RU2016147505A - Конъюгат, содержащий оксинтомодулин и фрагмент иммуноглобулина, и его применение - Google Patents
Конъюгат, содержащий оксинтомодулин и фрагмент иммуноглобулина, и его применение Download PDFInfo
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- RU2016147505A RU2016147505A RU2016147505A RU2016147505A RU2016147505A RU 2016147505 A RU2016147505 A RU 2016147505A RU 2016147505 A RU2016147505 A RU 2016147505A RU 2016147505 A RU2016147505 A RU 2016147505A RU 2016147505 A RU2016147505 A RU 2016147505A
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Claims (54)
1. Конъюгат, содержащий оксинтомодулин, Fc-область иммуноглобулина и непептидильный полимер, где конъюгат получают путем ковалентного связывания оксинтомодулина с Fc-областью иммуноглобулина через непептидильный полимер.
2. Конъюгат по п. 1, где оксинтомодулин имеет аминокислотную последовательность SEQ ID NO. 1.
3. Конъюгат по п. 1, где оксинтомодулин представляет собой производное оксинтомодулина, полученное путем вставки, делеции или замены аминокислот нативного оксинтомодулина.
4. Конъюгат по п. 3, где производное оксинтомодулина содержит аминокислотную последовательность следующей формулы 1:
где R1 представляет собой гистидин, дезамино-гистидил, диметил-гистидил (N-диметил-гистидил), бета-гидроксиимидазопропионил, 4-имидазоацетил, бета-карбокси-имидазопропионил или тирозин;
X1 представляет собой Aib (аминоизомасляная кислота), d-аланин, глицин, Sar (N-метилглицин), серии или d-серин;
Х2 представляет собой глутаминовую кислоту или глутамин;
Х3 представляет собой лейцин или тирозин;
Х4 представляет собой серии или аланин;
Х5 представляет собой лизин или аргинин;
Х6 представляет собой глутамин или тирозин;
Х7 представляет собой лейцин или метионин;
Х8 представляет собой аспарагиновую кислоту или глутаминовую кислоту;
Х9 представляет собой глутаминовую кислоту, серии, альфа-метил-глутаминовую кислоту или делетирован;
Х10 представляет собой глутамин, глутаминовую кислоту, лизин, аргинин, серии или делетирован;
X11 представляет собой аланин, аргинин, валин или делетирован;
Х12 представляет собой аланин, аргинин, серии, валин или делетирован;
Х13 представляет собой лизин, глутамин, аргинин, альфа-метил-глутаминовую кислоту или делетирован;
Х14 представляет собой аспарагиновую кислоту, глутаминовую кислоту, лейцин или делетирован;
Х15 представляет собой фенилаланин или делетирован;
Х16 представляет собой изолейцин, валин или делетирован;
Х17 представляет собой аланин, цистеин, глутаминовую кислоту, лизин, глутамин, альфа-метил-глутаминовую кислоту или делетирован;
Х18 представляет собой триптофан или делетирован;
Х19 представляет собой аланин, изолейцин, лейцин, серии, валин или делетирован;
Х20 представляет собой аланин, лизин, метионин, глутамин, аргинин или делетирован;
Х21 представляет собой аспарагин или делетирован;
Х22 представляет собой аланин, глицин, треонин или делетирован;
Х23 представляет собой цистеин, лизин или делетирован;
Х24 представляет собой пептид из 2-10 аминокислот, состоящий из комбинаций аланина, глицина и серина, или делетирован; и
R2 представляет собой KRNRNNIA (SEQ ID NO. 35), GPSSGAPPPS (SEQ ID NO. 36), GPSSGAPPPSK (SEQ ID NO. 37), HSQGTFTSDYSKYLD (SEQ ID NO. 38), HSQGTFTSDYSRYLDK (SEQ ID NO. 39), HGEGTFTSDLSKQMEEEAVK (SEQ ID NO. 40) или делетирован (за исключением случая, когда аминокислотная последовательность формулы 1 идентична последовательности SEQ ID NO. 1).
5. Конъюгат по п. 4, где производное оксинтомодулина обладает способностью активировать рецептор GLP-1 (глюкагоноподобный пептид-1) и рецептор глюкагона.
6. Конъюгат по п. 4, демонстрирующий эффекты против ожирения.
7. Конъюгат по п. 4, где одна или более аминокислот в положениях 10, 14, 16, 20, 24 и 28 аминокислотной последовательности формулы 1 заменены на аминокислоты или производные аминокислот, выбранные из группы, состоящей из Tyr(4-Me), Phe, Phe(4-Me), Phe(4-Cl), Phe(4-CN), Phe(4-NO2), Phe(4-NH2), Phg (фенилглицин), Pal (β-пиридил-аланин), Nal (β-нафтил-аланин), Ala(2-тиенил) и Ala(бензотиенил).
