WO2001047544A1

WO2001047544A1 - Protein-prefilled syringe preparation and method of stabilizing the same

Info

Publication number: WO2001047544A1
Application number: PCT/JP2000/009309
Authority: WO
Inventors: Yoshimi Imaeda; Daisuke Kameoka
Original assignee: Chugai Seiyaku Kabushiki Kaisha
Priority date: 1999-12-28
Filing date: 2000-12-27
Publication date: 2001-07-05
Also published as: AU2227501A

Abstract

A stabilized protein-prefilled syringe preparation equipped with a stopper which has a plastic film laminated at least to the surface in contact with the protein solution.

Description

Specification

TECHNICAL FIELD The present invention relates to a protein prefilled syringe preparation and a method for stabilizing the same.

The present invention relates to a protein prefilled syringe preparation which is easy to handle and stable for a long period of time. More specifically, the present invention relates to a stable protein prefilled syringe preparation comprising an elastic body coated with a resin on at least a protein solution contact surface. The invention further relates to a method for stabilizing a protein prefilled syringe formulation. Viewing technology

With the development of genetic recombination technology, various protein preparations have been provided in a stable supply amount. To ensure stability, these preparations are packaged separately with lyophilized protein component powder and a water-soluble diluent to dissolve it, and are provided in a form that can be dissolved at the time of use, or have improved stability It is provided in the form of a protein solution formulation to which additives are added. When comparing the two formulations, the solution formulation is advantageous because of the convenience in use. In particular, by dispensing a disposable syringe in advance and supplying it as a prefilled syringe solution preparation, the time required for dissolution operation and aspiration of the drug solution can be reduced, and quick response can be achieved. However, in the case of solution preparations, it is difficult to ensure stability.

Normally, protein prefilled syringe preparations contain diluents, solubilizers, tonicity agents, excipients, pH adjusters, buffers, sulfur-containing reducing agents, antioxidants, etc., in addition to the active ingredient protein. And provided in the market in a syringe. Satisfactory conditions for protein prefilled syringe preparations include (1) that protein products are stable under long-term storage conditions, (2) that they have heat resistance and pressure resistance that can withstand the conditions used for sterilization, 3) It has chemical resistance, (4) it has excellent sealing properties, and (5) it has good plunger slidability.

When preserving protein for a long period of time when preparing a prefilled syringe, the residual ratio of the protein decreased, and the stability was not always sufficient One of the factors of such a lack of stability is the material of the stopper of the disposable syringe used for the prefilled syringe preparation.

Conventionally, a syringe has been required to have a hermetic seal, and in order to satisfy this requirement, a rubber sealing stopper made of a single rubber material having the highest hermeticity has been used. However, in a prefilled syringe, the solution formulation and the rubber stopper are always in contact with the drug in the vial formulation, and the chemical effect of the drug from the rubber stopper is large, resulting in deterioration of the drug product. There is concern that maintaining long-term stability is difficult. Therefore, there is a demand for the development of a protein prefilled syringe formulation that can be stored for a long period of time, but a product that satisfies all the above requirements has not been developed. Disclosure of the invention

The present inventors have studied the above problems from various factors, and have found that a rubber stopper of a syringe used for a prefilled syringe formulation has an adverse effect on protein stabilization. In particular, there are variations in the surface treatment of rubber stoppers, which greatly affects the residual rate of protein solution preparations depending on the production lot of rubber stoppers, and the variation in residual rates among individuals within the same lot is large. I found that.

The present inventors have found that laminating a rubber stopper with Teflon can greatly contribute to the improvement of protein stability, particularly when the protein is granulocyte colony stimulating factor (G-CSF). That is, the present inventors have completed the present invention by finding that when a plastic film is laminated on the surface of a stopper made of a rubber material alone, a high content of protein can be retained for an extremely long time.

That is, the present invention provides a stable protein prefilled syringe preparation comprising an elastic body coated with a resin on at least the protein solution contact surface. The present invention provides the above-mentioned protein prefilled syringe preparation, comprising a stopper having a resin film laminated on at least a protein solution contact surface. The present invention provides the protein prefilled syringe preparation, wherein the resin is selected from polyethylene, polypropylene, polycarbonate, polyester, and fluororesin. I will provide a.

The present invention provides the protein prefilled syringe preparation, wherein the resin is a fluororesin.

