Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
  • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
  • A61K35/36—Skin; Hair; Nails; Sebaceous glands; Cerumen; Epidermis; Epithelial cells; Keratinocytes; Langerhans cells; Ectodermal cells
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
  • A61K35/66—Microorganisms or materials therefrom
  • A61K35/76—Viruses; Subviral particles; Bacteriophages
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P11/00—Drugs for disorders of the respiratory system
  • A61P11/02—Nasal agents, e.g. decongestants
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P17/00—Drugs for dermatological disorders
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P17/00—Drugs for dermatological disorders
  • A61P17/04—Antipruritics
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P19/00—Drugs for skeletal disorders
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P19/00—Drugs for skeletal disorders
  • A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P25/00—Drugs for disorders of the nervous system
  • A61P25/04—Centrally acting analgesics, e.g. opioids
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P37/00—Drugs for immunological or allergic disorders
  • A61P37/08—Antiallergic agents

Definitions

• the present invention relates to a vaccinia virus-inoculated rabbit inflammation skin extract having a more stable quality by confirming that it contains a predetermined amount of sulfated tyrosine by a test or inspection, a preparation containing the extract as an active ingredient, and the like.
• the raw materials used in the production of pharmaceuticals which are the active ingredients of pharmaceuticals, are called “Drugs”.
• the quality of the drug substance is also required to be ensured. This is because the quality of the drug depends on the quality of the drug substance.
• the drug substance is exclusively used for the manufacture of other drugs, and by definition, drug substances are included in drugs.
• the drug and the drug substance may be referred to separately, and the drug in this case means a drug excluding the drug substance.
• Vaccinia virus inoculated rabbit inflamed skin extract (hereinafter sometimes referred to as “this extract”) is an extract containing non-protein active substances extracted and isolated from inflamed rabbit tissue inoculated with vaccinia virus. It is a thing.
• the extract is liquid in the extracted state, but can also be made solid by drying.
• the preparation containing the present extract as an active ingredient (hereinafter sometimes referred to as “the present preparation”) is very useful as a pharmaceutical product as described later.
• this extract is the drug substance of this formulation.
• the specific product manufactured and sold by the applicant in Japan as this preparation is “Vaccinia virus inoculated rabbit inflammation skin extract-containing preparation” (trade name: Neurotropin / NEUROTROPIN (registered trademark), hereinafter referred to as “NTP preparation”.
• NTP preparations include injections (hereinafter referred to as “NTP injections”) and tablets (hereinafter referred to as “NTP tablets”), both of which are ethical drugs.
• NTP extract Vaccinia virus inoculated rabbit inflammation skin extract
• NTP preparations NTP injections and NTP tablets
• Indications for NTP injections are: “Low back pain, neck and shoulder syndrome, symptomatic neuralgia, pruritus associated with skin diseases (eczema, dermatitis, urticaria), allergic rhinitis, cold feeling / abnormality of SMON aftereffects” “Perception / Pain”.
• the indications for NTP tablets are “postherpetic neuralgia, low back pain, cervical-arm syndrome, peri-shoulderitis, osteoarthritis”.
• the NTP formulation was created by the applicant and developed as a pharmaceutical product. NTP preparations have been evaluated for their excellent effectiveness and safety, have been sold for many years, and have established a firm position in the Japanese pharmaceutical market. Currently, NTP products are exported to China and sold under the trade name “NEUROTROPIN”. The indications for NTP products in China are the same as in Japan.
• this preparation is very useful as a pharmaceutical product, and this extract is still very useful as a drug substance for this preparation.
• this extract was extracted and isolated from the inflamed skin tissue of rabbits inoculated with vaccinia virus. Therefore, the present extract contains a very large number of substances (components), and the present preparation produced using the extract also contains a very large number of substances (components). Therefore, how to control the quality of the extract and the preparation to be stable is a very important matter.
• the method is a biological test method for obtaining an analgesic factor using a SART stress (repeated cold load) animal, which is a chronic stress animal whose pain threshold is lower than that of a normal animal (“Nichi Pharmacology”, Vol. 72). No. 5, pp. 573-584, 1976).
• an analgesic coefficient is obtained by performing an analgesic test by a Randall-Selitto method using a SART stress (repeated cold load) animal which is a chronic stress animal whose pain threshold is lower than that of a normal animal.
• pressure is applied to the tail of the mouse, and the analgesic effect is measured using the pressure weight until the mouse shows an escape reaction as an index.
• the analgesic coefficient is a value obtained by dividing the pressurized weight measured after administering a drug by the value before drug administration.
• the analgesic coefficient exceeds a certain value determined by the applicant, the analgesic effect is positive, the percentage of animals determined to be positive is obtained, and the analgesic effective rate (%) To do. From this value, the ED 50 value is determined from the results of measurement of standard products diluted to various concentrations.
• “Neurotropin unit (NU)” the unit of biological activity (titer) used by applicants for NTP preparations, refers to 1 mg of this extract with an ED 50 value of 100 mg / kg (mouse body weight) The activity is defined as 1 neurotropin unit.
• the indication of “unit” is used as a measure of the amount of active ingredient (titer) of the present extract and the present preparation, but substantially, it is used in the NTP extract and NTP preparation. It has the same meaning as “neurotropin unit”.
