RU2002120486A - FREE FROM URATOXIDASE UNITS FOR PREPARATION OF NON-IMMUNOGENIC POLYMER CONJUGATES - Google Patents

FREE FROM URATOXIDASE UNITS FOR PREPARATION OF NON-IMMUNOGENIC POLYMER CONJUGATES

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Publication number
RU2002120486A
RU2002120486A RU2002120486/13A RU2002120486A RU2002120486A RU 2002120486 A RU2002120486 A RU 2002120486A RU 2002120486/13 A RU2002120486/13 A RU 2002120486/13A RU 2002120486 A RU2002120486 A RU 2002120486A RU 2002120486 A RU2002120486 A RU 2002120486A
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RU
Russia
Prior art keywords
uricase
poly
conjugate
ethylene glycol
ethylene oxide
Prior art date
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RU2002120486/13A
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Russian (ru)
Other versions
RU2281954C2 (en
Inventor
Мерри Р. Шерман
Марк Дж. П. СЭЙФЕР
Л. Дэвид Вильямс
Original Assignee
Маунтэйн Вью Фармасьютикл, Инк.
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Priority claimed from US09/501,730 external-priority patent/US6783965B1/en
Application filed by Маунтэйн Вью Фармасьютикл, Инк. filed Critical Маунтэйн Вью Фармасьютикл, Инк.
Publication of RU2002120486A publication Critical patent/RU2002120486A/en
Application granted granted Critical
Publication of RU2281954C2 publication Critical patent/RU2281954C2/en

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Claims (33)

