Package and method for manufacturing same

Abstract

Provided is a package in which a pharmaceutical composition containing a glycolytic enzyme is accommodated in a container, said package being capable of suppressing a decrease in titer due to a small amount of enzyme. The packaging body contains a pharmaceutical composition and a container, the pharmaceutical composition is a lyophilized preparation containing a glycolytic enzyme as an active ingredient, the container contains the pharmaceutical composition, and the inner surface of the container is provided with at least one material selected from the group consisting of fine ceramics, silicone resins, and fluororesins.

Description

[0001] This application is a divisional application based on the Chinese invention patent application with application number 201980014035.1, application date February 27, 2019, and invention name “Packaging body and its manufacturing method”. Technical Field

[0002] The present invention relates to a package body containing a pharmaceutical composition containing an enzyme as an active ingredient in a container, and a method for producing the same. Background Art

[0003] Pharmaceutical compositions containing glycolytic enzymes as active ingredients can be used in various disease areas. For example, pharmaceutical compositions for treating lysosomal storage diseases containing about 3 mg / bottle to about 10 mg / bottle of glycolytic enzymes as active ingredients, such as Aldurazyme (registered trademark), Elaprase (registered trademark), Naglazyme (registered trademark), Replagal (registered trademark), and Vimizim (registered trademark), are commercially available as liquid preparations for injection. In addition, for example, International Publication No. 2012 / 081227 describes a therapeutic agent for herniated intervertebral disc containing glycolytic enzymes (particularly chondroitinase ABC) as active ingredients. Summary of the invention

[0004] Compared with liquid preparations, freeze-dried preparations are excellent in reducing logistics costs. On the other hand, the titer of the enzyme is sometimes reduced due to freeze-drying, so when manufacturing freeze-dried preparations, efforts are made to produce products with desired enzymatic activity. For example, in the embodiment of International Publication No. 2012 / 081227, considering that the titer caused by a small amount of enzyme in freeze-drying is reduced, the enzyme solution prepared by the amount that greatly exceeds the amount required for administration once, that is, the unit dosage, is stored in a container to obtain freeze-dried preparations. And, in this document, after the freeze-dried preparation obtained is dissolved and diluted, the necessary amount is divided to prepare the administration liquid containing the active ingredient required for a single administration.

[0005] Here, in the method of preparing an enzyme solution in advance in an amount exceeding the unit dosage and taking a portion of it as a single dosage, the remaining enzyme not used for administration is discarded. Therefore, the remaining part of the expensive enzyme is sometimes wasted. In addition, since a small amount of operation is required to be temporarily taken from the prepared enzyme solution, it is also necessary to pay attention to the reduction of activity in this operation.

[0006] On the other hand, the present inventors have found that, in the manufacture of a package containing a freeze-dried preparation, the reduction in potency caused by a small amount of glycolytic enzyme contained is unexpectedly large. The present inventors have also found that such a reduction in potency is particularly significant when the amount of glycolytic enzyme contained is lower than that of a conventional pharmaceutical composition.

[0007] Therefore, an object of the present invention is to provide a package body in which a pharmaceutical composition containing a glycolytic enzyme as an active ingredient is contained in a container, wherein the package body can suppress a decrease in potency caused by a small amount of the contained glycolytic enzyme.

[0008] One aspect of the present invention relates to a package comprising a pharmaceutical composition and a container, wherein the pharmaceutical composition is a freeze-dried preparation containing a glycolytic enzyme as an active ingredient, the container accommodates the pharmaceutical composition, and the inner surface of the container has at least one material selected from fine ceramics, silicone resins, and fluororesins.

[0009] Another aspect of the present invention relates to a method for manufacturing a packaging body, wherein the packaging body contains a pharmaceutical composition and stores the pharmaceutical composition, and the method for manufacturing the packaging body comprises: a step of storing a solution containing a glycolytic enzyme in a container, wherein the inner surface of the container has at least one material selected from fine ceramics, silicone resin and fluororesin; and a step of freeze-drying the solution to obtain the pharmaceutical composition. DETAILED DESCRIPTION

[0010] According to one aspect of the present invention, there is provided a package body in which a pharmaceutical composition containing a glycolytic enzyme is contained in a container, wherein the package body can suppress a decrease in potency due to a small amount of the contained glycolytic enzyme.

[0011] Hereinafter, the present invention will be described in detail, but the present invention is not limited to these embodiments. In this specification, the term "process" is not only an independent process, but also is included in the term as long as the desired purpose of the process can be achieved even if it cannot be clearly distinguished from other processes. In addition, the content of each component in the composition, when there are multiple substances belonging to each component in the composition, refers to the total amount of the multiple substances present in the composition unless otherwise specified.

[0012] (1) Pharmaceutical composition and packaging The pharmaceutical composition is a freeze-dried preparation containing glycolytic enzyme as an active ingredient. The pharmaceutical composition can be a unit dosage preparation. In this specification, "unit dosage" is the necessary amount prepared for a single administration, and the unit dosage preparation is prepared by formulating the pharmaceutical composition in a unit dosage. In addition to the effective amount, the unit dosage may also include the additional input amount required for preparing a single administration solution. The package includes at least a container and a pharmaceutical composition contained in the container.

[0013] "Glycolytic enzyme" is not particularly limited as long as it is an enzyme that can be used in medicine. For example, examples of glycolytic enzymes include glucosaminoglycan glycolytic enzymes; glycosidases; peptides: N-glycanases (PNGaseF, endoglycosidase H, etc.), α-L-iduronidase, α-galactosidase, β-galactosidase, β-glucuronidase, β-glucocerebrosidase, idursulfase, iduronic acid-2-sulfatase, N-acetylgalactosamine-6-sulfatase, N-acetylgalactosamine-4-sulfatase, etc. As the glycosaminoglycan glycolytic enzyme, for example, cutinase I, cutinase II and other cutinase; heparinase I, heparinase II, heparinase III and other heparinase; heparinase IV, heparinase V, heparinase TI, heparinase T-II, heparinase T-III, heparinase T-IV and other heparinase; chondroitinase ABC, chondroitinase ACI, chondroitinase ACII, chondroitinase ACIII, chondroitinase B, chondroitinase C and other chondroitinase; hyaluronidase from actinomycetes, hyaluronidase from streptococcus and other hyaluronidase, etc. In addition, as glycosidase, for example, β-galactosidase and α-galactosidase from microorganisms can be mentioned.

[0014] In one embodiment, the glycosaminoglycan glycolytic enzyme can be used as a glycolytic enzyme. As the glycosaminoglycan glycolytic enzyme, hyaluronidase, chondroitinase, heparanase, cutanase, acetylheparanase, heparanase, etc. can be cited. As a glycolytic enzyme, wherein, preferably chondroitinase, more preferably chondroitinase ABC, chondroitinase B, chondroitinase ACI, chondroitinase ACII, particularly preferably chondroitinase ABC (also referred to as Hernicotinase (condoliase)). It should be noted that chondroitinase ABC is an enzyme that decomposes hyaluronic acid, chondroitin sulfate, chondroitin, dermatan sulfate, etc. into disaccharides or oligosaccharides containing unsaturated sugars.