• this extract and this preparation manufactured by the applicant should be subjected to the following multiple confirmation tests in addition to the above analgesic activity quantification and should be adapted to these.
• the following lots of confirmation tests are conducted instead of ensuring that the production lot is appropriate only by the biological activity (titer). In order to meet these requirements, the extract and preparation should be used and shipped.
• the amino acids, UV absorbing substances, phosphorus, nucleobases, etc. that are the test subjects are not the core active ingredients of the extract or the preparation.
• the confirmation tests for amino acids, UV-absorbing substances, phosphorus, and nucleobases are qualitative tests, as long as the test target amino acid is present, and how much is contained. It doesn't matter whether or not. Nonetheless, the applicants approved the manufacture of pharmaceuticals from the authorities as a measure to reduce the variation in quality among production lots and to ensure homogeneity in both the extract and the preparation with unknown active ingredients. It is the one that was determined in the get.
• the extract and the preparation are appropriately produced. It is treated as a product, that is, used or shipped. Thereby, the dispersion
• the applicant does not set a standard for the amount of sulfated tyrosine contained in NTP extracts and NTP preparations produced so far, and measures the content, and the content is constant. After confirming that it is within the range, the NTP extract or NTP preparation is not used or shipped.
• Patent Documents 1 to 3 exist as documents disclosing the present extract or the present preparation. These include a description of the content of amino acids and nucleobases in the extract or preparation. Similarly, regarding the present extract or the present preparation, Patent Document 4 describes the content of silicon. However, these Patent Documents 1 to 4 do not describe how much the extract or the preparation contains a specific substance called sulfated tyrosine. Furthermore, these references also describe that the content of sulfated tyrosine in the extract or the preparation is used as an index for managing the quality of the extract or the preparation more stably. There is no suggestion.
• the present invention ensures the quality of the extract and the preparation with the amount of sulfated tyrosine contained in the extract and the preparation as an index. That is, the present invention provides the present extract, the present preparation, and the like, which are appropriately manufactured by confirming that a prescribed amount of sulfated tyrosine is contained.
• the extract and the preparation according to the present invention contain a predetermined amount of sulfated tyrosine. Thereby, this extract and this formulation which concern on this invention can be handled as what was manufactured appropriately.
• the present invention contributes to ensuring that the quality of the extract and the preparation is more stably secured, and that the effectiveness and safety of the extract and the preparation are more constant.
• Protein plays an important role in vivo.
• the sequence information of amino acids constituting the protein is encoded in genomic DNA.
• Proteins are synthesized by synthesizing mRNA using DNA as a template, and then combining amino acids based on the mRNA information (translation). Some proteins synthesized in this way function themselves, but many have post-translational activity expressed or controlled by addition of sugar, partial cleavage, phosphorylation, etc. (post-translational modification: Post -translational modification).
• Tyrosine sulfation is one of the post-translational modifications of proteins, and sulfo groups are added to protein tyrosine residues.
• Sulfated tyrosine is a very stable substance that is produced in the process of metabolizing tyrosine sulfated protein and is not easily degraded by mammalian sulfatase.
• leukocyte rolling a phenomenon in which leukocytes roll on the vascular endothelium around the tissue (leucocyte rolling) is observed.
• Leukocytes are repeatedly contacted with endothelial cells by adhesion molecules such as selectins on the surface, and search for migration factors such as chemokines presented on the vascular endothelium.
• chemokines chemokines presented on the vascular endothelium.
• leukocytes recognize chemokines, they firmly adhere to vascular endothelial cells and stop moving.
• Leukocytes infiltrate into the inflamed tissue through gaps created by locally destroying the junctions of the endothelial cells.
• tyrosine sulfation has been shown to be involved in this adhesion reaction. That is, the tyrosine near the amino terminus of PSGL-1 (P-selectin glycoprotein ligand-1), which is a receptor for selectin, is sulfate-modified, which is why the binding of selectin to PSGL-1 is based on high affinity. Shown to be mandatory.
• chemokine receptors CCR5, CXCR4, etc.
• CCR5 CXCR4, etc.
• sulfated tyrosine originating from various sulfated proteins in inflamed tissue can be used as an indicator of the inflammatory state of skin tissue. is there.
• This extract is extracted from skin tissue inflamed by inoculating rabbits with vaccinia virus, and it is necessary to induce a sufficient inflammatory reaction. Therefore, sulfated tyrosine is a reasonable substance (component) as an index for managing the quality of the extract and the preparation as more stable.
• This extract was obtained by inoculating rabbit skin with vaccinia virus and collecting the inflamed skin tissue, crushing it, adding extraction solvent, treating it, removing the tissue fragment, and deproteinizing it. It can be obtained by adsorbing to an adsorbent in acidic conditions and then eluting the active ingredient in basic conditions.
• the vaccinia virus may be of any strain. Examples include Lister, Dairen, Ikeda, EM-63, and New York City Public Health Board (Health) strains.
• any rabbit may be used as long as it belongs to the order of rabbit eyes.
• examples include rabbits, rabbits (rabbits made from rabbits), rabbits (Japanese rabbits), rabbits, snow rabbits, and the like. Of these, chira rabbits are suitable for use.
