JP2015078177A - Lyophilized products for room temperature preservation - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide stable lyophilized products for room temperature preservation.SOLUTION: The invention provides lyophilized products and the like which contain an effective amount of a peptide drug, and about 0.50-1.85 mg/mmof sucrose to the internal cross section of a container, and are stable when preserving at room temperature.

Description

  The present invention relates to a lyophilized formulation that is stable at room temperature storage.

  Conventional injectable hump preparations are freeze-dried preparations containing mannitol as an additive. This formulation needs to be stored in a cold place from the viewpoint of stability, and a medical site that is easier to handle and that can be stored at room temperature has been demanded. For formulations that store at room temperature, the quality is guaranteed even when stored under constant temperature fluctuations, so it is possible even under dynamic semi-harsh conditions (stored at 40-50 ° C for 1-3 months) The quality must be maintained as much as possible.

  In lyophilized preparations, disaccharides such as sucrose (purified sucrose) are known as additives that improve the chemical storage stability of the active ingredients (peptides, proteins) (Patent Document 1). However, only a small amount at the laboratory level was used to evaluate the chemical stability of the main drug, and it was unclear from the viewpoint of the quality of the freeze-dried preparation. In particular, refined sucrose is one of the typical stabilizers, and has a large amount of actual use as a pharmaceutical additive, and can be said to have a high degree of freedom in the amount used in formulation design. However, when the amount and concentration of a dry object are increased in the production of a freeze-dried preparation, there is a risk that important quality including stability and other preparation characteristics may be impaired.

Japanese Patent No. 3559596

  The present invention provides a freeze-dried preparation containing a physiologically active peptide as an active ingredient, containing a certain amount or more of purified sucrose as a main component, the active ingredient being chemically stable during storage at room temperature, and other preparation quality being stable. The issue is to provide.

  Since the redissolved solution of lyophilized carperitide may be directly administered to a patient, it needs to be prepared so as to be isotonic with water for injection. When the amount of redissolved solution is small, the effect of a decrease in the guaranteed dosage due to the drug solution remaining in the vial becomes large. Therefore, it is desirable to use a certain amount or more of the redissolved solution. In order to obtain isotonicity, it is necessary to use more than a certain amount.

  In the freeze-dried preparation, the appearance shape and the water content of the freeze-dried product are preparation characteristics that serve as indices for ensuring the important quality of the product. A freeze-dried product having a poor external shape may cause a poor dissolution at the time of re-dissolution or a long dissolution time, which may cause a problem in the medical field. In addition, if the lyophilized product contains a large amount of water, the glass transition temperature of the lyophilized product may be lowered to impair the chemical stability of the active ingredient, or to cause changes in the external shape of the lyophilized product during storage. There is a possibility.

  In order to appropriately secure the external shape of the freeze-dried body, it is necessary that the freeze-dried body does not appear abnormal in production (during the process of vacuum freeze-drying) and product storage after freeze-drying.

  In the course of the manufacturing process (vacuum freeze-drying), a typical cause of appearance abnormality in the freeze-dried body is collapse of the already dried portion by water vapor (melting-like collapse of the microstructure). Moreover, as a typical phenomenon example of the appearance change of the freeze-dried body in product storage, there is a shrinkage (shrinkage of shape) caused by the limit of the heat resistance of the freeze-dried body with respect to the exposure temperature.

  Collapsing at the time of manufacture is promoted by the sublimation process of the vacuum freeze-drying process due to obstruction of water vapor passage (sublimation) due to narrowing or blockage of the water vapor passage path in the dried part, that is, abnormality of the water removal process. In the vacuum freeze-drying process, sublimation proceeds from the top surface to the bottom surface (depth direction) of the lyophilization solution filled in the preparation container. Thus, the water content of the freeze-dried product is increased. That is, a freeze-dried product having an abnormal appearance shape and a high water content may be obtained at the production stage. At this time, the high water content is a result of a change in the appearance shape at the production stage and is freeze-dried. It can reduce the glass transition temperature of the body, reduce chemical stability during formulation storage, and cause shrinkage.

  Shrinking due to heat exposure during product storage is likely to occur when the glass transition temperature of the lyophilized product is not properly designed for the storage temperature. If the temperature to which the lyophilized product is exposed exceeds the glass transition point of the components contained in the lyophilized product, there is a probability that the chemical reaction rate in the lyophilized product will increase or the shape of the lyophilized product will change. Get higher. For example, in the case of a lyophilized product whose main component is sucrose, when the temperature at which the lyophilized product is exposed is below the glass transition temperature of sucrose, sucrose exists as an amorphous solid, but near the glass transition temperature Or when exposed to higher temperatures, the phase state is more likely to become a rubber zone. When such a phase change occurs, in the lyophilizate containing a pharmaceutically active ingredient such as a peptide, the chemical reaction rate is significantly increased, and the chemical stabilization effect of the active ingredient by sucrose is lost. That is, the sucrose phase change contributes to the above-described physical property change such as the shape change of the lyophilizate, and at the same time, causes a decrease in the chemical stabilization effect of the active ingredient.

  Thus, in a lyophilized preparation using a certain amount or more of purified sucrose, it is necessary to carry out a preparation design that can appropriately maintain the quality of the preparation. The water content can be used as an index.

  As a result of intensive studies on a lyophilized preparation of a physiologically active peptide that is stable at room temperature storage, the present inventors varied the content of purified sucrose from 100 to 1000 mg using a cylindrical glass vial with an inner diameter of 22.1 mm. As a result of producing freeze-dried preparations and conducting property evaluations during production and stability tests after severe heat storage, good appearance and moisture content for preparations with purified sucrose of 300 to 500 mg and sucrose concentration of 125 mg / mL or less in the solution It was found that the content was maintained, and in the preparation with 100 mg or 1000 mg of refined sucrose, it was found that a good appearance shape and water content could not be maintained at the time of production or in a thermal severe test, and further investigation was advanced to complete the present invention. It came.

