WO2000051626A1 - Preparations de g-csf modifiees chimiquement - Google Patents

Preparations de g-csf modifiees chimiquement Download PDF

Info

Publication number
WO2000051626A1
WO2000051626A1 PCT/JP2000/001204 JP0001204W WO0051626A1 WO 2000051626 A1 WO2000051626 A1 WO 2000051626A1 JP 0001204 W JP0001204 W JP 0001204W WO 0051626 A1 WO0051626 A1 WO 0051626A1
Authority
WO
WIPO (PCT)
Prior art keywords
chemically modified
preparation
solution
stimulating factor
granulocyte colony
Prior art date
Application number
PCT/JP2000/001204
Other languages
English (en)
Japanese (ja)
Inventor
Yuji Ueno
Yasuki Kato
Kunio Ito
Noboru Konishi
Motoo Yamasaki
Original Assignee
Kyowa Hakko Kogyo Co., Ltd.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Kyowa Hakko Kogyo Co., Ltd. filed Critical Kyowa Hakko Kogyo Co., Ltd.
Priority to AU28247/00A priority Critical patent/AU2824700A/en
Publication of WO2000051626A1 publication Critical patent/WO2000051626A1/fr

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/56—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
    • A61K47/59—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes
    • A61K47/60—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol

Definitions

  • the present invention relates to a preparation comprising a chemically modified polypeptide having granulocyte colony-stimulating factor (G-CSF) activity and a method for stabilizing the preparation.
  • G-CSF granulocyte colony-stimulating factor
  • the present invention provides a preparation comprising a polypeptide having stabilized G-CSF activity by suppressing the elimination reaction of a chemical modifying group.
  • An object of the present invention is to provide a method for stabilizing a chemically modified polypeptide having G-CSF activity and a preparation comprising the chemically modified polypeptide having G-CSF activity. Is to provide.
  • the degradation reaction of elimination of the high molecular compound from the protein greatly affects the stability of the preparation. It has been found that the suppression of the elimination reaction of the polymer compound from the isolated protein is indispensable for the stabilization of the preparation.
  • the present inventors have proposed a G-CSF activity chemically modified with a polymer compound such as PEG. To The elimination reaction of a high molecular compound from a polypeptide having a strong pH dependence is strongly dependent, and the ⁇ ⁇ of a chemically-modified polypeptide solution having G-CSF activity is 5 or less, preferably 3 to 5. Thus, the present inventors have found that they are remarkably suppressed by the above and that they are further suppressed by adding sodium chloride to the solution, thereby completing the present invention.
  • the present invention relates to a preparation containing a chemically modified polypeptide having G—CSF activity and having a pH of 5 or less.
  • the present invention also relates to a method for stabilizing a chemically modified polypeptide having G—CSF activity in a solution by adjusting the pH of the solution to 5 or less.
  • the chemically modified polypeptide having G-CSF activity of the present invention may be any as long as a polymer compound is chemically bonded to a group in the polypeptide having G-CSF activity.
  • the molecular weight, chemical structure, bond molar ratio, and the like of the high molecular compound and the polypeptide having G-CSF activity are not particularly limited.
  • the polymer compound for example, PEG is suitably used.
  • Examples of polypeptides having G-CSF activity chemically modified by PEG include G-CSF described in Japanese Patent Application No. 316400, W090 / 06952, JP-A-5-32559, W095 / 23165, W098 / 55500 and the like.
  • Examples include G-CSF in which at least one amino group, hydroxyl group, carboxyl group, mercapto group or guanidyl group in the molecule of an active polypeptide is chemically modified with PEG.
  • the polypeptide having G-CSF activity used in the present invention can be obtained by using a natural extract or a recombinant DNA technique if it is a peptide included in the definition of granulocyte co-stimulatory factor.
  • Derivatives such as those in which a part of the amino acid sequence of natural G-CSF is substituted with another amino acid, or in which a part is deleted or added, is included. Specific derivatives include, for example, the Thr! Of amino acid at the N-terminal side of the amino acid sequence of natural G-CSF. , Leu 3, G 1 y P ro 5 and Cy s 17 each Al a Thr 3, ⁇ yr eight 5 Oyobi 36 7 (Japanese Patent Application Laid-Open No. 63-267292, hereinafter abbreviated as ND28).
