used in the lyophilized preparation of the present invention does not adversely affect the quality and safety of the lyophilized preparation of the present invention and / or reacts with excipients and other additives. It is preferable that the additive does not cause / can hardly occur. Unfavorable reactions can include cake collapse or shrinkage that can be observed immediately after manufacture of the lyophilized formulation or after storage thereof. The cake is sometimes referred to as a porous, dried “lyo cake”. When the cake collapses or shrinks, its dissolution (reconstitution) becomes difficult or impossible, and therefore the cake collapse or shrinkage is not preferable.

  The excipients and additives used in the lyophilized preparation of the present invention are not limited to one kind of excipient or additive, and may be two or more kinds of excipients and additives.

  Examples of excipients include sugar alcohols (mannitol, inositol, sorbitol, etc.), monosaccharides (glucose, fructose, etc.), disaccharides (maltose, lactose, sucrose, trehalose, etc.), amino acids (glycine, glutamic acid or salts thereof, asparagine, etc. Acid or a salt thereof, glutamine or a hydrochloride thereof, arginine or a hydrochloride thereof, etc., cellulose (ethyl cellulose, crystalline cellulose, carboxymethyl cellulose, etc.), citric acid or a salt thereof (calcium citrate, disodium citrate, etc.), chloride Sodium, etc. can be mentioned. Here, the amino acid may be D-form or L-form, but L-form is preferred. Here, the sugar alcohol may be D-form or L-form, but D-form is preferred. These may be hydrates.

  As excipients, among others, inositol, glycine, sodium glutamate, maltose or hydrate thereof, mannitol, lactose or hydrate thereof, trehalose or hydrate thereof, arginine hydrochloride, and sucrose (eg, purified sucrose) Glycine, sodium glutamate, trehalose or a hydrate thereof, arginine hydrochloride, and sucrose (eg, purified sucrose) can be further preferably mentioned.

  Examples of additives include amino acids or salts thereof (eg, histidine, cysteine, methionine, or salts thereof), sugar alcohols (eg, mannitol, inositol, sorbitol), citric acid, sodium bicarbonate, ascorbic acid, succinic acid, Sodium acetate, α-thioglycerin, phosphoric acid, sodium dihydrogen phosphate, sodium carbonate, trometamol, sodium dihydrogen phosphate, tartaric acid, sodium thioglycolate, sodium hydrogen sulfite, benzalkonium chloride, surfactant, etc. Can be mentioned. Here, the amino acid may be D-form or L-form, but L-form is preferred. Here, the sugar alcohol may be D-form or L-form, but D-form is preferred. These may be hydrates.

(2-2) Glycine:
When glycine is used as an excipient, it is preferable that one or more polar amino acids are further added to the preparation as an additive. Examples of polar amino acids include histidine, cysteine, or their hydrochlorides or their salts. Can be mentioned. Here, the polar amino acid may be D-form or L-form, but L-form is preferred. In the case of using glycine as an excipient for the purpose of suppressing collapse or shrinkage of a lyo cake during storage, an embodiment containing no additive is also preferred.

(2-3) Glutamic acid or a salt thereof:
When glutamic acid or a salt thereof is used as an excipient, it is preferable to further include one or more components selected from ascorbic acid, succinic acid, and sodium acetate as additives. Or as another aspect, the preparation which uses glutamic acid or its salt as an excipient | filler and does not use an additive can also be mentioned preferably.

(2-4) Maltose or a hydrate thereof:
When maltose or a hydrate thereof is used as an excipient for the purpose of suppressing the collapse or shrinkage of a lyo cake during storage, α-thioglycerin, phosphoric acid and phosphorus are further added as additives. It is preferable to include in the preparation one or more components selected from sodium dihydrogen acid.

(2-5) Mannitol:
In the case of using mannitol as an excipient for the purpose of suppressing collapse or shrinkage of lyo cake during storage, the additive further includes one or two selected from sodium bicarbonate, trometamol and methionine. The above ingredients are preferably included in the preparation. Here, methionine may be D-form or L-form, but L-form is preferred.

(2-6) Lactose or hydrates thereof:
In the case of using lactose or a hydrate thereof as an excipient for the purpose of suppressing collapse or shrinkage of a lyo cake during storage, sodium dihydrogen phosphate, tartaric acid and thiol are further added as additives. It is preferable to include one or more components selected from sodium glycolate in the preparation.

(2-7) Trehalose or a hydrate thereof:
When trehalose or a hydrate thereof is used as an excipient for the purpose of suppressing the increase in NFP over time in the preparation, D-sorbitol and / or sodium bisulfite may be further added to the preparation as an additive. preferable. In the case of using trehalose or a hydrate thereof as an excipient for the purpose of suppressing collapse or shrinkage of the lyo cake during storage, an embodiment containing no additive is preferred.

(2-8) Arginine or its hydrochloride:
When arginine or its hydrochloride is used as an excipient, it is preferable to include benzalkonium or its chloride and / or a surfactant in the preparation. Moreover, when using arginine or its hydrochloride as an excipient | filler in order to suppress collapse or shrinkage | contraction of the lyophilized cake (lyo cake) at the time of a preservation | save, the aspect which does not contain an additive can also be illustrated preferably.

(2-9) Sucrose (eg, purified sucrose):
In the case of using sucrose (eg, purified sucrose) as an excipient for the purpose of suppressing the increase in NFP over time in the preparation, succinic acid, sodium acetate or a hydrate thereof, D-sorbitol, It is preferable that one or more components selected from sodium chloride and sodium chloride are contained in the preparation.

  When using sucrose (eg, purified sucrose) as an excipient for the purpose of suppressing collapse or shrinkage of lyo cake during storage, addition of succinic acid or D-sorbitol as an additive It is not preferable to include a product in the preparation. 1) Histidine, a salt thereof, or a hydrate thereof, 2) Ascorbic acid, 3) Sodium acetate or a hydrate thereof, 4) Sodium bisulfite, 5) Benzalkco It is preferable to include in the preparation one or more additives selected from nium or its chloride and 6) methionine. Here, methionine and histidine may be D-form or L-form, respectively, but L-form is preferred.

  In the case of using sucrose (eg, purified sucrose) as an excipient for the purpose of suppressing collapse or shrinkage of a lyo cake during storage, an embodiment in which no additive is included in the preparation can also be preferably exemplified.

(2-10) Excipients and additives when the medicinal component is thrombomodulin:
When the medicinal ingredient contained in the freeze-dried preparation of the present invention is thrombomodulin, the excipient to be contained in the preparation is not particularly limited, but one or more amino acids are preferable, among which glutamine or a hydrochloride thereof, And / or arginine or its hydrochloride is preferred, and arginine or its hydrochloride is most preferred.

  When the medicinal component contained in the freeze-dried preparation of the present invention is thrombomodulin, the additive to be included in the preparation is not particularly limited, but it is preferable to include a surfactant also from the viewpoint of dissolution aid and anti-aggregation. .

(2-11) Excipients and additives when the medicinal component is teriparatide or a salt thereof:
When the medicinal ingredient contained in the freeze-dried preparation of the present invention is teriparatide or a salt thereof, the excipient to be contained in the preparation is not particularly limited, but glycine, glutamic acid or a salt thereof, trehalose or a hydrate thereof, arginine Or it is preferable that it is 1 or 2 or more types of excipient | fillers chosen from the hydrochloride and its sucrose (example: refined sucrose), Especially, it is glutamic acid or its salt, or sucrose (example: refined sucrose). It is more preferable. A preferred example of glutamic acid or a salt thereof is sodium glutamate.

  When the medicinal component contained in the freeze-dried preparation of the present invention is teriparatide or a salt thereof, the additive to be included in the preparation is not particularly limited, but for the purpose of suppressing the increase in NFP over time in the preparation, succinic acid, It is preferable to include one or more additives selected from sodium acetate hydrate, sorbitol and sodium chloride in the preparation, and the purpose is to suppress collapse or shrinkage of lyo cake during storage 1) histidine, its salt, or hydrate thereof, 2) ascorbic acid, 3) sodium acetate or hydrate thereof, 4) sodium bisulfite, 5) benzalkonium or chloride thereof, 6) It is preferable to include one or more additives selected from methionine in the preparation.

(2-12) Content of excipients and additives:
The content of excipients and additives contained in the freeze-dried preparation of the present invention is not particularly limited, and can be designed according to, for example, the size and physical properties of medicinal ingredients, medical containers, and the like.