The present invention provides the following.
(1) containing an effective amount of a peptide drug and about 0.50 to 1.85 mg / mm ² of purified sucrose with respect to the inner cross-sectional area of the container, and being stable at room temperature storage, A freeze-dried preparation characterized by
(2) The freeze-dried preparation according to (1), wherein the freeze-dried product exhibits a good appearance shape after storage at 40 ° C. for one month, and the water content is 1% or less.
(3) The freeze-dried preparation according to (1), wherein the freeze-dried body has a good appearance shape after storage at 50 ° C. for one month, and the water content is 1% or less.
(4) The freeze-dried preparation according to (1), wherein the content of purified sucrose is about 0.75 to 1.35 mg / mm ^{2 with} respect to the internal cross-sectional area of the container.
(5) The lyophilized preparation according to (1), wherein the container is a cylindrical vial having an inner diameter of about 20 to 29 mm, and contains about 300 to 500 mg of purified sucrose.
(6) The freeze-dried preparation according to (5), wherein the inner diameter of the cylindrical vial is about 22.1 mm.
(7) The lyophilized preparation according to any one of (1) to (6), which is produced by lyophilizing a solution having a purified sucrose concentration of 140 mg / mL or less.
(8) The freeze-dried preparation according to any one of (1) to (7), which contains 1 mg of carperitide as a peptide drug.
(9) The freeze-dried preparation according to any one of (1) to (8), which substantially comprises purified sucrose and a peptide drug as solid components.
(10) An aqueous solution containing purified sucrose of 140 mg / mL or less and an effective amount of a peptide drug is added to a container at about 0.50 to 1.85 mg / mm ^{2 with} respect to the internal cross-sectional area of the container. A method for producing a freeze-dried preparation characterized by being stable at room temperature storage, comprising a step of lyophilizing the filled container and freeze-drying the filled container.
(11) The production method according to (10), wherein the ratio of the preparation having good preparation quality characteristics is about 80% or more in the freeze-dried preparation to be produced.

  The freeze-dried preparation according to the present invention has not only the chemical stability of the active ingredient but also the physical stability of the freeze-dried body in room temperature storage. In addition to the need for such a facility, there is no need to return to room temperature in preparation of a drug for use, and a uniform solution can be obtained in a short time when re-dissolving with an injection solution.

FIG. 1 is a photograph of an appearance example corresponding to a ◯ criteria (good shape) in the criteria for the appearance evaluation adopted in this example. FIG. 2 is a photograph of an appearance example corresponding to the Δ judgment criterion (generally good shape) in the appearance evaluation judgment criteria employed in this example. FIG. 3 is a photograph of an appearance example corresponding to the x judgment criterion (defective shape) in the judgment criteria of the appearance evaluation employed in this example.

  Hereinafter, the present invention will be described in detail.

The present invention contains an effective amount of a peptide drug and about 0.50 to 1.85 mg / mm ² of purified sucrose with respect to the internal cross-sectional area of the container, and is stable at room temperature storage. A lyophilized formulation is provided.

  In the present invention, the term “lyophilized preparation” refers to a solid substance (“freeze-dried”) that is prepared by injecting a predetermined amount of a solution of components having a predetermined composition (referred to as “dissolved solution”) into a container and then drying by a vacuum freeze-drying method. Body "or" lyophilized cake ") and a closed container closure system containing the same. At the time of use at a medical site, a predetermined amount of an aqueous component is added to a predetermined amount of a container, and a lyophilized body is dissolved (referred to as “re-dissolution”) and administered as a solution to a subject.

  In the present invention, the “internal cross-sectional area of a container” refers to an internal cross-sectional area in a horizontal cross section of a trunk straight portion of a container used for freeze-drying. Here, the trunk straight portion of the container is a portion having a uniform horizontal cross section within a certain range in the vertical direction at the bottom of the container. An internal cross-sectional area means the area inside a container wall in the horizontal cross section of a container.

  In the present invention, “room temperature storage” means that the product is stored without using temperature control equipment such as a refrigerator, a freezer, and a heat storage during the period from manufacture to use of the preparation. Such pharmaceuticals that can be stored at room temperature must be confirmed to maintain stable quality for a predetermined period of time under the storage conditions. In preparations with room temperature conditions as storage methods, fluctuations in the exposure temperature within a certain range are assumed, and in particular, at a high temperature such as 40 ° C., 50 ° C., 60 ° C., for a certain period (for example, for 1 month to 6 months). It is recommended that the quality of the stored so-called thermal harsh storage has been confirmed to be stable.

  In the present invention, “stable formulation”, “stable quality” or “stable formulation quality” means that important formulation characteristics and / or product quality are maintained well, for example, contained in the formulation Means that the active ingredient is chemically stable and that the preparation is physically stable, such as good solubility when the lyophilized product is redissolved with water for injection. To do. The fact that the active ingredient is stable means that the active ingredient in the lyophilized preparation containing the active ingredient has little content reduction, activity reduction, purity reduction, generation of degradation products (generation of related substances), etc. To do.