  • the preparation of the present invention is obtained by dissolving an effective amount of a chemically modified polypeptide having G-CSF activity in an aqueous solvent and adjusting the pH of the solution to 5 or less, preferably 3 to 5 or less. It can be manufactured by adjusting to.
  • As the water-soluble solvent an acetate buffer, a phosphate buffer, a citrate buffer, a lactate buffer, a tartrate buffer, and the like are used, and preferably, an acetate buffer and a phosphate buffer are used. More preferably, a phosphate buffer is used.
  • the acid to be added for adjusting the pH is not particularly limited, and examples thereof include hydrochloric acid, acetic acid, phosphoric acid, and citric acid. These acids can be used alone or in combination.
  • the amount of these acids to be added to the solution is not particularly limited as long as the pH in the preparation can be kept at 5 or less, preferably 3 to 5.
  • the preparation of the present invention can stabilize a chemically modified polypeptide having G-CSF activity in the preparation, but by adding salts to the preparation of the present invention, the chemical in the preparation can be stabilized.
  • the stability of the modified polypeptide having G-CSF activity can be further improved.
  • any of organic salts and inorganic salts can be used as long as they can further improve the stability of the chemically modified polypeptide having G-CSF activity in the preparation.
  • Inorganic salts are preferably used. Examples of the organic salts include sodium citrate, sodium acetate, sodium lactate, and sodium succinate.
  • the inorganic salts include ammonium chloride, sodium chloride, potassium chloride, sodium sulfate, ammonium sulfate, and phosphorus.
  • Sodium acid, potassium phosphate, potassium hydrogen phosphate, sodium hydrogen phosphate, dipotassium hydrogen phosphate, sodium hydrogen phosphate, sodium carbonate and the like can be mentioned, and sodium chloride and the like are particularly preferably used.
  • the amount of the salt added is 0.02 mol / L to 1 mol / L, and preferably 0.1 mol / L to 0.5 mol / L.
  • the formulations of the present invention are preferably in unit dosage form suitable for administration by injection.
  • An injection can be prepared as a liquid preparation using a chemically modified polypeptide having G-CSF activity and a carrier comprising a salt solution, a glucose solution, or a mixture of saline and a glucose solution. Further, the liquid preparation can be freeze-dried to prepare a freeze-dried preparation.
  • the lyophilization conditions are not particularly limited, but are usually frozen at —50 ° C or less for 1 to 5 hours, shelf temperature is ⁇ 20 ° C to 0 ° C, and the degree of vacuum is 1 to 1 OPa. Dry for ⁇ 48 hours, and then dry for 16 to 24 hours at a shelf temperature of 10 to 30 ° C and a degree of vacuum of 1 to 10 Pa to obtain a freeze-dried product.
  • the freeze-dried preparation of the present invention can be obtained by, after freeze-drying, sealing with a rubber stopper or an aluminum cap or the like.
  • a solution in which at least one selected from an electrolyte, a water-soluble polymer, a polyhydric alcohol, and a surfactant is used as a stabilizer should be used as a re-dissolution liquid.
  • the electrolyte include sodium chloride, potassium chloride, and phosphate
  • examples of the water-soluble polymer include polyethylene glycol, polyvinyl pyridone, polyvinyl alcohol, and dextran.
  • polyhydric alcohols examples include glycerin, mannitol, xylitol, and sorbitol.
  • surfactants include Tween80 (polysorbate 80), Tween20 (polysorbate 20), pluronic, sodium dodecyl sulfate, and HC060 (polysorbate).
  • Oxyethylene hydrogenated castor oil Cremophor EL (Cremophor EL) and the like.
  • the preparation of the present invention can contain various ordinary pharmaceutical carriers, excipients, diluents, stabilizers or adsorption inhibitors.
  • Pharmaceutical carriers include, for example, ribosomes, fat emulsions, microcapsules, inclusion compounds, and the like, and excipients include, for example, lactose, mannitol, glycine, and the like.
  • examples of the diluent include ethanol water, glucose solution, and the like.
  • the stabilizer examples include ascorbic acid, sodium sulfite, erythorbic acid, and ethylenediaminetetraacetic acid (EDTA ), Proteins such as gelatin and albumin, amino acids such as glycine, arginine and glutamic acid, and sugars such as glucose, sucrose and lactose.
  • EDTA ethylenediaminetetraacetic acid