The present invention provides the above protein prefilled syringe formulation, wherein the resin is polytetrafluoroethylene.

The present invention provides the above-mentioned protein pre-filled syringe preparation, wherein the protein is a recombinant protein.

The present invention provides the above protein prefilled syringe formulation, wherein the protein is erythropoietin.

The present invention provides the above-mentioned protein pre-filled syringe formulation, wherein the protein is a granulocyte colony stimulating factor.

The present invention provides the above protein pre-filled syringe preparation, wherein the protein is a protein having a sugar chain.

The present invention provides a method for stabilizing a protein prefilled syringe formulation, which comprises at least filling and storing a protein solution formulation in a syringe provided with an elastic body coated with a resin on the protein solution contact surface. BEST MODE FOR CARRYING OUT THE INVENTION

The material of the syringe used in the present invention may be a material known in the art, and may be glass or resin. Suitable resins include polymethylpentene resin, polypropylene resin, propylene-ethylene copolymer resin, polyester resin, polyethersulfone resin, polysulfone resin, or any of the above resins and polyamide resin, polyvinyl alcohol resin, ethylene vinyl acetate. It is preferable to use a resin obtained by laminating two or more layers of a resin selected from a resin and a resin. Particularly preferred resin materials for protein solution preparations include those described in Japanese Patent Application No. 11-254896. That is, it includes well-known medical container materials such as polyethylene (PE), polypropylene (PP), polyethylene terephthalate (PET), polycarbonate, and polyethylene methacrylate, and is preferably made of, for example, silicone such as lacyclododecene or a derivative thereof. Cyclopentyl residue or substituted cyclopentyl residue was inserted into the molecular chain by polymerization of ethylene- or propylene with ring-opening polymer and its hydrogenated product, cycloolefin such as norbornene or tetracyclododecene or its derivative, and ethylene or propylene. It is a resin that is a copolymer. Here, the cycloolefin includes monocyclic and polycyclic ones. Preferred is a thermoplastic norportene resin or a thermoplastic tetracyclododecene resin. Examples of the thermoplastic norbornene-based resin include a ring-opened polymer of a norbornene-based monomer, a hydrogenated product thereof, an addition-type polymer of a norbornene-based monomer, and an addition-type polymer of a norbornene-based monomer and an olefin. I can do it. Examples of the thermoplastic tetracyclododecene resin include a ring-opened polymer of a tetracyclododecene monomer, a hydrogenated product thereof, an addition polymer of a tetracyclododecene monomer, and a tetracyclododecene monomer. An addition type polymer of a decene monomer and an olefin is exemplified.

The elastic body used in the present invention is used as a stopper (slider) of a syringe and a rubber stopper for sealing, and is preferably made of a rubber body. The elastic body can also be manufactured using any known rubber elastic body used for pharmaceutical containers and medical equipment, and further copolymerized with natural rubber, isoprene rubber, butadiene rubber, butyl rubber, and divinyl benzene. Butyl rubber, chlorinated or brominated butyl rubber, ethylene-propylene copolymer rubber, isoprene-isobutylene copolymer rubber, isoprene-isobutylene copolymer halogenated rubber, thermoplastic elastomer and the like. As a thermoplastic elastomer, there is a technique of using a styrene-butadiene copolymer and a hydrogenated product of the copolymer as a plug for a pharmaceutical container (Japanese Patent Application Laid-Open No. 59-289695, Publication No. 57-266782, Japanese Patent Publication No. Hei 2-424296). Also, a technology in which a mixture of a thermoplastic elastomer and an isoprene-isobutylene copolymer is used as a rubber stopper (Japanese Patent Application Laid-Open No. 59-28965), and a fluoroelastomer is used for an isoprene-isobutylene copolymer. There is a laminated rubber stopper (Japanese Patent Publication No. 63-314104, Japanese Utility Model Publication No. 55-47950).

