ME00183B - Novi postupak za deregulaciju amiloida - Google Patents

Novi postupak za deregulaciju amiloida

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ME00183B
ME00183B MEP-2008-304A MEP2008304A ME00183B ME 00183 B ME00183 B ME 00183B ME P2008304 A MEP2008304 A ME P2008304A ME 00183 B ME00183 B ME 00183B
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cell
analog
epitope
app
nucleic acid
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Peter Birk
Martin Roland M E Biotech A/S Jensen
Klaus Gregorius Nielsen
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Pharmexa As
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Abstract

Opisani su novi postupci za borbu protiv bolesti koje karakteriše taloženje amiloida. Ovi postupci se obično oslanjaju na imunizaciju protiv amiloidogenih proteina (proteini koji doprinose stvaranju amiloida), kao što je beta amiloid (Aß). Poželjno je da se imunizacija obavlja ordiniranjem analoga svojstvenih amiloidogenih polipeptida. Naročito poželjan kao imunogen je svojstveni Aß, koji se modifi kuje uvođenjem samo jednog ili više stranih imunodominantnih i promiskuitetnih epitopa T-ćelije, uz bitno zadržavanje većine Aß epitopa B-ćelije. Opisana je takođe vakcinacija sa nukleinskom kisjelinom protiv amiloidogenih polipeptida, i vakcinacija koja koristi žive vakcine, kao i korisni postupci i načini ovih vakcinacija. Ti postupci i načini obuhvataju metode za identi fikaciju korisnih imunogenih analoga amiloidogenih proteina, postupke za dobijanje analoga i farmaceutskih formulacija, kao i fragmenata nukleinskih kisjelina, vektora, transformisanih ćelija, polipeptida i farmaceutskih formulacija.

Claims (45)

1.Postupak za in viva deregulaciju svojstvenog proteina beta amiloida (A ) ili svojstvenog prekursorskog proteina amiloida (APP) u životinji, uključujući humano biće, postupak koji se sastoji od ostvarivanja prezentacije imuno sistemu životinje imunogeno efikasne količine najmanje jednog analoga svojstvenog Aβ ili svojstvenog APP životinje, naznačen time, što se uvodi najmanje jedan izolovani strani epitop T helpera (TH epitop), pomoću umetanja, adicije, izbacivanja ili supstitucije, ili pomoću odvojenog kuplovanja na polihidroksipolimer kao osnovu nosača TH epitopa i sekvencije peptida izvedene iz Aβ ili APP, tako da imunizovanje životinje sa ovim analogom izaziva u životinji stvaranje antitela protiv svojstvenog Aβ ili svojstvenog APP, pri čemu se strani TH epitop uvodi u Aβ ili APP, kao što je shematski pokazano za epitope P2 i P30 na Slici 1, ili se strani T H epitop kupluje u polihidroksipolimer kao osnovu nosača, koji takođe nosi sekvenciju peptida izvedenu iz Aβ ili APP.
2. Postupak prema Zahtevu 1, naznačen time, što je rezultat modifikacije očuvanje bitne frakcije epitopa B-ćelije A ili APP, tako što se - uvodi najmanje jedan prvi ostatak koji ostvaruje targetiranje analoga na antigen koji prezentira ćelija (APC) ili B-limfocit, i/ili -uvodi najmanje jedan drugi ostatak koji stimuliše imuno sistem, i/ili - uvodi najmanje jedan treći ostatak koji optimizuje prezentaciju analoga imuno sistemu.
3.Postupak prema Zahtevu 2, naznačen time, što se analog modifikuje kovalentnim ili ne-kovalentnim vezivanjem za pogodne hemijske grupe u Al3, APP ili njihovoj subsekvenciji, uvođenjem kao bočne grupe prvog, i/ili drugog, i/ili trećeg ostatka.
4.Postupak prema bilo kom od prethodnih Zahteva, naznačen time, što uvođenje supstitucije, i/ili izbacivanja, i/ili umetanja, i/ili adicije aminokiseline vodi bitnom očuvanju ukupne tercijarne strukture amiloidogenog polipeptida.