  When the medicinal component contained in the freeze-dried preparation of the present invention is teriparatide or a salt thereof, the content of the additive is the mass of the freeze-dried preparation (here, medical containers and packaging such as rubber stoppers, vials and ampoules). 10 times the amount or less, preferably 5.0 times or less, equal times or less, or 0.5 times or less, and more preferably 0.001 times or more. It is preferable that it is 0.005 times or more or 0.01 times or more.

  When the medicinal ingredient contained in the freeze-dried preparation of the present invention is teriparatide or a salt thereof, the content of the additive is 1000 times or less with respect to the mass of the medicinal ingredient (component 1) contained in the freeze-dried preparation. Among them, it is preferably 100 times or less, preferably 50 times or less, or 10 times or less, more preferably 0.1 times or more, and particularly preferably 0.5 times or more, or 1 time or more. .

  When the medicinal component contained in the lyophilized preparation of the present invention is teriparatide or a salt thereof, the content of the additive can be, for example, 100 mg or less, and among them, it can be 50 mg or less, 10 mg or less, or 5 mg or less. In addition, it may be 0.01 mg or more, and among them, 0.05 mg or more, 0.08 mg or more, or 0.1 mg or more is preferable.

  When the medicinal ingredient contained in the freeze-dried preparation of the present invention is thrombomodulin, the content of the additive is the weight of the freeze-dried preparation (here, the weight of medical containers such as rubber stoppers, vials and ampoules, and packaging) And the like, and in particular, it may be 0.5 times or less, 0.1 times or less, 0.05 times or less, 0.01 times or less, or 0.005 times or less. Preferably, it is 0.0001 times or more, 0.0005 times or more, 0.001 times or more, or 0.002 times or more. The ratio of the mass of the lyophilized formulation of the present invention to the mass of the surfactant amount contained in the lyophilized formulation is preferably, for example, 1: 0.001 to 0.05.

  When the medicinal ingredient contained in the freeze-dried preparation of the present invention is thrombomodulin, the content of the additive is 0.5 times or less with respect to the mass of the medicinal ingredient (component 1) contained in the freeze-dried preparation. In particular, it is preferably 0.1 times or less, 0.05 times or less, or 0.02 times or less, and 0.001 times or more, 0.005 times or more, or 0.01 times or more It is preferable that The ratio of the mass of the medicinal component (component 1) of the present invention to the mass of the surfactant is preferably 1: 0.01 to 0.2, for example.

  When the medicinal component contained in the freeze-dried preparation of the present invention is teriparatide or a salt thereof, the content of the excipient can be 1 mg or more, and preferably 5 mg or more, or 10 mg or more, It may be 100 mg or less, and among them, 50 mg or less, or 20 mg or less is preferable.

  When the medicinal component contained in the freeze-dried preparation of the present invention is thrombomodulin, the content of the excipient can be 1 mg or more, preferably 10 mg or more, or 20 mg or more, and preferably 100 mg or less. Among them, it is preferably 80 mg or less or 50 mg or less.

(2-13) Surfactant:
Surfactants can be broadly classified into ionic surfactants and nonionic surfactants, and ionic surfactants are classified into anionic surfactants, cationic surfactants, and amphoteric surfactants. obtain. Preferred examples of the surfactant to be included in the lyophilized preparation of the present invention include an anionic surfactant and a nonionic surfactant.

  As a nonionic surfactant, a polyoxyethylene (hereinafter also referred to as POE) -polyoxypropylene (hereinafter also referred to as POP) block copolymer, a POE-POP block copolymer adduct, Examples include POE sorbitan fatty acid ester, POE hydrogenated castor oil, POE castor oil, POE alkyl ether, POE / POP alkyl ether, POE alkyl phenyl ether, polyethylene glycol monostearate, glycerin fatty acid ester, sorbitan fatty acid ester and the like.

  Examples of the anionic surfactant include alkyl allyl sulfonate, alkyl sulfate, POE alkyl ether phosphate, fatty acid or salt thereof, and sodium alkylbenzene sulfonate, sodium lauryl sulfate, POE cetyl ether phosphate, respectively. Preferred examples include sodium and ursodeoxycholic acid.

  As the nonionic surfactant, POE sorbitan fatty acid ester is preferable. Among them, monolauric acid POE (20) sorbitan (polysorbate 20), POE sorbitan monostearate (polysorbate 60), POE sorbitan tristearate (polysorbate 65), Monooleic acid POE (20) sorbitan (polysorbate 80) is suitable. Polysorbate 80 is a substance described in the Japanese Pharmacopoeia, and polysorbates 20, 40, 60, and 65 are substances described in pharmaceutical additive standards, and these are all widely used as pharmaceutical additives.

Further, as the nonionic surfactant, POE hydrogenated castor oil is preferable. Among them, POE (5) hydrogenated castor oil, POE (10) hydrogenated castor oil, POE (20) hydrogenated castor oil (for example, NIKKOL HCO-20 ^{(registered) trademark)),} POE (40) hydrogenated castor oil (e.g. NIKKOL HCO-40 ^{(registered trademark),} etc.), POE (50) hydrogenated castor oil (e.g. NIKKOL HCO-50 ^{(registered trademark),} etc.), POE (60) hardened Castor oil (for example, NIKKOL HCO-60 ^{(registered trademark),} etc.) and POE (100) hydrogenated castor oil are suitable.

Alternatively, a nonionic surfactant having an HLB (Hydrophilic-Lipophilic Balance) value of 10.5 or more is suitable according to the following Griffin method formula, and among them, nonionic surfactants having 11 or more or 14 or more are preferred. A surfactant is preferred. Although the upper limit of HLB will not be specifically limited if it is 20 or less, Among these, the nonionic surfactant which is 19 or less, 18 or less, or 17 or less is preferable.

Further, an anionic surfactant having an HLB value of 5 or more according to the following Davis method formula is suitable, and among them, an anionic interface having 10 or more, 20 or more, 30 or more, or 35 or more. Activators are preferred. The upper limit of HLB is not particularly limited as long as it is 40 or less.
(Formula for determining the balance between hydrophilicity and lipophilicity based on 40 sodium dodecyl sulfate and 1 oleic acid)

  The amount of the surfactant contained in the freeze-dried preparation of the present invention is not particularly limited, but relative to the weight of the freeze-dried preparation (excluding the weight of medical containers such as rubber stoppers, vials and ampoules, and packaging). Or less, preferably 0.5 times or less, 0.1 times or less, 0.05 times or less, 0.01 times or less, or 0.005 times or less, It is preferably 0.0001 times or more, 0.0005 times or more, 0.001 times or more, or 0.002 times or more. The ratio of the mass of the lyophilized formulation of the present invention to the mass of the surfactant amount contained in the lyophilized formulation is preferably, for example, 1: 0.001 to 0.05.

  The amount of the surfactant contained in the freeze-dried preparation of the present invention is not particularly limited, but may be 0.5 times or less with respect to the mass of the medicinal ingredient (component 1) contained in the freeze-dried preparation, It is preferably 0.1 times or less, 0.05 times or less, or 0.02 times or less, and 0.001 times or more, 0.005 times or more, or 0.01 times or more. preferable. The ratio of the mass of the medicinal component (component 1) of the present invention to the mass of the surfactant is preferably 1: 0.01 to 0.2, for example.

  The amount of the surfactant contained in the freeze-dried preparation of the present invention is not particularly limited. For example, 0.003 mg or more, 0.01 mg or more, 0.03 mg or more, 0.1 mg or more, 0.11 mg or more, 0.12 mg As mentioned above, it can be 0.13 mg or more, or 1.0 mg or more. Among them, 0.1 mg or more is preferable, and it can be 0.1 to 1.0 mg.

  The method of including the surfactant in the freeze-dried preparation of the present invention is not particularly limited, but an embodiment in which the surfactant is also dissolved in advance in a solution containing Component 1 and the like subjected to the freeze-drying treatment is preferable. That is, it is preferable that the surfactant is contained in the solution together with other components, and the solution is freeze-dried to produce the freeze-dried preparation of the present invention.

(3) Rubber stopper:
The freeze-dried preparation of the present invention is preferably sealed with a rubber stopper. Here, sealing is easily understood by those skilled in the art, and means that the mouth of a container filled with a lyophilized preparation is sealed (see Patent Document 6).