[0015] The source of glycolytic enzyme is not particularly limited. In a preferred embodiment, glycolytic enzyme from microorganism can be used. As non-limiting examples of microorganisms, for example, Bacillus, Escherichia coli, Pseudomonas, Flavobacterium, Proteus, Arthrobacter, Streptococcus, Bacteroides, Aspergillus, Elizabethkingia and Streptomyces can be cited. For example, in the case where the glycolytic enzyme is chondroitinase ABC, an enzyme from Proteus vulgaris (e.g., chondroitinase ABC from Proteus vulgaris) can be exemplified.

[0016] The method for producing the glycolytic enzyme is not particularly limited. An exemplary method for producing the glycolytic enzyme includes a step of obtaining a culture of a microorganism or animal cell that produces the glycolytic enzyme, and a step of recovering the glycolytic enzyme from the culture.

[0017] The glycolytic enzyme produced by the microorganism can be the enzyme originally possessed by the microorganism, or can be the enzyme obtained by modifying the microorganism by the genetic engineering method described later so that it produces the enzyme as the target. For example, when the glycolytic enzyme is chondroitinase ABC, microorganisms such as common Proteus can be cultivated to produce glycolytic enzyme, or the DNA encoding chondroitinase ABC can be used to produce glycolytic enzyme using genetic engineering methods. In addition, the glycolytic enzyme can be the same amino acid sequence as the enzyme originally possessed by the organism, but as long as the expected purpose as a pharmaceutical can be achieved, it is also possible to lack, replace and / or add part of amino acids, etc.

[0018] As microorganisms, for example, Bacillus, Escherichia coli, Pseudomonas, Flavobacterium, Proteus, Arthrobacter, Streptococcus, Bacteroides, Aspergillus, Elizabeth and Streptomyces can be exemplified. The growth conditions of the microorganism (e.g., culture medium, culture conditions, etc.) can be appropriately selected according to the microorganism used, and can be arbitrarily set by those skilled in the art. By using microorganisms to manufacture glycolytic enzymes, compared with the situation of using animal cells to manufacture glycolytic enzymes, it can be economically and mass-produced.

[0019] The method for producing a glycolytic enzyme may include the step of introducing a recombinant vector expressing a gene encoding a target glycolytic enzyme into a host. As a vector, for example, an appropriate expression vector (preferably comprising a regulatory sequence such as a promoter) capable of expressing the introduced gene (e.g., a phage vector, a plasmid vector, etc.) may be used. The vector is appropriately selected according to the host cell. More specifically, as such a host-vector system, a combination of Escherichia coli (E. coli) and expression vectors for prokaryotic cells such as pET series, pTrcHis, pGEX, pTrc99, pKK233-2, pEZZ18, pBAD, pRSET, and pSE420 may be exemplified; a combination of mammalian cells such as COS-7 cells and HEK293 cells and expression vectors for mammalian cells such as pCMV series, pME18S series, and pSVL, and, as a host cell, insect cells, yeast, Bacillus subtilis, etc. may be exemplified, and various vectors corresponding to these may be exemplified.

[0020] In addition, the above-mentioned vector can also be constructed by expressing a fusion protein of a recombinant gene-encoded protein and a marker peptide, a signal peptide, etc. As the peptide, for example, protein A, insulin signal sequence, histidine tag (HisTag), FLAG, CBP (calmodulin binding protein), GST (glutathione-S-transferase), etc. can be cited. No matter which vector is used, it can be treated by restriction endonuclease according to a conventional method so that the inserted nucleic acid sequence can be linked to the vector, and smoothing, sticky end connection, etc. can be performed as needed, and then the inserted nucleic acid sequence can be linked to the vector.

[0021] Transformation using a host vector can be performed by conventional methods. For example, the vector can be introduced into a host for transformation by methods such as using a commercially available transfection reagent, DEAE-dextran, electroporation, or a gene gun.

[0022] The growth conditions (e.g., culture medium, culture conditions, etc.) of the microorganism or animal cell producing the glycolytic enzyme can be appropriately selected according to the microorganism or cell used. For example, when using Escherichia coli, a culture medium appropriately prepared using LB culture medium as a main component can be used. In addition, for example, when using COS-7 cells as host cells, DMEM culture medium containing about 2% (v / v) fetal bovine serum can be used and cultured at 37°C.

[0023] Glycolytic enzymes can be recovered from the growth by known protein extraction and purification methods according to the form of the produced glycolytic enzymes. For example, when the glycolytic enzymes are produced in a soluble form secreted in the culture medium (the supernatant of the culture solution), the culture medium can be taken and used directly as glycolytic enzymes. In addition, when the glycolytic enzymes are produced in a soluble form or an insoluble (membrane-bound) form secreted in the cytoplasm, they can be extracted by the following treatment operations, that is, extraction by breaking cells using a nitrogen cavitation device, homogenization, glass bead grinding, sonication, osmotic shock, freeze-thaw method, etc., extraction by a surfactant, or a combination thereof. Glycolytic enzymes can also be purified by the following conventionally known methods, that is, salting out, ammonium sulfate fractionation, centrifugal separation, dialysis, ultrafiltration, adsorption chromatography, ion exchange chromatography, hydrophobic chromatography, reverse phase chromatography, gel permeation chromatography, affinity chromatography, electrophoresis, etc., and combinations thereof.

[0024] Glycolytic enzymes can be used alone or in combination of two or more. In addition, glycolytic enzymes can also be enzymes with chemical modification groups known in the past such as acetylation, polyalkylene glycolation (e.g., pegylation), alkylation, acylation, biotinylation, labeling (e.g., labeling with fluorescent substances, luminescent substances, etc.), phosphorylation, and sulfation added.

[0025] The pharmaceutical composition may contain a pharmaceutically acceptable carrier. In the present specification, examples of "pharmaceutically acceptable carrier" include conventional excipients, adhesives, buffers, water for injection, isotonic agents, preservatives, analgesics, and other components generally used in medicine.

[0026] Examples of the buffer include a buffer containing one or more selected from the group consisting of hydrochloric acid, sodium hydroxide, sodium carbonate, sodium bicarbonate, phosphoric acid, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, sodium dihydrogen phosphate, disodium hydrogen phosphate, aminoacetic acid, sodium benzoate, citric acid, sodium citrate, acetic acid, sodium acetate, tartaric acid, sodium tartrate, lactic acid, sodium lactate, ethanolamine, arginine, and ethylenediamine, preferably at least one of sodium dihydrogen phosphate and disodium hydrogen phosphate.

[0027] Examples of the isotonic agent include sodium chloride, potassium chloride, glycerin, mannitol, sorbitol, boric acid, borax, glucose, and propylene glycol.

[0028] As other pharmaceutically acceptable carriers, specifically, for example, dextran, sucrose, lactose, maltose, xylose, trehalose, mannitol, xylitol, sorbitol, inositol, serum albumin, gelatin, creatinine, polyalkylene glycol, nonionic surfactant (e.g., polyoxyethylene sorbitan fatty acid ester, polyoxyethylene hardened castor oil, sucrose fatty acid ester, polyoxyethylene polyoxypropylene glycol), etc., preferably at least one of sucrose and polyalkylene glycol, more preferably at least one of sucrose and polyethylene glycol. The average molecular weight of polyethylene glycol is preferably 200 or more and 25000 or less. In addition, polyethylene glycol is solid at room temperature, for example, the average molecular weight is more preferably 2000 or more and 9000 or less, and further preferably 3000 or more and 4000 or less. As polyethylene glycol, ethylene glycol with an average molecular weight of 3250, 3350 and 4000 can be exemplified. When a mixture of polyethylene glycol and sucrose is used as a pharmaceutically acceptable carrier, the weight ratio of polyethylene glycol / sucrose is generally preferably in the range of 1 / 10 to 10 / 1, and the weight ratio of polyethylene glycol / sucrose is more preferably about 2 / 1.