• Japan there is a so-called rabbit that has been bred from the past and used widely as domestic animals or experimental animals.
• varieties breeding of the rabbit, but varieties such as Japanese white varieties and New Zealand white varieties (New Zealand white) can be suitably used.
• the following processes are used.
• A The skin tissue of rabbits inoculated with vaccinia virus intradermally is collected, the germed tissue is crushed, and an extraction solvent such as water, phenol water, physiological saline, or phenol-added glycerin water is added to the number. After performing the extraction process for one day, a crude extract (filtrate or supernatant) from which tissue pieces have been removed is obtained by filtration or centrifugation.
• the crude extract obtained in (A) is adjusted to an acidic pH, heated, filtered or centrifuged to remove protein.
• the deproteinized solution is adjusted to a basic pH, heated, and further filtered or centrifuged to obtain a deproteinized filtrate or supernatant.
• the filtrate or supernatant obtained in (B) is adjusted to an acidic pH and adsorbed on an adsorbent such as activated carbon or kaolin.
• an extraction solvent such as water to the adsorbent obtained in (C), adjust to basic pH, and elute the adsorbed component to remove the inflammatory skin extract (this extract) inoculated with vaccinia virus. obtain.
• Inflamed skin tissue obtained by inoculating the skin of rabbits with vaccinia virus intradermally is collected.
• the collected skin tissue is washed and disinfected with a phenol solution.
• This inflamed skin tissue is crushed and 1 to 5 times the amount of extraction solvent is added.
• crushing means crushing finely into a mince using a mincing machine or the like.
• extraction solvents distilled water, physiological saline, weakly acidic to weakly basic buffer solutions, and the like can be used as extraction solvents.
• Sterilizers and preservatives such as phenol, stabilizers such as glycerin, sodium chloride, potassium chloride
• salts such as magnesium chloride may be appropriately added.
• extraction can be facilitated by disrupting the cell tissue by treatment with freeze-thaw, ultrasound, cell membrane lytic enzyme, surfactant or the like.
• the resulting suspension is left for 5-12 days. Meanwhile, the mixture may be heated to 30 to 45 ° C. with or without stirring as appropriate.
• the obtained liquid is subjected to solid-liquid separation (filtration, centrifugation, etc.) to remove the tissue piece to obtain a crude extract (filtrate or supernatant).
• a protein removal process is performed about the crude extract obtained by (A).
• Deproteinization can be carried out by known methods commonly used, such as heat treatment, treatment with a protein denaturant (for example, an organic solvent such as acid, base, urea, guanidine, acetone, etc.), isoelectric precipitation, salting out. Etc. can be applied.
• a protein denaturant for example, an organic solvent such as acid, base, urea, guanidine, acetone, etc.
• Etc. can be applied.
• the insoluble protein that has been precipitated by a usual method for removing insoluble matters for example, filtration using filter paper (cellulose, nitrocellulose, etc.), glass filter, celite, zeit filtration plate, etc., ultrafiltration, centrifugation, etc.
• a removed filtrate or supernatant is obtained.
• the filtrate or supernatant obtained in (B) is adjusted to acidic, preferably pH 3.5 to 5.5, and the adsorption operation to the adsorbent is performed.
• adsorbents include activated carbon, kaolin, and the like. Add the adsorbent to the extract and stir, or pass the extract through an adsorbent-filled column to add the active ingredient to the adsorbent. Can be adsorbed. When an adsorbent is added to the extract, the adsorbent can be obtained by adsorbing the active ingredient by removing the solution by filtration or centrifugation.
• an elution solvent is added to the adsorbent, and the mixture is adjusted to basic, preferably pH 9-12, at room temperature or appropriately. Elution is carried out by heating or stirring, and the adsorbent is removed by a usual method such as filtration or centrifugation.
• a basic solvent such as water adjusted to a basic pH, methanol, ethanol, isopropanol or the like, or an appropriate mixed solution thereof can be used, and preferably water adjusted to pH 9-12.
• the amount of the elution solvent can be appropriately set.
• the pH is appropriately adjusted to near neutral, etc. to finally obtain a vaccinia virus-inoculated rabbit inflammation skin extract (this extract). Can do.
• this extract is liquid at the time of completion, it can be made to have a desired concentration by appropriately concentrating and diluting.
• sodium chloride or the like can be added to prepare a solution that is isotonic with physiological saline.
• oral solid preparations such as a tablet, can also be manufactured by performing operation, such as concentration and drying suitable for this extract in a liquid state. Specific methods for producing such an oral solid preparation from this extract are described in the specifications of Japanese Patent Nos. 3818657 and 4883798. This preparation is an injection or a solid preparation for oral use thus obtained.
• the content of sulfated tyrosine in the present extract and the present preparation can be measured by a commonly used quantitative method. Specifically, for example, a measurement method using a liquid chromatograph mass spectrometer (LC-MS) or a capillary electrophoresis mass spectrometer (CE-MS), high performance liquid chromatography (HPLC), or the like can be used. . In any method, sulfated tyrosine in the present extract or the present preparation can be quantified using a calibration curve prepared with a sulfated tyrosine sample for calibration.
• LC-MS liquid chromatograph mass spectrometer
• CE-MS capillary electrophoresis mass spectrometer
• HPLC high performance liquid chromatography
• the amount of sulfated tyrosine contained in the present extract and the present preparation produced by the applicant by the method as described above was measured.