In the present invention, at least a protein solution contact surface of the elastic body is coated with a resin. As a coating method, various methods known to those skilled in the art, such as lamination (lamination) with a resin film or spraying (coating) of a resin, can be used. As the resin used in the present invention, any resin which is inert to the protein solution preparation filled in the prefilled syringe can be used. For example, polyethylene, polypropylene, polycarbonate, polyester, fluororesin and the like can be mentioned. Particularly preferred are fluororesins, such as polytetrafluoroethylene (PTF E: Teflon (registered trademark)), tetrafluoroethylene-ethylene copolymer (E TFE), and tetrafluoroethylene-hexane. Fluoropropylene copolymer (FEP), tetrafluoroethylene-fluoroalkylvinylether copolymer (PFA), polyvinylidene fluoride (PVDF), polychloro trifluoroethylene (PCTFE), etc. Can be Most preferred is polytetrafluoroethylene (PTFE). These resins are preferably used as a sheet or a film, and the thickness is not particularly limited, but is usually 1 to 500.

The elastic body used for the protein prefilled syringe preparation of the present invention is usually produced by compression molding using the above rubber composition and a resin film. Such a stopper is disclosed in, for example, JP-A-10-248929 and JP-A-10-314305.

Proteins used as an active ingredient in the present invention include, for example, hematopoietic factors such as granulocyte colony-stimulating factor (G-CSF), granulocyte macrophage colony-stimulating factor (GM-CSF), erythropoietin (EP〇), and tropopoetin; Cytochromes such as IL-1 and IL-6, monoclonal antibodies, tissue plasminogen-activating factor (TPA), perokinase, serum albumin, blood coagulation factor VIII, lebutin, insulin , Including but not limited to, stem cell growth factor (SCF). Among proteins, hematopoietic factors such as EP〇, G-CSF, and thrombopoietin, and monoclonal antibodies are preferable, and EP〇, G-CSF and monoclonal antibodies are more preferable.

The protein used as an active ingredient in the present invention is a protein having substantially the same biological activity as that of a biologically active protein of a mammal, particularly a human, and is obtained from a naturally-occurring protein or a recombinant gene. Preferred are those obtained by the gene recombination method. In particular, the protein obtained by the genetic recombination method is unstable compared to the natural protein, and it is preferable to use the preparation of the present invention. Good. Proteins obtained by the genetic recombination method include those having the same amino acid sequence as the natural protein, or those having one or more of the amino acid sequences deleted, substituted, or added and having the biological activity. Including. Furthermore, proteins include those chemically modified with PEG and the like.

Examples of the protein used as an active ingredient in the present invention include a protein having a sugar chain. The origin of the sugar chain is not particularly limited, but a sugar chain added to mammalian cells is preferable. Mammalian cells include, for example, Chinese hamster ovary cells (CHO cells), BHK cells, COS cells, human-derived cells and the like, among which CHO cells are most preferred.

When the protein used as the active ingredient in the present invention is EPO, EP〇 may be produced by any method, extracted from human urine by various methods, separated and purified, using a genetic engineering technique ( For example, according to Japanese Unexamined Patent Publication No. Sho 61-12288), hamster ovary cells (CHO), BHK cells, COS cells, human-derived cells, and the like, which are extracted and separated and purified by various methods, are used. Furthermore, it includes EPO chemically modified by PEG or the like (see International Patent Application Publication No. W090 / 12874). Furthermore, EPO with no sugar chain is chemically modified with PEG or the like. Also included are EPO analogs that have been modified to increase the number of one or more glycosylation sites at the N-linked carbohydrate chain binding site or the O-linked carbohydrate chain binding site in the amino acid sequence of EPO (eg, (See Kaihei 8—151398, Tokuhei 8-506023). Further, the amount of sugar chains may be increased by increasing the content of sialic acid or the like without changing the number of sugar chain binding sites.

When the protein used as an active ingredient in the present invention is G-CSF, any G-CSF can be used as long as it is highly purified G-CSF. The G-CSF in the present invention may be produced by any method, such as those obtained by culturing a cell line of human tumor cells and extracting and separating and purifying it from various cells, or E. coli by genetic engineering techniques. Bacteria; Listeria monocytogenes; Chinese hamster ovary (CH 、) cells, C127 cells, COS cells, and other animal-derived culture cells that are extracted and purified by various methods. Can be Like Alternatively, it is produced by E. coli, yeast, or CHO cells using a genetic recombination method. Most preferably, it is produced by a genetic modification method using CH 組換え cells. Furthermore, G-CSF chemically modified with PEG or the like is also included (see International Patent Application Publication No. WO 90/12874).