[0029] In this specification, "container" is not particularly limited as long as it can accommodate the pharmaceutical composition. As a container, a syringe, a vial, an ampoule, a syringe, etc. can be exemplified, preferably a vial. The base material of the container can be exemplified by glass, plastic, etc., preferably glass. Glass can include, for example, borosilicate glass, soda-lime glass, etc. The container more preferably further has a plug member or a lid, and more preferably has a rubber stopper. The size of the container is not particularly limited, and can be exemplified by more than 0.5mL and less than 100mL, preferably more than 1mL and less than 10mL, more preferably more than 2mL and less than 4mL, and more preferably 3mL. The package formed by the pharmaceutical composition being accommodated in the container can be enclosed with inactive gases such as nitrogen and argon, and can also be degassed.

[0030] The container may have at least one material selected from fine ceramics, silicone resins and fluororesins on its inner surface. As fine ceramics, for example, metal oxides, metal nitrides, metal oxynitrides, and the like such as silicon oxide, silicon nitride, silicon oxynitride, aluminum oxide, aluminum nitride, and aluminum oxynitride may be cited. As fluororesins, for example, polytetrafluoroethylene, polyvinylidene fluoride, polychlorotrifluoroethylene, tetrafluoroethylene / perfluoroalkyl vinyl ether copolymers, tetrafluoroethylene-hexafluoropropylene copolymers, tetrafluoroethylene-ethylene copolymers, chlorotrifluoroethylene-ethylene copolymers, and the like may be cited. As silicone resins, organopolysiloxanes (for example, dimethylpolysiloxane, etc.) and their modified products (for example, polyether modification, phenol modification, amine modification, silanol modification, trimethylsilyl modification, methanol modification, carboxyl modification, epoxy modification, acrylic acid modification, methacrylic acid modification, etc.) may be cited. As long as at least the portion of the inner surface of the container in contact with the pharmaceutical composition has the above-mentioned materials, it is sufficient. At least a portion of the inner surface of the container has the above-mentioned materials. In one embodiment, the entire inner surface of the container is provided with the above-mentioned material. In other embodiments, the container itself is formed of the above-mentioned material. In a preferred embodiment, the inner surface of the container is provided with a film containing the above-mentioned material. In the case where the inner surface of the container has a film containing the above-mentioned material, its thickness is, for example, greater than 0.1 μm and less than 0.2 μm. In a more preferred embodiment, all parts of the inner surface of the container that can be contacted by the pharmaceutical composition are provided with a film containing the above-mentioned material.

[0031] From the perspective of storage stability of the lyophilized enzyme, for example, from the perspective of the residual potency, in a preferred embodiment, the material is selected from silicon oxide, silicon nitride, silicon oxynitride, aluminum oxide, aluminum nitride, and aluminum oxynitride. The material is more preferably selected from silicon oxide, silicon nitride, and silicon oxynitride, further preferably silicon oxide, and particularly preferably silicon dioxide.

[0032] The film containing the above-mentioned materials can be made by a conventionally known wet or dry film-forming method. The wet film-forming method includes, for example, a coating method. The film containing the above-mentioned materials is preferably formed by dry film-forming methods such as chemical vapor deposition (CVD), physical vapor deposition, spray pyrolysis, sputtering, and more preferably by CVD or spray pyrolysis. From the storage stability of the freeze-dried enzyme, for example, from the viewpoint of the titer residual rate, it is further preferably formed by CVD, wherein it is particularly preferably formed by plasma chemical vapor deposition (plasma CVD).

[0033] As a container, for example, there can be exemplified a film having an inner surface containing a material selected from silicon oxide, silicon nitride and silicon oxynitride and formed by a CVD method; a film having an inner surface containing silicon dioxide and formed by a CVD method; a film having an inner surface containing a material selected from silicon oxide, silicon nitride and silicon oxynitride and formed by a plasma CVD method; and a film having an inner surface containing silicon dioxide and formed by a plasma CVD method, but the container is not limited to these.

[0034] As the container having a film containing the above-mentioned material on the inner surface, a commercially available silica-coated vial, silicone-coated vial, fluororesin-coated vial, or the like can be used.

[0035] It is believed that the inner surface of the container can inhibit the mixing of impurities such as metal ions from the container base material into the pharmaceutical composition by having the material as described above. It is believed that the inhibition of the mixing of impurities may be related to the inhibitory effect of the enzyme activity reduction of the present invention. In addition, the above presumed mechanism of action does not limit the technical scope of the present invention in any way.

[0036] The water content of the lyophilized preparation is, for example, 5% (w / w) or less, preferably 3% (w / w) or less, and more preferably 2% (w / w) or less. In this specification, the water content is a value measured by coulometric titration.

[0037] In a certain embodiment, the storage amount of glycolytic enzymes per container is, for example, less than 10 μg. The storage amount of glycolytic enzymes per container can be set to a numerical range consisting of a combination of any numerical values ​​of 0.1 μg, 0.5 μg, 1 μg, 2 μg, 2.5 μg, 4 μg, 5 μg, 6 μg, 7 μg, 8 μg, 9 μg, 9.5 μg, 9.8 μg and less than 10 μg. That is, the storage amount of glycolytic enzymes per container can be 0.1 μg or more, 0.5 μg or more, 1 μg or more, 2 μg or more, 2.5 μg or more, or 4 μg or more, and can be 5 μg or less, 6 μg or less, 7 μg or less, 9.5 μg or less, or 9.8 μg or less. In a preferred embodiment, the storage amount of glycolytic enzymes per container is 0.1 μg or more and 9.8 μg or less. In a more preferred embodiment, the amount of glycolytic enzyme stored in one container is 2 μg or more and 7 μg or less. In a particularly preferred embodiment, the amount of glycolytic enzyme stored in one container is 2.5 μg or more and 5 μg or less.

[0038] "Titer" refers to the activity (unit) of the enzyme against 1 μg of glycolytic enzyme, expressed in units of units / μg. In one embodiment, the titer of the lyophilized glycolytic enzyme is 0.3 (units / μg). In other embodiments, the titer of the lyophilized glycolytic enzyme is greater than 0.3 (units / μg) and less than 1 (units / μg). In other embodiments, the titer of the lyophilized glycolytic enzyme is greater than 0.32 (units / μg) and less than 1 (units / μg). In other embodiments, the titer of the lyophilized glycolytic enzyme is greater than 0.33 (units / μg) and less than 1 (units / μg). In other embodiments, the titer of the lyophilized glycolytic enzyme is greater than 0.36 (units / μg) and less than 1 (units / μg).

[0039] In one embodiment, the titer of the freeze-dried glycolytic enzyme is 0.3 (unit / μg) or more and 0.5 (unit / μg) or less. In other embodiments, the titer of the freeze-dried glycolytic enzyme is 0.33 (unit / μg) or more and 0.5 (unit / μg) or less, or 0.36 (unit / μg) or more and 0.5 (unit / μg) or less.

