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TR201802933T4 - Özellikleri değiştirilmiş faktör h bağlama proteinleri (fhbp) ve bunların kullanım yöntemi. - Google Patents

Özellikleri değiştirilmiş faktör h bağlama proteinleri (fhbp) ve bunların kullanım yöntemi. Download PDF

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TR201802933T4
TR201802933T4 TR2018/02933T TR201802933T TR201802933T4 TR 201802933 T4 TR201802933 T4 TR 201802933T4 TR 2018/02933 T TR2018/02933 T TR 2018/02933T TR 201802933 T TR201802933 T TR 201802933T TR 201802933 T4 TR201802933 T4 TR 201802933T4
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fhbp
composition
amino acid
naturally occurring
seq
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T Beernink Peter
M Granoff Dan
Pajon Feyt Rolando
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Childrens Hospital & Res Center At Oakland
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
    • C07K14/22—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Neisseriaceae (F)
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    • A61P31/04—Antibacterial agents
    • A—HUMAN NECESSITIES
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    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
    • A61K2039/55505—Inorganic adjuvants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
    • A61K2039/55511—Organic adjuvants
    • A61K2039/55566—Emulsions, e.g. Freund's adjuvant, MF59
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/195—Assays involving biological materials from specific organisms or of a specific nature from bacteria
    • G01N2333/22—Assays involving biological materials from specific organisms or of a specific nature from bacteria from Neisseriaceae (F), e.g. Acinetobacter

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Abstract

N. meningitidis'in en az bir suşu için bakterisidal olan antikorlar meydana getirebilen faktör H bağlama proteinleri ve bu proteinlerin kullanım yöntemleri temin edilir.