• the extract and the preparation contained 125 ng or more of sulfated tyrosine per unit, although there were variations.
• the extract and the preparation contained 125 to 425 ng of sulfated tyrosine per unit. Therefore, when the quality of this extract and this preparation was managed using the content of sulfated tyrosine in this extract and this preparation as an index, it was determined that such content could be provided as a reference.
• “per unit” means, as understood from the above description, per content of the active ingredient in the present extract or the present preparation.
• the present extract (NTP extract) produced by the applicant contains 1.2 units / mL of active ingredient.
• the present preparation for injection (NTP injection) produced using this also contains 1.2 units / mL of the active ingredient. NTP injections are available in 3 mL and 1 mL volumes. Therefore, 3.6 mL of the active ingredient is contained in the 3 mL preparation, and 1.2 units of the active ingredient is contained in the 1 mL preparation.
• the present oral preparation (NTP tablet) manufactured by the applicant contains 4 units of active ingredient per tablet.
• this extract can be concentrated or diluted.
• the present preparation can also be prepared containing various units.
• the amount of the active ingredient contained in the extract or the amount of the preparation (per mL, per mg, per tube, per tablet, etc.) can also vary. Therefore, basically, it is meaningful to define the content of sulfated tyrosine in relation to the amount of the active ingredient in the extract or the preparation. This is because it leads to a relationship with the effectiveness and safety of the extract and the preparation. Therefore, the applicant defined the content of sulfated tyrosine in the present extract or the present preparation per content of active ingredient ("per unit").
• the applicant currently manufactures and sells NTP preparations it is possible to specify specific injections per mL and per tube, and per tablet, depending on the dose. Such provisions were also made because of their significance.
• NTP injections are generic products (generic drugs) or similar formulations (similar drugs) manufactured by companies other than the applicant (hereinafter referred to as “other companies”) in Japan and China (hereinafter referred to as “other company injections”). ”)”.
• other companies in Japan and China
• other company injections also subjected to quantification of the inflammatory skin extract of rabbit skin inoculated with vaccinia virus (this extract), which is an active ingredient, using SART stress mice as an index of analgesic effect.
• the active ingredient content is 1.2 units or 1.2 Analgecine (1.2AGC) units per mL as in the case of the NTP injection, and 3.6 units or 3.6 Analgecine ( 3.6AGC) unit.
• “Neurotropin units” used in NTP preparations and "Units” and “Analgecine units” used by other companies differ only in the display method, and the same scale is used to regulate the content of active ingredients. It has become. Therefore, in the present application, “unit” is used as a display indicating the content of the present extract, which is an active ingredient, including not only NTP preparations but also injections of other companies. In this way, a preparation comprising a plurality of vaccinia virus-inoculated rabbit inflammation skin extracts as an active ingredient displays the amount of the active ingredient in “units”, and the “unit” indication for the preparation is clear to those skilled in the art.
• this preparation is a concept including NTP preparations (NTP injections and NTP tablets) and injections from other companies.
• other company tablets when another company manufactures and sells tablets (hereinafter referred to as “other company tablets”) as generic products or similar products of NTP tablets, it is a concept that includes other company tablets.
• Example 1 (Production of the present extract) The skin of healthy mature rabbits was inoculated intradermally with vaccinia virus, and the cut skin was cut out and collected. The collected skin is washed and disinfected with a phenolic solution, then the excess phenolic solution is removed, crushed, the phenolic solution is added and mixed, allowed to stand for 3-7 days, and then stirred for an additional 3-4 days Warmed to 35-40 ° C. Thereafter, the extract obtained by solid-liquid separation was adjusted to pH 4.5 to 5.2 with hydrochloric acid, heat-treated at 90 to 100 ° C. for 30 minutes, and then filtered to remove proteins. Further, the filtrate was adjusted to pH 9.0 to 9.5 with sodium hydroxide, heated at 90 to 100 ° C. for 15 minutes, and then separated into solid and liquid.
• the obtained deproteinized solution was adjusted to pH 4.0 to 4.3 with hydrochloric acid, and 2% of activated carbon was added to the amount of deproteinized solution and stirred for 2 hours, followed by solid-liquid separation.
• Example 2 (Method for measuring sulfated tyrosine content) The sulfated tyrosine content of this extract and this preparation was measured with a high performance liquid chromatograph mass spectrometer (LC-MS) as follows.
• the extract prepared according to Example 1 (1.2 units / mL) was diluted 10-fold with water and injected into LC-MS.
• This preparation (NTP injection) produced using this extract produced according to Example 1 was similarly diluted 10-fold with water and injected into LC-MS.
• This preparation (NTP tablet) produced using this extract produced according to Example 1 was washed 3 times with 3 mL of methanol / chloroform (1: 1) 3 times to remove the film coat layer and dried.
• a calibration curve was prepared by preparing a standard solution of sulfated tyrosine as an aqueous solution.