  Furthermore, physical properties such as the appearance (color, shape, etc.) and moisture content of the lyophilized product may affect the chemical stability, physical properties, and stability of such lyophilized formulations. These physical properties can also be regarded as important formulation characteristics. That is, it is desirable that at least one, and preferably all of these physical characteristics are maintained well even after the stability test of the lyophilized preparation. For example, the appearance shape affects the re-solubility of the lyophilizate. In addition, the shape change that occurs in the freeze-drying process affects the water removal efficiency. If the change occurs, the water is not properly removed, and the water content at the time of manufacture increases. In addition, when the water content increases, the glass transition temperature of the main base material of the lyophilizate is lowered, the chemical stability of the active ingredient is likely to be lowered, and the possibility of shape change during storage of the preparation is increased. Thus, the appearance and water content of the lyophilized product are closely related properties that can affect the critical quality of the formulation, ensuring important product quality such as stability and solubility. It becomes an index in doing.

  In order to ensure storage stability at room temperature (25 ° C.), it is desirable that the glass transition temperature of the product is about 20 ° C. higher than room temperature, 45 ° C. or higher (F. Franks et al., “Protein / Nucleic acid”). "Enzyme" p.810-816, 41 (6), 1996). As described above, the glass transition temperature of the purified sucrose is considered to be around 70 ° C. when the moisture content is 0%, 40 to 50 ° C. when the retained moisture content is 1%, and 30 to 40 ° C. when the moisture content is 2%. Therefore, it is desirable that the freeze-dried preparation containing purified sucrose as a main component has a moisture content of 1% or less in order to ensure storage stability at room temperature (25 ° C.).

  In the present specification, when “about” is added to the numerical value description, it means that a range of 30% above and below the specified numerical value is allowed. Here, when “about” is added to the description of the numerical range, the above-described allowable range means that from the lower 30% to the upper 30% is allowed. The acceptable range is preferably 20%, more preferably 15%, even more preferably 10%, even more preferably 5%.

  Next, each component of the freeze-dried preparation of the present invention will be described in detail.

  In the freeze-dried preparation of the present invention, various peptide drugs can be used as the active ingredient, but preferably carperitide. In the present specification, “carperitide” is a peptide hormone consisting of 28 amino acids: SLRRSSCFGG RMRIGAQSG LGCNSFRY (SEQ ID NO: 1), α-human atrial natriuretic peptide (“α-hANP”) or simply Also referred to as “hANP”). Carperitide can be produced by various known methods such as extraction / purification from tissues derived from living organisms, biotechnological synthesis by genetic recombination methods, and chemical synthesis such as solid phase synthesis of peptides. (For example, see Table 01/85945). In addition, a freeze-dried preparation stored in a cool dark place containing 1 mg of carperitide in one vial is sold as “1000 for Hump Injection”. Also in the freeze-dried preparation of the present invention, when carperitide is used as the physiologically active peptide, the content per vial is preferably 1 mg.

The freeze-drying agent of the present invention contains a predetermined amount of purified sucrose as a stabilizer. As the purified sucrose of the present invention, a purified sucrose listed in the 16th revision Japanese Pharmacopoeia can be preferably used. Purified sucrose provides the active ingredient peptide of the freeze-dried preparation of the present invention with chemical stability during storage at room temperature, and can achieve an appearance and final moisture content that do not adversely affect other preparation quality and preparation characteristics. The amount must be added. In the lyophilization process, the water content of the solution evaporates only from the liquid surface (upper portion of the lyophilized body), so the efficiency of water removal is greatly affected by the depth of the solid component in the container. When the solid component in the container is shallow, moisture is easily removed efficiently. On the other hand, when the solid component in the container is deep, it is difficult to remove moisture, and the final moisture content of the freeze-dried product may be increased. That is, the physical stability of the freeze-dried product depends on the content of purified sucrose with respect to the internal cross-sectional area of the body of the container used for freeze-drying. In the present invention, the content of purified sucrose relative to the inner cross-sectional area of the container is about 0.50 to 1.85 mg / mm ² , preferably about 0.65 to 1.60 mg / mm ² , more preferably About 0.75 to 1.35 mg / mm ² , even more preferably about 1.30 mg / mm ² .

  The content of purified sucrose per freeze-dried body is appropriately selected according to the size of the container. For example, when a cylindrical vial having an inner diameter of 22.1 mm is adopted as the container, the amount of purified sucrose per freeze-dried body is usually about 200 to 700 mg, preferably about 250 to 600 mg, more preferably about 300. ˜500 mg, even more preferably about 500 mg.

  The container used for the lyophilized preparation of the present invention includes a trunk straight part in the lower part of the container, and can be used for vials, ampoules, injections, etc., as long as it does not cause alteration or deformation due to temperature changes in the lyophilization operation. A variety of syringes can be used. The cross section of the trunk straight portion may have various shapes, for example, a polygon (polygonal column vial) such as a quadrangle and a hexagon, or a circle (cylindrical vial). The size of the container is not particularly limited as long as it can be lyophilized, but the inner diameter of a commonly used cylindrical vial is about 12 mm to 50 mm, and is appropriate depending on the amount of purified sucrose used. Internal diameter vials can be selected. For example, when 300 mg of purified sucrose is used, a cylindrical vial having an inner diameter of about 14.6 to 29 mm (preferably about 16 to 24 mm) is employed. When 500 mg is used as the purified sucrose, a cylindrical vial having an inner diameter of about 19 to 34 mm (preferably about 21 to 31.4 mm) can be used.

<Additives other than refined sucrose>
The lyophilized preparation of the present invention may be a preparation substantially consisting of carperitide and purified sucrose, and if necessary, other components (stabilizers other than purified sucrose, buffers, pH adjusters) , Surfactants, solubilizers, antioxidants, soothing agents, tonicity agents, and the like). The content of these components other than the main drug and purified saccharose has substantially no effect on the glass transition temperature of the lyophilizate, or has a small effect, and / or the effect of stabilizing the main drug by purified sucrose. It is necessary that the amount is not reduced, and the content in the case of using the component is preferably 20% by weight or less, more preferably 10% by weight or less, more preferably 10% by weight or less, based on the total amount. It is 5% by weight or less, and more preferably 1% by weight or less.