[0040] "Unit" means the activity of glycolytic enzyme, and 1 unit means the amount of 1 micromole of degradation product released from substrate per unit time under the conditions of optimal temperature and optimal pH. For example, when the glycolytic enzyme is chondroitinase ABC, 1 unit means the amount of 1 micromole of unsaturated disaccharide released from sodium chondroitin sulfate (sodium chondroitin sulfate conforming to the Japanese Pharmacopoeia Standards for Non-Japanese Drugs 2002) per minute under the conditions of pH 8.0 and 37°C.

[0041] In one embodiment, the titer of the glycolytic enzyme after the amount of the pharmaceutical composition required for one administration is stored in the container is, for example, 75% or more, when the titer before being stored in the container is set to 100%. In a preferred embodiment, the titer of the glycolytic enzyme after the amount of the pharmaceutical composition required for one administration is stored in the container is 80% or more, when the titer before being stored in the container is set to 100%.

[0042] In one embodiment, the enzyme activity (enzyme activity of glycolytic enzymes) per container is, for example, less than 5 units. In another embodiment, the enzyme activity per container is, for example, less than 4.5 units. In another embodiment, the enzyme activity per container is, for example, less than 4.1 units. In another embodiment, the enzyme activity per container is, for example, less than 4 units. In another embodiment, the enzyme activity per container is, for example, more than 0.1 units and less than 4.5 units. In another embodiment, the enzyme activity per container is, for example, more than 0.1 units and less than 4.1 units. In another embodiment, the enzyme activity per container is, for example, more than 0.1 units and less than 4 units. In another embodiment, the enzyme activity per container is, for example, more than 0.9 units and less than 2.5 units, more than 0.9 units and less than 2 units, 1.25 units and less than 2 units, or 1.5 units.

[0043] According to one embodiment of the present invention, for example, a package having a storage stability of more than 12 months can be provided. In a preferred embodiment, the package has a storage stability of more than 24 months, and in a more preferred embodiment, the package has a storage stability of more than 36 months. The upper limit of the storage stability is not particularly limited, and for example, it can be less than 48 months or less than 36 months.

[0044] Here, "having storage stability" means that the potency (%) after being stored in a sealed and light-proof state under prescribed conditions (for example, more than 12 months at 5°C ± 3°C, more than 6 months at 25°C ± 2°C, or more than 3 months or more than 6 months at 40°C ± 2°C) is maintained at a pharmaceutically acceptable level. In this specification, "storage stability" is evaluated, for example, by the potency residual rate (%). For example, the potency residual rate after the sample is stored at 5°C ± 3°C for more than 12 months is, for example, more than 90%, preferably more than 95%. The potency residual rate after the sample is stored at 5°C ± 3°C for more than 24 months is, for example, more than 90%, preferably more than 95%. The potency residual rate after the sample is stored at 5°C ± 3°C for more than 36 months is, for example, more than 90%, preferably more than 95%. The potency residual rate after the sample is stored at 25°C ± 2°C for more than 6 months is, for example, more than 90%, preferably more than 95%. The titer residual rate after the sample is stored at 40°C ± 2°C for more than 3 months is, for example, 90% or more, preferably 95% or more. The titer residual rate after the sample is stored at 40°C ± 2°C for more than 6 months is, for example, 65% or more, preferably 70% or more, and more preferably 75% or more. It should be noted that "titer residual rate (%)" refers to the titer (%) value calculated after the package in the present invention is stored under the conditions of a specified temperature (for example, 5°C ± 3°C, 25°C ± 3°C or 40°C ± 2°C), with the titer at the start of storage being set as 100%. If this value is 100%, the value of the enzyme activity is equal before and after the start of storage.

[0045] According to one embodiment of the present invention, a package having an expiration date of more than 12 months can be provided. In a preferred embodiment, the package has an expiration date of more than 24 months, and in a more preferred embodiment, the package has an expiration date of more than 36 months. The upper limit of the expiration date is not particularly limited, and for example, it can be less than 48 months or less than 36 months.

[0046] The "expiration date" as used in this specification refers to the time from the moment when the usefulness of a drug is determined, when it is stored using a certain storage method (for example, at 5°C±3°C, sealed / protected from light, at 25°C±2°C, sealed and protected from light, or left standing at 40°C±2°C, sealed and protected from light), during which the drug can be expected to have the same usefulness as the drug at the time of confirmation.

[0047] In this specification, as the purpose of the above-mentioned pharmaceutical composition, known various purposes can be selected as the purpose of glycolytic enzyme. For example, the purpose of the pharmaceutical composition comprising glycolytic enzyme as an effective ingredient is not particularly limited, and the treatment of hernia, lysosomal storage disease, keloid, hypertrophic scar, muscular dystrophy or spinal cord injury can be exemplified. In a preferred embodiment, the pharmaceutical composition can be used for the treatment of hernia. In a more preferred embodiment, it is used for the treatment of intervertebral disc herniation (for example, lumbar disc herniation).

[0048] In this specification, "treatment" is not only a complete cure, but also includes improving part or all of the symptoms of the disease, inhibiting the progression of the disease (including maintaining and reducing the rate of progression), and preventing the disease. Prevention here means that in the case where the symptoms of the accompanying disease have not yet occurred, it includes preventing the occurrence of these symptoms before they occur. In addition, prevention means, for example, in the case where no clear organic lesions have been confirmed, but the symptoms of the accompanying disease have already occurred, it includes preventing the occurrence of the organic lesions and inhibiting the development of symptoms that are not obvious among these symptoms before they occur.

[0049] In this specification, "as an active ingredient" and "effective amount" refer to an amount that is reasonable and balanced with a risk / benefit ratio, and does not have excessive harmful side effects (toxicity, irritation, etc.), and is sufficient to obtain the desired response. The "as an active ingredient" and "effective amount" can be changed according to the symptoms, physique, age, sex and other factors of the patient who is the subject of administration. However, those skilled in the art can determine the effective amount for other situations based on the results of one or more specific test examples and technical common sense, without having to test the combination of various factors separately.

[0050] In the present specification, the "patient" refers to an animal, preferably a mammal (eg, human, mouse, rat, hamster, guinea pig, rabbit, dog, cat, horse, etc.), more preferably a human.

[0051] In a preferred embodiment, the pharmaceutical composition contained in the container can be provided in a sterilized state. The sterilization method of the pharmaceutical composition is not particularly limited, and it can be carried out by conventionally known methods such as filtration sterilization and dry heat sterilization.

[0052] The mode of administration of the pharmaceutical composition is not particularly limited and can be appropriately selected according to the disease to be treated, symptoms, severity, attributes of the patient (e.g., age, etc.). The lyophilized preparation can be dissolved in any solvent (e.g., water for injection, saline, etc.) for use. As the mode of administration, for example, any administration route such as intradiscal injection, intravenous injection, intramuscular injection, subcutaneous injection, and drip can be cited. The dosage of the pharmaceutical composition can also be appropriately set by those skilled in the art according to the disease to be treated, symptoms, severity, attributes of the patient (e.g., age, etc.).

[0053] The following are preferred specific embodiments of the present invention, but they do not limit the technical scope of the present invention. Although the pharmaceutical composition is described below using the example of an indication for intervertebral disc herniation, it can of course be a "pharmaceutical composition" for any applicable disease.