  The material of the rubber plug according to the present invention is not particularly limited. For example, butyl rubber such as butyl rubber, chlorinated butyl rubber, brominated butyl rubber, divinylbenzene terpolymer butyl rubber, polyisoprene rubber (high to low cis 1,4 Bond), polybutadiene rubber (high to low cis 1,4 bond), conjugated diene rubbers such as styrene-butadiene copolymer rubber, and ethylene-propylene-diene terpolymer rubber (EPDM).

  The rubber stopper used in the freeze-dried preparation of the present invention has no observed increase in NFP over time during storage of the preparation, or the observed increase in NFP over time when other rubber stoppers are used. It is preferable that the rubber plug is suppressed as compared with the increase.

  Here, as the rubber plug according to the present invention, a rubber plug whose material is halogenated butyl rubber can be preferably exemplified, and among them, chlorinated butyl rubber and brominated butyl rubber are more suitable. In a freeze-dried preparation sealed with such a rubber stopper, an increase in the amount of NFP in the preparation over time is suppressed.

  Further, the rubber plug according to the present invention can be a rubber plug having a low dehydration rate. For example, a rubber plug having a low dehydration rate and made of butyl rubber is preferable. The dehydration rate of the rubber plug can be determined, for example, by placing the rubber plug to be tested in a high humidity or high humidity / high temperature environment for a predetermined period of time, followed by a drying process for a predetermined time. The change rate of the amount of water contained in the rubber plug can be measured by a method known per se. In a freeze-dried preparation sealed with such a rubber stopper, an increase in the amount of NFP in the preparation over time is suppressed.

  For example, the moisture treatment is sterilized at 121 ° C. for 30 minutes, and after drying for 1 hour, the rate of change in the amount of water contained in the rubber plug (the moisture content of the rubber plug after the drying treatment / after the sterilization treatment) The water content of the rubber plug × 100 (% / hour)) can be measured by the Karl Fischer titration method as the dehydration rate.

  More specifically, the water content immediately after sterilization treatment at high humidity or high humidity / high temperature is 5.32 mg / rubber plug, and the water content after 1 hour of drying treatment is 1.92 mg / rubber plug. In this case, the dehydration rate of the single rubber stopper is 1.92 / 5.32 × 100 (% / hour), that is, about 36 (% / hour). Alternatively, if the moisture content immediately after sterilization treatment at high humidity or high humidity / high temperature is 2.54 mg / rubber plug, and the moisture content after 1 hour of drying treatment is 0.83 mg / rubber plug, The dehydration rate of one rubber plug is 0.83 / 2.54 × 100 (% / hour), that is, about 33 (% / hour).

  When water content is sterilized at 121 ° C. for 30 minutes and dried for 1 hour, the rate of change in the amount of water contained in the rubber plug is calculated. The dehydration rate of the rubber plug is 33 (% / hour). Among them, it is preferably 32 (% / hour) or less, 31 (% / hour) or less, or 30 (% / hour) or less.

The rubber plug according to the present invention is preferably a rubber plug made of butyl rubber or polyisobutylene having a low density. The density can be measured by, for example, the JISK6268 test method (a vulcanized rubber density measurement method defined by Japanese Industrial Standards). The density can be less than 1.3 (g / cm ³ ), among them 1.2 (g / cm ³ ) or less, 1.1 (g / cm ³ ) or less, or 1.0 (g / cm ³ ). cm ³ ) or less is preferable. The lower limit of the density is not particularly limited, but can be, for example, 0.5 (g / cm ³ ) or more, and more preferably 0.7 (g / cm ³ ) or 0.9 (g / cm ^3). ) Or more. In a freeze-dried preparation sealed with such a rubber stopper, an increase in the amount of NFP in the preparation over time is suppressed.

Alternatively, the rubber plug according to the present invention may be a rubber plug having a low water vapor permeability, made of butyl rubber, and having a top surface laminated (for example, with a material such as polytetrafluoroethylene (PTFE)). The water vapor permeability can be measured by, for example, a JISZ0208 test method (commonly referred to as a cup method; Non-Patent Document 1). The water vapor permeability can be less than 0.57 (g / m ² · 24 h), among them, 0.46 (g / m ² · 24 h) or less, 0.4 or less, 0.33 (g / m ² · 24h) or less, 0.3 (g / m ² · 24h) or less, 0.1 (g / m ² · 24h) or less, 0.05 (g / m ² · 24h) or less, or 0.04 It is preferably (g / m ² · 24h) or less. In a freeze-dried preparation sealed with such a rubber stopper, an increase in the amount of NFP in the preparation over time is suppressed.

  The lyophilized preparation of the present invention filled with a medical container, in which the rubber plug material according to the present invention is butyl rubber, suppresses the increase in NFP in the preparation over time when the preparation is stored. It is preferable to contain one or two or more excipients and / or additives. In particular, when the material is butyl rubber and the rubber plug is a rubber plug that satisfies at least one of the following conditions 1 to 3, the use of the excipients and / or additives is more preferable.

Condition 1: not laminated, or the water vapor permeability of the rubber plug is 0.57 (g / m ² · 24 h) or more.
・ Condition 2: Dehydration rate of rubber plug exceeds 33% / h (however, the dehydration rate is 1 hour when sterilization is performed for water plug at 121 ° C. for 30 minutes) The rate of change in the amount of water contained in the rubber plug obtained after performing the drying treatment of: water content of the rubber plug after the drying treatment / water content of the rubber plug after the water treatment × 100 (% / hour) ).
Condition 3: The density of the rubber plug is 1.3 g / cm ³ or more.

(4) Medical container:
The freeze-dried preparation of the present invention is preferably filled in a medical container. The medical container is not particularly limited, but an infusion bag, a syringe, a vial, an ampoule, and the like are preferable, and among these, a vial is preferable. The material of the medical container is not particularly limited, and may be made of glass or plastic, for example.

  When the lyophilized preparation of the present invention is used for human treatment, prevention or diagnosis, the lyophilized preparation of the present invention is preferably sterilized. The sterilization method is not particularly limited, and examples thereof include a hot water immersion sterilization method, a hot water shower sterilization method, and a high-pressure steam sterilization (autoclave) method. If desired, the lyophilized preparation of the present invention (specifically, a chemical solution corresponding to the lyophilized preparation before being lyophilized) can be filtered with a dustproof filter or the like when filling a medical container.

  When the medical container is a vial, the medicinal solution can be dispensed into the vial, and the opening can be sealed with a rubber stopper after lyophilization, and additionally, sealed with an aluminum cap or the like, The lyophilized formulation of the present invention can be manufactured.

(5) NFP content:
The freeze-dried preparation of the present invention is preferably a preparation in which the NFP content is reduced at the time of production, over the storage period, or at the time of use after storage.

  The amount of NFP contained in the freeze-dried preparation of the present invention is not particularly limited, but the amount of NFP contained in the freeze-dried preparation of the present invention administered per day is reduced to be, for example, 1 to 10 μg. Is preferred.

The amount of NFP contained in the freeze-dried preparation of the present invention is not particularly limited, but relative to the mass of the freeze-dried preparation of the present invention (excluding the weight of medical containers such as rubber stoppers, vials and ampoules, and packaging). 2 × 10 ⁻³ % or less, preferably 1 × 10 ⁻³ % or less, 5 × 10 ⁻⁴ % or less, 3 × 10 ⁻⁴ % or less, or 2 × 10 ⁻⁴ % or less. .

The amount of NFP contained in the freeze-dried preparation of the present invention is not particularly limited, but may be 2 × 10 ⁻² % or less with respect to the mass of the medicinal ingredient (component 1) contained in the freeze-dried preparation of the present invention. × 10 ⁻² % or less, 8 × 10 ⁻³ % or less, 5 × 10 ⁻³ % or less, 3 × 10 ⁻³ % or less, or 2 × 10 ⁻³ % or less are preferable.

  The amount of NFP contained in the freeze-dried preparation of the present invention is not particularly limited, but may be, for example, 20 μg or less. Among them, 10 μg or less, 8 μg or less, 5 μg or less, 1 μg or less, 0.5 μg or less, 0.2 μg or less, 0. It is preferably 15 μg or less, 0.12 μg or less, 0.1 μg or less, 0.08 μg or less, 0.07 μg or less, 0.05 μg or less, 0.02 μg or less, or 0.01 μg or less. Of course, an embodiment in which NFP is not included at all in the freeze-dried preparation of the present invention is most preferable.

