COMBINATION THERAPIES USING ANTI-PSEUDOMONAS PSL AND PCRV BINDING MOLECULES

• US 20150284450A1
• Filed: 11/06/2013
• Published: 10/08/2015
• Est. Priority Date: 11/06/2012
• Status: Abandoned Application

• Current Assignee: Medimmune LLC (Astrazeneca PLC)
• Original Assignee: Medimmune LLC (Astrazeneca PLC)
• Inventors: Digiandomenico, Antonio, Warrener, Paul, Stover, Charles, Sellman, Bret, Minter, Ralph, Rust, Steven, Tomich, Mladen, Venkatraman, Vignesh, Varkey, Reena, Peng, Li, Damschroder, Melissa, Chowdhury, Partha, Dimasi, Nazzareno, Fleming, Ryan, Bezabeh, Binyam, Gao, Changshou, Guillard, Sandrine
• Application Number:
  US14/440,215
• Publication Number:
  US 20150284450A1
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 2039/507 Comprising a combination of two or more separate antibodies
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 39/40 bacterial
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 16/1214 from Pseudomonadaceae (F)
• CPC Class Codes: C07K 2317/21 from primates, e.g. man
• CPC Class Codes: C07K 2317/31 multispecific
• CPC Class Codes: C07K 2317/53 Hinge
• CPC Class Codes: C07K 2317/565 Complementarity determining region [CDR]
• CPC Class Codes: C07K 2317/622 Single chain antibody (scFv)
• CPC Class Codes: C07K 2317/76 Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value

Abstract

The disclosure relates to combination therapies comprising anti-Pseudomonas Psl and PcrV binding molecules and related compositions, for use in prevention and treatment of Pseudomonas infection.

Claims

1. An isolated binding molecule which specifically binds to Pseudomonas PcrV, comprising an immunoglobulin VH and an immunoglobulin VL, wherein the VH comprises an amino acid sequence selected from the group consisting of SEQ ID NO:

255 and SEQ ID NO;

257, and wherein the VL comprises the amino acid sequence of SEQ ID NO;

256.

2. The binding molecule of claim 1, wherein the VH comprises SEQ ID NO:

255.

3. The binding molecule of claim 1, wherein the VH comprises SEQ ID NO:

257.

8. The binding molecule of any one of claims 1 to 7, which comprises an anti-PcrV antibody or antigen-binding fragment thereof.

9. The binding molecule of claim 8, wherein the VH is part of an antibody heavy chain which further comprises one or more heavy chain constant regions, and wherein the VL is part of an antibody light chain which further comprises a light chain constant region.

10. The binding molecule of claim 9, comprising two antibody heavy chains and two antibody light chains.

11. The binding molecule of any one of claims 8 to 10, which comprises a bispecific antibody.

12. The binding molecule of claim 11, further comprising a binding domain which specifically binds to Pseudomonas Psl.

13. The binding molecule of claim 12, wherein the binding domain which specifically binds to Pseudomonas Psl comprises an anti-Psl ScFv molecule.

14. The binding molecule of claim 13, wherein the anti-Psl ScFv molecule comprises an amino acid sequence of SEQ ID NO:

240-SEQ ID NO;

254, or any combination of two or more amino acid sequences of SEQ ID NO;

240-SEQ ID NO;

254.

15. The binding molecule of claim 13 or claim 14, wherein an anti-Psl ScFv molecule is inserted into the hinge region of each heavy chain of the anti-PcrV antibody or fragment thereof.

16. The binding molecule of claim 15, wherein each antibody heavy chain comprises the formula VH-CH1-H1-L1-S-L2-H2-CH2-CH3, wherein CH1 is a heavy chain constant region domain-1, H1 is a first heavy chain hinge region fragment, L1 is a first linker, S is an anti-PcrV ScFv molecule, L2 is a second linker, H2 is a second heavy chain hinge region fragment, CH2 is a heavy chain constant region domain-2, and CH3 is a heavy chain constant region domain-3.

17. The binding molecule of claim 16, wherein VH comprises the amino acid sequence of SEQ ID NO:

255, SEQ ID NO;

257, or SEQ ID NO;

317.

18. The binding molecule of claim 17, wherein CH1 comprises SEQ ID NO:

319.

19. The binding molecule of any one of claims 16 to 18, wherein L1 and L2 are the same or different, and independently comprise (a) [GGGGS]n, wherein n is 0, 1, 2, 3, 4, or 5, (b) [GGGG]n, wherein n is 0, 1, 2, 3, 4, or 5, or a combination of (a) and (b).