• ⁇ DP Voltage applied to orifice plate
• ⁇ FP Voltage applied to focus ring
• CE Collagen energy
• CXP Voltage applied to Q2 outlet
• NEB Nebulizer gas pressure
• CUR Curtain gas pressure
• Ion spray -CAD pressure of the collage gas -TEM: turbo gas temperature
• Example 3 (Measurement result of sulfated tyrosine content of this extract) Table 2 shows the results of measuring the content of sulfated tyrosine in this extract by the method described in Example 2 above.
• the active ingredient content in this extract is 1.2 units / mL.
• the content of sulfated tyrosine in the extract was expressed both per unit of the extract (“/ unit”) and per mL of the extract (“/ mL”).
• the symbols A to C in the lot number represent differences in the manufacturing location (facility) of the applicant.
• the measured values were aligned with 3 significant figures (hereinafter the same for all measured values).
• Example 4 (Measurement result 1 of sulfated tyrosine content of this preparation) Table 3 shows the results of measuring the sulfated tyrosine content in the NTP injection (containing 1.2 units per mL) among the present preparations by the method described in Example 2 above. The results are shown as the content per unit (“/ unit”) of the active ingredient of this preparation, 1 mL of this preparation (“/ mL”), and 1 ampule of 3 mL (“/ tube”). It was.
• Example 5 (Measurement result 2 of sulfated tyrosine content of this preparation) Table 4 shows the results of measuring the sulfated tyrosine content in NTP tablets (containing 4 units per tablet) by the method described in Example 2 above. The results are shown in terms of each content of active ingredient per unit (“/ unit”) and per tablet (“/ tablet”).
• Comparative Example 1 (Measurement result of sulfated tyrosine content of other company's injections)
• Table 5 shows the results of measuring the sulfated tyrosine content of other companies' injections in the same manner as NTP injections. “Rose Morgen Injection” has been sold at the time of filing of the present application and does not exist in the market. The following measurement results are the results obtained and measured by the applicant in the market in the past. .
• sulfated tyrosine contained 125 ng / unit or more, more specifically 150 ng / unit or more (Tables 2 to 4).
• the extract and the preparation produced by the applicant contained sulfated tyrosine in the range of 125 to 425 ng / unit, more specifically 150 to 400 ng / unit.
• none of the preparations produced by other companies had such a sulfated tyrosine content (Table 5). Although it is not clear what causes the difference in the content of sulfated tyrosine in this preparation produced by each company, it is strongly estimated that it is caused by the difference in the production method of this preparation in each company.
• tyrosine sulfation is closely related to inflammation of rabbit skin, and sulfated tyrosine released from various sulfated proteins is an indicator of skin tissue inflammation (inflammatory state). Therefore, as an applicant, this substance (component) is contained in the present preparation (NTP injection) manufactured by the applicant more than the injections of other companies. It is understood that this is a desirable feature.
• the presence of sulfated tyrosine in the extract and the preparation in an amount of 125 ng / unit or more, preferably 150 ng / unit or more, is a feature of the extract and the preparation.
• the presence of sulfated tyrosine in the range of 125 to 425 ng / unit, preferably in the range of 150 to 400 ng / unit, is a feature of the extract and the preparation.
• an injection containing 1.2 units per mL contains sulfated tyrosine of 150 ng / mL or more, preferably 180 ng / mL or more. This is a feature of the injection.
• sulfated tyrosine is contained in the range of 150 to 510 ng / mL, preferably 180 to 480 ng / mL, is a feature of the present injection.
• the sulfated tyrosine is 450 ng / tube or more per tube, preferably The inclusion of 540 ng / tube or more is a feature of this 3 mL injection.
• tablets containing 4 units per tablet contain sulfated tyrosine of 500 ng / tablet or more, preferably 600 ng / tablet or more. It is a characteristic of tablets. Similarly, the fact that sulfated tyrosine is contained in the range of 500 to 1700 ng / tablet, preferably 600 to 1600 ng / tablet, is a feature of this tablet.
• the amount of sulfated tyrosine contained in the extract and the preparation it is possible to measure the amount of sulfated tyrosine contained in the extract and the preparation, confirm whether the extract and the preparation are properly manufactured, and control the quality of these extracts. Is possible. That is, the amount of sulfated tyrosine contained in the present extract and the present preparation is measured, and if it is 125 ng / unit or more, preferably 150 ng / unit or more, it can be properly produced. is there. Also, confirm whether the amount of sulfated tyrosine contained in the extract and the preparation is in the range of 125 to 425 ng / unit, preferably 150 to 400 ng / unit, and the extract and the preparation are appropriate. It is also possible to be manufactured.
• the amount of sulfated tyrosine contained in this injection was measured, and if it was 150 ng / mL or more, preferably 180 ng / mL or more, the injection was appropriately produced. Can be.
• the amount of sulfated tyrosine contained in this injection is measured, and it is confirmed that it is in the range of 150 to 510 ng / mL, preferably 180 to 480 ng / mL.
• the amount of sulfated tyrosine contained is measured and is 450 ng / tube or more, preferably 540 ng / tube or more, or 450 to 1530 ng / tube, It is possible to confirm that the dose is preferably in the range of 540 to 1440 ng / tube, and to make the 3 mL injection preparation appropriately manufactured.
• the amount of sulfated tyrosine contained in this tablet is measured, and if it is 500 ng / tablet or more, preferably 600 ng / tablet or more, this tablet is properly manufactured. It is possible. It is also possible to confirm that the amount of sulfated tyrosine contained in the tablet is in the range of 500 to 1700 ng / tablet, preferably 600 to 1600 ng / tablet, and that the tablet is appropriately manufactured. Is possible.