When the bioactive protein used as an active ingredient in the present invention is a monoclonal antibody, the monoclonal antibody may be produced by any method. Monoclonal antibodies are basically immunized with a sensitizing antigen according to a usual immunization method using a known technique, and the obtained immune cells are fused with a known parent cell by a normal cell fusion method. Can be prepared by screening monoclonal antibody-producing cells by the screening method described above. Further, the monoclonal antibody is not limited to the monoclonal antibody produced by the hybridoma, and includes a chimeric antibody artificially modified for the purpose of, for example, reducing the xenoantigenicity to humans. Alternatively, a reshaped (reshaped) humanized antibody can also be used in the present invention. This is obtained by replacing the complementarity-determining region of a human antibody with the complementarity-determining region of a non-human mammal, for example, a mouse antibody, and a general gene recombination technique is also known. Using the known method, a reshaped humanized antibody useful in the present invention can be obtained.

If necessary, amino acids in the framework (FR) region of the variable region of the antibody may be substituted so that the complementarity determining region of the reshaped humanized antibody forms an appropriate antigen-binding site (S ato et al., Cancer Res. 53: 1-6, 1993). A preferred example of such a reshaped humanized antibody is a humanized PM-1 (hPM-1) antibody which is an anti-IL-16 receptor humanized antibody (International Patent Application Publication No. WO 92-19759).

Furthermore, the antibodies of the present invention also include human antibodies produced by known techniques using transgenic animals and the like.

The present invention can be applied to antibody fragments (for example, Fab, F (ab) _2, etc.) and modified antibodies (for example, single-chain antibodies, etc.).

In particular, chimeric antibodies, humanized antibodies, antibody fragments, and modified antibodies are more unstable than natural antibodies, and therefore it is preferable to use the formulations of the present invention. The protein preparation of the present invention may contain a diluent, a solubilizing agent, a tonicity agent, an excipient, a pH adjuster, a soothing agent, a buffer, a sulfur-containing reducing agent, an antioxidant, etc. Good. For example, as the tonicity agent, polyethylene glycol; dextran, mannitol, sorbitol, inositol, glucose, flak, and lac! You can use sugars such as sucrose, xylose, mannose, maltose, sucrose, and raffinose. Examples of the sulfur-containing reducing agent include N-acetyl cysteine, N-acetyl homocystine, thioctic acid, thiodiglycol, thioethanolamine, thiodaricerol, thiosorbitol, thioglycolic acid and salts thereof, sodium thiosulfate, and dal. And those having a sulfhydryl group such as thiothione and thioalkanoic acid having 1 to 7 carbon atoms. Examples of antioxidants include erythorbic acid, dibutylhydroxytoluene, butylhydroxyanisole, α-tocopherol, tocopherol acetate, L-ascorbic acid and its salts, L-ascorbic acid palmitate, and L-ascorbic acid Chelating agents such as stearate, sodium bisulfite, sodium sulfite, triamyl gallate, propyl gallate or disodium ethyl diamine tetraacetate (EDTA), sodium pyrophosphate, sodium metaphosphate and the like. In addition, inorganic salts such as sodium chloride, potassium chloride, calcium chloride, sodium phosphate, potassium phosphate, and sodium hydrogen carbonate; and solution preparations such as organic salts such as sodium citrate, potassium citrate, and sodium acetate. It may contain components that are usually added.

The protein solution preparation of the present invention may further contain an appropriate stabilizer for each protein, and the stabilizer includes a surfactant (for example, sorbitan fatty acid ester or glycerin fatty acid ester which is a nonionic surfactant). , Polyglycerin fatty acid ester, polyoxyethylene sorbitan fatty acid ester, polyoxyethylene sorbite fatty acid ester, polyoxyethylene glycerin fatty acid ester, polyethylene glycol fatty acid ester, polyoxyethylene alkyl ether, polyoxyethylene polyoxypropylene alkyl ether, Polyoxyethylene alkylphenyl ether, polyoxyethylene hydrogenated castor oil, polyoxyethylene beeswax derivative, polyoxyethylene lanolin derivative, polyoxyethylene fatty acid Amide; alkyl sulfate as anionic surfactant, polio: Alkyl ether sulfates and alkyl sulfosuccinates; natural surfactants such as lecithin, glycerol phospholipids, sphingophospholipids, and sucrose fatty acid esters, and particularly preferred are polyoxyethylene sorbitan fatty acid esters, Polyoxyethylene sorbitan monooleate (polysorbate 80) and polyoxyethylene sorbitan monolaurate (polysorbate 20) are preferred, and amino acids (eg, D-, L- and DL-forms) Of leucine, tryptophan, serine, glutamic acid, arginine, histidine, lysine, methionine, fenylalanine and acetyltributophane and salts thereof, and more preferred are L-leucine, L-tributophan, and L-dalminic acid. , L Arginine, L- histidine and a L- lysine, and their salts) including such as, but not limited thereto.