20. The binding molecule of any one of claims 16 to 19, wherein H1 comprises EPKSC (SEQ ID NO:

320).

21. The binding molecule of any one of claims 16 to 20, wherein H2 comprises DKTHTCPPCP (SEQ ID NO:

321).

22. The binding molecule of any one of claims 16 to 21, wherein S comprises an amino acid sequence selected from the group consisting of SEQ ID NO:

240 to SEQ ID NO;

254, and any combination thereof.

23. The binding molecule of any one of claims 16 to 22, wherein CH2-CH3 comprises APELLGGPSVFLFPPKPKDTLX1IX2RX3PEVTCVVVDVSHEDPEVKFNWY VDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTI SKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTP PSLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO:

322), wherein X1 is M or Y, X2 is S or T, and X3 is T or E.

24. The binding molecule of any one of claims 16 to 23, wherein each antibody light chain comprises VL-CL, wherein CL is an antibody light chain kappa constant region or an antibody light chain lambda constant region.

25. The binding molecule of claim 24, wherein VL comprises the amino acid sequence of SEQ ID NO:

256 or SEQ ID NO;

318.

26. The binding molecule of claim 24, wherein CL comprises the amino acid sequence of SEQ ID NO:

323.

27. The binding molecule of any one of claims 15 to 26, comprising two identical heavy chains and two identical light chains, wherein each antibody heavy chain comprises SEQ ID NO:

264, and wherein each antibody light chain comprises SEQ ID NO;

263.

56. The binding molecule of claim 1, wherein the VH CDR3 is SEQ ID NO:

279, and the VL CDR3 is SEQ ID NO;

287.

57. The binding molecule of claim 56, wherein the VH comprises SEQ ID NO:

308, and the VL comprises SEQ ID NO;

325.

69. A cell comprising or producing the binding molecule of any one of claim 1-27, or 29-67, or a subunit thereof, the bispecific antibody of claim 28 or a subunit thereof, the polypeptide of claim 68, or any combination thereof.

70. An isolated polynucleotide comprising a nucleic acid which encodes the binding molecule of any one of claim 1-27, or 29-67, or a subunit thereof, the bispecific antibody of claim 28 or a subunit thereof, the polypeptide of claim 68, or any combination thereof.

71. A vector comprising the polynucleotide of claim 70.

72. A cell comprising the polynucleotide of claim 69 or the vector of claim 70.

73. A composition comprising the binding molecule of any one of claim 1-27, or 29-67, or an antigen-binding subunit thereof, and a pharmaceutically acceptable carrier.

75. The composition of claim 73 or claim 74, which binds to at least 80%, at least 85%, at least 90% or at least 95% of P. aeruginosa strains isolated from infected patients.

76. The composition of claim 75, wherein the P. aeruginosa strains are isolated from one or more of lung, sputum, eye, pus, feces, urine, sinus, a wound, skin, blood, bone, or knee fluid.

77. The composition of any of claims 73 to 76, wherein the binding molecule or a subunit thereof, or the bispecific antibody or a subunit thereof, is conjugated to an agent selected from the group consisting of antimicrobial agent, a therapeutic agent, a prodrug, a peptide, a protein, an enzyme, a lipid, a biological response modifier, pharmaceutical agent, a lymphokine, a heterologous antibody or fragment thereof, a detectable label, polyethylene glycol (PEG), and a combination of two or more of any said agents.

78. The composition of claim 77, wherein the detectable label is selected from the group consisting of an enzyme, a fluorescent label, a chemiluminescent label, a bioluminescent label, a radioactive label, or a combination of two or more of any said detectable labels.

79. The composition of any of claims 73 to 78, further comprising an antibiotic.

80. The composition of claim 79, wherein the antibiotic is selected from the group consisting of Ciprofloxacin, Meropenem, and a combination thereof.

81. A method of preventing or treating a Pseudomonas infection in a subject in need thereof, comprising administering to a subject an effective amount of the composition of any one of claims 73-80.

85. The method of any one of claims 81 to 84, wherein said composition is administered for two or more prevention/treatment cycles.

86. A method of preventing or treating a Pseudomonas infection in a subject in need thereof, comprising administering to a subject an effective amount of the composition of any one of claim 79 or 80, wherein said administration provides a synergistic therapeutic effect in the prevention or treatment of the Pseudomonas infection in the subject, and wherein said synergistic effect is greater than the sum of the individual effects of administration of equimolar quantities of one or more of (a) just the bispecific antibody, (b), the monospecific binding molecules with the same Pseudomonas Psl and Pseudomonas PcrV binding specificities as the bispecific antibody and (c) the antibiotic.