• the amount of sulfated tyrosine is measured is usually that the amount of sulfated tyrosine is measured for each production lot of the extract or the preparation. Used in meaning.
• a rabbit inflammatory skin extract inoculated with vaccinia virus wherein the amount of sulfated tyrosine contained in the extract is measured, and the content is 125 to 425 ng per unit of the extract Vaccinia virus-inoculated rabbit inflamed skin extract that was confirmed to be properly manufactured.
• a rabbit inflammatory skin extract inoculated with vaccinia virus which is a liquid the amount of sulfated tyrosine contained in the extract is measured, and the content is 150 to 510 ng per 1 mL of the extract
• a preparation containing an extract of inflammatory skin of rabbits inoculated with vaccinia virus, the amount of sulfated tyrosine contained in the preparation being measured, and the content of the extract of inflammatory skin of rabbits inoculated with vaccinia virus in the preparation A preparation containing an inflammatory skin extract of rabbit inflammation inoculated with vaccinia virus that has been properly manufactured by confirming that it is 125 ng or more per unit.
• Formulation containing vaccinia virus-inoculated rabbit inflammation skin extract shipped as properly manufactured by confirming that it is 125 ng or more per unit.
• Formulation containing vaccinia virus inoculated rabbit inflammation skin extract shipped as properly manufactured by confirming that it is 125-425 ng per unit.
• Formulation containing vaccinia virus inoculated rabbit inflammation skin extract shipped to meet the standard of 125 to 425 ng per product.
• An injection containing 3 mL of vaccinia virus-inoculated rabbit inflammation skin extract, the amount of sulfated tyrosine contained in the injection is measured, and the content is per tube of the injection 3 mL injection containing vaccinia virus-inoculated rabbit inflammation skin extract shipped to meet the criteria of 450 ng or more.
• a rabbit inflammatory skin extract inoculated with vaccinia virus which is a liquid, the amount of sulfated tyrosine contained in the extract is measured, and the content is 150 to 510 ng per 1 mL of the extract
• a preparation containing an extract of inflammatory skin of rabbits inoculated with vaccinia virus, the amount of sulfated tyrosine contained in the preparation being measured, and the content of the extract of inflammatory skin of rabbits inoculated with vaccinia virus in the preparation A preparation containing a inflammatory skin extract of rabbit inoculated with vaccinia virus, which is confirmed to be 125 ng or more per unit.
• a preparation containing an extract of inflammatory skin of rabbit inoculated with vaccinia virus, the amount of sulfated tyrosine contained in the preparation being measured, and the content of the extract of inflammatory skin of rabbit inoculated with vaccinia virus in the preparation A preparation containing inflammatory skin extract of rabbit inflammation inoculated with vaccinia virus, which is confirmed to be 125 to 425 ng per unit.
• An injection containing an inflammatory skin extract of rabbits inoculated with vaccinia virus, the amount of sulfated tyrosine contained in the injection being measured, and the content being 150 to 510 ng per 1 mL of the injection An injection containing an inflammatory skin extract of rabbits inoculated with vaccinia virus, characterized in that (40) A 3 mL injection containing a inflammatory skin extract of rabbits inoculated with vaccinia virus, the amount of sulfated tyrosine contained in the injection being measured, and the content per tube of the injection A 3 mL injection containing a inflammatory skin extract of rabbit inflammation inoculated with vaccinia virus, characterized in that it was confirmed to be 450 ng or more.
• a 3 mL-containing injection containing a inflammatory skin extract of rabbit inflammation inoculated with vaccinia virus, the amount of sulfated tyrosine contained in the injection being measured, and the content per tube of the injection A 3 mL injection containing a inflammatory skin extract of rabbit inflammation inoculated with vaccinia virus, characterized in that it was confirmed to be 450 to 1530 ng.
• (46) The tablet according to (42) or (43), which is an analgesic.
• a vaccinia virus-inoculated rabbit inflammation preparation containing an extract of inflammation skin, the amount of sulfated tyrosine contained in the preparation was measured before shipment, and the content was determined to be vaccinia virus-inoculated rabbit in the preparation Formulation containing inflammatory skin extract inoculated with vaccinia virus, characterized in that it is confirmed to be 125 ng or more per unit of inflammatory skin extract.
• a vaccinia virus inoculated rabbit preparation containing an inflammatory skin extract, the amount of sulfated tyrosine contained in the preparation was measured before shipment, and the content was determined to be the vaccinia virus inoculated rabbit in the preparation.
• Formulation containing inflammatory skin extract inoculated with vaccinia virus characterized in that it is confirmed to be 125 to 425 ng per unit of inflammatory skin extract.
• An injection containing a inflammatory skin extract of rabbit inflammation inoculated with vaccinia virus, measuring the amount of sulfated tyrosine contained in the injection before shipment, and the content per mL of the injection An injection containing an inflammatory skin extract of a rabbit inoculated with vaccinia virus, characterized in that it was confirmed to be 150 ng or more.