  Stabilizers other than purified sucrose include sugars belonging to the same disaccharides as purified sucrose such as trehalose, lactose and maltose, and amino acids such as glycine, alanine, arginine, lysine, histidine and methionine that have been used as stabilizers in the past. Polysaccharides such as heparin and dextran sulfate, sugar alcohols such as sorbitol and mannitol, proteins such as egg albumin and serum albumin can be preferably used. Of these, disaccharides are preferable, and trehalose is particularly preferable. The amount of the stabilizer other than the purified sucrose is not particularly limited, but is preferably 20% by weight or less of the purified sucrose, and more preferably 10% by weight or less.

  Examples of the buffer used in the present invention include phosphate buffer, citrate buffer, acetate buffer, glycine / hydrochloric acid buffer, and the like. The buffer can be used to adjust the pH of the aqueous solution after re-dissolution as necessary, and to maintain the solubility and stability of carperitide. The buffering agent preferably has a pH of the aqueous solution after re-dissolution of 4.0 to 6.5 and does not contain an electrolyte such as sodium salt or potassium salt in order to prevent salting out of carperitide. A particularly preferable buffer is, for example, glycine / hydrochloric acid buffer. The amount of the buffer added is preferably such that the concentration in the lysate immediately before lyophilization when producing a lyophilized preparation is in the range of 1 to 100 mM.

  Examples of the surfactant used in the present invention include polysorbate 20, polysorbate 80, pluronic F-68, and polyethylene glycol, and two or more kinds may be used in combination. As the surfactant, a polysorbate-based surfactant is preferable, and polysorbate 80 is particularly preferable. The surfactant can be added as necessary, for example, in order to prevent adsorption of carperitide in the aqueous solution to the container. As for the addition amount of the surfactant, the concentration in the solution immediately before lyophilization when producing a lyophilized preparation is preferably in the range of 0.001 to 2.0% by weight.

  In order to adjust the pH of the redissolved solution, the freeze-dried composition of the present invention may further contain an acid or a base as a pH adjuster. Examples of acids that may be further added to the lyophilized preparation of the present invention include inorganic acids (for example, hydrochloric acid, phosphoric acid, hydrobromic acid, sulfuric acid), organic acids (for example, formic acid, acetic acid, propionic acid, fumaric acid). Acid, maleic acid, succinic acid, tartaric acid, citric acid, malic acid, succinic acid, benzoic acid, methanesulfonic acid, benzenesulfonic acid) and the like.

Of these, hydrochloric acid and acetic acid are preferred. The amount of these acids is not particularly limited, and the acid may be added to the freeze-drying composition in an amount capable of adjusting the pH of the redissolved solution of the freeze-drying composition to a predetermined range.
Examples of the antioxidant used in the present invention include sodium sulfite, sodium bisulfite, sodium pyrosulfite, L-cysteine hydrochloride, thioglycolic acid and other substances having autooxidation (reducing), and amino acids such as methionine. Can be mentioned. However, when carperitide and a reducing substance coexist, the peptide may be decomposed by reductive cleavage of a disulfide bond. Therefore, amino acids such as methionine are preferably used as substances that prevent peptide oxidation, and it is preferably used in the range of 0.001 to 0.1 parts by weight with respect to 1 part by weight of purified sucrose.

Examples of the isotonic agent used in the present invention include monosaccharides such as glucose, sugar alcohols such as mannitol, inorganic salts such as sodium chloride, and preferably monosaccharides such as glucose or sugar alcohols such as mannitol. It is preferable to use within a range of 0.001 to 0.2 parts by weight with respect to 1 part by weight of purified white sugar.
Examples of the soothing agent used in the present invention include alcohols such as benzyl alcohol and chlorobutanol, lidocaine hydrochloride and the like, and the concentration in the solution when producing a lyophilized preparation is 0.001 to 2. It is preferable to use in the range of 0.0% by weight.

<Freeze-dried preparation>
The freeze-dried preparation of the present invention can be produced by a known method.
For example, a predetermined amount of carperitide or a salt thereof and a predetermined amount of purified sucrose are dissolved in a predetermined amount of water or a suitable aqueous solvent (for example, a mixture of water and alcohol) to prepare a solution, and an acid is optionally added. Adjust the pH of the aqueous solution by addition, and then inject a sterile solution obtained by sterilization filtration using a filter (for example, a sterilization filter having a pore size of 0.22 μm) into a vial or ampoule. The freeze-drying agent can be obtained by freeze-drying the container to form a solid.

  In the preparation of the lyophilizing agent, the active ingredient peptide and purified sucrose are dissolved in water or an aqueous solvent (for example, a mixture of water and alcohol) according to a known method, and an acid is added as necessary. The pH may be adjusted. Further, the order of dissolving the active ingredient peptide and the purified sucrose is not particularly limited.

  In the preparation of the lyophilizate, the concentration of the active ingredient peptide in the aqueous liquid is appropriately set so as to be a dose approved as a pharmaceutical product in the final product, but is usually 0.01 mg / mL to 5 mg / mL. . In a freeze-dried preparation containing 1 mg of carperitide, when the filling amount of the dissolving solution per vial is 5 mL, the concentration of carperitide in the dissolving solution is adjusted to 0.2 mg / mL.