8. Конъюгат по п. 4, где одна или более чем одна пара аминокислот в положениях 10 и 14, 12 и 16, 16 и 20, 20 и 24 и 24 и 28 аминокислотной последовательности формулы 1 заменена на глутаминовую кислоту или лизин с образованием колец.
9. Конъюгат по п. 4, где одна или более чем одна пара аминокислот в положениях 10 и 14, 12 и 16, 16 и 20, 20 и 24 и 24 и 28 аминокислотной последовательности формулы 1 образует кольца.
10. Конъюгат по п. 4, где производное оксинтомодулина выбрано из группы, состоящей из пептидов с SEQ ID NO. 2-34.
11. Конъюгат по п. 1, где непептидильный полимер выбран из группы, состоящей из полиэтиленгликоля, полипропиленгликоля, сополимеров этиленгликоля и пропиленгликоля, полиоксиэтилированных полиолов, поливинилового спирта, полисахаридов, декстрана, поливинил-этилового простого эфира, полимолочной кислоты (PLA), полимолочной-гликолевой кислоты (PLGA), липидных полимеров, хитинов, гиалуроновой кислоты, полисахарида и их комбинаций.
12. Конъюгат по п. 1, где аминогруппа и тиоловая группа Fc-области иммуноглобулина и оксинтомодулина присоединены к обоим концам непептидильного полимера, соответственно.
13. Конъюгат по п. 1, где непептидильный полимер содержит реакционноспособные группы, обладающие способностью связываться с Fc-областью иммуноглобулина и белковым лекарственным средством по обоим концам.
14. Конъюгат по п. 13, где реакционноспособная группа выбрана из группы, состоящей из альдегидной группы, пропиональдегидной группы, бутиральдегидной группы, малеимидной группы и производного сукцинимида.
15. Конъюгат по п. 13, где реакционноспособные группы на обоих концах являются одинаковыми или разными по отношению друг к другу.
16. Конъюгат по п. 1, где Fc-область иммуноглобулина представляет собой негликозилированную Fc-область.
17. Конъюгат по п. 1, где Fc-область иммуноглобулина выбрана из группы, состоящей из СН1-домена, СН2-домена, СН3-домена и СН4-домена; СН1-домена и СН2-домена; СН1-домена и СН3-домена; СН2-домена и СН3-домена; комбинации одного или более чем одного домена и шарнирной области иммуноглобулина (или части шарнирной области); и димера каждого домена константных областей тяжелой цепи и константной области легкой цепи.
18. Конъюгат по п. 1, где Fc-область иммуноглобулина представляет собой производное, в котором делетирован участок, способный образовывать дисульфидную связь, удалены определенные аминокислотные остатки на N-конце нативной формы Fc, на N-конец нативной формы Fc добавлен остаток метионина, делетирован комплемент-связывающий сайт или делетирован сайт антитело-зависимой клеточно-опосредованной цитотоксичности (ADCC).
19. Конъюгат по п. 1, где Fc-область иммуноглобулина представляет собой Fc-область, происходящую из иммуноглобулина, выбранного из группы, состоящей из IgG, IgA, IgD, IgE и IgM.
20. Конъюгат по п. 19, где Fc-область иммуноглобулина представляет собой Fc-область IgG4.
21. Конъюгат по п. 1, где Fc-область иммуноглобулина представляет собой происходящую из IgG4 человека негликозилированную Fc-область.
22. Фармацевтическая композиция для предупреждения или лечения ожирения, содержащая конъюгат по любому из пп. 1-21.
23. Фармацевтическая композиция по п. 22, дополнительно содержащая фармацевтически приемлемый носитель.
24. Фармацевтическая композиция по п. 22, где композицию вводят по отдельности или в комбинации с другими фармацевтическими композициями, демонстрирующими профилактические или терапевтические эффекты в отношении ожирения.
25. Фармацевтическая композиция по п. 24, где фармацевтическая композиция выбрана из группы, состоящей из агониста рецептора GLP-1, агониста лептинового рецептора, ингибитора DPP-IV (дипептидилпептидаза-IV), антагониста Y5-рецептора, антагониста рецептора меланин-концентрирующего гормона (МСН), агониста Y2/3-рецептора, агониста МС3/4-рецептора, ингибитора желудочной/панкреатической липазы, агониста 5НТ2с, агониста β3А-рецептора, агониста рецептора амилина, антагониста грелина и антагониста рецептора грелина.
26. Способ предупреждения или лечения ожирения, включающий стадию введения субъекту конъюгата по любому из пп. 1-21 или композиции по любому из пп. 22-25.
27. Применение конъюгата по любому из пп. 1-21 или композиции по любому из пп. 22-25 в изготовлении лекарственных средств для предупреждения или лечения ожирения.
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