• An injection containing an inflammatory skin extract of rabbit inflammation inoculated with vaccinia virus, the amount of sulfated tyrosine contained in the injection is measured before shipment, and the content is 1 mL of the injection, An injection containing an inflammatory skin extract of rabbit inoculated with vaccinia virus, characterized in that it was confirmed to be 150 to 510 ng.
• the amount of sulfated tyrosine contained in the preparation containing the inflammatory skin extract of rabbit inoculated with vaccinia virus was measured, and the content was 125 per 1 unit of the inflammatory skin extract inoculated with vaccinia virus in the preparation.
• the method for testing the preparation wherein the preparation is appropriately manufactured when the amount is ⁇ 425 ng.
• (62) When the amount of sulfated tyrosine contained in the injection is measured in an injection containing the inflammatory skin extract of rabbit inflammation inoculated with vaccinia virus, and the content is 150 ng or more per mL of the injection A method for testing the injection, wherein the injection is appropriately manufactured.
• the preparation A method for managing the production of the preparation, wherein the production of the preparation is appropriately performed.
• the amount of sulfated tyrosine contained in the preparation containing the inflammatory skin extract of rabbit vaccinia virus inoculated rabbit is 125 to 425 ng per unit of the inflammatory skin extract of rabbit vaccinia virus inoculated rabbit A method for managing the production of the preparation, wherein the preparation of the preparation is appropriately made.
• the tablet containing the inflammatory skin extract of rabbit inflammation inoculated with vaccinia virus the tablet is appropriately manufactured when the amount of sulfated tyrosine contained in the tablet is 500 ng or more per tablet.
• a method for managing the manufacture of the tablets In a tablet containing a inflammatory skin extract extracted from vaccinia virus-inoculated rabbit, when the amount of sulfated tyrosine contained in the tablet is 500 to 1700 ng per tablet, the tablet is appropriately produced. A method for managing the manufacture of the tablets, which has been made.
• the amount of sulfated tyrosine contained in the preparation containing a vaccinia virus-inoculated rabbit inflammation skin extract was measured, and the content thereof was determined as the vaccinia virus-inoculated rabbit inflammation skin extract 1 in the preparation.
• a method for shipping the preparation wherein when it is confirmed that the amount is 125 ng or more per unit, the preparation is determined to be shippable as appropriately manufactured.
• the amount of sulfated tyrosine contained in the preparation containing a vaccinia virus-inoculated rabbit inflammation skin extract was measured, and the content of the preparation was determined as vaccinia virus-inoculated rabbit inflammation skin extract 1 in the preparation.
• a method for shipping the preparation wherein it is determined that the preparation can be shipped as properly manufactured when it is confirmed that the amount is 125 to 425 ng per unit.
• the amount of sulfated tyrosine contained in the injection containing the inflammatory skin extract inoculated with vaccinia virus was measured, and the content was 150 ng or more per mL of the injection Is confirmed, it is determined that the injection can be shipped as properly manufactured.
• the amount of sulfated tyrosine contained in the injection containing the inflammatory skin extract inoculated with vaccinia virus was measured, and the content was 150 to 510 ng per mL of the injection When it is confirmed that the injection is shipped, it is determined that the injection can be shipped as properly manufactured.
• (101) The preparation according to (93) or (94), wherein the effect is an analgesic effect.
• the method for producing an injection wherein the variation of the effect of the injection is reduced for each production lot, wherein the production of the injection is appropriately performed when the amount is ⁇ 510 ng.
• the amount of sulfated tyrosine contained in the injection is measured for each production lot, and the content is determined by the injection 1 Management of the production of injections containing 3 mL, which reduces the variability of the effects of the injections from lot to lot, assuming that the production of the injections was appropriately made when the amount is 450 ng or more per tube how to.
• a method for managing the manufacture of the tablet in which, in some cases, the manufacture of the tablet is appropriately performed, and the variation of the effect of the tablet is reduced for each production lot.
• the amount of sulfated tyrosine contained in the tablet was measured for each production lot, and the content was 500 to 1700 ng per tablet. In this case, it is assumed that the manufacture of the tablet is appropriately performed, and the method for managing the manufacture of the tablet with the variation of the effect of the tablet being reduced for each production lot is reduced.
• the method for inspecting an injection wherein the injection has been appropriately manufactured, and variation in the effect of the injection has been reduced for each production lot.
• the amount of sulfated tyrosine contained in the injection is measured for each production lot, and the content is 150 per 1 mL of the injection.
• the method for inspecting an injection, wherein the variation of the effect of the injection is reduced for each production lot, wherein the production of the injection is appropriately performed when the amount is ⁇ 510 ng.
• a method for inspecting a tablet in which, in some cases, manufacturing of the tablet is appropriately performed, and variation in the effect of the tablet is reduced for each production lot.
• (129) In a tablet containing an extract of inflammatory skin of rabbits inoculated with vaccinia virus, the amount of sulfated tyrosine contained in the tablet was measured for each production lot, and the content was 500 to 1700 ng per tablet. In this case, the tablet is manufactured appropriately, and the variation of the effect of the tablet for each production lot is reduced.
• the examination method according to any one of (122) to (129), wherein the effect is an analgesic effect.
• a method for shipping the tablet in which, when it is confirmed that the tablet is properly manufactured, it is determined that the tablet can be appropriately manufactured, and the variation of the effect of the tablet in each production lot is reduced.