  The concentration of the purified sucrose in the aqueous solution subjected to the above lyophilization depends on the composition of other additives, but the lower limit is usually about 10 mg / mL or more. The concentration of solid components contained in the solution subjected to lyophilization affects the water removal efficiency of the lyophilization process, so the purified sucrose concentration must be low to achieve a low water content at the end of lyophilization. desirable. The upper limit of the purified sucrose concentration in the lysate is usually 140 mg / mL or less, preferably 125 mg / mL or less.

  The filter used for sterilization is preferably a sterilization filter having a pore size of 0.22 μm or less. Examples of the filter for sterilization include Durapore (registered trademark, manufactured by Nippon Millipore Corporation) or Zaltopore 2 (registered trademark, manufactured by Sartorius Corporation).

  A fixed amount of the solution prepared by the above process is filled in a container, and the filled container is subjected to freeze-drying. The filling amount of the dissolving liquid per container at this time varies depending on the size of the container, but the depth (liquid depth) of the filling liquid is usually about 3 mm to 20 mm (preferably about 5 mm to 17 mm). . If the liquid depth is too small, a lyophilized product is not formed. Conversely, if the liquid depth is too large, there is a possibility that water removal at the lower part of the container does not proceed efficiently.

  The freeze-drying method includes, for example, a freezing process that cools and freezes under normal pressure, a primary drying process that sublimates and drys free water that is not restricted by the solute, and a secondary drying process that removes adsorbed water and crystallization water inherent to the solute. The method by three unit operation is mentioned. The cooling temperature in the freezing process is preferably −60 ° C. to −40 ° C., the temperature in the primary drying process is preferably −50 ° C. to 0 ° C., and the temperature in the secondary drying process is preferably 4 ° C. to 40 ° C. The vacuum pressure in the primary drying process is preferably controlled to 0.1 to 50 Pa, particularly preferably 0.5 to 30 Pa. Restore the pressure in the drying cabinet after freeze-drying. As a method of restoring pressure, aseptic air or inert gas (for example, sterile nitrogen gas, sterile helium gas, etc.) is sent into the chamber to perform primary decompression to about 60 to 90 kPa, and then further if necessary. A method of restoring pressure (secondary restoring pressure) is preferable. The vial is preferably stoppered after the primary decompression.

<Use of formulation>
The lyophilized preparation of the present invention can be redissolved in water for injection or infusion (for example, physiological saline, glucose solution, etc.) before use and used for injection, infusion and the like. However, when carperitide is dissolved in physiological saline, gelation may occur if the dissolution conditions are not appropriate. Therefore, it is necessary to adjust the drug concentration after redissolving and pay attention to the state during use. Glucose fluid may also be restricted in use when the patient has diabetes or is at risk. Therefore, it is desirable to use water for injection for redissolving the lyophilized preparation of the present invention.

  The lyophilizate of the present invention includes various injectable compositions such as a composition for subcutaneous injection, a composition for intravenous injection, a composition for intramuscular injection, a composition for drip injection, and a composition for needleless injection. It can be used as a product. In this case, the concentration of the active ingredient peptide at the time of injection in the injectable composition (in the case of a lyophilized agent, the reconstitution concentration) is usually about 0.01 mg / mL to 60 mg / mL, preferably about 0.05 mg / mL. mL to 20 mg / mL.

  When the freeze-dried agent of the present invention is redissolved at the time of use and used for injection, it is preferable to prepare the aqueous solution by a known aseptic preparation method such as filtration sterilization. In addition, before preparing the lyophilizing agent of the present invention, a saccharide and a surfactant, or a mixture of the saccharide, the surfactant and other additives as necessary is subjected to a sterilization treatment such as a depyrogenic treatment in advance. It can also be used after.

Furthermore, the lyophilized preparation of the present invention can be advantageously used not only as a prophylactic or therapeutic agent for acute heart failure, but also as a prophylactic or therapeutic agent for diseases associated with an increase in blood pressure or a load on the heart.
The lyophilized preparation of the present invention has an excellent blood pressure lowering action and cardioprotective action as a single agent, and exhibits an effective action as a preventive or therapeutic agent for heart disease, but is further used in combination with other pharmaceutical ingredients (multiple drugs) Can also be used together).

  Hereinafter, the present invention will be specifically described with reference to examples. What is shown in the examples is an example of an embodiment of the present invention, and the present invention is not limited to this. In this example, a cylindrical glass vial for “Hump Injection 1000” (outer diameter 24.5 mm, inner diameter 22.1 mm) was employed as a container for the lyophilized preparation.

<Example 1: Examination of compounding conditions of purified sucrose (sucrose placebo preparation)>
In order to find a pharmaceutical design of a freeze-dried drug product that contains purified sucrose as the main drug component and has good product characteristics, the compounding conditions for purified sucrose in the drug product formulation and manufacturing process (liquid for freeze-drying) Condition ").
The examination range of the amount of purified sucrose in this test was determined for the following reasons. In the current dosage of Carperitide (product name: “Hump Injection 1000”), one formulation (one vial) is redissolved with 10 mL of water for injection. From the viewpoint of maintaining or improving the efficiency of the preparation, the type of the re-dissolved solution is not changed, and the amount is preferably less than the current product (10 mL or less). On the other hand, in dispensing at the medical site, the residual rate of solution (residual liquid rate) due to adhesion to the formulation container after extracting the solution after re-dissolution with a syringe etc. increases as the amount of solution decreases. In designing, it is necessary to set the amount of the re-dissolved liquid so that the residual liquid ratio becomes as small as possible. Therefore, in this product design, the amount of the re-dissolved solution was set in the range of 1 to 10 mL.
Moreover, since a carpertide formulation may be directly administered without reconstitution with an infusion solution after re-dissolution with water for injection in a medical field, it is necessary to be isotonic after dissolution. In the case of a preparation containing purified sucrose as a main component, the amount of the solution required to ensure isotonicity is about 10 times the amount of purified sucrose (about 1 mL of water for injection per 100 mg of purified sucrose). In this product design, since the amount of water for injection was 1 to 10 mL, the examination range of the amount of purified white sugar was 100 to 1000 mg.
In this study, the amount of purified sucrose per formulation (one vial) (hereinafter “sucrose amount”) and the concentration of purified sucrose in the lyophilized solution (hereinafter “sucrose concentration”) were used as variables. Moreover, the external appearance shape and water | moisture content of the freeze-dried body were evaluated as a formulation quality characteristic.
Since this test focuses on the physical properties such as the appearance and moisture of the lyophilized product, the effect of carperitide on these physical properties is negligible from the ratio of the content of carperitide to be examined and the amount of purified white sugar. Judging that it was negligible, this study was carried out using a formulation containing no carperitide (sucrose placebo formulation).