[0054] (Packaging body 1) Active ingredient: Chondroitinase ABC Material of the inner surface of the container: Silicon dioxide Amount of active ingredient per container: 2.5 μg or more and less than 10 μg Enzyme titer: 0.32 (unit / μg) or more and 1 (unit / μg) or less Enzyme activity per container: 0.4 units or more and 4.5 units or less Indications: Herniated disc

[0055] (Packaging 2) Active ingredient: Chondroitinase ABC Material of the inner surface of the container: Silicone resin Amount of active ingredient per container: 2.5 μg or more and less than 10 μg Enzyme titer: 0.32 (unit / μg) or more and 1 (unit / μg) or less Enzyme activity per container: 0.4 units or more and 4.5 units or less Indications: Herniated disc

[0056] (Packaging 3) Active ingredient: Chondroitinase ABC Material of the inner surface of the container: Fluorine resin Amount of active ingredient per container: 2.5 μg or more and less than 10 μg Enzyme titer: 0.32 (unit / μg) or more and 1 (unit / μg) or less Enzyme activity per container: more than 1 unit and less than 4.5 units Indications: Herniated disc

[0057] (Packaging body 4) Active ingredient: Chondroitinase ABC The material of the inner surface of the container: at least one selected from silicon dioxide, silicone resin and fluororesin Amount of active ingredient per container: 2 μg or more and 5 μg or less Enzyme titer: 0.32 (unit / μg) or more and 0.5 (unit / μg) or less Enzyme activity per container: 1.25 units or more and 2.5 units or less Indications: Herniated disc (Packaging body 5) Active ingredient: Chondroitinase ABC The material of the inner surface of the container: at least one selected from silicon dioxide, silicone resin and fluororesin Amount of active ingredient per container: 2.5 μg or more and 5 μg or less Enzyme titer: 0.32 (unit / μg) or more and 0.5 (unit / μg) or less Enzyme activity per container: 1.5 units Indications: Herniated disc

[0058] (2) Test kit In one embodiment, a kit is provided comprising the above-mentioned package and additional information or labeling for use of the above-mentioned pharmaceutical composition, wherein the above-mentioned pharmaceutical composition is used to treat hernia, lysosomal storage disease, keloid, hypertrophic scar, muscular dystrophy or spinal cord injury.

[0059] The kit may be any kit as long as it includes a package in which the pharmaceutical composition is stored in a container and additional information or a label explaining the use of the pharmaceutical composition, wherein the pharmaceutical composition is used to treat hernia, lysosomal storage disease, keloid, hypertrophic scar, muscular dystrophy or spinal cord injury. That is, it may further include other components.

[0060] (3) Manufacturing method One aspect of the present invention relates to a method for manufacturing a packaging body comprising a pharmaceutical composition and a container for storing the pharmaceutical composition, comprising: a first step, wherein a solution containing a glycolytic enzyme is stored in a container, and the inner surface of the container has at least one material selected from fine ceramics, silicone resins and fluororesins; and a second step, freeze-drying the solution to obtain the pharmaceutical composition.

[0061] In the first step, the solvent used in the preparation of the solution containing the glycolytic enzyme is not particularly limited, and for example, a buffer such as water, physiological saline, or phosphate buffer can be used. In addition, the solution may contain a pharmaceutically acceptable carrier as described above. The pH value of the solution containing the glycolytic enzyme contained in the container is not particularly limited, but is preferably in the range of 6.5 or more and 7.5 or less.

[0062] In one embodiment, in the first step, the solution containing glycolytic enzymes is stored in a container in such a way that the enzyme activity per container is less than 5 units. In other embodiments, in the first step, the solution containing glycolytic enzymes is stored in a container in such a way that the enzyme activity per container is less than 4.5 units. In other embodiments, in the first step, the solution containing glycolytic enzymes is stored in a container in such a way that the enzyme activity per container is less than 4.1 units. In other embodiments, in the first step, the solution containing glycolytic enzymes is stored in a container in such a way that the enzyme activity per container is 0.5 units and less than 4.5 units. In other embodiments, in the first step, the solution containing glycolytic enzymes is stored in a container in such a way that the enzyme activity per container is more than 0.5 units and less than 4.1 units. In a preferred embodiment, in the first step, the solution containing glycolytic enzymes is stored in a container in such a way that the enzyme activity per container is less than 4 units. In a more preferred embodiment, the solution is stored in a container in such a way that the enzyme activity per container is more than 0.5 units and less than 4 units. In another embodiment, the solution is contained in the container so that the enzyme activity per container becomes 0.5 units to 3.5 units, 1.1 units to 3 units, 1 unit to 2 units, 1.25 units to 1.6 units, or 1.5 units.

[0063] The second step includes a freeze-drying step, wherein the solution containing the glycolytic enzyme is frozen, and water is removed by sublimation in the frozen state and dried. In the second step, drying is performed until the moisture content of the freeze-dried pharmaceutical composition becomes, for example, 5% (w / w) or less. The drying in the second step is preferably performed until the moisture content of the freeze-dried pharmaceutical composition becomes 3% (w / w) or less, and more preferably until the moisture content becomes 2% (w / w) or less.

[0064] The description, examples, preferred ranges, etc. of "the above-mentioned (1) composition and package" and "the above-mentioned (2) kit" can be directly applied to the production method of the present invention.

[0065] As another embodiment, the present invention also includes the use of a package in the treatment of hernia, lysosomal storage disease, keloid, hypertrophic scar, muscular dystrophy or spinal cord injury, wherein the package is formed by storing a freeze-dried preparation containing at least one of glycolytic enzymes and matrix metalloproteinases as an active ingredient, i.e., a pharmaceutical composition, in a container, and the inner surface of the container has at least one material selected from fine ceramics, silicone resins and fluororesins. In addition, as another embodiment, the present invention also includes a package for the treatment of hernia, lysosomal storage disease, keloid, hypertrophic scar, muscular dystrophy or spinal cord injury, wherein the package is formed by storing a freeze-dried preparation containing at least one of glycolytic enzymes and matrix metalloproteinases as an active ingredient, i.e., a pharmaceutical composition, in a container, and the inner surface of the container has at least one material selected from fine ceramics, silicone resins and fluororesins.

[0066] Although embodiments of the present invention are described below, the present invention is not limited thereto. <1> A packaging body comprises a pharmaceutical composition and a container, wherein the pharmaceutical composition is a freeze-dried preparation containing at least one enzyme of glycolytic enzyme and matrix metalloproteinase as an active ingredient, the container stores the pharmaceutical composition, and the inner surface of the container has at least one material selected from fine ceramics, silicone resin and fluororesin.

[0067] <2> according to <1> The package body described above, wherein the material is selected from silicon oxide, silicon nitride, silicon oxynitride, aluminum oxide, aluminum nitride and aluminum oxynitride.

[0068] <3> according to <2> The package body described above, wherein the silicon oxide is silicon dioxide.

[0069] <4> according to <1> ~ <3> The packaging body described in any one of the above, wherein the inner surface of the container comprises a film containing the material.

[0070] <5> according to <4> The package described herein, wherein the film containing the above material is formed by chemical vapor deposition, physical vapor deposition, spray pyrolysis or sputtering.

[0071] <6> according to <5> The package described above, wherein the chemical vapor deposition method is a plasma chemical vapor deposition method.