  NFP in the preparation can be detected or quantified by high performance liquid chromatography, mass spectrometry or the like.

(6) Suppression of collapse or shrinkage of freeze-dried cake:
The freeze-dried preparation of the present invention is preferably a preparation in which collapse or shrinkage of the freeze-dried cake is suppressed during its production, its storage period, or its use after storage.

  About the collapse or shrinkage | contraction of a freeze-dried cake, the presence or absence and a grade can be easily judged by observing the external appearance of a freeze-dried cake.

  In order to suppress collapse or shrinkage of the freeze-dried cake, it is preferable to appropriately select excipients and additives used in the freeze-dried preparation of the present invention.

  When trehalose or a hydrate thereof is used as an excipient, an embodiment in which sorbitol or sodium bisulfite is not used as an additive is preferable. When the medicinal ingredient is teriparatide or a salt thereof, no additive is contained. Embodiments are more preferred.

  When sodium bisulfite is used as an additive, it is preferable not to use trehalose or a hydrate thereof as an excipient. When the medicinal ingredient is teriparatide or a salt thereof, sucrose (for example: An embodiment using purified sucrose) is more preferred.

  When succinic acid is used as an additive, an embodiment in which sucrose (eg, purified sucrose) is not used as an excipient is preferable, and an embodiment in which sodium glutamate is used as an excipient is more preferable. When succinic acid is used as an additive and sodium glutamate is used as an excipient, an embodiment in which ascorbic acid or sodium acetate hydrate is further added to the preparation as an additive is more preferable.

  When sucrose (eg, purified sucrose) is used as an excipient, an embodiment in which cysteine, a salt thereof, a hydrate thereof, succinic acid and sorbitol are not used as an additive is preferable 1) histidine, a salt thereof, Or a hydrate thereof, 2) ascorbic acid, 3) sodium acetate or a hydrate thereof, 4) sodium bisulfite, 5) benzalkonium, and 6) one or more additives selected from methionine are contained in the preparation. Preferably.

  In Patent Document 7, it is reported that shrinkage of a freeze-dried cake was not observed in almost all preparations using saccharides such as sucrose and mannitol. On the other hand, in the lyophilized preparation of the present application, when sucrose or trehalose (or a hydrate thereof) is used as an excipient, a specific additive can be selected from the viewpoint of suppressing collapse or shrinkage of the lyophilized cake or The use of no additives is preferred.

  Patent Document 8 discloses that the freeze-dried cake contracts by replacing sodium chloride in the preparation with potassium chloride, which is another salt. On the other hand, in the freeze-dried preparation of the present application, when a salt such as histidine hydrochloride, sodium acetate, sodium hydrogen sulfite and benzalkonium chloride is used, shrinkage of the freeze-dried cake is observed in visual appearance. There wasn't.

  When additives are added to excipients such as sucrose and trehalose (or hydrates thereof) for the purpose of suppressing collapse or shrinkage of the lyophilized cake, sucrose or trehalose (or hydrates thereof) It is preferable that the amount ratio of the excipient and the additive such as (product) is 1: 0.005 to 0.2. That is, it is preferable to add about 0.5 to 20% of an additive such as sucrose or trehalose (or a hydrate thereof) to the preparation. Moreover, it is preferable that the mass of the additive in a formulation shall be 0.05-2.0 mg.

  In Patent Document 7, when the content of sodium chloride in the preparation is 25% or more by weight with respect to the saccharide in the preparation, shrinkage is observed in the cake after lyophilization. On the other hand, in the freeze-dried preparation of the present application, shrinkage of the cake is observed even when the succinic acid additive is about 5% by mass with respect to the excipient sucrose. Therefore, even when the mass ratio of the excipient amount and the additive is established, when sucrose or trehalose (or its hydrate) is used as an excipient, the collapse or shrinkage of the lyophilized cake is suppressed. From this point of view, the selection of specific additives is preferred.

(7) Use:
The freeze-dried preparation of the present invention is a freeze-dried preparation that is used after being dissolved in water for injection, glucose injection solution, physiological saline or the like before use (treatment, prevention, diagnosis for mammals including humans). Can do.

  When a dissolved lyophilized preparation (medical solution) is administered to a human, it can be administered parenterally, for example, intravenous administration, intramuscular administration, or subcutaneous administration. In the case of intravenous administration, it is possible to administer a desired amount at once (rapid intravenous administration) or intravenous drip administration.

2. A method for suppressing an increase in the amount of NFP over time:
In one embodiment of the freeze-dried preparation of the present invention, the rubber plug material used for sealing the preparation is a halogenated butyl rubber. In one embodiment of the freeze-dried preparation of the present invention, the dehydration rate of the rubber stopper used for sealing the preparation is below a certain threshold value. In one embodiment of the freeze-dried preparation of the present invention, the density of the rubber stopper used for sealing the preparation is not more than a certain threshold value, and the material thereof is butyl rubber or polyisobutylene. In one aspect of the freeze-dried preparation of the present invention, the water vapor permeability of the rubber stopper used for sealing the preparation is below a certain threshold value, and the rubber stopper is laminated (top surface, foot, or both). The material is butyl rubber.

  In these lyophilized preparations, an increase in the amount of NFP in the preparation over time can be suppressed.

  In one embodiment of the freeze-dried preparation of the present invention, the specific excipient and additive exemplified above are contained. In these lyophilized preparations, an increase in the amount of NFP in the preparation over time can be suppressed.

  Here, the suppression of the increase over time refers to the time-dependent increase in the NFP content over time compared to the increase over time in the NFP content in the lyophilized preparation not using the rubber plug or the specific excipient or additive. This means that the NFP content does not change with time, or the NFP content decreases with time. Here, the time scale is not particularly limited, but may be, for example, suppression of increase over time when stored for several days to several months at room temperature to about 80 ° C.

  For the purpose of suppressing the increase in NFP content over time in the freeze-dried preparation of the present invention, a method for incorporating a specific excipient or additive into the preparation is not particularly limited, but it is one of the manufacturing steps of the preparation. (E.g., preparing an aqueous solution containing medicinal ingredients, excipients, and additives, pre-treating the aqueous solution with a pretreatment such as filtration, and then adding the aqueous solution to a medical container) Filling).

  The presence and / or NFP content of NFP in the lyophilized preparation can be confirmed and quantified by a method known per se such as high performance liquid chromatography. The conditions for high performance liquid chromatography are, for example, as follows.

(1) Detector: UV absorptiometer (measurement wavelength: 214 nm).
(2) Column: A stainless steel tube having an inner diameter of 4.6 mm and a length of 150 mm is filled with 5 μm of octadecylsilylated silica gel for liquid chromatography (YMC-Pack ODS-AM manufactured by YMC Corporation or equivalent).
(3) Column temperature: a constant temperature around 40 ° C.
(4) Mobile phase:
Mobile phase A: Prepared by adding 1 mL of acetic acid (100) to 1000 mL of water.
Mobile phase B: Prepared by adding 1 mL of acetic acid (100) to 1000 mL of acetonitrile.
(5) Mobile phase feeding:
The mixing ratio of mobile phase A and mobile phase B is set to 81:19.
(6) Flow rate: Adjust so that the NFP retention time is about 4.5 minutes.
(7) Detection time: About twice the retention time of NFP.

  A sample to be donated to high-performance liquid chromatography can be prepared by dissolving a lyophilized preparation with an appropriate solvent, but this preparation method is not particularly limited. For example, a solution obtained by dissolving a lyophilized preparation with physiological saline can be used as a sample to be provided to high performance liquid chromatography.

  The mechanism of the increase in NFP content over time in lyophilized formulations is not clear. However, the inventor gradually transfers NFP from the rubber stopper used for the sealing to the surface of the preparation or the inside thereof (eg, the inside of the cake, etc.) during the period of storage or standing of the freeze-dried preparation of the present invention. This takes into account the possibility that the NFP content in the formulation will increase over time.