  • Proteins such as gelatin and albumin
  • amino acids such as glycine, arginine and glutamic acid
  • sugars such as glucose, sucrose and lactose.
  • the adsorption inhibitor examples include surfactants such as Tween (polysorbate) and pull nicks, and proteins such as albumin and gelatin.
  • the dose and frequency of administration of the preparation of the present invention are determined according to the dosage form, the age and weight of the patient, the disease to be affected, the condition of the patient, and the like.
  • the formulation is administered at a dose of 15 / g to 5 mg, preferably 25 to 5000 / g, 1 to 7 times per week as the amount of the polypeptide having CSF activity.
  • intravenous injection As an administration method, intravenous injection, subcutaneous injection and the like are used.
  • Examples, Reference Examples and Test Examples of the present invention will be described.
  • the pH was adjusted by adding 6 mol / L hydrochloric acid to a PEGylated ND28 solution (20 t / l acetate buffer containing 50 t / l sodium chloride, pH 6) at a concentration of about 3 mg / mL,
  • the PEGylated ND 28 solution can be produced by purifying the reaction solution obtained in Example 13 of W098 / 55500 by a conventional method).
  • this solution was sterile-filtered in a clean pliers, dispensed in 1 mL portions into 4 mL vials, stoppered with a Teflon-laminated rubber stopper, and wound tightly with an aluminum cap to obtain a preparation.
  • Example 2 The pH was adjusted by adding 6 mol / L hydrochloric acid to a PEGylated ND 28 solution (20 t / l acetate buffer containing 50 t / l sodium chloride, pH 6) at a concentration of about 3 mg / mL. A pH4 solution was prepared. Next, this solution was sterile-filtered in a clean bench, dispensed 1 mL each into a 4 mL vial, and stoppered with a Teflon laminated rubber stopper. The product was tightened with an aluminum cap.
  • the pH was adjusted by adding 6 mol / L hydrochloric acid to a PEGylated ND 28 solution (concentration of about 30 mg / mL acetate buffer containing 50 mL / L sodium chloride, pH 6) with a concentration of about 3 mg / mL.
  • the solution was prepared.
  • this solution was aseptically filtered in a clean bench, dispensed in 1 mL portions into 4 mL vials, stoppered with a Teflon laminated rubber stopper, and wound with an aluminum cap to form a preparation.
  • PEGylated ND 28 solution (20 mL / L acetate buffer, pH 4) was aseptically filtered in a clean bench, dispensed in 1 mL portions into 4 mL vials, and stoppered with a Teflon laminated rubber stopper. The product was rolled up with an aluminum cap to obtain a preparation.
  • PEG 28 ND 28 solution 33 t ol A phosphate buffer, pH 4
  • sodium chloride and 154 t ol / L and polysorbate 80 (Tween 80) become 50 g / mL.
  • soy sauce sterile-filtered in a clean bench, dispensed lmL into 4mL vials, stoppered with Teflon laminate rubber stopper, and then aluminum cap To form a preparation.
  • a PEG-modified ND 28 solution (20 mmol / L acetate buffer containing 50 mmol / L sodium chloride, pH 6) at a concentration of about 3 mg / mL was aseptically filtered in a clean bench, and dispensed in 4 mL vials in 1 mL aliquots of Teflon. After stoppering with a laminated rubber stopper, it was tightly wound with an aluminum cap to obtain a preparation.
  • the vial was opened, and the pH was measured directly using a pH meter (Toa Denpa Kogyo, HM-26S).
  • Free PEG in the preparation was quantified by the HPLC method.
  • the conditions of HPLC are as follows.
  • Injection volume 100 zL or 200 / L Column temperature: room temperature
  • ⁇ Free PEG ( ⁇ g / mL) refers to the free PEG concentration ( ⁇ g / mL) in the initial (before storage) formulation from the free PEG concentration (zg / mL) in the formulation after storage. / mL).
  • Example 6 87 51
  • Example 7 87 49
  • Example 2 As shown in Table 2, the free PEG in Example 1 (pH 3) and Example 2 (pH 4) was 1/20 or less as compared with Comparative Example 2 (pH 7). Also, in Example 3 (pH5), it was 1/5 or less as compared with Comparative Example 2 (pH7).
  • Example 5 which contained a high concentration, the amount of free PEG was lowest, and the effect of sodium chloride to suppress PEG elimination was observed. Also in Examples 6 and 7 using a phosphate buffer containing sodium chloride, an effect of suppressing PEG detachment was observed.
  • the present invention provides a method for stabilizing a chemically modified polypeptide having G_CSF activity and a preparation comprising the stabilized chemically modified polypeptide having G-CSF activity.