1. Очищенная уратоксидаза (уриказа), практически свободная от агрегатов больших, чем октамеры.1. Purified uratoxidase (uricase), practically free of aggregates larger than octamers. 2. Уриказа по п.1, в которой уриказа является уриказой млекопитающего.2. The uricase according to claim 1, wherein the uricase is a mammalian uricase. 3. Уриказа по п.2, в которой уриказа является уриказой свиной печени, бычьей печени или овечьей печени.3. The uricase according to claim 2, wherein the uricase is the uricase of a pork liver, bovine liver or sheep liver. 4. Уриказа по п.1, в которой уриказа является рекомбинантной.4. The uricase according to claim 1, in which the uricase is recombinant. 5. Уриказа по п.4, в которой уриказа в значительной степени имеет последовательность уриказы свиной, бычьей, овечьей печени или печени бабуина.5. The uricase according to claim 4, in which the uricase largely has the sequence of uricase of pork, bovine, sheep’s or baboon’s liver. 6. Уриказа по п.4, в которой уриказа является химерической.6. Uricase according to claim 4, in which uricase is chimeric. 7. Уриказа по п.6, в которой химерическая уриказа содержит части уриказы свиной печени и уриказы печени бабуина.7. The uricase according to claim 6, in which the chimeric uricase contains parts of the uricase of the pork liver and uricase of the liver of the baboon. 8. Уриказа по п.7, в которой химерическая уриказа является PKS уриказой.8. The uricase of claim 7, wherein the chimeric uricase is PKS uricase. 9. Уриказа по п.4, в которой уриказа в значительной степени имеет последовательность уриказы печени бабуина, в которой тирозин 97 заменен гистидином.9. The uricase according to claim 4, in which the uricase largely has a baboon liver uricase sequence in which tyrosine 97 is replaced by histidine. 10. Уриказа по п.4, в которой уриказа содержит аминотерминал и карбокситерминал, и в которой уриказа усечена на одном терминале или на обоих терминалах.10. The uricase according to claim 4, in which the uricase contains an amino terminal and a carboxyterminal, and in which the uricase is truncated at one terminal or both terminals. 11. Уриказа по п.1, в которой уриказа является грибной или микробной уриказой.11. The uricase according to claim 1, in which the uricase is a fungal or microbial uricase. 12. Уриказа по п.11, в которой грибная или микробная уриказа выделяется из Aspergillus flavus, Arthrobacter globiformis, Bacillus sp. или Candida utilis, или является рекомбинантным ферментом, в значительной степени имеющим последовательность одной из упомянутых уриказ.12. The uricase according to claim 11, in which the fungal or microbial uricase is isolated from Aspergillus flavus, Arthrobacter globiformis, Bacillus sp. or Candida utilis, or is a recombinant enzyme substantially having the sequence of one of the mentioned uricases. 13. Уриказа по п.1, в которой уриказа является уриказой беспозвоночного.13. The uricase according to claim 1, wherein the uricase is an invertebrate uricase. 14. Уриказа по п.13, в которой уриказа беспозвоночного выделяется из Drosophila melanogaster или Drosophila pseudoobscura, или является рекомбинантным ферментом, в значительной степени имеющим последовательность одной из упомянутых уриказ.14. The uricase of claim 13, wherein the invertebrate uricase is isolated from Drosophila melanogaster or Drosophila pseudoobscura, or is a recombinant enzyme substantially having the sequence of one of the mentioned uricases. 15. Уриказа по п.1, в которой уриказа является растительной уриказой.15. The uricase according to claim 1, wherein the uricase is a plant uricase. 16. Уриказа по п.15, в которой уриказа растительная выделяется из корневых клубеньков Glycine max или является рекомбинантным ферментом, в значительной степени имеющим последовательность упомянутой уриказы.16. The uricase according to claim 15, wherein the plant uricase is isolated from the root nodules of Glycine max or is a recombinant enzyme substantially having the sequence of said uricase. 17. Уриказа по п.1, конъюгированная с поли(этиленгликолем) или поли(этиленоксидом), в которой уриказа в упомянутом конъюгате практически свободна от агрегатов больших, чем октамеры.17. The uricase according to claim 1, conjugated with poly (ethylene glycol) or poly (ethylene oxide), in which the uricase in said conjugate is practically free of aggregates larger than octamers. 18. Уриказный конъюгат по п.17, в котором упомянутый поли(этиленгликоль) является монометокси поли(этиленгликолем).18. The uricase conjugate of claim 17, wherein said poly (ethylene glycol) is monomethoxy poly (ethylene glycol). 19. Уриказа по п.17, в которой упомянутая уриказа конъюгируется с упомянутым поли(этиленгликолем) или поли(этиленоксидом) через связь, выбранную из группы, состоящей из уретановой (карбаматной), вторичной аминной и амидной связей.19. The uricase according to claim 17, wherein said uricase is conjugated to said poly (ethylene glycol) or poly (ethylene oxide) via a bond selected from the group consisting of urethane (carbamate), secondary amine and amide bonds. 