  That is, a rubber plug made of halogenated butyl rubber, a rubber plug whose dehydration rate is below a certain threshold, or a rubber plug whose density is less than a certain threshold and whose material is butyl rubber or polyisobutylene, Alternatively, the freeze-dried preparation in which the water vapor permeability of the rubber plug is equal to or less than a certain threshold value, laminated, and sealed with a rubber plug whose material is butyl rubber is different in physical properties from these rubber plugs, for example. Compared to a freeze-dried preparation sealed with a rubber stopper made of butyl rubber, NFP that migrates from the rubber stopper to the freeze-dried preparation can be suppressed over a period in which the preparation is stored or left standing. The inventor thinks.

  Although the mechanism by which the NFP content in the freeze-dried preparation is suppressed by adding a specific excipient or additive to the freeze-dried preparation is not clear, one theory is that some interaction ( The inventor considers the possibility of (eg: chemical interaction inside the freeze-dried cake) or the mechanism of collapse or shrinkage of the freeze-dried cake.

3. Storage method of lyophilized formulation:
As one aspect of the present invention, there is provided a storage method for a freeze-dried preparation sealed with a rubber stopper, in which an increase in the amount of NFP in the freeze-dried preparation over time is suppressed.

  As described above, the suppression of the increase in the amount of NFP over time can be achieved by using a rubber plug used for sealing or a surfactant incorporated in a freeze-dried preparation with a specific rubber plug (eg, rubber plug with specific physical properties, specific material) It is possible to carry out by using a rubber plug manufactured in (1) or a surfactant.

  The storage conditions are not particularly limited and can be stored at room temperature or under refrigeration, but refrigeration is preferred. The storage period is not particularly limited, and examples include a storage period of 1 month, 3 months, 6 months, 12 months, 18 months, or 24 months.

4). Quality inspection method for freeze-dried preparations:
It is known that the quality of an injection is tested from the viewpoint of the size and number of insoluble foreign substances and fine particles (Patent Document 1).

  As one aspect of the present invention, there is provided a method for inspecting the quality of the lyophilized preparation of the present invention, which comprises the steps of confirming the presence of NFP in the preparation and / or quantifying the abundance. The

  The presence of NFP can be confirmed by donating a sample derived from the lyophilized preparation of the present invention to high performance liquid chromatography. The quality inspection method of the present invention can be usefully used for the purpose of inspecting the presence or absence and degree of quality deterioration of a preparation when the freeze-dried preparation is stored for a long period of time.

5). Preparation method of chemicals:
As one embodiment of the present invention, a method for preparing a chemical solution including the following steps is provided.
1) A step of adding a solvent to the freeze-dried preparation of the present invention (where the freeze-dried preparation is filled in a medical container).
2) A step of twisting a medical container filled with a freeze-dried preparation, the number of times being 2 to 4 times.

  The solvent used here is not particularly limited, and may be water for injection or physiological saline, for example. Further, here, the medical container is not particularly limited, and an infusion bag, a syringe, a vial, an ampoule and the like can be suitably used.

  The specific mode of torsion is not particularly limited, and for example, the medical container can be grasped by hand, the elbow is substantially fixed, and the wrist can be rotated while twisting clockwise or counterclockwise. When the wrist is turned clockwise or counterclockwise, it is forced to turn over due to restrictions on the physical structure of the upper arm or wrist, but the turnover number is about 1 to 5 times, especially about 2 to 4 times. preferable. The speed of torsion is not particularly limited, but it is preferable that the torsion is performed so as to reverse about once per second.

  The lyophilized preparation used in the preparation method of the present invention may be a preparation containing no surfactant.

6). Method for producing a lyophilized formulation comprising teriparatide acetate, wherein collapse or shrinkage of the lyo cake is suppressed:
As one aspect of the present invention, a method for producing a freeze-dried preparation containing teriparatide acetate, which is sealed with a rubber stopper and filled in a medical container, wherein teriparatide acetate, purified sucrose, and (1) to The manufacturing method including the process of freeze-drying the chemical | medical solution containing at least any one additive among (6) is provided.
(1) Histidine, a salt thereof, or a hydrate thereof.
(2) Ascorbic acid.
(3) Sodium acetate or a hydrate thereof.
(4) Sodium bisulfite.
(5) Benzalkonium chloride.
(6) Methionine.

One aspect of the present invention is a method for producing a lyophilized preparation containing teriparatide acetate, which is sealed with a rubber stopper and filled in a medical container, wherein teriparatide acetate and (1) to (4) There is provided a production method comprising a step of freeze-drying a chemical solution containing at least one of the excipients and not containing an additive.
(1) Glycine.
(2) Sodium glutamate.
(3) Trehalose.
(4) Purified white sugar.

  In these manufacturing methods, for example, the above-described chemical solution is prepared, the medical solution is filled into a medical container, the filled chemical solution half-plugged with a rubber stopper is freeze-dried with a freeze dryer, and after freeze-drying is completed After re-pressure with nitrogen, the cap and the aluminum cap can be tightened.

  Freeze-drying can pass through three processes, for example, freezing, primary drying, and secondary drying. The freezing process includes a freezing process in which the temperature is lowered over 1 to 3 hours, and the reduced temperature is 1 Freezing steps maintained for up to 3 hours can be formed in combination.

  The rubber stopper to be used is preferably washed and sterilized in advance before use, and is preferably preliminarily dried for the purpose of suppressing collapse or shrinkage of the freeze-dried cake. Although drying conditions are not specifically limited, For example, it is preferable to dry a rubber stopper over 2-10 hours under the temperature conditions of 90-120 degreeC.

  The medical container to be used is preferably washed and sterilized in advance before use, and is preferably dried in advance for the purpose of suppressing collapse or shrinkage of the freeze-dried cake. Although drying conditions are not specifically limited, For example, it is preferable to dry a medical container over 10 to 120 minutes on the temperature conditions of 250-400 degreeC.

  Hereinafter, the present invention will be described in more detail with reference to examples. However, the present invention is not limited to the following examples, and can be implemented in any form without departing from the spirit of the present invention.

  In the following examples, “prescription” and “prescription preparation” may be described as terms corresponding to the “lyophilized preparation” of the present invention.

Example 1 (Production of thrombomodulin):
DNA encoding the amino acid sequence of SEQ ID NO: 2 (specifically, consisting of the base sequence of SEQ ID NO: 3) is transfected into Chinese hamster ovary (CHO) cells, and the culture solution of the transformed cells is purified. The active fraction was collected with 20 mmol / L phosphate buffer (pH 7.3) containing 50 mmol / L sodium chloride (NaCl). The recovered liquid was further concentrated using an ultrafiltration membrane to obtain a 11.0 mg / mL thrombomodulin solution.

Example 2 (Production of freeze-dried preparation and solution):
(1) Preparation of freeze-dried formulations 1 to 4:
For lyophilized preparations 1 to 4, first, each chemical solution was prepared by the following procedure.

  In the production of the lyophilized preparation 1, a 60 mg / mL thrombomodulin aqueous solution prepared in advance, a 1 mg / mL polysorbate 80 aqueous solution, a 250 mg / mL L-arginine hydrochloride aqueous solution adjusted to a pH of about 7 with sodium hydroxide, After adding 5 mL of each, medicinal solution 1 was prepared by measuring up to 50 mL with water for injection.

  In the production of the lyophilized preparation 2, after adding 5 mL each of a 1 mg / mL polysorbate 80 aqueous solution prepared in advance and a 250 mg / mL L-arginine hydrochloride aqueous solution adjusted to pH about 7 with sodium hydroxide, The medicinal solution 2 was prepared by measuring up to 50 mL with water for injection.

  In the production of the freeze-dried preparation 3, 5 mL each of a 60 mg / mL thrombomodulin aqueous solution prepared in advance and a 250 mg / mL L-arginine hydrochloride aqueous solution adjusted to pH 7 with sodium hydroxide were added, followed by 50 mL with water for injection. The medicinal solution 3 was prepared by measuring up.

  In the production of the freeze-dried preparation 4, the medicinal solution 4 was prepared by measuring 5 mL of an aqueous L-arginine hydrochloride solution of 250 mg / mL adjusted to pH 7 with sodium hydroxide prepared in advance to 50 mL with water for injection.

  Each chemical solution 1 to 4 was filtered and sterilized, and then filled into glass vials by 1.2 mL. Thereafter, the rubber stopper was half-capped and freeze-dried in a freeze dryer. After completion of lyophilization, each lyophilized preparation was produced by restoring pressure with nitrogen, sealing the rubber stopper completely, and winding the aluminum cap.

Along with the above production method, freeze-dried preparations 1 to 4 shown in Table 1 below were obtained.