The stable protein prefilled syringe preparation of the present invention is usually administered by parenteral administration route, for example, injection (subcutaneous or intravenous injection), transdermal, transmucosal, nasal, etc., but oral administration is also possible.

The amount of protein contained in the stable protein prefilled syringe formulation of the present invention can be determined according to the protein used, the type of disease to be treated, the severity of the disease, the age of the patient, and the like.

Generally, 0.01 g / m1 or more, preferably 0.1 gZm1 or more, more preferably 1 gZm1 or more, more preferably 10 / ig Zm1 or more, further preferably 50 g / m Contains 1 or more proteins. In particular, in the case of a prefilled syringe solution preparation, it contains 0.1 gZml or more, preferably 1 / xg noml or more, more preferably 10 g / m1 or more, and still more preferably 50 gZm1 or more.

For example, in the case of EPO, the amount of EP〇 in the solution formulation is generally between 100 and 500 000 I UZm, preferably between 200 and 100000 I UZm and more preferably between 750 and 72000 1 U / m 1. In the case of G-CSF, it is generally l to 1000 g / m 1, preferably 10 to 800 // g Zm, and more preferably 50 to 500 / ig / m 1. In addition, in the case of an anti-IL-16 receptor humanized antibody, the final administration concentration is generally 0.1 to 20 OmgZm, preferably 1 to 20 OmgZm. ~ 12 Omg / m1.

The protein prefilled syringe formulation of the present invention can be prepared by dissolving these components in an aqueous buffer known in the field of solution formulation, such as phosphate and Z or citrate buffers. The phosphate buffer is preferably a sodium monohydrogen phosphate sodium dihydrogen phosphate system, and the citrate buffer is preferably a sodium citrate buffer solution. Further, depending on the type of protein, a buffer such as a glycine buffer or a histidine buffer may be used.

When the evening protein prefilled syringe preparation of the present invention is a erythrocyte poetin solution preparation, it contains EP〇, a nonionic surfactant (for example, polysorbate 80, polysorbate 20), an isotonic agent ( It preferably contains, for example, sodium chloride) and, if necessary, a stabilizer (eg, an amino acid, preferably L-histidine), and has a pH of 5.0 to 8.0, preferably 5.5 to 7.0.

When the protein prefilled syringe preparation of the present invention is a G-CSF solution preparation, the preparation contains G-CSF, a nonionic surfactant (for example, polysorbate 80, polysorbate 20), and if necessary, a diluent. , A solubilizing agent, a tonicity agent, an excipient, a pH adjuster, a soothing agent, a buffer, a sulfur-containing reducing agent, an antioxidant, etc., and a pH of 5.0-7.0, preferably It is preferred to be 6-6.8. When the protein prefilled syringe preparation of the present invention is an anti-IL16 receptor-humanized antibody solution preparation, the anti-IL16 receptor humanized antibody is preferably used in a glycine buffer and a Z or histidine buffer. Dissolved in a buffer solution such as a liquid, and other nonionic surfactants (for example, polysorbate 80, polysorbate 20), and if necessary, diluents, dissolution aids, isotonic agents, excipients, and pH adjusters. It preferably contains a soothing agent, a buffer, a sulfur-containing reducing agent, an antioxidant, and the like, and has a pH of 5 to 8.

A stable protein pre-filled syringe formulation pre-filled in a syringe equipped with a stopper in which the plastic film of the present invention was laminated at least on the protein solution contact surface was tested using G-CSF, as shown in Examples described later. Even after a 2-week accelerated test at 40 ° C, the G-CSF retention rate is much better than that of a non-laminated stopper. Or 6 months at 25 ° C, 1 Even after long-term storage at 0 ° C for 1 year, it shows good G-CSF retention. Also, if the protein is EPO, even after accelerated tests at 50 ° C-2 weeks or 40 ° C-1 month, or at 6 months at 25 ° C and 1 year at 10 ° C Very good EPO persistence after storage.