5.Postupak prema bilo kome od Zahteva 2 do 7, naznačen time, što analog obuhvata dupliranje najmanje jednog epitopa B-ćelije amiloidogenog polipeptida i/ili uvođenje haptena.
6.Postupak prema bilo kom od prethodnih Zahteva, naznačen time, što je u životinji imunodominantan strani epitop T-ćelije.
7.Postupak prema bilo kom od prethodnih Zahteva, naznačen time, što je strani epitop T-ćelije promiskuitetan, kao što je strani epitop T-ćelije koji se bira između prirodnog promiskuitetnog epitopa T-ćelije i veštačke sekvencije koja vezuje MHC-11 peptid.
8.Postupak prema Zahtevu 7, naznačen time, što se prirodni epitop T-ćelije bira između epitopa anatoksina tetanusa, kao što su P2 ili P30, epitopa anatoksina difterije, epitopa hemaglutinina virusa gripa, i epitopa P. falciparum CS.
9.Postupak prema bilo kom od Zahteva 2 do 8, naznačen time, što je prvi ostatak bitno specifičan vezujući partner specifične površine antigena 8-limfocita, ili specifične površine antigena APC, kao što je hapten ili ugljeni hidrat, za koji postoji receptor na B-limfocitu ili na APC.
10.Postupak prema bilo kom od Zahteva 2 do 9, naznačen time, što se drugi ostatak bira između citokina, kao što je interferon y (IFN-y) ili njegov efikasni deo, Flt3L ili njegov efikasni deo, interleukin 1 (IL-1) ili njegov efikasni deo, interleukin 2 (IL-2) ili njegov efikasni deo, interleukin 4 (IL-4) ili njegov efikasni deo, interleukin 6 (IL-6) ili njegov efikasni deo, interleukin 12 (IL-12) ili njegov efikasni deo, interleukin 13 (IL-13) ili njegov efikasni deo, interleukin 15 (IL-15) ili njegov efikasni deo, stimulacioni faktor kolonije granulocit-makrofaga (GM-CSF) ili njegov efikasni deo, hormon, i protein toplotnog šoka, kao što je HSP70 ili njegov efikasni deo, HSC70 ili njegov efikasni deo, GRP94 ili njegov efikasni deo, i kalretikulin (CRT) ili njegov efikasni deo.
11.Postupak prema bilo kom od Zahteva 2 do 1O, naznačen time, što je treći ostatak po prirodi lipid, kao što je palmitoilna grupa, miristilna grupa, farnezilna grupa, geranil-geranilna grupa, sidrište GPI i N-acildigliceridna grupa, ili što je treći ostatak polihidroksipolimer, kao što je polisaharid.
12.Postupak prema Zahtevu 11, naznačen time, što polisaharid služi kao noseća osnova za koju se odvojeno vežu peptid izveden iz Al3 ili APP i strani epitop T-ćelije.
13.Postupak prema Zahtevu 12, naznačen time, što se peptid izveden iz Al3 ili APP i strani epitop T-ćelije vezuju za polisaharid preko amidne veze.
14.Postupak prema bilo kom od prethodnih Zahteva, naznačen time, što je svojstveni Al3 ili APP tako modifikovan da se očuvaju epitopi B-ćelije koji nisu izloženi vanćelijskoj fazi, kada se prezentuju u obliku vezanom za ćeliju svojstvenog APP.
15.Postupak prema Zahtevu 14, naznačen time, što se amiloidogeni polipeptid tako modifikuje da mu nedostaje najmanje jedan epitop B-ćelije koji je izložen vanćelijskoj fazi, kada se prezentuje u obliku vezanom za ćeliju svojstvenog APP.