[0072] <7> according to <1> ~ <6> The package body according to any one of the above, wherein the amount of the enzyme stored per container is less than 10 μg.

[0073] <8> according to <1> ~ <7> The package body according to any one of the above, wherein the enzyme has a titer of 0.3 (unit / μg) or more.

[0074] <9> according to <1> ~ <8> The package body according to any one of the above, wherein the enzyme activity per container is less than 5 units.

[0075] <10> according to <1> ~ <9> The package body according to any one of claims , wherein the enzyme activity of the enzyme is 75% or more, when the value before lyophilization is set to 100%.

[0076] <11> according to <1> ~ <10> The package body according to any one of the above, wherein the package body has storage stability at 5°C±3°C for 12 months or more.

[0077] <12> according to <1> ~ <11> The package body according to any one of the above, wherein the enzyme is a glycosaminoglycan glycolytic enzyme.

[0078] <13> according to <12> The packaged body described above, wherein the glycosaminoglycan glycolytic enzyme is chondroitinase.

[0079] <14> according to <13> The packaged body described above, wherein the chondroitinase is chondroitinase ABC.

[0080] <15> according to <1> ~ <14> The package body described in any one of the above, wherein the matrix metalloproteinase is matrix metalloproteinase-7.

[0081] <16> according to <1> ~ <15> The package body described in any one of the above, wherein the pharmaceutical composition contains a pharmaceutically acceptable carrier.

[0082] <17> according to <16> The packaged body described above, wherein the carrier comprises at least one of polyalkylene glycol and sucrose.

[0083] <18> according to <1> ~ <17> The packaged body according to any one of items , wherein the pharmaceutical composition is used to treat hernia, lysosomal storage disease, keloid, hypertrophic scar, muscular dystrophy or spinal cord injury.

[0084] <19> according to <1> ~ <18> The package body according to any one of the above, wherein the container is a vial, a syringe or an ampoule.

[0085] <20> A pharmaceutical composition is obtained by freeze-drying a solution containing at least one enzyme selected from fine ceramics, silicone resins and fluororesins in a container having an inner surface provided with at least one material selected from fine ceramics, silicone resins and fluororesins.

[0086] <21> according to <20> The pharmaceutical composition described above, wherein the titer of the enzyme is 0.3 unit / μg or more.

[0087] <22> according to <20> ~ <21> The pharmaceutical composition according to any one of the above, wherein the pharmaceutical composition is a unit dosage preparation in which the amount of the enzyme is less than 10 μg.

[0088] <23> according to <20> ~ <22> The pharmaceutical composition according to any one of claims , wherein the enzyme activity is less than 5 units.

[0089] <24> according to <20> ~ <23> The pharmaceutical composition according to any one of the above, wherein the pharmaceutical composition has a storage stability of 12 months or more at 5°C±3°C.

[0090] <25> according to <20> ~ <24> The pharmaceutical composition according to any one of claims , wherein the enzyme is a glucosaminoglycan glycolytic enzyme.

[0091] <26> according to <25> The pharmaceutical composition described above, wherein the glycosaminoglycan glycolytic enzyme is chondroitinase.

[0092] <27> according to <26> The pharmaceutical composition described above, wherein the chondroitinase is chondroitinase ABC.

[0093] <28> according to <20> ~ <27> The pharmaceutical composition according to any one of claims , wherein the matrix metalloproteinase is matrix metalloproteinase-7.

[0094] <29> according to <20> ~ <28> The pharmaceutical composition according to any one of the above, wherein the pharmaceutical composition comprises a pharmaceutically acceptable carrier.

[0095] <30> according to <29> The pharmaceutical composition described above, wherein the carrier contains at least one of polyalkylene glycol and sucrose.

[0096] <31> according to <20> ~ <30> The pharmaceutical composition according to any one of the above, wherein the pharmaceutical composition is used to treat hernia, lysosomal storage disease, keloid, hypertrophic scar, muscular dystrophy or spinal cord injury.

[0097] <32> A kit comprising <1> ~ <19> The packaging body described in any one of the above items and the additional document or label explaining the use of the above pharmaceutical composition, wherein the above pharmaceutical composition is used to treat hernia, lysosomal storage disease, keloid, hypertrophic scar, muscular dystrophy or spinal cord injury.

[0098] <33> A method for manufacturing a packaging body comprising a pharmaceutical composition and a container for accommodating the pharmaceutical composition, comprising: accommodating a solution comprising at least one enzyme of glycolytic enzyme and matrix metalloproteinase in a container, wherein the inner surface of the container has at least one material selected from fine ceramics, silicone resin and fluororesin, and freeze-drying the solution to obtain the pharmaceutical composition.

[0099] <34> according to <33> The described manufacturing method, wherein the material is selected from silicon oxide, silicon helium oxide, silicon oxynitride, aluminum oxide, aluminum nitride and aluminum oxynitride.

[0100] <35> according to <34> The production method described above, wherein the silicon oxide is silicon dioxide.

[0101] <36> according to <33> to <35> The production method described in any one of claims , wherein the inner surface of the container comprises a film containing the material.

[0102] <37> according to <36> The described production method, wherein the film containing the above-mentioned material is formed by chemical vapor deposition, physical vapor deposition, spray pyrolysis or sputtering.

[0103] <38> according to <37> The production method described above, wherein the chemical vapor deposition method is a plasma chemical vapor deposition method.

[0104] <39> according to <33> ~ <38> The production method according to any one of claims , wherein the amount of the enzyme stored is less than 10 μg.

[0105] <40> according to <33> ~ <39> The production method according to any one of claims , wherein the titer of the enzyme in the pharmaceutical composition is 0.3 (unit / μg) or more.

[0106] <41> according to <33> ~ <40> The production method according to any one of claims , wherein the enzyme activity of the solution contained in the container is less than 5 units.

[0107] <42> according to <33> ~ <41> The production method according to any one of claims , wherein the enzyme activity of the enzyme after lyophilization is 75% or more, when the enzyme activity before lyophilization is assumed to be 100%.

[0108] <43> according to <33> ~ <42> The production method according to any one of claims , wherein the enzyme is a glucosaminoglycan glycolytic enzyme.

[0109] <44> according to <43> The production method described above, wherein the glycosaminoglycan glycolytic enzyme is chondroitinase.

[0110] <45> according to <44> The production method described above, wherein the chondroitinase is chondroitinase ABC.

[0111] <46> according to <33> ~ <45> The production method according to any one of claims 1 to 5, wherein the matrix metalloproteinase is matrix metalloproteinase-7.

[0112] <47> according to <33> ~ <46> The production method according to any one of claims , wherein the pharmaceutical composition comprises a pharmaceutically acceptable carrier.

[0113] <48> according to <47> The production method described above, wherein the carrier comprises at least one of polyalkylene glycol and sucrose.

[0114] <49> according to <33> ~ <48> The production method according to any one of items , wherein the pharmaceutical composition is used to treat hernia, lysosomal storage disease, keloid, hypertrophic scar, muscular dystrophy or spinal cord injury.

[0115] <50> according to <33> ~ <49> The production method according to any one of the above, wherein the container is a vial, a syringe or an ampoule. Example

[0116] The present invention will be described in more detail below, but the technical scope of the present invention is not limited thereto.