(1) Preparation of sucrose placebo preparation (Formulation Examples 1 to 17)
Purified sucrose (Japanese Pharmacopoeia, Shisui Minato Seika Co., Ltd.) was dissolved in purified water so as to have a predetermined sucrose concentration in each formulation example in Table 1. After filtering the sucrose solution with a filter (Stericup-GV 0.22 μm, PVDF, Millipore Corporation), use a theta dispenser (FH-10s, Hirasawa Co., Ltd.) or a microscale pipette (PIPETMAN, GILSON, Inc., etc.) In the same manner, glass vials (cylinder outer diameter 24.5 mm), Daiwa Special Glass Co., Ltd.) were filled so as to have a predetermined filling amount. After half-plugging a rubber stopper (V10-F8 D713 RB2-40, Daikyo Seiko Co., Ltd.) into the filled vial, it is carried into a freeze dryer (RL-30KPs, Kyowa Vacuum Co., Ltd.) and pre-frozen (- 50 ° C setting), followed by freeze-drying under vacuum conditions (20 Pa setting) (primary drying: -20 to -5 ° C setting, secondary drying: 40 ° C setting), and after the completion of drying, nitrogen was introduced into the vial. The pressure was restored. After all the rubber stoppers have been plugged into the vial, the vial is wrapped with a flip-off cap (AP cap, 20AP7.0H, Ishida Press Industry Co., Ltd.) using a winding machine (GAC-01, Inaba Machinery Co., Ltd.) Thus, sucrose placebo preparations of Formulation Examples 1 to 17 were obtained.

(2) Analysis of formulation quality characteristics after freeze-drying of freeze-dried preparations Constant temperature and humidity chamber (LH31-15M type, Nagano Science Co., Ltd.) set to 40 ° C and 75% RH Stored in a thermostat (LH20-01 type, Nagano Scientific Machinery Co., Ltd.) set to 50 ° C. for one month. At the time of preparation of the preparation and before and after storage under each condition, the appearance shape and water content of the lyophilized product were evaluated by the following methods.

[Method: Appearance evaluation of freeze-dried product]
The appearance of the freeze-dried preparation was visually observed, and the sign was determined in three stages as follows. At the time of manufacture, 23-25 randomly selected were observed. In the heat harsh storage, three samples having a good appearance shape were randomly selected before storage at each temperature, and the same preparation was observed before and after storage. These results are shown in Tables 2 and 3.
○: Good shape (normal shape)
Δ: generally good shape (only slight changes are observed compared to normal shape)
X: Poor shape (shape defect: a large change such as significant shrinkage is recognized as compared with a normal shape).

[Method: Moisture measurement of lyophilized product]
10 mg or more of the lyophilized product is weighed into a collection tube with a glass stopper in a humidity environment to reduce moisture absorption, and the water content (CA-100, Mitsubishi Chemical Corporation) The moisture content (%) of the lyophilized body weight was measured. Two or three samples randomly selected at each measurement were measured, and the average water content is shown in Tables 2 and 3.

[Results] Table 2 shows the external shape and water content after preparation for Formulation Examples 1 to 7 (white sugar content 1000 mg or 500 mg / vial) after storage at 40 ° C 75% RH for 1 month and after storage at 50 ° C for 1 month. It was. Similarly, Table 3 shows the external shape and water content of the preparations of Formulation Examples 8 to 17 (amount of white sugar of 300 mg or 100 mg / vial). For each formulation example, it was determined that the appearance shape was ○ or △ and the moisture content was 1% or less at the time of production and thermal severe storage at the time of judging that the preparation quality was good. The criterion was used.
The preparations of Formulation Examples 5-12 (sucrose amount 500 mg / vial (sucrose concentration 125 mg / mL or less) or 300 mg / vial) meet the criteria for appearance and moisture both at the time of manufacture and after severe heat storage. It shows that when a carperitide preparation is produced under these conditions, good product characteristics such as thermal stability can be secured.
On the other hand, the preparations of Formulation Examples 1 to 4 (sucrose amount of 1000 mg / vial or 500 mg / vial (sucrose concentration of 160 mg / mL or more)) all cause an apparent poor appearance (x judgment of 10% or more) during production. In addition, the moisture at the time of production was 1% or more, and both the appearance and moisture were incompatible with the judgment criteria. Evaluation after heat-harmful storage revealed specimens that had poor appearance (nonconformance to the criteria) when stored at 50 ° C. This is uncertain about the thermal stability of formulations whose moisture does not meet the criteria. It shows that there is.
The preparations of Formulation Examples 13 to 17 (sucrose amount 100 mg / vial) satisfy the criteria for both appearance and moisture at the time of production, but the moisture content increased to 1% or more after 1 month of heat-harvest storage (to the criteria) Non-conformity). It was suggested that the formulation with a small amount of sucrose was affected by the water content of the rubber stopper used to seal the container of the formulation, and the water content increased during storage, which could change the product characteristics.