16.Postupak prema bilo kom od prethodnih Zahteva, naznačen time, što se sastoji od supstitucije najmanje jedne sekvencije aminokiselina unutar svojstvenog Al3 ili APP sa sekvencijom aminokiselina jednake ili različite dužine, koja tom analogu daje strani epitop TH·
17.Postupak prema bilo kom od Zahteva, naznačen time, što analog sadrži sekvenciju aminokiselina koja odgovara aminokiselinama 672-714 u SEQ ID NO: 2, pri čemu umetnuta sekvencija aminokiselina daje analogu strani epitop TH, ili što analog sadrži sekvenciju aminokiselina koja odgovara aminokiselinama 672-714 u SEQ ID NO: 2, pri čemu se najmanje jedna sekvencija aminokiselina supstituiše sekvencijom jednake ili različite dužine, tako da daje strani epitop TH·
18.Postupak prema bilo kom od prethodnih Zahteva, naznačen time, što se prezentacija imuno sistemu ostvaruje tako što se najmanje dve kopije analoga kovalentno ili ne-kovalentno vezuju za molekul nosača koji je u stanju da ostvari prezentaciju višestrukih kopija antigenskih determinanti
19.Postupak prema bilo kom od prethodnih Zahteva, naznačen time, što se analog formuliše sa adjuvantom koji olakšava raskidanje autotolerancije prema auto-antigenima.
20.Postupak prema bilo kom od prethodnih Zahteva, naznačen time, što efikasna količina analoga ordinira životinji putem koji se bira između parenteralnog puta, kao što su intrakutani, subkutani i intramuskularni putevi; peritonealnog puta; oralnog puta; bukalnog puta; sublingvalnog puta; epiduralnog puta; spinalnog puta; analnog puta i intrakranijalnog puta.
21.Postupak prema Zahtevu 20, naznačen time, što je efikasna količina analoga između 0,5 !Jg i 2000 !Jg.
22.Postupak prema Zahtevu 20 ili 21, naznačen time, što se analog nalazi u uređaju virtuelnog limfnog čvora (VLN).
23.Postupak prema bilo kom od Zahteva 1 do 17, naznačen time, što se prezentacija modifikovanog amiloidogenog polipeptida imuno sistemu ostvaruje uvođenjem nukleinske kiseline (ili kiselina) koja kodira analog u ćelije životinje, čime se dobija da ćelije in vivo izlučuju uvedenu nukleinsku kiselinu (ili kiseline).
24.Postupak prema Zahtevu 23, naznačen time, što se uvedena nukleinska kiselina (ili kiseline) bira između ogoljene DNA, DNA formulisane sa naelektrisanim ili nenaelektrisanim lipidima, DNA formulisane u lipozome, DNA uključene u vektor virusa, DNA formulisane sa proteinom ili polipeptidom koji olakšava transfekciju, DNA formulisane sa targetiranim proteinom ili polipeptidom, DNA formulisane sa agensima za taloženje kalcijuma, DNA kuplovane u molekul inertnog nosača, DNA enkapsulirane u hitin ili hitosan i DNA formulisane sa adjuvantom.
25.Postupak prema Zahtevu 24, naznačen time, što se nukleinska kiselina (ili kiseline) nalazi u uređaju VLN.
26.Postupak prema bilo kom od Zahteva 20 do 25, naznačen time, što se sastoji od najmanje jednog ordiniranja/uvođenja godišnje, kao što je najmanje 2, najmanje 3, najmanje 4, najmanje 6 i najmanje 12 ordiniranja/uvođenja.
27.Postupak za tretiranje i/ili prevenciju i/ili ublažavanje Alzheimer-ove bolesti ili drugih bolesti i stanja koje karakterišu naslage A . naznačen time, što se taj postupak sastoji u deregulaciji svojstvenog A ili APP, u skladu sa postupkom bilo kog od zahteva 1 do 26, do te mere da se ukupna količina amiloida smanjuje, ili da se smanjuje brzina stvaranja amiloida, sa kliničkim značajem.
28.Analog amiloidogenog polipeptida koji se izvodi iz Af) ili APP životinje, naznačen time, što se uvodi najmanje jedan izolovani strani THepitop, kao što je shematski pokazano za epitope P2 i P30 na Slici 1, ili što se kupiuje najmanje jedan strani TH epitop na polihidroksipolimer kao osnovu nosača, koji takođe nosi sekvenciju peptida izvedenu iz A f) ili APP, tako da imunizacija životinje sa analogom izaziva stvaranje antitela protiv amiloidogenog polipeptida.
29.Analog prema Zahtevu 28, naznačen time, što je modifikacija definisana u bilo kom od Zahteva 2-17.