The present invention also provides a method for stabilizing a protein prefilled syringe formulation, which comprises filling and storing a protein solution formulation in a syringe provided with a stopper having a plastic film laminated on at least the protein solution contact surface. The stabilization in the present invention depends on the type of protein to be filled. For example, in the case of a G-CSF solution preparation (containing 12 G-CSF), this is stored at, for example, 40 ° C for 2 weeks or more. At that time, it means that the residual ratio of G-CSF is maintained at 75% or more, preferably 80% or more, and more preferably 85% or more. Alternatively, after storing a G-CSF solution preparation (containing 250 g / ml of G-CSF) at 25 ° C for 6 months, the residual ratio of G-CSF is 80% or more, preferably 85% or more, more preferably Means to keep it above 90%.

According to the present invention, a protein prefilled syringe preparation can be stably stored at room temperature for a long period of time. Industrial applicability

A stable protein prefilled syringe formulation comprising a stopper in which the plastic film of the present invention is laminated at least on the protein solution contacting surface is a stable protein prefilled syringe formulation. More stable than syringe formulations. According to the present invention, a protein prefilled syringe preparation can be stably stored at room temperature for a long period of time. In particular, when the protein is obtained by a genetic recombination method, it is unstable compared to natural proteins, and should be stably stored at room temperature for a long period of time by using the prefilled syringe preparation of the present invention. Can be. Example

In the following examples, granulocyte colony stimulating factor (G-CSF) is used as a representative example. The results of an accelerated test are described below, but the scope of the present invention is not limited to this. Various changes and modifications can be made by those skilled in the art.

In the following examples, the evaluation of the preparation was performed by determining the content of G-CSF using RP-HPLC analysis.

Example 1: G-CSF solution at 40 ° C for 2 weeks accelerated test

Preparation of G-CSF solution preparation

The following ingredients in 1 ml of the preparation solution:

G-C S F 1 25 g (Polysorbate 20) 0.lmg

Sodium chloride 7.5mg

And adjusted to pH 6.5 with 1mo 1 ZL hydrochloric acid.

Test method

After the G-CSF solution preparation prepared as described above was aseptically filtered, 0.5 mL each was aseptically filled into the container form shown in Table 1 below and stoppered.

The G-CSF used in this example is a recombinant protein having a sugar chain (rG-CSF) produced in CHO cells. Container form

A PTFE-laminated glass syringe is filled with the above dispensing solution, and the stopper is sealed and stoppered by attaching a PTFE sheet to the liquid contacting surface of a WEST stop stopper (Material: bromobutyl rubber PH4023 / 50). .

Untreated stopper A syringe filled with the above preparation liquid and sealed with a stopper (material: bromobutyl rubber PH4023 / 50) manufactured by WEST.

PTFE laminating stock A PTFE sheet with a PTFE sheet attached to the liquid contacting surface of a WEST stopper (material: PTFE is peeled from bromobutyper), and the liquid contacting surface is used. Was peeled off. A glass syringe filled with the above dispensing solution and sealed and stoppered using this stopper.

Glass vial An untreated white glass vial filled with the above preparation liquid and stoppered with a rubber stopper S5-F3 manufactured by Daikyo Seiki. Each container form containing the G-CSF preparations shown in Table 1 aseptically prepared and filtered was subjected to two-week acceleration in a constant-temperature bath at 40 ° C. For the unaccelerated sample and the sample accelerated for 2 weeks at 40 ° C, reverse phase using pure water, acetonitrile, and trifluoroacetic acid as the mobile phase using a C4 reversed-phase column (4.6 iMi X 250 imn, 300 Å). G-CSF content was measured by high-performance liquid chromatography.

A considerable amount of G-CSF was injected, and G-CSF was eluted with an acetonitrile gradient, and the G-CSF content was measured by spectroscopic detection at a wavelength of 215ηπι. Using the G-CSF content measured by this method, the survival rate (%) after acceleration at 40 ° C for 2 weeks was calculated based on the following equation.

(G-CSF content of accelerated sample at 40 ° C for 2 weeks)

Survival rate (%) = X 100

(G-CSF content of unaccelerated sample)

The results obtained are shown in Table 2 below.