Description

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° BEERNINK PT ; GRANOFF DM.
° BORROW et al. Infect Iinmun., ° PAJON R et al. Vaccine, 2010, vol.
° WESTERINK MA et al. Proc. Natl. 4025 [0078] ° LOWELL GH et al. J. Exp. Med., ° PAGOTTO et al. Gene, 2000, vol.
° Short Protocols in Molecular Biology.
Wiley & Sons, 1999 [0097] ° RUSSELL et al. Emerging Infect o SACCHI CT et al. Clin Diagn Lab ° SACCHI et al. J. Infect Dis, 2000, - DOLAN-LIVENGOOD et al. J. 0 F lSSEl-IA et al. lnfect. lmmun., 2005, - ANDERSSON et al. J. o FLETCHER et al. Infection and ° FRASCH, C. E. ; CHAPMAN. J.
° SACCHI et al. Clin. Diag. Lab.
- URWIN et al. Epidem. and Infect., ° MASIGNANI et al. J Exp Med, ° WELSCH et al. J Immunol, 2004, ° ALTSCHUL et al. J. Mol. Biol., o ALTSCHUL et al. Nucleîc Acids ° HIGGINS D. et al. Nucleic Acids ° SAMBROOK et al. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor, 1989 [0092] - MOLDOVEAMI. Vaccine, 1988, ° KRIEG et al. Nature, 1995, vol. 374, 9 KLINMAN et al. PNAS USA, 1996, 0 BALLAS et al. J. Immunol, 1996, ° COWDERY et al. J. Inununol, 1996, ° YAMAMOTO et al. J pn. J. Cancer 0 STACEY et al. J. lmmunol., 1996, o MESSINA et al. J. Immunol, 1991, ° STEPHENS et al. Infect Iminun, 0 FROSCH et al. Mol Microbiol., ° SWARTLEY et al. J Bacteriol., - MOE et al. lnfeet. lmmun., 1999, vol. o ADU-BOBIE et al. lnfect lminun., ° STEEGHS et al. Infect Immun, 1999, ° VAN DER LEY et al. Infect Immun, o STEEGHS et al. J Endotoxin Res, o FISSHA et al. lnfect lmmun, 2005, ° GUNN J. S. ; KI-IENG, B. L. ; 0 VAN DER LEY et al. Proceedings of the ninth international pathogenic Neisseria conference. The Guildhall, 1994 [0125] ° RUSTICI et al. Science, 1993, vol.
° PORRO et al. Prog Clin Biol Res., - MARTIN et 21.1. Exp. Med., 1997, ° Vaccine design: the subunit and ad juvant approach, eds. Powell & Newman. Plenum Press, 1995 [0136] - KRIEG. Vaccine, 2000, vol. 19, 618- 622 [0136] ° KRIEG. Curr Opin Mol Ther, 2001, ° ROMAN et al. Nat. Med, 1997, vol. o WEINER et al. PNAS USA, 1997, ° DAVIS et al. J. lmmunol, 1998, vol.
- CHU et al. .1. Exp. Med, 1997, vol. o YI et al. J. Iinmunol, 1996, vol. 157, ° YI et al. J. lmmunol, 1998, vol. 160, - YI et al. 1. lmmunol, 1998, vol. 160, ° SZOKA et al. Ann. Rev. Biophys.
- Remington”s Pharmaceutical Science.
Mack Publishing Company, 1980 . NIWA et al. Gene, 1991, vol. 108, 193-9 [0165] ° BEERNINK PT et al. Infect Immun, ° SCHNEIDER MC et al. Nature, o PAJON et al. Vaccine, 2009, vol. 28, ° PAJON et al. Vaccine, 2009, V01. 28, 2122 [0185] . BEERNINK PT. M0] lmmunol, - BEERNINK et al. Microbiology, . BEERNINK ; GRANOFF. lnfect. 201 1 [0223] o GIULIANI et al. Infect. Iinmun., ° GRANOFF et al. .1. lmmunol., 1998, 0 LIPFORD et al. Ear. J. Immunol., 0 WELSCH et al. J. Infect. Dis., 2008, ° WELSCH et al. J. Immunol., 2004, - LEWIS et al. PLoS Pathog., 2010, 0 PLESTED ; GRANOFF. Clin.