(2) Preparation of freeze-dried formulations 5-10:
Regarding freeze-dried preparations 5 to 10, first, each chemical solution was prepared by the following procedure.

  In the production of freeze-dried preparation 5, 5 mL of 250 mg / mL L-arginine hydrochloride aqueous solution adjusted to pH 7 with sodium hydroxide prepared in advance was added, and then the volume was increased to 50 mL with water for injection. Was prepared.

  In the production of the freeze-dried preparation 6, after adding 15 μL of a 10 mg / mL polysorbate 80 aqueous solution prepared in advance and 5 mL of a 250 mg / mL L-arginine hydrochloride aqueous solution adjusted to a pH of about 7 with sodium hydroxide, respectively, The medicinal solution 6 was prepared by measuring up to 50 mL.

  In the production of the freeze-dried preparation 7, 50 μL of a 10 mg / mL polysorbate 80 aqueous solution prepared in advance and 5 mL of a 250 mg / mL L-arginine hydrochloride aqueous solution adjusted to a pH of about 7 with sodium hydroxide were added respectively. The medicinal solution 7 was prepared by measuring up to 50 mL.

  In the production of the lyophilized preparation 8, 150 μL of a 10 mg / mL polysorbate 80 aqueous solution prepared in advance and 5 mL of a 250 mg / mL L-arginine hydrochloride aqueous solution adjusted to a pH of about 7 with sodium hydroxide were added, respectively. The liquid 8 was prepared by measuring up to 50 mL.

  In the production of the freeze-dried preparation 9, 500 μL of a 10 mg / mL polysorbate 80 aqueous solution prepared in advance and 5 mL of a 250 mg / mL L-arginine hydrochloride aqueous solution adjusted to a pH of about 7 with sodium hydroxide were added, respectively. The medicinal solution 9 was prepared by measuring up to 50 mL.

  In the production of the freeze-dried preparation 10, 5 mL each of 10 mg / mL polysorbate 80 aqueous solution prepared in advance and 250 mg / mL L-arginine hydrochloride aqueous solution adjusted to pH 7 with sodium hydroxide were added, and then water for injection was used. The medicinal solution 10 was prepared by measuring up to 50 mL.

  Each chemical solution 5 to 10 was sterilized by filtration and then filled into glass vials by 1.2 mL. Thereafter, the rubber stopper was half-capped and freeze-dried in a freeze dryer. After completion of lyophilization, each lyophilized preparation was produced by restoring pressure with nitrogen, sealing the rubber stopper completely, and winding the aluminum cap.

Along with the above production method, freeze-dried preparations 5 to 10 described in Table 2 below were obtained.

(3) Preparation of solution formulations 11-13:
The solution preparation 11 was produced by adding 1.2 mL of water for injection to the freeze-dried preparation 1 shown in Table 1 without opening a rubber stopper. The solution formulation 12 was produced by adding 1.2 mL of water for injection with a syringe to the lyophilized formulation 3 in Table 1 without opening the rubber stopper.

Along with the above production method, solution formulations 11 to 12 shown in Table 3 below were obtained.

(4) Preparation of freeze-dried formulations 13-19:
Regarding the freeze-dried preparations 13 to 19, first, each chemical solution was prepared by the following procedure.

  In the production of the lyophilized preparation 13, 5 mL of 250 mg / mL L-arginine hydrochloride aqueous solution whose pH was adjusted to about 7 with sodium hydroxide prepared in advance was added, and then diluted to 50 mL with water for injection. Was prepared.

  In the production of the freeze-dried preparation 14, 5 mL each of 1 mg / mL polysorbate 80 aqueous solution prepared in advance and 250 mg / mL L-arginine hydrochloride aqueous solution adjusted to pH 7 with sodium hydroxide were added, and then water for injection was used. The medicinal solution 14 was prepared by measuring up to 50 mL.

  In the production of the freeze-dried formulation 15, 5 mL each of a 1 mg / mL polysorbate 20 aqueous solution prepared in advance and a 250 mg / mL L-arginine hydrochloride aqueous solution adjusted to a pH of about 7 with sodium hydroxide were added. The medicinal solution 15 was prepared by measuring up to 50 mL.

  In the production of the freeze-dried preparation 16, after adding 5 mL each of 1 mg / mL polysorbate 60 aqueous solution prepared in advance and 250 mg / mL L-arginine hydrochloride aqueous solution adjusted to pH 7 with sodium hydroxide, The medicinal solution 16 was prepared by measuring up to 50 mL.

  In the production of the freeze-dried preparation 17, 5 mL each of 1 mg / mL polysorbate 65 cloudy solution prepared in advance and 250 mg / mL L-arginine hydrochloride aqueous solution adjusted to pH 7 with sodium hydroxide were added, followed by water for injection. The medicinal solution 17 was prepared by measuring up to 50 mL.

In the production of the freeze-dried preparation 18, 250 mg / mL L prepared by adjusting the pH to about 7 with 1 mg / mL POE (60) hydrogenated castor oil (NIKKOL HCO-60 ^{(registered trademark)} ) prepared in advance and sodium hydroxide. -After adding 5 mL each of arginine hydrochloride aqueous solution, the chemical | medical solution 19 was prepared by measuring up to 50 mL with water for injection.

  In the production of the lyophilized preparation 19, 5 mL each of a 1 mg / mL aqueous sodium dodecyl sulfate solution prepared in advance and a 250 mg / mL aqueous L-arginine hydrochloride solution adjusted to a pH of about 7 with sodium hydroxide were added. The medicinal solution 20 was prepared by measuring up to 50 mL.

  Each chemical solution 13-19 was sterilized by filtration and then filled into glass vials by 1.2 mL. Thereafter, the rubber stopper was half-capped and freeze-dried in a freeze dryer. After completion of lyophilization, each lyophilized preparation was produced by restoring pressure with nitrogen, sealing the rubber stopper completely, and winding the aluminum cap.

Along with the above production method, freeze-dried preparations 13 to 19 described in Table 4 below were obtained.

Example 3 (Production of teriparatide lyophilized formulation):
After adding 50 g of purified sucrose and 2.5 g of sodium chloride to about 1.0 kg of water for injection and dissolving, weight adjustment was made to 2.5 kg with water for injection to obtain Preliminary Solution A. After 267 mg of teriparatide acetate was added and dissolved in 250 mL of this preconditioning solution A, the total amount was corrected to 2.2 kg in the preconditioning solution A to obtain a drug solution.

  After this chemical solution was sterilized by filtration, each glass vial was filled with 0.56 g. Thereafter, the rubber stopper was half-capped and freeze-dried in a freeze dryer. After completion of lyophilization, each formulation was produced by restoring pressure with nitrogen, sealing with a rubber stopper all the way, and winding with an aluminum cap.

In accordance with the above manufacturing method, the lyophilized preparation 20 described in Table 5 below was obtained.

Example 4 (Production of lyophilized preparation containing various excipients and additives):
Each lyophilized formulation shown in Table 6 below is prepared by preparing a chemical solution having the composition shown in Table 6 below, filling a glass vial with 0.56 g of the chemical solution, and then half-plugging the rubber stopper to freeze. It was manufactured by lyophilization in a drier. After completion of lyophilization, the pressure was restored with nitrogen, and the rubber plug was fully plugged and the aluminum cap was tightened.

  In addition, all preparation amounts of a chemical | medical solution were 20 mL. At the time of preparation of each chemical solution, the additive weighed amount is a weighed amount as described in Table 6 below, and the excipient weighed amount was 1090.9 mg (formulation 29) or 407.3 mg (formulation 21). 28, 30-31).

Example 5 (Production of teriparatide lyophilized preparation containing various excipients and additives):
Each lyophilized preparation described in the following table is prepared by preparing a chemical solution having the composition described in the following table, filling a glass vial with 0.56 g of the chemical solution, and then half-plugging the rubber stopper. It was manufactured by freeze-drying. After completion of lyophilization, the pressure was restored with nitrogen, and the rubber plug was fully plugged and the aluminum cap was tightened.

  The chemical solutions of formulations 32-34 were prepared by weighing 203.6 mg of each excipient, adding distilled water to 5 mL, and adding 5 mL of teriparatide acetic acid solution and stirring. The teriparatide acetic acid solution here was prepared by dissolving 19.8 mg of teriparatide acetate in 80 mL with distilled water.