87. The method of claim 86, wherein the composition is administered for two or more prevention/treatment cycles.

88. The method of any of claims 81 to 87, wherein the Pseudomonas infection is a P. aeruginosa infection.

89. The method of any of claims 81 to 88, wherein the subject is a human

90. The method of any one of claims 81 to 89, wherein the infection is an ocular infection, a lung infection, a burn infection, a wound infection, a skin infection, a blood infection, a bone infection, or a combination of two or more of said infections.

91. The method of any one of claims 81 to 90, wherein the subject has acute pneumonia, burn injury, corneal infection, cystic fibrosis, or a combination thereof.

92. A kit comprising the composition of any one of claims 73 to 80.

4. An isolated binding molecule which specifically binds to Pseudomonas PcrV, comprising an immunoglobulin VH and an immunoglobulin VL, each comprising a CDR1, CDR2, and CDR3,

(a) wherein the VH CDR1 is SEQ ID NO:

311) or a variant thereof comprising 1, 2, 3, or 4 conservative amino acid substitutions, the VH CDR2 is SEQ ID NO;

312, or a variant thereof comprising 1, 2, 3, or 4 conservative amino acid substitutions, and the VHCDR3 is SEQ ID NO;

313, or a variant thereof comprising 1, 2, 3, or 4 conservative amino acid substitutions;

or

(b) wherein the VL CDR1 is SEQ ID NO;

314, or a variant thereof comprising 1, 2, 3, or 4 conservative amino acid substitutions, the VL CDR2 is SEQ ID NO;

315, or a variant thereof comprising 1, 2, 3, or 4 conservative amino acid substitutions, and the VL CDR3 is SEQ ID NO;

316, or a variant thereof comprising 1, 2, 3, or 4 conservative amino acid substitutions;

or

(c) a combination of (a) and (b);

wherein the VH and VL CDRs are according to the Kabat numbering system.

5. The binding molecule of claim 4,

(a) wherein the VH CDR1 is SEQ ID NO:

311) the VH CDR2 is SEQ ID NO;

312, and the VHCDR3 is SEQ ID NO;

313;

or

(b) wherein the VL CDR1 is SEQ ID NO;

314, the VL CDR2 is SEQ ID NO;

315, and the VL CDR3 is SEQ ID NO;

316;

or

(c) a combination of (a) and (b).

6. An isolated binding molecule which specifically binds to Pseudomonas PcrV, comprising an immunoglobulin VH and an immunoglobulin VL,

(a) wherein the VH comprises an amino acid sequence at least 90% identical to SEQ ID NO:

317;

or

(b) wherein the VL comprises an amino acid sequence at least 90% identical to SEQ ID NO;

318;

or

(c) a combination of (a) and (b).

7. The binding molecule of claim 6, wherein the VH comprises SEQ ID NO:

317 and the VL comprises SEQ ID NO;

318.

28. A bispecific antibody which specifically binds to Pseudomonas Psl and Pseudomonas PcrV, comprising an immunoglobulin heavy chain and an immunoglobulin light chain, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO:

264, and the light chain comprises the amino acid sequence of SEQ ID NO;

263.

74. A composition comprising the bispecific antibody of claim 28 or an antigen-binding subunit thereof, and a pharmaceutically acceptable carrier.

82. A method of preventing or treating a Pseudomonas infection in a subject in need thereof, comprising administering to a subject an effective amount of the bispecific antibody of claim 28 or the composition of any one of claims 74-80 wherein the composition comprises a bispecific antibody.

83. The method of claim 82, wherein the administration provides a synergistic therapeutic effect in the prevention or treatment of the Pseudomonas infection in the subject, and wherein the synergistic effect is greater than the sum of the individual effects of administration of equimolar quantities of monospecific binding molecules with the same Pseudomonas Psl and Pseudomonas PcrV binding specificities as the bispecific antibody.

84. The method of claim 83, wherein the synergistic therapeutic effect results in greater percent survival than the additive percent survival of subjects to which only one of the binding domains has been administered.