[0117] <Preparation Example 1> 1) Preparation of Chondroitinase ABC Chondroitinase ABC was prepared according to the method described in Japanese Patent Application Laid-Open No. 6-153947. That is, it was produced by purifying from the culture supernatant of Proteus vulgaris. The titer of the obtained chondroitinase ABC was 0.40 U / μg.

[0118] 2) Determination of enzyme activity and concentration of chondroitinase ABC The enzymatic activity of chondroitinase ABC was measured by the following method. The enzyme sample (chondroitinase ABC) was diluted 4000 times using 0.01% (w / v) casein reagent (20mM phosphate buffer). 400μL of substrate solution (3mg / ml chondroitin sulfate sodium (Japanese Pharmacopoeia prescription drug specifications), 50mM 2-amino-2-hydroxymethyl-1,3-propanediol, 50mM sodium acetate, pH8) was added to 100μL of the diluted enzyme sample and mixed. After the solution was reacted at 37°C for 20 minutes, it was heated in a 100°C water bath for 1 minute. After cooling to room temperature, 5.0mL of 0.05M hydrochloric acid was added to prepare the sample solution. The chondroitinase ABC standard substance was diluted 400 times using 0.01% (w / v) casein reagent. For 100μL of the diluted chondroitinase ABC standard substance solution, the same operation as the preparation of the sample solution was performed to prepare the standard solution. In addition, 100 μL of 0.01% (w / v) casein reagent was subjected to the same operation as the sample solution to prepare a control solution. The absorbance A at a wavelength of 232 nm was measured for the sample solution, standard solution, and control solution by ultraviolet visible light absorbance measurement. T , A S and A B The enzyme solution activity (U / mL) was calculated by the following mathematical formula. Here, the enzyme solution activity is the enzyme activity per unit volume of liquid.

[0119] Enzyme solution activity (U / mL) = (A T -A B ) / (A S -A B )×4000 / 400×Us A T : Absorbance of sample solution A B :Absorbance of control solution A S :Absorbance of standard solution U S :Enzyme solution activity of chondroitinase ABC standard substance (U / mL)

[0120] It should be noted that 1U (unit) is defined as the value of the enzyme activity that catalyzes the reaction of releasing 1 micromole of unsaturated disaccharide in 1 minute under the above reaction conditions. The value of the enzyme activity used in this specification is determined based on the activity of the enzyme solution.

[0121] The enzyme amount (protein amount, μg) of chondroitinase ABC is determined by the following semi-limited injection method (Lowry Method). That is, 2.5mL of alkaline copper reagent is added to 0.5mL of enzyme sample (chondroitinase ABC) diluted 50 times with pure water and mixed, and placed at room temperature (above 20°C and below 25°C) for 10 minutes. 0.25mL of 1mol / L phenol reagent is added to the liquid and mixed, and placed at room temperature for 30 minutes as a sample solution. Bovine serum albumin is dissolved in water to prepare a solution in a manner of reaching 30μg / mL, 40μg / mL, 50μg / mL, 60μg / mL or 70μg / mL. For these solutions 0.5mL, the same operation as the enzyme sample diluted 50 times above is performed as a standard solution. In addition, for 0.5mL of water, the same operation as the enzyme sample diluted 50 times above is performed as a blank solution. For these liquids, the absorbance at a wavelength of 750nm is measured. The absorbance of the standard solution and the protein concentration are plotted by linear regression to obtain a standard curve that best approximates each point. The amount of protein in the sample solution is determined from the obtained standard curve and the absorbance of the sample solution.

[0122] 3) Preparation of enzyme buffer An enzyme buffer was prepared to have the following composition. (Composition: 1L distilled water for injection) pH: 6.5 or higher and 7.5 or lower.

[0123] <Reference example> The enzyme sample (chondroitinase ABC) was dissolved in an enzyme buffer. The obtained enzyme solution was placed in a glass vial (vial 3010; manufactured by Fuji Nitro Corporation) and the enzyme amounts were as follows: Sample 1: 15.0 μg / vial (6.0 U / vial) Sample 2: 30.0 μg / vial (12.0 U / vial) Sample 3: 60.0 μg / vial (24.0 U / vial).

[0124] The enzyme solutions stored in these vials were lyophilized (water content after lyophilization was 2% (w / w) or less). After lyophilization, the vials were repressurized with nitrogen gas and plugged with rubber stoppers to obtain packages.

[0125] For each sample, the enzyme activity (U / vial) after lyophilization was measured (n=3). In addition, based on the obtained enzyme activity value, the enzyme titer (U / μg) after lyophilization was calculated. The results are shown in Table 1. It should be noted that the titer is the enzyme activity per enzyme amount.

[0126] [Table 1] Sample 1 Sample 2 Sample 3 Enzyme amount (μg / vial) 15.0 30.0 60.0 Enzyme activity before lyophilization (U / vial) 6.0 12.0 24.0 Enzyme activity after freeze-drying (U / vial) 4.6 10.5 21.8 Lyophilized titer (U / μg) 0.307 0.350 0.363

[0127] As shown in Table 1, the smaller the amount of enzyme contained in the vial, the lower the titer.

[0128] <Example 1> The chondroitinase ABC solution prepared using enzyme buffer was placed in a 3 ml glass vial to give 9.1 μg / vial (3.63 U / vial; samples 4 to 6) or 1.3 μg / vial (0.50 U / vial; sample 7). As glass vials, vials 3010 (manufactured by Fuji Nitro Corporation; sample 4), vials 3010 silicon-coated (the inner surface of the vial has a silicon dioxide film formed by spray pyrolysis (SPD method); manufactured by Fuji Nitro Corporation; sample 5) and SCHOTT (registered trademark) Type 1 plus (the inner surface of the vial has a silicon dioxide film formed by plasma chemical vapor deposition (plasma CVD method); manufactured by SCHOTT; samples 6 and 7) were used.

[0129] The enzyme solutions stored in these vials were lyophilized (water content after lyophilization was 2% (w / w) or less). After drying, the vials were repressurized with nitrogen gas and plugged with rubber stoppers to obtain packages storing a unit amount.

[0130] The enzyme activity (U / vial) after lyophilization was measured for each sample (n=10). In addition, the titer (U / μg) of the enzyme after lyophilization was calculated based on the obtained enzyme activity value. The results are shown in Table 2.

[0131] [Table 2] Sample 4 Sample 5 Sample 6 Sample 7 Enzyme amount (μg / vial) 9.1 9.1 9.1 1.3 Enzyme activity before lyophilization (U / vial) 3.63 3.63 3.63 0.50 Enzyme activity after freeze-drying (U / vial) 2.60 2.99 3.13 0.43 Lyophilized titer (U / μg) 0.286 0.328 0.344 0.331

[0132] <Example 2> The packages of Samples 4 and 6 were tightly sealed with aluminum caps and then dry-heat sterilized at 250° C. for 5 hours. The enzyme activity after dry-heat sterilization was 2.53 U / vial in Sample 4 and 3.12 U / vial in Sample 6.

[0133] <Example 3> The enzyme solution was placed in vials 3010 (manufactured by Fuji Nitro Co., Ltd.; sample 8) and SCHOTT (registered trademark) Type 1 plus (manufactured by SCHOTT; sample 9), and freeze-dried (water content 2% (w / w) or less). After freeze-drying, the vials were re-pressurized with nitrogen gas and plugged with rubber stoppers to obtain packages. The obtained packages were stored at 40±2°C and protected from light for 1 month, 3 months or 6 months, and the titer after each storage time was calculated (n=3). The titer residual rate (%) after each storage period was calculated when the titer at the beginning of storage was set to 100%, and the calculated results are shown in Table 3.