20. Уриказный конъюгат по п.17, в котором упомянутый поли(этиленгликоль) или поли(этиленоксид) имеет молекулярный вес приблизительно между 5 и 30 кДа.20. The uricase conjugate of claim 17, wherein said poly (ethylene glycol) or poly (ethylene oxide) has a molecular weight of between about 5 and 30 kDa. 21. Уриказный конъюгат по п.20, в котором упомянутый поли(этиленгликоль) или поли(этиленоксид) имеет молекулярный вес приблизительно между 10 и 20 кДа.21. The uricase conjugate of claim 20, wherein said poly (ethylene glycol) or poly (ethylene oxide) has a molecular weight of between about 10 and 20 kDa. 22. Уриказный конъюгат по п.17, в котором среднее количество нитей упомянутого поли(этиленгликоля) или поли(этиленоксида) составляет приблизительно от 2 до 12 на субъединицу уриказы.22. The uricase conjugate of claim 17, wherein the average number of strands of said poly (ethylene glycol) or poly (ethylene oxide) is from about 2 to 12 per uricase subunit. 23. Уриказный конъюгат по п.22, в котором среднее количество нитей упомянутого поли(этиленгликоля) или поли(этиленоксида) составляет приблизительно от 6 до 10 на субъединицу уриказы.23. The uricase conjugate of claim 22, wherein the average number of strands of said poly (ethylene glycol) or poly (ethylene oxide) is from about 6 to 10 per uricase subunit. 24. Уриказный конъюгат по п.23, в котором среднее количество нитей упомянутого поли(этиленгликоля) или поли(этиленоксида) составляет приблизительно от 7 до 9 на субъединицу уриказы.24. The uricase conjugate of claim 23, wherein the average number of strands of said poly (ethylene glycol) or poly (ethylene oxide) is from about 7 to 9 per uricase subunit. 25. Уриказный конъюгат по п.17, в котором поли(этиленгликоль) или поли(этиленоксид) является линейным.25. The uricase conjugate of claim 17, wherein the poly (ethylene glycol) or poly (ethylene oxide) is linear. 26. Уриказный конъюгат по п.17, в котором поли(этиленгликоль) или поли(этиленоксид) является разветвленным.26. The uricase conjugate of claim 17, wherein the poly (ethylene glycol) or poly (ethylene oxide) is branched. 27. Фармацевтический состав для понижения уровней мочевой кислоты в жидкости или ткани организма, содержащий конъюгат по п.17 и фармацевтически приемлемый носитель.27. A pharmaceutical composition for lowering uric acid levels in a body fluid or tissue, comprising the conjugate of claim 17 and a pharmaceutically acceptable carrier. 28. Фармацевтический состав по п.27, в котором упомянутый состав стабилизируется путем лиофилизации и растворяется после восстановления, обеспечивая растворы, пригодные для парентерального введения.28. The pharmaceutical composition according to item 27, in which said composition is stabilized by lyophilization and dissolved after reconstitution, providing solutions suitable for parenteral administration. 29. Способ очистки уриказы, имеющей пониженную иммуногенность, содержащий шаг отделения уриказных агрегатов больших, чем октамеры, во фракциях уриказы и исключения таких агрегатов из очищенной уриказы.29. A method for purifying uricase having reduced immunogenicity, comprising the step of separating uricase aggregates larger than octamers in uricase fractions and excluding such aggregates from purified uricase. 30. Способ по п.29, в котором упомянутый шаг отделения выбирается из группы, состоящей из ионообменной хроматографии, хроматографии с исключением по размеру и ультрафильтрации.30. The method according to clause 29, wherein said separation step is selected from the group consisting of ion exchange chromatography, size exclusion chromatography, and ultrafiltration. 31. Способ по п.30, в котором упомянутый шаг отделения содержит шаг обнаружения агрегатов больших, чем октамеры, во фракциях уриказы и исключения упомянутых фракций, содержащих упомянутые агрегаты.31. The method of claim 30, wherein said separation step comprises a step of detecting aggregates larger than octamers in uricase fractions and excluding said fractions containing said aggregates. 32. Способ по п.31, в котором упомянутый шаг обнаружения содержит измерение светорассеивания.32. The method of claim 31, wherein said detecting step comprises measuring light scattering. 33. Выделенная уриказа, приготовленная по способу по п.29.33. Isolated uricase prepared by the method according to clause 29.
RU2002120486/13A 2000-02-10 2001-02-07 Purified preparation of mammalian uricase, method for its preparing, preparation of active uricase conjugate and pharmaceutical composition RU2281954C2 (en)

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US09/501,730 US6783965B1 (en) 2000-02-10 2000-02-10 Aggregate-free urate oxidase for preparation of non-immunogenic polymer conjugates
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RU2009104003/10A RU2557318C9 (en) 2000-02-10 2009-02-06 Purified preparation of urate oxidase, purified recombinant urate oxidase, conjugate (versions) and pharmaceutical composition for reducing mammalian liquid or tissue uric acid, purified fragments of urate oxidase and method for urate oxidase purification

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