29. An isolated binding molecule which specifically binds to Pseudomonas Psl, comprising an immunoglobulin VH and an immunoglobulin VL, each comprising a complementarity determining region 1 (CDR1), CDR2, and CDR3, wherein the VH CDR1 is SEQ ID NO:

47, the VH CDR2 is SEQ ID NO;

48, the VH CDR3 is selected from the group consisting of SEQ ID NO;

258, SEQ ID NO;

267, SEQ ID NO;

268, SEQ ID NO;

269, SEQ ID NO;

270, SEQ ID NO;

271, SEQ ID NO;

272, SEQ ID NO;

273, SEQ ID NO;

274, SEQ ID NO;

275, SEQ ID NO;

276, SEQ ID NO;

277, SEQ ID NO;

278, and SEQ ID NO;

279, the VL CDR1 is SEQ ID NO;

50, the VL CDR2 is SEQ ID NO;

51, and the VL CDR3 is selected from the group consisting of SEQ ID NO;

280, SEQ ID NO;

281, SEQ ID NO;

282, SEQ ID NO;

52, SEQ ID NO;

283, SEQ ID NO;

284, SEQ ID NO;

285, SEQ ID NO;

286, and SEQ ID NO;

287, wherein the VH and VL CDRs are according to the Kabat numbering system.

30. The binding molecule of claim 29, wherein the VH CDR3 is SEQ ID NO:

258, and the VL CDR3 is SEQ ID NO;

280.

31. The binding molecule of claim 30, wherein the VH comprises SEQ ID NO:

288, and the VL comprises SEQ ID NO;

289.

32. The binding molecule of claim 29, wherein the VH CDR3 is SEQ ID NO:

267, and the VL CDR3 is SEQ ID NO;

281.

33. The binding molecule of claim 32, wherein the VH comprises SEQ ID NO:

290, and the VL comprises SEQ ID NO;

291.

34. The binding molecule of claim 29, wherein the VH CDR3 is SEQ ID NO:

268, and the VL CDR3 is SEQ ID NO;

282.

35. The binding molecule of claim 34, wherein the VH comprises SEQ ID NO:

292, and the VL comprises SEQ ID NO;

293.

36. The binding molecule of claim 29, wherein the VH CDR3 is SEQ ID NO:

269, and the VL CDR3 SEQ ID NO;

52.

37. The binding molecule of claim 36, wherein the VH comprises SEQ ID NO:

294, and the VL comprises SEQ ID NO;

11.

38. The binding molecule of claim 29, wherein the VH CDR3 is SEQ ID NO:

270, and the VL CDR3 is SEQ ID NO;

283.

39. The binding molecule of claim 38, wherein the VH comprises SEQ ID NO:

295, and the VL comprises SEQ ID NO;

296.

40. The binding molecule of claim 29, wherein the VH CDR3 is SEQ ID NO:

271, and the VL CDR3 is SEQ ID NO;

284.

41. The binding molecule of claim 40, wherein the VH comprises SEQ ID NO:

297, and the VL comprises SEQ ID NO;

298.

42. The binding molecule of claim 29, wherein the VH CDR3 is SEQ ID NO:

272, and the VL CDR3 is SEQ ID NO;

285.

43. The binding molecule of claim 42, wherein the VH comprises SEQ ID NO:

299, and the VL comprises SEQ ID NO;

300.

44. The binding molecule of claim 29, wherein the VH CDR3 is SEQ ID NO:

273, and the VL CDR3 is SEQ ID NO;

286.

45. The binding molecule of claim 44, wherein the VH comprises SEQ ID NO:

301, and the VL comprises SEQ ID NO;

302.

46. The binding molecule of claim 29, wherein the VH CDR3 is SEQ ID NO:

274, and the VL CDR3 is SEQ ID NO;

52.

47. The binding molecule of claim 46, wherein the VH comprises SEQ ID NO:

303, and the VL comprises SEQ ID NO;

11.

48. The binding molecule of claim 29, wherein the VH CDR3 is SEQ ID NO:

275, and the VL CDR3 is SEQ ID NO;

52.

49. The binding molecule of claim 48, wherein the VH comprises SEQ ID NO:

304, and the VL comprises SEQ ID NO;

11.

50. The binding molecule of claim 29, wherein the VH CDR3 is SEQ ID NO:

276, and the VL CDR3 is SEQ ID NO;

52.

51. The binding molecule of claim 50, wherein the VH comprises SEQ ID NO:

305, and the VL comprises SEQ ID NO;

11.

52. The binding molecule of claim 29, wherein the VH CDR3 is SEQ ID NO:

277, and the VL CDR3 is SEQ ID NO;

52.

54. The binding molecule of claim 29, wherein the VH CDR3 is SEQ ID NO:

278, and the VL CDR3 is SEQ ID NO;

52.