  The chemical solutions of formulas 35 to 46 were weighed in the amounts shown in the table below for each additive, 1 mL of purified sucrose solution was added, 5 mL was further added with distilled water, and then 5 mL of teriparatide acetic acid solution was added. Prepared by adding and stirring. The purified sucrose solution was prepared by dissolving 2.646 g of purified sucrose in 13 mL of distilled water.

Example 6 (Search for components that affect NFP content over time):
(1) Test method:
Using the freeze-dried preparations 1 to 4 produced in Example 2 “Manufacture of freeze-dried preparations and solutions”, the components that affect the increase in NFP content over time in each preparation were searched. Specifically, it is as follows.

  The lyophilized preparations 1 to 4 were stored in an 80 ° C. stability tester, sampled over time, and the NFP content was measured by high performance liquid chromatography.

(2) Test results:
The test results are shown in Table 8 below.

Example 7 (Confirmation of effects of components affecting NFP content increase with time):
(1) Test method:
In Example 6 described above, it was suggested that the component that affects the NFP content increase over time in the preparation is a surfactant. Therefore, using the freeze-dried preparations 1 and 3 produced in Example 2 “Manufacture of freeze-dried preparations and liquids”, the NFP content-increase suppressing effect suggested by the surfactant was shown to be a progress after the preparation production. I reconfirmed by changing the period and progress conditions. Specifically, it is as follows.

  The lyophilized preparations 1 and 3 were stored in a stability tester at 60 ° C., sampled over time, and the NFP content was measured by high performance liquid chromatography.

(2) Test results:
The test results are shown in Table 9 below.

Example 8 (Evaluation of the influence of the surfactant amount on the NFP content-time increase suppression effect):
(1) Test method:
In Examples 6 to 7 described above, it was revealed that the component that affects the increase in NFP content with time in the preparation is a surfactant. Therefore, using the lyophilized preparations 5 to 10 produced in Example 2 “Production of lyophilized preparation and solution”, the amount of the surfactant was reduced with respect to the NFP content increase inhibitory effect exhibited by the surfactant. The impact was evaluated. Specifically, it is as follows.

  After storing the freeze-dried preparations 5 to 10 in a stability tester at 60 ° C., sampling was performed over time, and the NFP content was measured by high performance liquid chromatography.

(2) Test results:
The test results are shown in Table 10 below.

The relationship between the polysorbate 80 content (surfactant amount (mg)) in the above table and the NFP content (μg / vial) after 2 weeks is shown in FIG.
Depending on the amount of polysorbate 80 added, the time-dependent increase in NFP in the lyophilized preparation was suppressed, and the addition of about 0.1 mg or more almost did not increase NFP over time.

Example 9 (Evaluation of the influence of the dosage form of the preparation on the inhibitory effect of NFP content over time):
(1) Test method:
In Examples 6 to 7 described above, it was revealed that the component that affects the increase in NFP content with time in the preparation is a surfactant. Therefore, the formulation of the preparation is given to the NFP content-inhibiting effect of the surfactant over time, using the solution preparations 11 to 12 manufactured in the above-described Example 2 “Manufacture of lyophilized preparation and solution”. The impact was evaluated. Specifically, it is as follows.

  The solution preparations 11 to 12 were stored in stability testers at 40 ° C., 60 ° C., and 80 ° C., respectively, sampled with time, and the NFP content was measured by high performance liquid chromatography.

(2) Test results:
The test results are shown in Table 11 below.

Example 10 (Evaluation of effect of rubber plug type on NFP content over time):
(1) Test method:
The effect of rubber plug type on the NFP content increase over time in the preparation was evaluated. The rubber plugs used in the test are as shown in Table 12 below.

  1 mL of water for injection was added to a glass vial, each was sealed with various rubber stoppers described in Table 13, and wound with an aluminum cap. After storing these samples in a stability tester at 80 ° C., sampling was performed one week later, and the NFP content was measured by high performance liquid chromatography.

(2) Test results:
The test results are shown in Table 13 below.

The rubber plug No. in Tables 12-13 above. 1 to 4 and 7, the relationship between the rubber plug dehydration rate (% / hour) and the NFP content (μg / vial) is shown in FIG. 1 to 3 and 7, the relationship between the rubber plug density (g / cm ³ ) and the NFP content (μg / vial) is shown in FIG. Regarding 1 to 6, the relationship between the rubber plug material and the NFP content (μg / vial) is shown in FIG. In FIG. The high performance liquid chromatography result of one sample is shown.

Example 11 (Evaluation of the effect of post-manufacture rubber plug replacement on the NFP content increase suppression effect over time):
(1) Test method:
In Examples 6 to 8 described above, it was revealed that the component that affects the transition with time of NFP contained in the preparation is a surfactant. Therefore, using the freeze-dried preparations 1 and 3 produced in Example 2 “Manufacture of freeze-dried preparations and solutions”, the rubber after the preparation was manufactured to suppress the NFP content increase over time exhibited by the surfactant. The effect of plug replacement was evaluated. Specifically, it is as follows.

  Remove the aluminum cap and rubber stopper of lyophilized formulations 1 and 3. Immediately thereafter, another rubber plug was completely plugged and wound with an aluminum cap. The lyophilized preparations 1 and 3 with the rubber plugs replaced were stored in a stability tester at 60 ° C., then sampled over time, and the NFP content was measured by high performance liquid chromatography.

(2) Test results:
The test results are shown in Table 14 below.

Example 12 (Evaluation of influence of surfactant on cake shape of preparation):
(1) Test method:
Using the freeze-dried preparations 1 and 3 produced in Example 2 “Production of freeze-dried preparations and solutions”, the cake of each preparation was observed with an electron microscope. Specifically, the aluminum cap and the rubber plug of the freeze-dried preparation 1 or 3 were removed, and a part of the cake was quickly taken out with a spatula. The taken-out cake was platinum-deposited with ION SPUTTER (E-1010, manufactured by HITACHI) for 120 seconds. The platinum-deposited sample was subjected to scanning Electron Microscope (S-2380N, manufactured by HITACHI), and an electron micrograph was taken.

(2) Test results:
The result of photographing the cake of the freeze-dried preparation 1 with an electron microscope is shown in FIG. 6 (60 times), FIG. 8 (300 times), and FIG. 10 (1000 times). Similarly, the results of photographing the cake of freeze-dried preparation 3 with an electron microscope are shown in FIG. 7 (60 times), FIG. 9 (300 times), and FIG. 11 (1000 times). From these results, it was found that the surfactant in the preparation did not substantially affect the shape of the cake of the preparation.

Example 13 (Evaluation of Influence of Surfactant Addition Timing on NFP Content Temporal Increase Suppression Effect):
(1) Test method:
Using the lyophilized preparations 1 and 3 produced in Example 2 “Production of lyophilized preparation and solution”, the addition of the same preparation to the preparation with respect to the inhibitory effect of NFP content over time exhibited by the surfactant The effect of timing was evaluated.

  Specifically, the aluminum cap and rubber stopper of lyophilized preparations 1 and 3 are removed. Thereafter, the cake was rapidly stirred with a spatula, and then the rubber stoppers were completely stoppered and wound with an aluminum cap to obtain freeze-dried preparations 1-1 and 3-1. Subsequently, the aluminum cap and the rubber stopper of the freeze-dried preparation 3 are removed. Thereafter, 0.1 mg of polysorbate 80 was dropped into the vial, and the cake was rapidly stirred with a spatula. Thereafter, all the rubber stoppers were stoppered and wound with an aluminum cap to obtain a freeze-dried preparation 3-2. The lyophilized preparations 1-1, 3-1 and 3-2 were stored in a stability tester at 60 ° C., then sampled over time, and the NFP content was measured by high performance liquid chromatography.

(2) Test results:
The test results are shown in Table 15 below.

  The NFP content increase suppression effect exhibited by the surfactant contained in the freeze-dried preparation is that the surfactant is added to the preparation (more specifically, to the chemical solution corresponding to the preparation) before freeze-drying. It was found that the effect was recognized by

Example 14 (Evaluation of Influence of Surfactant Type on NFP Content Temporal Increase Suppression Effect):
(1) Test method:
Using the freeze-dried preparations 14 to 20 produced in Example 2 “Production of freeze-dried preparations and liquids”, the surfactant type has the effect of suppressing the increase in NFP content over time exhibited by the surfactant. The impact was evaluated.