Table 2

As shown in Table 2, the instability of G-CSF observed when using the untreated stopper was not observed at all with the product using the PTFE laminate stopper, and showed the same good stability as the product using the glass spear. Was. In addition, when a PTFE laminate stopper with a PTFE sheet peeled was used, the same degree of instability as the untreated stopper was used, indicating the importance of the PTFE laminate. Example 2: Storage stability test of G-CSF solution formulation at 25 ° C and 10 ° C

In addition to the G—CSF solution preparation (125 / xgZlm) used in Example 1, the following high DOSE preparation was prepared.

Preparation of G-CS F (250/2 R / lm 1) solution preparation

The following ingredients in 1 ml of the preparation solution:

GC SF 250 g Port

(Polysorbate 20) 0 1 mg

Sodium chloride 6 Omg

And adjusted to pH 6.5 with lmo 1 / L hydrochloric acid,

Test method

After sterile filtration of the two G—CSF solution preparations prepared as described above,

5 mL each was aseptically filled into a container equipped with a PTFE laminate stopper (A to E) and stoppered.

In this way, each container form containing the solution preparation prepared by aseptic preparation and filtration was stored at 25 ° C and 10 ° C for a long time. Thereafter, the residual ratio was calculated in the same manner as in Example 1. Tables 3 and 4 show the results.

Table 3

125 / z / mL (0.5mL filling)

Container form Residual rate at 25 ° C (%) Residual rate at 10 ° C (%)

2 months 4 months 6 months 3 months 6 months 1 year

PTFE laminated stripper, 92.6 90.1 85.4 94.7 93.7 90.7 A

PTFE laminated stopper, 88.9 85.8 72.9 91.9 93.1 85.5 B

PTFE laminated stopper, 91.2 90.2 78.1 94.1 93.4 89.9 C

PTFE laminated stopper, 92.3 89.8 77.9 94.8 92.8 89.6 D

PTFE laminating stopper, 91.6 89.8 84.0 94.2 92.7 88.4 E

Table 4

As is clear from the table, the G-CSF preparation containing no stabilizer other than the surfactant and sodium chloride exhibited a good residual ratio. Example 3: Stability test of EPO solution preparation

EPO solution preparations of the following two concentrations (1500 IUZmL and 48000 IU / mL) were prepared, filled aseptically into containers equipped with PTFE laminate stoppers (BD), and stoppered. Accelerated tests were performed at 50 ° C and 40 ° C, and long-term storage tests were performed at 25 ° C and 10 ° C, and the residual rates were calculated respectively. The results are shown in Table 5-8. Formulation: 1500IU L 48000IU / mL

EPOCH 1500IU 48000IU Polysorbate 80 0.05 mg 0.05 mg L-Histidine hydrochloride 1.35 mg 1.35 mg Sodium chloride 7.74 mg 2.26 mg Phosphate buffer 10 mM (pH 6.0) 25 mM (pH 6.0) Table 5

Table 6

Table 7

8

All solution preparations showed extremely good residual rates.

Claims

The scope of the claims

1. A stable protein prefilled syringe formulation comprising an elastic body coated with a resin at least on the protein solution contact surface.

2. The stable protein prefilled syringe preparation according to claim 1, further comprising a stopper having a resin film laminated on at least a protein solution contact surface.

3. The protein prefilled syringe preparation according to claim 1, wherein the resin is selected from polyethylene, polypropylene, polycarbonate, polyester, and fluororesin.

4. The preprotein syringe preparation according to claim 3, wherein the resin is a fluororesin.

5. The protein prefilled syringe preparation according to claim 4, wherein the resin is polytetrafluoroethylene.

6. The protein prefilled syringe preparation according to any one of claims 1 to 5, wherein the protein is a recombinant protein.

7. The protein pre-filled syringe preparation according to claim 6, wherein the protein is erythropoietin.

8. The protein preeryred syringe preparation according to claim 6, wherein the protein is a granulocyte colony stimulating factor.

9. The protein prefilled syringe preparation according to any one of claims 1 to 8, wherein the protein is a protein having a sugar chain.

10. A method for stabilizing a protein prefilled syringe formulation, which comprises filling and storing a protein solution formulation in an injector having an elastic body coated with a resin on at least the protein solution contact surface.