55. The binding molecule of claim 54, wherein the VH comprises SEQ ID NO:

307, and the VL comprises SEQ ID NO;

11.

59. The binding molecule of any one of claims 29 to 58, which comprises an anti-Psl antibody or antigen-binding fragment thereof.

60. The binding molecule of claim 59, which comprises a single-chain Fv (ScFv) antibody molecule.

61. The binding molecule of claim 60, wherein the ScFv comprises the formula:

VH-L-VL, and wherein L is a linker.

62. The binding molecule of claim 61, wherein the ScFv comprises the formula:

VL-L-VH, and wherein L is a linker.

63. The binding molecule of claim 61 or claim 62, wherein L comprises (a) [GGGGS]n, wherein n is 0, 1, 2, 3, 4, or 5, (b) [GGGG]n, wherein n is 0, 1, 2, 3, 4, or 5, or a combination of (a) and (b).

64. The binding molecule of claim 63, wherein the linker further comprises ala-leu at the C-terminus of the linker.

65. The binding molecule of any one of claim 29, 30, 31, 59 to 61, 63, or 64, comprising the amino acid sequence of SEQ ID NO:

240.

66. The binding molecule of any one of claim 29, 30, 31, 59 to 61, 63, or 64, comprising the amino acid sequence of SEQ ID NO:

262.

67. The binding molecule of any one of claim 29, 32 to 61, 63, or 64, comprising an amino acid sequence selected from the group consisting of SEQ ID NO:

241, SEQ ID NO;

242, SEQ ID NO;

243, SEQ ID NO;

244, SEQ ID NO;

245, SEQ ID NO;

246, SEQ ID NO;

247, SEQ ID NO;

248, SEQ ID NO;

249, SEQ ID NO;

250, SEQ ID NO;

251, SEQ ID NO;

252, SEQ ID NO;

253, SEQ ID NO;

254, and any combination thereof.

53. The binding molecule of claim, wherein the VH comprises SEQ ID NO:

306, and the VL comprises SEQ ID NO;

11.

58. An isolated binding molecule which specifically binds to Pseudomonas Psl, comprising an immunoglobulin VH and an immunoglobulin VL, wherein the VH comprises the amino acid sequence of SEQ ID NO:

309, and wherein the VL comprises the amino acid sequence of SEQ ID NO;

310.

68. An isolated polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:

240, SEQ ID NO;

241, SEQ ID NO;

242, SEQ ID NO;

243, SEQ ID NO;

244, SEQ ID NO;

245, SEQ ID NO;

246, SEQ ID NO;

247, SEQ ID NO;

248, SEQ ID NO;

249, SEQ ID NO;

250, SEQ ID NO;

251, SEQ ID NO;

252, SEQ ID NO;

253, SEQ ID NO;

254, SEQ ID NO;

262, SEQ ID NO;

255, SEQ ID NO;

256, SEQ ID NO;

257, SEQ ID NO;

263, SEQ ID NO;

264, SEQ ID NO;

258, SEQ ID NO;

263, SEQ ID NO;

264, SEQ ID NO;

267, SEQ ID NO;

268, SEQ ID NO;

269, SEQ ID NO;

270, SEQ ID NO;

271, SEQ ID NO;

272, SEQ ID NO;

273, SEQ ID NO;

274, SEQ ID NO;

275, SEQ ID NO;

276, SEQ ID NO;

277 SEQ ID NO;

278 SEQ ID NO;

279, SEQ ID NO;

280, SEQ ID NO;

281, SEQ ID NO;

282, SEQ ID NO;

283, SEQ ID NO;

284, SEQ ID NO;

285, SEQ ID NO;

286, SEQ ID NO;

287, SEQ ID NO;

288, SEQ ID NO;

289, SEQ ID NO;

290, SEQ ID NO;

291, SEQ ID NO;

292, SEQ ID NO;

293, SEQ ID NO;

294, SEQ ID NO;

295, SEQ ID NO;

296, SEQ ID NO;

298, SEQ ID NO;

299,300, SEQ ID NO;

301, SEQ ID NO;

302, SEQ ID NO;

303, SEQ ID NO;

304, SEQ ID NO;

305, SEQ ID NO;

306, SEQ ID NO;

307, SEQ ID NO;

308, SEQ ID NO;

309, SEQ ID NO;

310, SEQ ID NO;

311, SEQ ID NO;

312, SEQ ID NO;

313, SEQ ID NO;

315, SEQ ID NO;

316, SEQ ID NO;

317, SEQ ID NO;

318, SEQ ID NO;

325, and any combination thereof.