  Specifically, the freeze-dried preparations 13 to 19 were stored in a stability tester at 60 ° C., sampled over time, and the NFP content was measured by high performance liquid chromatography.

(2) Test results:
The test results are shown in Table 16 below.

FIG. 12 shows the relationship between HLB and NFP content (μg / vial) after 2 weeks for the lyophilized preparation containing the non-surfactant in the above table.
From this result, it was revealed that the effect of suppressing the increase in NFP over time in a freeze-dried preparation with a surfactant is remarkable with a nonionic surfactant having an HLB of 10.5 or more.

Example 15 (Evaluation of the influence of the dissolution method on the NFP content-time increase inhibitory effect):
(1) Test method:
Using the lyophilized preparations 1 and 3 produced in Example 2 “Production of lyophilized preparation and liquid”, the effect of the method of dissolving the lyophilized preparation on the NFP content in the solution was evaluated.

  Specifically, 1.2 mL of water for injection was added to the lyophilized preparations 1 and 3 with a syringe without opening the rubber stopper. Thereafter, the cake was dissolved by shaking by hand for 60 seconds or 3 seconds at a speed of about once per second. The lysate was sampled and the NFP content was measured by high performance liquid chromatography.

(2) Test results:
The test results are shown in Table 17 below.

Example 16 (Evaluation of the effect of medicinal components on NFP content over time):
(1) Test method:
Using the freeze-dried preparation 20 produced in Example 3 “Production of teriparatide freeze-dried preparation”, the effect of the medicinal component on the NFP content over time was evaluated. Specifically, the freeze-dried preparation 20 was stored in a stability tester at 40 ° C. and 75% RH, then sampled over time, and the NFP content was measured by high performance liquid chromatography.

(2) Test results:
The test results are shown in Table 18 below.

  An increase in NFP content in the preparation over time was observed not only when the medicinal ingredient contained in the lyophilized preparation was thrombomodulin but also when the medicinal ingredient contained in the lyophilized preparation was teriparatide acetate.

Example 17 (Effect of excipient / additive type, etc. on NFP aging during formulation storage and lyophilized cake of formulation):
(1) Test method:
Using the freeze-dried preparations 21 to 31 produced in Example 4 “Manufacture of freeze-dried preparations containing various excipients / additives”, various excipients and additives were freeze-dried preparations. The effect on the NFP content after storage was evaluated.

  Specifically, the lyophilized preparations 21 to 31 were stored in a stability tester at 60 ° C. for 2 weeks, then sampled, and the NFP content was measured by high performance liquid chromatography.

(2) Test results:
In FIG. 13, the result of having measured the NFP content in the freeze-dried preparations 21-31 after a preservation | save is shown. The upper number shown in the lower part of the graph is the formulation number, and the lower number is the NFP content (NFP content in one vial; μg). In preparations 22, 23, 27, and 28, it was revealed that the increase in NFP after storage was suppressed.

  In FIG. 14, the photograph which image | photographed the freeze-dried preparations 25-28 after a preservation | save is shown. Each formulation is sealed with a rubber stopper and filled in a vial container. In the photograph, the preparation marked with “5” is the freeze-dried preparation after storage (prescription 25), and the preparation marked with “6” is the freeze-dried preparation after storage (prescription 26) with “7”. The preparations marked with indicate freeze-dried preparations (prescription 27) after storage, and the preparations marked with “8” indicate freeze-dried preparations (prescription 28) after storage. It can be seen that each lyo cake of formulations 25, 26 and 28 has been suppressed from collapsing or shrinking after storage. On the other hand, it turns out that the freeze-dried cake of the preparation 27 has collapsed after storage. In Formulation 27, it is highly probable that this large change in shape of the freeze-dried cake had an effect on the suppression of NFP increase after storage.

Example 18 (Effects of excipient / additive type, etc. on NFP aging during storage of teriparatide formulation and lyophilized cake of formulation):
(1) Test method:
Using the freeze-dried preparations 32-46 produced in Example 5 “Manufacture of teriparatide lyophilized preparations containing various excipients / additives”, various excipients and The effect of additives and the like on the NFP content after storage of the lyophilized preparation was evaluated.

  Specifically, the freeze-dried preparations 32-46 were stored in a stability tester at 60 ° C. for 2 weeks, then sampled, and the NFP content was measured by high performance liquid chromatography.

(2) Test results:
FIG. 15 shows the results of measuring the NFP content in teriparatide lyophilized preparations 32-46 after storage. The upper number shown in the lower part of the graph is the formulation number, and the lower number is the NFP content (NFP content in one vial; μg). In the preparations 33, 39, 40, and 41, it was revealed that the increase in NFP after storage was suppressed.

  FIG. 16 shows photographs taken of teriparatide lyophilized preparations 33, 39, 40, 41, and 46 after storage. Each formulation is sealed with a rubber stopper and filled in a vial container. In the photograph, the preparation marked with “2” is the freeze-dried preparation after storage (prescription 33), and the preparation marked with “8” is the freeze-dried preparation after storage (prescription 39) with “9”. The preparations marked with are freeze-dried preparations after storage (prescription 40), the preparations marked with “10” are freeze-dried preparations after storage (prescription 41) and the preparations marked with “14” are The freeze-dried preparation (prescription 45) after storage is shown respectively. It can be seen that the freeze-dried cakes of the preparations 33, 40 and 45 are suppressed from collapsing or shrinking after storage. On the other hand, it can be seen that the freeze-dried cakes of the preparations 39 and 41 have collapsed or contracted after storage. In preparations 39 and 41, it is highly likely that this large change in shape of the lyophilized cake had an effect on the suppression of NFP increase after storage.

  FIG. 17 shows photographs of teriparatide lyophilized preparations (prescriptions 32-46; total 15 preparations) after storage. It can be seen that each of the freeze-dried cakes of the preparations 32-34, 36-38, 40, 42-46 (total of 13 preparations) was suppressed from collapsing or shrinking after storage. On the other hand, it can be seen that the freeze-dried cakes of the preparations 35, 39, and 41 have collapsed or contracted after storage.

  The freeze-dried preparation of the present invention has a reduced N-formylpiperidine content and is useful as a high-quality pharmaceutical.

Claims (7)

A freeze-dried preparation containing teriparatide acetate, sealed with a rubber stopper and filled in a medical container, containing at least one excipient of (1) to (4) and added Freeze-dried preparation without food:
(1) Glycine;
(2) sodium glutamate;
(3) trehalose;
(4) Purified white sugar. A freeze-dried preparation containing teriparatide acetate, which is sealed with a rubber stopper and filled in a medical container, containing purified white sugar, and an additive of at least one of (1) to (6) Freeze-dried formulations containing:
(1) histidine, a salt thereof, or a hydrate thereof;
(2) ascorbic acid;
(3) sodium acetate or a hydrate thereof;
(4) sodium bisulfite;
(5) benzalkonium chloride;
(6) Methionine.   The freeze-dried preparation according to claim 1 or 2, wherein the amount of teriparatide acetate in the freeze-dried preparation is 67.9 µg.   The freeze-dried preparation according to any one of claims 1 to 3, wherein the medical container is a vial. A method for producing a freeze-dried preparation containing teriparatide acetate, which is sealed with a rubber stopper and filled in a medical container, wherein teriparatide acetate and at least one of (1) to (4) are added. A freeze-dried formulation comprising a step of freeze-drying a drug solution containing a form and containing no additives, wherein collapse or shrinkage of a freeze-dried cake (lyo cake) in the freeze-dried formulation is suppressed Production method:
(1) Glycine;
(2) sodium glutamate;
(3) trehalose;
(4) Purified white sugar. A method for producing a freeze-dried preparation containing teriparatide acetate, which is sealed with a rubber stopper and filled in a medical container, wherein teriparatide acetate, purified white sugar, and at least one of (1) to (6) A method for producing a freeze-dried preparation, comprising the step of freeze-drying a drug solution containing the additive of 1, wherein the collapse or shrinkage of the freeze-dried cake (lyo cake) in the freeze-dried preparation is suppressed:
(1) histidine, a salt thereof, or a hydrate thereof;
(2) ascorbic acid;
(3) sodium acetate or a hydrate thereof;
(4) sodium bisulfite;
(5) benzalkonium chloride;
(6) Methionine.   The manufacturing method according to claim 5 or 6, wherein the medical container is a vial.