Vaccine Immunol., 2008, vol. 15, 799 o BEERNINK et al. J. Infect. Dis., ° SCHNEIDER et al. Nature, 2009, -SCARguiJaaLJMMBmLmmi 0 BEERNINK et al. lnfect lmmun, o MOE et al. lnfect. lmmun., 2002, vol.
SEKILLERDEKI YAZILARIN ANLAMLARI A = Faktör H, ug/ml B : Fareler C = Insanlar D = l/Titre E : Antikapsüler F = WT Fareler G = fH Tg Fareler H = Neg. Kontrol J : Fare fH,si K = Bakteriyel GMT Orani, %95 CI L = Serum insan fH°si, ng/ml M : fH, J AR 4 ve J AR 5 'in fHbp No. 1 vahsi tip ve mutantlarina baglanmasi N = Konsantrasyon, ug/ml O = Standart P = Vahsi tip R = Yüzde inhibisyon S : Inhibitör, ug/ml SEKIL 7A T = Fazla isi kapasitesi U = Sicaklik (°C) SEKIL 7C V = Çalisma Y = 1/ I gG Anti-fHbp Titresi SEKIL 7D Z = Vahsi tipe karsi P degeri SEKIL 9B A1: Sekans kod no 81 = fH baglama, pre-immün Cl = fH baglama, post-immün El = % inhibisyon ± %95 C1 Fl = Alüminyum Gl = Trans jenik Hl : 1/ Serum Dilüsyonu ll : fH Baglama % Inhibisyonu J 1 = fH ve J AR 5”in fHbp No. 1 vahsi tip ve mutantina baglanmasi K1= fH ve J AR 5 ”in fHbp No. 15 vahsi tip ve mutantina baglanmasi Ll = fH”nin fHbp No. 1 mutantina baglanmasi M1 = J AR 5”in No. 1 mutantlarina baglanmasi Nl = J AR 4”i°1n No. 1 mutantlarina baglanmasi 01 = Modüler Segment Pl = Modüler Grup R1 : Varyant Grubu Sl : Kimera Tl = IH”nin iHbp No. 77 tekli mutantlarina baglanmasi Ul : JAR 31 'in fHbp NO. 77 tekli mutantlarina baglanmasi V] = fH'nin fHbp No. 77 ikili mutantlarina baglanmasi Yl = J AR 31 'in fHbp No. 77 ikili mutantlarina baglanmasi Zl = fH”nin fHbp No. 22 mutantlariiia baglanmasi A2 : Grafik 82 = JAR 31°in fHbp No. 22 vahsi tip ve mutantlarina baglanmasi C2 = JAR 31 bagi D2 : J AR 4”ün fHbp No. 22 vahsi tip ve mutantlarina baglanmasi E2 = JAR 4 bagi F2 : J AR 35”in fHbp No. 22 vahsi tip ve mutantlarina baglanmasi G2 = JAR 35 bagi H2 : J AR 31'in fHbp No. 22 mutantlarina baglanmasi 12 : J AR 35 ”in fHbp No. 22 mutantlarina baglanmasi J 2 = J AR 4”ün fHbp No. 22 mutantlarina baglanmasi K2 : fHbp NO. 1 mutantlari L2 : l/SBA Titresi M2 = fHbp No. 22 mutantlari N2 = fHbp No. 77 mutanti 02 : JAR 3 ve JAR 5 P2 = G121 ve K122 R2 = fH fragmani SEKIL 37A SZ: OD (Aralik) T2 = Cevap birimleri U2 : Zaman (S) SEKIL 37C V2 = Sayi SEKIL 38C Y2 = 60 dk.”da Yüzde SürVi SEKIL 41A 22 : 60 dk.”da Yüzde Sürvi (±2 SE) A3 = 60 dk.°da Yüzde Sürvi B3 = Grup A, Senegal 1/99 C3 : MAb, l/dilüsyon D3 = % fH lnhibisyon E3 : Ortalama, %95 C1 F3 : l/Serum Dilüsyonu G3 = %50 fH Inhibisyonu H3 : Peptit 00405 !mi 600 00 E 8 A 2A: <12 ali-III 105 IgG, anti~CRM 104 530“ ü.