• the amount of sulfated tyrosine contained in a tablet containing a vaccinia virus-inoculated rabbit inflammation skin extract was measured for each production lot, and the content was 500-1700 ng per tablet. When it is confirmed that the tablet is properly manufactured, it is determined that the tablet can be manufactured appropriately.
• the shipping method according to any one of (137) to (144), wherein the effect is an analgesic effect.
• a method for producing a inflammatory skin extract of rabbit inflammatory skin inoculated with vaccinia virus comprising adding an extraction solvent to a crushed sprouted tissue and allowing to stand for 5 to 12 days, .
• the extract according to (152), which is heated to 30 to 45 ° C. with or without stirring while adding an extraction solvent to the crushed germinated tissue and allowing to stand for 5 to 12 days A method for producing an inflammatory skin extract of rabbit inoculated with vaccinia virus.
• the vaccinia according to (153) wherein an extraction solvent is added to the crushed germinated tissue and the mixture is allowed to stand for 3 to 7 days, and further heated to 35 to 40 ° C. with stirring for 3 to 4 days.
• a method for producing a virus-inoculated rabbit inflammation skin extract (155) The method for producing an inflammatory skin extract inoculated with vaccinia virus according to any one of (152) to (154), wherein the extraction solvent added to the crushed germinated tissue is a phenol solution. (156) The method for producing an inflammatory skin extract of rabbit inflammation inoculated with vaccinia virus according to (155), wherein the amount of the phenol solution is 1 to 5 times the amount of the crushed germinated tissue. (157) A inflammatory skin extract inoculated with vaccinia virus produced by the production method according to any one of (152) to (156).
• the preparation according to (160), wherein the amount of sulfated tyrosine contained in the preparation is 125 ng or more per unit of inflammatory skin extract inoculated with vaccinia virus contained in the preparation.
• the preparation according to (160), wherein the amount of sulfated tyrosine contained in the preparation is 125 to 425 ng per unit of inflammatory skin extract inoculated with vaccinia virus contained in the preparation.
• the preparation according to (161), wherein the amount of sulfated tyrosine contained in the injection is 150 ng or more per mL of the injection.
• the heating step for 3 to 4 days in the step of producing a inflammatory skin extract of rabbit inflammation inoculated with vaccinia virus is a step of heating to 35 to 40 ° C. while stirring (172) or ( 173) A rabbit inflammatory skin extract inoculated with vaccinia virus according to 173).
• an extractant was added to the crushed wounded tissue and allowed to stand for 3 to 7 days, and then further maintained at 30 to 45 ° C. for 3 to 4 days.
• a inflammatory skin extract rabbit inoculated with vaccinia virus which is a liquid characterized by (176)
• the inflammatory skin extract of rabbit inflammation inoculated with vaccinia virus which is a liquid according to (175), wherein the content of sulfated tyrosine per mL of the extract is 150 to 510 ng.
• step of heating for 3 to 4 days in the step of producing a inflammatory skin extract of rabbit inflammation inoculated with vaccinia virus is a step of heating to 35 to 40 ° C. with stirring (175) or (176)
• a inflammatory skin extract of rabbit inoculated with vaccinia virus which is a liquid according to (176).
• An injection comprising the inflammatory skin extract of rabbit vaccinia virus inoculated rabbit which is a liquid according to (175) or (177), wherein the amount of sulfated tyrosine contained in the injection is the injection
• a 3 mL injection containing the inflammatory skin extract inoculated with vaccinia virus, which is a liquid according to (175) or (177), wherein the amount of sulfated tyrosine contained in the injection comprises 3 mL injection containing vaccinia virus inoculated rabbit inflammation skin extract, characterized in that variation in sulfated tyrosine content of each production lot is reduced so that it is 450 ng or more per tube of the injection .
• the amount of sulfated tyrosine contained in the extract is 125 ng or more per unit of the extract, so that the extract is contained as an active ingredient, Use of the extract for producing a preparation with reduced variation in the content of sulfated tyrosine.
• the heating step of 3 to 4 days in the step of producing a inflammatory skin extract of rabbit inflammation inoculated with vaccinia virus is a step of heating to 35 to 40 ° C.
• the amount of sulfated tyrosine contained in the extract is 125 ng or more per unit of the extract.
• Use of the preparation according to (196) or (197), wherein the content of sulfated tyrosine per unit of the extract is 125 to 425 ng.
• the heating step of 3 to 4 days in the step of producing a inflammatory skin extract of rabbit inflammation inoculated with vaccinia virus is a step of heating to 35 to 40 ° C. with stirring, according to (197) or (198) Use of the formulation.
• the present invention measures the content of sulfated tyrosine and confirms that it contains a predetermined amount, and the inflammatory skin extract of rabbit inflammation inoculated with vaccinia virus that has been appropriately manufactured or the said extract A preparation containing the product is provided.
• the present invention provides an inspection method in which the extract or preparation is appropriately manufactured by confirming that a predetermined amount of sulfated tyrosine is contained in each production lot of the extract or preparation. Is to provide. Since such extracts and preparations are produced using biological tissues, it is possible to more strictly ensure the constancy of quality for each production lot and are very useful.

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• 2014
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