[0134] [Table 3]

[0135] In addition, when sample 9 was stored at 5°C ± 3°C in the dark for 12 months, the residual potency ratio was 98%.

[0136] <Preparation Example 2> 1) Preparation of Chondroitinase ABC Chondroitinase ABC was prepared according to the method described in Japanese Patent Application Laid-Open No. 6-153947 in the same manner as in Preparation Example 1. The titer of the obtained chondroitinase ABC was 0.42 U / μg.

[0137] <Example 4> An enzyme solution was prepared by adding an enzyme buffer to the chondroitinase ABC obtained in Preparation Example 2. The enzyme solution was stored in a glass vial so that the enzyme content was 9.8 μg / vial (4.1 U / vial; Samples 10 to 12). As glass vials, SCHOTT (registered trademark) Type 1 plus (the inner surface of the vial had a silicon dioxide film formed by plasma chemical vapor deposition (plasma CVD); manufactured by SCHOTT; Sample 10), silicone-coated vials (the inner surface of the vial had a silicone resin film: manufactured by Iwata Glass Industries, Ltd.; Sample 11), and fluorine-coated vials (the inner surface of the vial had a polytetrafluoroethylene film; manufactured by Universal Corporation; Sample 12) were used.

[0138] The enzyme solutions stored in these vials were lyophilized (water content after lyophilization was 2% (w / w) or less). After drying, the vials were repressurized with nitrogen gas and plugged with rubber stoppers to obtain packages storing a unit amount.

[0139] The enzyme activity (U / vial) after lyophilization was measured for each sample (n=3). In addition, the titer (U / μg) of the enzyme after lyophilization was calculated based on the obtained enzyme activity value. The results are shown in Table 4. [Table 4] Sample 10 Sample 11 Sample 12 Enzyme amount (μg / vial) 9.8 9.8 9.8 Enzyme activity before lyophilization (U / vial) 4.10 4.10 4.10 Enzyme activity after freeze-drying (U / vial) 4.05 4.06 3.86 Lyophilized titer (U / μg) 0.415 0.416 0.395

[0140] The present invention has been described with respect to specific examples and various implementations, but it is easily understood by those skilled in the art that many changes and applications may be made to the implementations described in this specification without departing from the spirit and scope of the present invention. This application claims priority based on Japanese Patent Application No. 2018-35884 filed on February 28, 2018, and Japanese Patent Application No. 2018-141542 filed on July 27, 2018, filed on the Japan Patent Office, the contents of which are incorporated herein by reference in their entirety. All documents, patent applications, and technical standards described in this specification are incorporated herein by reference, the same as if each document, patent application, and technical standard were specifically and individually recorded as being incorporated into this specification by reference.

Claims

1. A packaging body, characterized in that: comprising a pharmaceutical composition and a container, The pharmaceutical composition is a freeze-dried preparation containing glycolytic enzymes as active ingredients. The container contains the pharmaceutical composition, and the inner surface of the container is made of fine ceramics. The glycolytic enzyme is chondroitinase ABC, The amount of enzyme contained in each container is 1 μg or more and less than 10 μg, The enzyme titer is 0.32 units / μg or more and 1 unit / μg or less.

2. The packaging body according to claim 1, wherein: The fine ceramic is selected from silicon oxide, silicon nitride, silicon oxynitride, aluminum oxide, aluminum nitride and aluminum oxynitride.

3. The packaging body according to claim 2, wherein: The silicon oxide is silicon dioxide.

4. The packaging body according to claim 1, wherein: The inner surface of the container is provided with a film containing the fine ceramic.

5. The packaging body according to claim 4, wherein: The film containing the fine ceramic is formed by chemical vapor deposition, physical vapor deposition, spray pyrolysis or sputtering.

6. The packaging body according to claim 5, wherein: The chemical vapor deposition method is a plasma chemical vapor deposition method.

7. The packaging body according to claim 1, wherein: The enzyme activity per container is greater than 0.1 units and less than 4.5 units.

8. The packaging body according to claim 1, wherein: When the value before lyophilization is set as 100%, the enzyme activity of the enzyme is 75% or more.

9. The packaging body according to claim 1, wherein: The package has storage stability at 5°C±3°C for more than 12 months.

10. The packaging body according to any one of claims 1 to 9, wherein The pharmaceutical composition comprises at least one of polyethylene glycol and sucrose.

11. The packaging body according to claim 10, wherein: The pharmaceutical composition comprises a mixture of polyethylene glycol and sucrose.

12. The package according to claim 1, wherein: The pharmaceutical composition is used to treat hernia, lysosomal storage disease, keloid, hypertrophic scar, muscular dystrophy or spinal cord injury.

13. The package according to claim 1, wherein: The container is a vial, a syringe or an ampoule.

14. A kit comprising the packaging body according to any one of claims 1 to 13 and an additional document or label indicating the use of the pharmaceutical composition, wherein the use of the pharmaceutical composition is for treating hernia, lysosomal storage disease, keloid, hypertrophic scar, muscular dystrophy or spinal cord injury.

15. A method for manufacturing a package, the package comprising a pharmaceutical composition and a container for accommodating the pharmaceutical composition, the method for manufacturing the package comprising: A step of storing a solution containing a glycolytic enzyme in a container, wherein the inner surface of the container has fine ceramics; and The solution is freeze-dried to obtain a pharmaceutical composition. The glycolytic enzyme is chondroitinase ABC, The amount of enzyme contained in each container is 1 μg or more and less than 10 μg, The enzyme titer is 0.32 units / μg or more and 1 unit / μg or less.

16. The manufacturing method according to claim 15, wherein: The fine ceramic is selected from silicon oxide, silicon nitride, silicon oxynitride, aluminum oxide, aluminum nitride and aluminum oxynitride.

17. The manufacturing method according to claim 16, wherein: The silicon oxide is silicon dioxide.

18. The manufacturing method according to claim 15, wherein: The inner surface of the container is provided with a film containing the fine ceramic.

19. The manufacturing method according to claim 18, wherein: The film containing the fine ceramic is formed by chemical vapor deposition, physical vapor deposition, spray pyrolysis or sputtering.

20. The manufacturing method according to claim 19, wherein: The chemical vapor deposition method is a plasma chemical vapor deposition method.

21. The manufacturing method according to claim 15, wherein: The enzyme activity of the solution contained in each container is greater than or equal to 0.5 units and less than 4.5 units.

22. The manufacturing method according to claim 15, wherein: When the enzyme activity before lyophilization is assumed to be 100%, the enzyme activity of the enzyme after lyophilization is 75% or more.

23. The production method according to any one of claims 15 to 22, wherein: The pharmaceutical composition comprises at least one of polyethylene glycol and sucrose.

24. The manufacturing method according to claim 23, wherein: The pharmaceutical composition comprises a mixture of polyethylene glycol and sucrose.

25. The manufacturing method according to claim 15, wherein: The pharmaceutical composition is used to treat hernia, lysosomal storage disease, keloid, hypertrophic scar, muscular dystrophy or spinal cord injury.

26. The manufacturing method according to claim 15, wherein: The container is a vial, a syringe or an ampoule.

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