D 10"' 105. '95. E D10“« &37° Dgâa C 105 SBA,E 1 2? J R fHprT B 9› 2 A” ”^A fH, pg/ml fH, pg/ml C 10"' 501". . . . 1 4 +ID1WT 4 E 3 uA-E218A E 3 g 2 ..su ° 2 v E218AIE239A 'A g - www 13_ SEKIL 5 0.1 pg/ml 10050 100 JAR4 100 JARS 40 60 80 100 SEKIL 7A EZîßAI E239A CD-1 BALB/c SEKIL 7C BALE/c, AI(OH)3 SEKIL 7D 00405nm SEKIL 9A /48 SEKIL 9B 00 405nm OD dOSnm OD ausnm 11/48 fH.i.igi'mI MAb.iigimi MAb,pglml 12/48 3 m fHbplD4 3 JAR4 3- fH fHbp ID 9 3 n-i fHbpIDM 3 JAR4 M. ug/ml Anti-fHbp mAb 13/48 3 JAR4 M. pglml Anli-fHbp mAb 3 m mbpiozz 81 .RMS fH. pglml Anli-fHbp mAb fH, pglml Anli-fHbp mAb 14/48 105' $ M D104 ^ 13: en D , 8<>o°°<> 4 o °o<> U) 10` (10, @0000 <› /48 .%3 +1› E 3 0 P K' 16/48 g 1.0 .WT 05 «R130A ,x fH.pg/ml 05 *H119A oR13OA Anti-fHbp MAb pglml 05 *H119A oR130A Anti-fHbp MAb ug/ml 17/48 No18/48 3 , ' 0891 2 pep15WT S41P O .m..uu. -o"'o pep fH.i.ig/ml B' JAR5 pep15541P 4 pep 15 WT . M”. ..... ...man-_ud pep 28 MAb, pg/ml 0 1 .i' MAb.ngmI 19/48 3 +1D1WT 3 2 -o-si R41S 2 4...: fH. pglml MAb. uglml /48 2 Emma 21/48 2 52%:: 2 Emma 2 ;Uma 22/48 a 92;& 2 ;9% OD 4050m OÜ 4050m 23/48 2_ 4-` K113A - 1 I 1 I | OD 405nm ÖD 405nm 24/48 1 I I I 1 /48 g 1 --A-vR41SIK113A/Dî21A B 3 JAR4 gz ~A~R41SIK113N0121A MAb, lig/ml MAb. uglml 26/48 ne" WT ..o -9- D211A p'" 03 -A- E218A , o 075 4-- E248A d" "13-- WT 3 + R41S fH.i.ig/ml 27/48 1-5- C2 g 10- --o--R8ÜA 1.0 C2 g 1.0 +5218A 9› +E248A Anli-fHbp MAb. pglml 28/48 E 1.0~ ' Ez 1.0- -A- EZ18A .11. g + E248A 'xiâfmomnû Anti-fHbp MAb pg/ml 29/48 1.5 GZ g 1.0 -o-RBOA 3 -e-D211A + E248A 1 I 7 1 I I Anii-fHbp MAb pg/ml /48 - T221AJH223A fH.pg/ml 0 5 - T221AV'H223A 062361 ' 001 01 1 ':0 'iOU Anti-!Htm MAb pglml E 10 li.' ' 1 0 7 .'l 0 4? v -0 n 0-5 F _ " +WT Anti-Mbp MAb pg/ml 1 3- J2 g *T2_›21A›i-`H223A_,__. k 8 01 “m“ E An(i›tHbp MAb pglml 31/48 fH,pg/ml 1.5 HZ E 10 ..... o 05 R41S Anii-fHbp MAb uglml 0 015 +R41S ~1A'O38A AntiJHbp MAb pglml 32/48 g 10~ ----- 8 +0126A , ûDZOîA 05” 0E202A fH,iJg/ml g 1.0~ 05.. ûDZOîA Anti-fHbp MAb uglml 33/48 09 0_ F _0 So _ooo . :& @Ö o :& 0.6 . (marx .nv O 0._ ms. v ..z mN 34/48 H44/76 /48 "; Ghana 7.04 '. ' L2 : I ' _ (10': I ' 'Q xKL; G'iiliiaî'O-î - _7_ A .
LZ 10"; .0 t.' '-r 1C's . (15': i i. w'i #isi-A 24 :Bu- 36/48 K113A/ D121A 37/48 38/48 39/48 52 1.0 SEKIL 37A 100 FL1-H SEKIL 37C 40/48 TZ 1000 SEKIL 37B ' 10` îO'` 10' 10': FL1-H SEKIL 37D 41/48 FL1-H= SEKIL 38A SEKIL 38B > 195' o-o-c›\0 150' VJAR 3 ° \": 1 1'0 160 mAh. pg/ml SEKIL 38C 42/48 mAbs, pg/ml SEKIL 39A FL1-H SEKIL 39C 43/43 FL1-H SEKIL 40 A FL1 -H FL1 -H SEKIL 40C 44/48 m; 15:?, m: ;Uma 45/48 ami-NspA mAb AL12 A3 150' anti-PorA mAh P1.9 46/48 3_ 10'J 6 A .1. ^ A NOMV OE fHbD NOMV KO B 100 e NOMV OE 80 .. NOMV KO D3 E3 6020 ....... 3_ 10' A1 G3 g 0 9 _i_ < o10" ' ' 47/48 48/48