30.lmunogeni preparat, naznačen time, što se sastoji od imunogeno efikasne količine analoga prema Zahtevu 28 ili 29, a preparat još sadrži farmaceutski i imunološki prihvatljiv nosač, i/ili tečni nosač i opciono adjuvant.
31.Fragment nukleinske kiseline, naznačen time, što kodira analog prema Zahtevu 28 ili 29.
32.Vektor, naznačen time, što nosi fragment nukleinske kiseline prema Zahtevu 31, kao što je vektor koji je sposoban za autonomnu replikaciju.
33.Vektor prema Zahtevu 32, naznačen time, što se bira iz grupe koju čine plazmid, faga, kosmid, minohromozom i virus.
34.Vektor prema bilo kom od Zahteva 32 ili 33, naznačen time, što u smeru 5'_,3' i u operabilnoj vezi sadrži promoter za izazivanje izlučivanja fragmenta nukleinske kiseline prema Zahtevu 31, opciono sekvencije nukleinske kiseline koja kodira vodeći peptid koji omogućava izlučivanje ili integraciju u membranu polipeptidnog fragmenta, fragment nukleinske kiseline prema Zahtevu 31, opciono terminator.
35.Vektor prema bilo kom od Zahteva 32 do 34, naznačen time, što je, kada se uvede u ćeliju domaćina, u stanju ili nije u stanju da se integriše u genom ćelije domaćina.
36.Vektor prema Zahtevu 34 ili 35, naznačen time, što promoter izaziva izlučivanje u eukariotsku ćeliju i/ili u prokariotsku ćeliju.
37.Transformisana ćelija, naznačena time, što nosi vektor prema bilo kom od Zahteva 32 do 36, kao što je transformisana ćelija koja je u stanju da replicira fragment nukleinske kiseline prema Zahtevu 31.
38.Transformisana ćelija prema Zahtevu 37, naznačena time što predstavlja mikroorganizam koji se bira između bakterije, kvasca, protozoe, ili ćelije izvedene iz višećelijskog organizma koji se bira između gljivice, ćelije insekta, kao što je s2 ili SF ćelija, ćelije biljke i ćelije sisara.
39.Transformisana ćelija prema Zahtevu 37 ili 38, naznačena time, što izlučuje fragment nukleinske kiseline prema Zahtevu 31, kao što je transformisana ćelija koja na njenoj površini izlučuje ili nosi analog prema Zahtevu 28 ili 29.
40.Postupak prema bilo kom od zahteva 1-17, naznačen time, što se prezentacija imuno sistemu ostvaruje ordiniranjem ne-patogenog mikroorganizma ili virusa, koji nosi fragment nukleinske kiseline koji kodira i izlučuje analog.
41.Preparat za izazivanje stvaranja antitela protiv A ili APP, naznačen time, što taj preparat sadrži: -fragment nukleinske kiseline prema Zahtevu 31 ili vektor prema bilo kom od Zahteva 32 do 36, i - farmaceutski i imunološki prihvatljiv nosač, i/ili tečni nosač, i/ili adjuvant.
42.Stabilan ćelijski soj koji nosi vektor prema bilo kom od Zahteva 32-36, i koji izlučuje fragment nukleinske kiseline prema Zahtevu 31, i koji opciono na njegovoj površini izlučuje ili nosi analog prema Zahtevu 28 ili 29.
43.Postupak za dobijanje ćelije prema bilo kom od Zahteva 37 do 39, naznačen time, što se taj postupak sastoji od transformisanja ćelije domaćina sa fragmentom nukleinske kiseline prema Zahtevu 31 ili sa vektorom prema bilo kom od Zahteva 32 do 36.
44.Upotreba analoga prema Zahtevu 28 ili 29 za dobijanje imunogenog preparata, koji opciono sadrži adjuvant, za deregulaciju amiloida u životinji.
45.Upotreba analoga prema Zahtevu 28 ili 29 za dobijanje imunogenog preparata, koji opciono sadrži adjuvant, za tretman, profilaksu ili ublažavanje Alzheimer-ove bolesti ili drugih stanja koja karakterišu naslage amiloida.
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HUP0300067A2 (en) 2003-05-28
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