*: Randomly selected formulations with an appearance evaluation of ○ (no change in appearance) at the time of manufacture and stored at each temperature. Appearance shows the measured value (judgment code for 3 preparations). -: Not measured.

*: Randomly selected formulations with an appearance evaluation of ○ or △ (no change or slight change in appearance) and stored at each temperature. Appearance shows the measured value (judgment code for 3 preparations). -: Not measured.

From these results, in the production of a freeze-dried preparation containing purified sucrose as the main solid component using a glass vial with a barrel outer diameter of 24.5 mm (inner diameter 22.1 mm) as a container for lyophilization, the appearance shape, The sucrose blending conditions satisfying the water judgment standard are 300 mg (0.78 mg / mm ² per container internal cross-sectional area) to 500 mg / vial (1.30 mg / mm ² per container internal cross-sectional area) for the amount of sucrose. The sucrose concentration was 48 to 125 mg / mL (filling amount was 2.5 to 6.25 mL / vial).

When focusing on quality characteristics at the time of production under these conditions, when the amount of purified white sugar per vial is large or the white sugar concentration of the filling liquid is high, the ratio of appearance defects at the time of production is high, There was also a high tendency. In these freeze-dried bodies, it is considered that water removal in the freeze-drying process did not proceed normally. Moisture removal in the freeze-drying process proceeds as the frozen water sublimes and is released from the top surface of the freeze-dried body, so the smaller the amount of white sugar per area of the top surface of the freeze-dried body (inner cross-sectional area of the container) The water removal efficiency is considered to be high. Also in this examination, in the same size vial, the moisture content at the time of manufacture tended to be lower as the amount of white sugar was smaller and the concentration of white sugar was lower. Therefore, in order to ensure good pharmaceutical quality characteristics at the time of manufacture, it is necessary to reduce the amount of purified sucrose per container internal cross-sectional area (usually about 1.85 mg / mm ² or less). For the same purpose, the concentration of the filling liquid used for lyophilization is about 140 mg / mL or less, considering that the quality characteristics in the case of Formulation Example 4 (160 mg / mL) did not reach the standard slightly. It is considered desirable.

In addition, in order to ensure good quality characteristics after severe heat storage, it is important that the moisture content during production is low, but even if the moisture content during production is sufficiently low, If the amount of white sugar is too small, the effect of the freeze-dried product absorbing water contained in rubber stoppers, etc. cannot be ignored, and the water content after storage will exceed the specified value. Was suggested. Since the size of the rubber stopper increases in proportion to the vial diameter, it is necessary to consider the amount of white sugar per internal cross-sectional area of the container even when considering the influence of the water content of the rubber stopper. It is thought that it is necessary to set it to 50 mg / mm ² or more.

<Example 2: Production and evaluation of active drug formulation>
A carperitide-containing preparation (containing 1 mg of carperitide) was produced according to the sucrose blending conditions judged to be preferable in Example 1 (300 to 500 mg per vial having an inner diameter of 22.1 mm) and the sucrose concentration (125 mg / mL or less). The characteristics during production and the thermal stability were evaluated.
(1) Preparation of freeze-dried preparation of Formulation Example A (carperitide 1 mg / vial, sucrose amount 500 mg / vial (1.30 mg / mm ² ), sucrose concentration 100 mg / mL (10%)) carperitide 81.6 mg ( Carperitide designed content of 1 mg was increased by 2%) and 40 g of purified white sugar (Japanese Pharmacopoeia, Shisui Minato Seika Co., Ltd.) was dissolved in purified water, and the total volume was adjusted to 400 mL with purified water. This solution is filtered through a filter (Stericup-GV 0.22 μm, PVDF, Millipore Corporation), and then a Tahr dispenser (FH-10s, Hirasawa Co., Ltd.) or a microscale pipette (PIPETMAN, GILSON, Inc., etc.) is used. 5 mL was filled into a glass vial (outer diameter 24.5 mm, inner diameter 22.1 mm), Daiwa Special Glass Co., Ltd.). After half-plugging a rubber stopper (V10-F8 D713 RB2-40, Daikyo Seiko Co., Ltd.) into the filled vial, it is carried into a freeze dryer (RL-30KPs, Kyowa Vacuum Co., Ltd.) and pre-frozen (- 50 ° C setting), followed by freeze-drying under vacuum conditions (20 Pa setting) (primary drying: -20 to -5 ° C setting, secondary drying: 40 ° C setting). The pressure was restored. After all the rubber stoppers have been plugged into the vial, the vial is wrapped with a flip-off cap (AP cap, 20AP7.0H, Ishida Press Industry Co., Ltd.) using a winding machine (GAC-01, Inaba Machinery Co., Ltd.) Thus, a carperitide-containing preparation of Formulation Example A was obtained.

(2) Production of freeze-dried preparation of Formulation Example B (carperitide 1 mg / vial, sucrose amount 300 mg / vial (0.78 mg / mm ² ), sucrose concentration 75 mg / mL (7.5%)) carperitide 102 After dissolving 30 mg of purified sucrose (Japanese Pharmacopoeia, Shisui Minato Seika Co., Ltd.) and mg (designed content of carperitide with a 2% increase with respect to 1 mg design), the total volume was adjusted to 400 mL with purified water. This solution is filtered through a filter (Stericup-GV 0.22 μm, PVDF, Millipore Corporation), and then a Tahr dispenser (FH-10s, Hirasawa Co., Ltd.) or a microscale pipette (PIPETMAN, GILSON, Inc., etc.) is used. 4 mL was filled into a glass vial (outer diameter 24.5 mm, inner diameter 22.1 mm), Daiwa Special Glass Co., Ltd.). Thereafter, the carperitide-containing preparation of Formulation Example B was obtained by the same method as Formulation Example A.