Landscapes

  • Health & Medical Sciences (AREA)
  • Epidemiology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Engineering & Computer Science (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Medicinal Preparation (AREA)

Abstract

On décrit des préparations pharmaceutiques qui contiennent des polypeptides chimiquement modifiés avec des composés à poids moléculaire élevé tels que PEG et qui présentent des activités du facteur stimulant les colonies de granulocytes (G-CSF) et dont le pH est régulé à 5 ou moins; et un procédé de stabilisation de polypeptides modifiés chimiquement avec des composés à poids moléculaire élevé tels que PEG et qui présentent des activités de G-CSF dans des solutions lorsqu'on ajuste le pH des solutions à 5 ou à moins de 5.
PCT/JP2000/001204 1999-03-01 2000-03-01 Preparations de g-csf modifiees chimiquement WO2000051626A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU28247/00A AU2824700A (en) 1999-03-01 2000-03-01 Chemically modified g-csf preparations

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP5275199 1999-03-01
JP11/52751 1999-03-01

Publications (1)

Publication Number Publication Date
WO2000051626A1 true WO2000051626A1 (fr) 2000-09-08

Family

ID=12923616

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2000/001204 WO2000051626A1 (fr) 1999-03-01 2000-03-01 Preparations de g-csf modifiees chimiquement

Country Status (2)

Country Link
AU (1) AU2824700A (fr)
WO (1) WO2000051626A1 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005025600A1 (fr) * 2003-09-11 2005-03-24 Kyowa Hakko Kogyo Co., Ltd. Medicament inhibant la mobilisation de cellules adhesives dans le sang peripherique
KR101810191B1 (ko) 2016-09-21 2017-12-19 연세대학교 산학협력단 세포 접착의 저해를 위한 줄기세포 성장 인자 또는 그의 단편의 용도

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1994014466A1 (fr) * 1992-12-18 1994-07-07 Boehringer Mannheim Gmbh Preparations pharmaceutiques aqueuses de g-csf stables au stockage
EP0822199A2 (fr) * 1994-10-12 1998-02-04 Amgen Inc. Polypeptides monopegylés à l'extrémité N-terminale, et procédé pour leur préparation

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1994014466A1 (fr) * 1992-12-18 1994-07-07 Boehringer Mannheim Gmbh Preparations pharmaceutiques aqueuses de g-csf stables au stockage
EP0822199A2 (fr) * 1994-10-12 1998-02-04 Amgen Inc. Polypeptides monopegylés à l'extrémité N-terminale, et procédé pour leur préparation

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005025600A1 (fr) * 2003-09-11 2005-03-24 Kyowa Hakko Kogyo Co., Ltd. Medicament inhibant la mobilisation de cellules adhesives dans le sang peripherique
JPWO2005025600A1 (ja) * 2003-09-11 2006-11-16 学校法人東海大学 接着性を有する細胞の末梢血への動員を阻害する薬剤
KR101810191B1 (ko) 2016-09-21 2017-12-19 연세대학교 산학협력단 세포 접착의 저해를 위한 줄기세포 성장 인자 또는 그의 단편의 용도

Also Published As

Publication number Publication date
AU2824700A (en) 2000-09-21

Similar Documents

Publication Publication Date Title
FI112169B (fi) Menetelmä ihmisproteiinipitoisten infuusio- tai injektointitarkoituksiin tarkoitettujen säilöttyjen lääkkeiden valmistamiseksi
AU733592B2 (en) A pharmaceutical formulation containing growth hormone, an amino acid and a non-ionic detergent
KR101482304B1 (ko) 에타너셉트의 안정한 용액 제형
US8642532B2 (en) Excipients for protein stabilization
EP2618814B1 (fr) Composition à base de caspofongine
EP2586459A1 (fr) Formulations antagonistes du VEGF
JPH03503764A (ja) ヒト成長ホルモン製剤
JP2002187852A (ja) 生理活性ペプチド含有粉末
US8841252B2 (en) Pharmaceutical formulation
EP1855661B1 (fr) Formulation pour l'aviptadil
JP2006137678A (ja) インターロイキン−２組成物
TWI243057B (en) Formulations for protection of peg-interferon alpha conjugates
EP2422770A1 (fr) Formulations de conjugués d'interféron alpha PEG
JPS62164631A (ja) インタ−ロイキン−２組成物
CN1802171B (zh) 不包含血清白蛋白的人红细胞生成素的稳定水溶液
EP2049148B1 (fr) Formulation liquide stable à base d'hormone de croissance humaine
WO2000051626A1 (fr) Preparations de g-csf modifiees chimiquement
JP3501471B2 (ja) カルシトニン類の安定化組成物および安定化法
KR100537260B1 (ko) 인간 성장 호르몬의 안정화된 액상 제제용 조성물
US20030162711A1 (en) Pharmaceutical formulation
EP3744319B1 (fr) Formulation lyophilisée stable pour g-csf fusionné fc hybride
JPS62223129A (ja) インタ−ロイキン−２組成物

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AE AL AM AT AU AZ BA BB BG BR BY CA CH CN CR CU CZ DE DK DM EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX NO NZ PL PT RO RU SD SE SG SI SK SL TJ TM TR TT TZ UA UG US UZ VN YU ZA ZW

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): GH GM KE LS MW SD SL SZ TZ UG ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE BF BJ CF CG CI CM GA GN GW ML MR NE SN TD TG

121 Ep: the epo has been informed by wipo that ep was designated in this application
DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
ENP Entry into the national phase

Ref country code: JP

Ref document number: 2000 602293

Kind code of ref document: A

Format of ref document f/p: F

REG Reference to national code

Ref country code: DE

Ref legal event code: 8642

122 Ep: pct application non-entry in european phase