Claims (20)

ISTEMLER
1. fHbp No. l”den (SEKANS KOD NO. l”de belirtilen sekansa sahip) türetilmis ve SEKANS KOD NO. l”le en az %90 sekans özdesligine sahip olan, dogal olusumlu olmayan bir faktör H baglama proteini (fHbp) içeren ve dogal olusumlu olmayan fHbp°nin konum R41 ”de bir amino asit ikamesine sahip oldugu; söz konusu dogal olusumlu olmayan fHpriin insan faktör H'si (fH) için fHbp No. l”den daha düsük bir afiniteye sahip oldugu ve N. meningitidis'e karsi bakterisidal antikorlar meydana getirdigi; R41 ”deki amino asit ikamesinin R4IS veya R4lA oldugu; ve alüminyum hidroksit, alüminyum fosfat, MF59T'V', bir “su içinde yag” emülsiyonu, monofosforil lipid A veya bir saponin içeren; veya fHbp eksprese edecek sekilde genetik olarak degistirilmis Neisseria meningitidis dis membran vezikülleri içeren; veya fHbp eksprese edecek sekilde genetik olarak degistirilmis Neisseria meningitidis mikrovezikülleri içeren bir immünojenik bilesim.
2. fHbp No. l”den (SEKANS KOD NO. 1°de belirtilen sekansa sahip) türetilmis ve SEKANS KOD NO. 1°1e en az %90 sekans özdesligine sahip olan; konum R4l”de bir amino asit ikamesine sahip olan; ikamenin R4IS veya R41A oldugu; ve insan faktör H°si (fH) için fHbp No. l”den daha düsük bir afiniteye sahip olan; ve N. ineningitidis”e karsi bakterisidal antikorlar meydana getiren, dogal olusumlu olmayan bir faktör H baglama proteini (fHbp).
3. Dogal olusumlu olmayan amino asit sekansinin SEKANS KOD NO. 1'deki amino asit sekansindan 1 ila 10 amino asit kadar farkli oldugu, Istem l°de tarif edilen bilesim veya Istem 2°de tarif edilen
4. Istem 1 ila 3”ten herhangi birinde tarif edilen ve dogal Olusumlu olinayan ainino asit sekansinin SEKANS KOD NO. 1°deki ainino asit içerdigi bilesim veya fHbp.
5. Istein 1 ila 4sten herhangi birinde tarif edilen ve söz konusu dogal olusumlu olinayan fHbp°nin insan fH,si için fl-lbp No. l°in afinitesinin yaklasik %60°indan fazla olinayan bir afiniteyle insan fH”sine baglandigi bilesiin veya fHbp.
6. Istem 1 ila 5”ten herhangi birinde tarif edilen ve R4l°deki amino asit ikainesiniii R4IS oldugu bilesim veya fHbp.
7. Istem 1 ila 5 'ten herhangi birinde tarif edilen ve R4l°deki ainino asit ikamesinin R41A oldugu bilesim veya fHbp.
8. Istein 6 veya 7”de tarif edilen ve dogal olusumlu olinayaii fHbp”nin, SEKANS KOD NO. 17 amino asit sekansina en az %90 özdes bir amino asit sekansi içeren bir degisken A (VA) segmenti içerdigi bilesim veya fHbp.
9. Onceki istemlerden herhangi birinde tarif edilen ve dogal olusuinlu olmayan fHbpîiin SEKANS KOD NO. 1°1e en az %95 sekans özdesligi içerdigi bilesim veya fHbp.
10. Istem 9°da tarif edilen ve dogal olusuinlu olinayan fHbp°nin SEKANS KOD NO. l”le en az %98 sekans Özdesligi içerdigi bilesim veya fHbp.
11. Istem 1 veya 2”de tarif edilen ve dogal olusumlu olmayan fHbp'nin R41 ”deki amino asit ikamesi haricinde SEKANS KOD NO. 1°le sekans özdesligi içerdigi bilesim veya fHbp.
12. Istem 1 veya 3 ila ll°de tarif edilen ve alüminyum hidroksit, alüminyum fosfat, MF59TM, bir “su içinde yag› emülsiyonu, monofosforil lipid A veya bir saponin arasindan seçilen bir adjuvan içeren bilesim.
13. Istem 1 veya 3 ila llsde tarif edilen ve tHbp eksprese edecek sekilde genetik olarak degistirilmis Neisseria meningitidis dis membran vezik'ülleri içeren bilesim,
14. Istem 1 veya 3 ila ll”de tarif edilen ve tHbp ek3prese edecek sekilde genetik olarak degistirilmis Neisseria meningitidis mikrovezikülleri içeren bilesim.
15. Istem 1 veya 3 ila 14°ten herhangi birinde tarif edilen ve bir memelide bir antikor cevabi meydana getirme yönteminde kullanim için olan ve söz konusu meinelinin bir insan oldugu bir bilesim.
16. Istem 15°te tarif edilen ve söz konusu antikor cevabinin Neisseria ineningitidis”e karsi bakterisidal antikorlar üretilmesini içerdigi bir bilesim.
17. Istem 1 veya 3 ila 11”den herhangi birinde tarif edilen bilesimin dogal olusumlu olmayan bir fHbp°sini kodlayan veya Istem 2 ila ll'den herhangi birinde tarif edilen bir dogal olusumlu olmayan fHbp,yi kodlayan bir nükleik asit.
18. Istem l7”de tarif edilen nükleik asidi içeren bir rekombinant ekSpresyon vektörü.
19. Istem l7°de tarif edilen nükleik asidi içeren bir genetik olarak degistirilmis konak hücre veya Istem 18”de tarif edilen rekombinant ekspresyon vektörü.
20. Neisseria hastaligini 'Önleme yönteminde kullanim için Istem 1 veya 3 ila 1 1'den herhangi birinde tarif edilen bir bilesim veya Istem 2 ila ll”den herhangi birinde tarif edilen bir fHbp.
TR2018/02933T 2010-03-30 2011-03-29 Özellikleri değiştirilmiş faktör h bağlama proteinleri (fhbp) ve bunların kullanım yöntemi. TR201802933T4 (tr)

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