RU2509776C2 - МОДИФИЦИРОВАННЫЕ ВАРИАНТЫ ИНГИБИТОРОВ ПРОТЕАЗ Bowman Birk - Google Patents

МОДИФИЦИРОВАННЫЕ ВАРИАНТЫ ИНГИБИТОРОВ ПРОТЕАЗ Bowman Birk Download PDF

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RU2509776C2
RU2509776C2 RU2011119481/10A RU2011119481A RU2509776C2 RU 2509776 C2 RU2509776 C2 RU 2509776C2 RU 2011119481/10 A RU2011119481/10 A RU 2011119481/10A RU 2011119481 A RU2011119481 A RU 2011119481A RU 2509776 C2 RU2509776 C2 RU 2509776C2
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bbpi
amino acid
variant
seq
binding peptide
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RU2011119481/10A
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Неелам С. АМИН
Кэтрин Д. КОЛЛИЕР
Мелоди ИСТАБРУК
Дэвид А. ЭСТЕЛЛ
Брайан ФОКС
Скотт Д. ПАУЭР
Брайан Ф. ШМИДТ
Гудрун ФОГТЕНТАНЦ
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ДАНИСКО ЮЭс ИНК.
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    • C—CHEMISTRY; METALLURGY
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    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/62—DNA sequences coding for fusion proteins
    • 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
    • A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
    • 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
    • A61P17/06—Antipsoriatics
    • 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
    • A61P17/12—Keratolytics, e.g. wart or anti-corn preparations
    • 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
    • A61P17/14—Drugs for dermatological disorders for baldness or alopecia
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/81—Protease inhibitors
    • C07K14/8107—Endopeptidase (E.C. 3.4.21-99) inhibitors
    • C07K14/811—Serine protease (E.C. 3.4.21) inhibitors

Abstract

Изобретение относится к области биохимии, в частности к выделенному варианту ингибитора протеаз Bowman Birk (BBPI), где указанный вариант BBPI содержит каркас, содержащий аминокислотные последовательности 1-40 и 50-66 последовательности SEQ ID NO: 187 и остатки цистеина в аминокислотных положениях 41 и 49 последовательности SEQ ID NO: 187, между которыми заключен связывающий пептид, выбранный из связывающего VEGF пептида, связывающего FGF-5 пептида, связывающего TGFβ пептида и связывающего TNFα пептида, где указанный вариант BBPI дополнительно содержит замененную аминокислоту в аминокислотном положении, выбранном из положений, эквивалентных положениям 1, 4, 5, 11, 13, 18, 25, 27, 29, 31, 38, 40, 50, 52, 55 и 65 последовательности SEQ ID NO: 187, где указанная замещенная аминокислота в положении 1 представляет собой A, в положении 4 представляет собой V, в положении 5 выбрана из P и A, в положении 11 представляет собой 11G, в положении 13 выбрана из 13Y, 13I, 13F, 13M, 13L, 13V, 13K и 13R, в положении 18 выбрана из 18I и 18V, в положении 25 выбрана из 25K, 25N, 25W, 25I, 25A и 25R, в положении 27 выбрана из 27H, 27K, 27V, 27A и 27Q, в положении 29 выбрана из 29R, 29K и 29P, в положении 31 выбрана из 31Q, 31H, 31E, 31A, 31R, 31W, 31K и 31T, в положении 38 выбрана из 38N, 38K и 38R, в положении 40 выбрана из 40H, 40K, 40Q, 40R и 40Y, в положении 50 выбрана из 50R, 50Q, 50K, 50T, 50V, 50M и 50S, в положении 52 выбрана из 52K, 52T, 52R, 52Q, 52L, 52H, 52A, 52M, 52S и 52E, в положении 55 представляет собой 55M, в положении 65 выбрана из 65E и 65D. Изобретение позволяет эффективно ингибировать трипсин по сравнению с немодифицированным вариантом. 5 з.п. ф-лы, 18 ил., 7 табл., 18 пр.

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Claims (6)

1. Выделенный вариант ингибитора протеаз Bowman Birk (BBPI), где указанный вариант BBPI содержит каркас, содержащий аминокислотные последовательности 1-40 и 50-66 последовательности SEQ ID NO: 187 и остатки цистеина в аминокислотных положениях 41 и 49 последовательности SEQ ID NO: 187, между которыми заключен связывающий пептид, где указанный вариант BBPI дополнительно содержит замененную аминокислоту в аминокислотном положении, выбранном из положений, эквивалентных положениям 1, 4, 5, 11, 13, 18, 25, 27, 29, 31, 38, 40, 50, 52, 55 и 65 последовательности SEQ ID NO: 187, где указанная замещенная аминокислота в положении 1 представляет собой A, в положении 4 представляет собой V, в положении 5 выбрана из P и A, в положении 11 представляет собой 11G, в положении 13 выбрана из 13Y, 13I, 13F, 13M, 13L, 13V, 13K и 13R, в положении 18 выбрана из 18I и 18V, в положении 25 выбрана из 25K, 25N, 25W, 25I, 25A и 25R, в положении 27 выбрана из 27H, 27K, 27V, 27A и 27Q, в положении 29 выбрана из 29R, 29K и 29P, в положении 31 выбрана из 31Q, 31H, 31E, 31A, 31R, 31W, 31K и 31T, в положении 38 выбрана из 38N, 38K и 38R, в положении 40 выбрана из 40H, 40K, 40Q, 40R и 40Y, в положении 50 выбрана из 50R, 50Q, 50K, 50T, 50V, 50M и 50S, в положении 52 выбрана из 52K, 52T, 52R, 52Q, 52L, 52H, 52A, 52M, 52S и 52E, в положении 55 представляет собой 55M, в положении 65 выбрана из 65E и 65D, где указанный выделенный вариант BBPI обладает большей активностью ингибитора трипсина, чем соответствующий BBPI, лишенный указанной аминокислотной замены, где указанный связывающий пептид выбран из связывающего VEGF пептида, связывающего FGF-5 пептида, связывающего TGFβ пептида и связывающего TNFα пептида.
2. Выделенный вариант BBPI по п.1, где указанный модифицированный вариант BBPI содержит комбинацию из двух аминокислотных замен в положениях аминокислот, эквивалентных положениям 50 и 52 в SEQ ID NO: 187.
3. Выделенный вариант BBPI по п.1, где указанный модифицированный вариант BBPI имеет аминокислотную последовательность SEQ ID NO: 595.
4. Выделенный вариант BBPI по п.1, где указанный модифицированный вариант BBPI связывает VEGF.
5. Выделенный вариант BBPI по п.1, где указанный связывающий VEGF пептид выбран из SEQ ID NO: 9, 458, 459, 460, 468, 469, 470, 471, 472 и 473.
6. Выделенный вариант BBPI по п.1, где указанный модифицированный вариант BBPI представляет собой VEGF-BBPI, выбранный из SEQ ID NO: 601, 602, 627, 628, 630, 631, 643, 491, 632, 633, 634, 635 и 636.
RU2011119481/10A 2008-10-15 2008-10-15 МОДИФИЦИРОВАННЫЕ ВАРИАНТЫ ИНГИБИТОРОВ ПРОТЕАЗ Bowman Birk RU2509776C2 (ru)

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