(3) Evaluation of heat-severe storage stability of carperitide-containing preparations One month in a thermo-hygrostat (LH31-15M type, Nagano Kagaku Kikai Seisakusho Co., Ltd.) in which the preparations of Formulation Examples A and B were set to 40 ° C. and 75% RH It was stored for 1 month in a constant temperature bath (CH-P 30-01 type, Nagano Kagaku Kikai Seisakusho Co., Ltd.) set at 50 ° C. for 3 months or 3 months. About these preparations, visual appearance determination (manufacturing time: 9 products, after storage: 3 products) was performed according to the same criteria as in Example 1 at the time of manufacture, before each storage and after storage. Moreover, about the formulation after storage, content of carperitide and related substances was measured by the following method.

[Method: Determination of carperitide content and related substances in pharmaceutical preparations]
For each preparation, dilute acetic acid was added to dissolve the contents, and the total volume was adjusted to 10 mL to prepare a sample solution. Further, this sample solution was diluted 20 times with dilute acetic acid to obtain a standard solution for related substances. A separately prepared standard solution for quantifying carperitide was used.
Using a high-performance liquid chromatograph (Shimadzu Corporation), a sample solution, a standard solution for quantitative determination of carperitide, and a standard solution for testing related substances are measured using a reversed-phase separation system that uses an ODS column as the stationary phase and water / acetonitol as the mobile phase. (Measurement number: 3).
The content was calculated as follows. Calculate the amount of carperitide per vial from the measurement results of the sample solution and standard solution for quantitation of carperitide by high performance liquid chromatography (detection: ultraviolet absorbance method), and calculate the ratio of carperitide to the designed content (1 mg). Carperitide content (%).
The related substances were calculated as follows. From the measurement results of the sample solution and related substance standard solution by high performance liquid chromatography (ultraviolet absorbance detection), the ratio of the related substance peak area of the sample solution to the 20 times value of the carperitide peak area of the related substance standard solution is obtained. This was defined as the amount of related substances (%).

[Results: Formulation characteristics and thermal stability of carperitide-containing preparations]
Table 4 shows the appearance shape at the time of production of the preparations of Formulation Examples A and B, and the evaluation results of the appearance shape and components contained at 40 ° C., 75% RH for 1 month and 3 months, and at 50 ° C. for 1 month.
Formulation example A (sucrose amount: 500 mg / vial, 1.30 mg / mm ² , sucrose concentration: 100 mg / mL), and formulation example B (sucrose amount: 300 mg / vial: 0.78 mg / mm ² , sucrose Concentration: 75 mg / mL) Carperitide-containing preparations are all maintained well from the normal shape without change from the normal shape throughout the heat-harvest period of 40 ° C 75% RH 3 months and 50 ° C 1 month Is clear. Further, during these heat and severe storage periods, the decrease in the content of carperitide and the increase in the related substances are hardly observed, and it can be seen that the chemical stability of the main drug is very good. Moreover, the same test was separately conducted under the same refined sucrose blending conditions as in Blending Examples A and B, and the water content at the time of production and storage under severe heat was 0.5% or less. From the above, it was shown that lyophilized preparations prepared under these blending conditions have quality characteristics suitable for preparations that store at room temperature.

*: Describes the actual measurement (judgment code for 3 preparations).

Claims (11)

It contains an effective amount of a peptide drug and about 0.50 to 1.85 mg / mm ² of purified sucrose with respect to the internal cross-sectional area of the container, and is stable at room temperature storage. Freeze-dried preparation.   The freeze-dried preparation according to claim 1, wherein the freeze-dried product has a good external shape after storage at 40 ° C for 1 month, and the water content is 1% or less.   The freeze-dried preparation according to claim 1, wherein the freeze-dried product has a good external shape after storage at 50 ° C for 1 month, and the water content is 1% or less. The freeze-dried preparation according to claim 1, wherein the content of the purified sucrose is about 0.75 to 1.35 mg / mm ^{2 with} respect to the internal cross-sectional area of the container.   The lyophilized formulation according to claim 1, wherein the container is a cylindrical vial having an inner diameter of about 20 to 29 mm and contains about 300 to 500 mg of purified white sugar.   6. The lyophilized formulation of claim 5, wherein the cylindrical vial has an inner diameter of about 22.1 mm.   The lyophilized preparation according to any one of claims 1 to 6, produced by lyophilizing a lysate having a purified sucrose concentration of 140 mg / mL or less.   The lyophilized preparation according to any one of claims 1 to 7, comprising 1 mg of carperitide as a peptide drug.   The lyophilized preparation according to any one of claims 1 to 8, which substantially comprises purified sucrose and a peptide drug as solid components. An aqueous solution containing purified white sugar of 140 mg / mL or less and an effective amount of peptidic drug is placed in a container so that the amount is about 0.50 to 1.85 mg / mm ² with respect to the internal cross-sectional area of the container. A method for producing a freeze-dried preparation characterized by being stable at room temperature storage, comprising a step of lyophilizing the filled container and freeze-drying the filled container. 11. The production method according to claim 10, wherein in the lyophilized preparation produced, the ratio of the preparation having good preparation quality characteristics is about 80% or more.