Thrombopoietic compounds

• US 20080070840A1
• Filed: 06/18/2007
• Published: 03/20/2008
• Est. Priority Date: 06/19/2006
• Status: Active Grant

• Current Assignee: Amgen Inc.
• Original Assignee: Amgen Inc.
• Inventors: Tsai, Mei-Mei, Sun, Jeonghoon, Min, Hosung, Galimi, Francesco, Gegg, Colin, Walker, Kenneth
• Granted Patent: US 7,981,425 B2
• Time in Patent Office: Days
• US Class Current: 514/12
• CPC Class Codes: A61K 38/17 from animals; from humans enzyme inhibitors A61K38/005
• CPC Class Codes: A61P 7/02 Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
• CPC Class Codes: C07K 14/001 by chemical synthesis
• CPC Class Codes: C07K 14/524 Thrombopoietin, i.e. C-MPL ligand
• CPC Class Codes: C07K 2319/30 Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
• CPC Class Codes: C07K 7/06 having 5 to 11 amino acids
• CPC Class Codes: C07K 7/08 having 12 to 20 amino acids gastrins C07K14/595; somatostatins C07K14/655; melanotropins C07K14/68

Abstract

The invention relates to the field of compounds, especially peptides or polypeptides, that have thrombopoietic activity. The peptides and polypeptides of the invention may be used to increase platelets or platelet precursors (e.g., megakaryocytes) in a mammal.

Claims

1. A compound that binds to an mp1 receptor comprising a structure set out in Formula I,

[(X¹)_a—

(F¹)_z—

(X²)_b]-(L¹)_c-WSP_d 

 

Formula I

and multimers thereof, wherein;

F¹ is a vehicle;

X¹ is independently selected from;

P¹-(L²)_e-

P²-(L³)_f-P¹-(L²)_e-

P³-(L⁴)_g-P²-(L³)_f-P¹-(L²)_e- and

P⁴-(L⁵)_h-P³-(L⁴)_g-P²-(L³)_f-P¹-(L²)_e-

X² is independently selected from;

-(L²)_e-P¹,

-(L²)_e-P¹-(L³)_f-P²,

-(L²)_e-P¹-(L³)_f-P²-(L⁴)_g-P³, and

-(L²)_e-P¹-(L³)_f-P²-(L⁴)_g-P³-(L⁵)_h-P⁴

wherein P¹, P², P³, and P⁴ are each independently sequences of pharmacologically active peptides;

L¹, L², L³, L⁴, and L5 are each independently linkers;

a, b, c, d, e, f, g, and h are each independently 0 or 1;

z is 0, 1, 2, or more; and

WSP is a water soluble polymer, the attachment of which is effected at any reactive moiety in F¹;

wherein P is a molecule comprising the structure U¹-Y¹-Y²-Y³-Y⁴-Y⁵-Y⁶-Y⁷-U² wherein

U¹-Y¹(Cys, Leu, Met, Pro, Gln, Val, or X₁)-Y² (Phe, Lys, Leu, Asn, Gln, Arg, Ser, Thr, Val, or X₂)-Y³(Cys, Phe, Ile, Leu, Met, Arg, Ser, Val, Trp, or X₃)-Y⁴-Y⁵(Ala, Asp, Glu, Gly, Lys, Met, Gln, Arg, Ser, Thr, Val, Tyr, or XS)-Y⁶(Cys, Phe, Gly, Leu, Met, Ser, Val, Trp, Tyr, or X₆)-Y⁷(Cys, Gly, Ile, Lys, Leu, Met, Asn, Arg, Val, or X₇)-U²,

wherein at least one of Y¹-Y³ and Y⁵-Y⁷ corresponds to a respective X₁-X₃ and X₅-X₇;

wherein U¹ or U² is any amino acid or peptide;

wherein when Y¹ is not an amino acid selected from the group consisting of Cys, Leu, Met, Pro, Gln, and Val, then X₁ is selected from the group consisting of Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Asn, Arg, Ser, Thr, Trp, and Tyr;

wherein when Y² is not an amino acid selected from the group consisting of Phe, Lys, Leu, Asn, Gln, Arg, Ser, Thr, and Val, then X₂ is selected from the group consisting of Ala, Cys, Asp, Glu, Gly, His, Ile, Met, Pro, Trp, and Tyr;

wherein when Y³ is not an amino acid selected from the group consisting of Cys, Phe, Ile, Leu, Met, Arg, Ser, Val, and Trp, then X₃ is selected from the group consisting of Ala, Asp, Glu, Gly, His, Lys, Asn, Pro, Gln, Thr, and Tyr;

wherein when Y⁴ is any amino acid;

wherein when Y⁵ is not an amino acid selected from the group consisting of Ala, Asp, Glu, Gly, Lys, Met, Gln, Arg, Ser, Thr, Val, and Tyr, then X₅ is selected from the group consisting of Cys, Phe, His, Ile, Leu, Asn, Pro, and Trp;

wherein when Y⁶ is not an amino acid selected from the group consisting of Cys, Phe, Gly, Leu, Met, Ser, Val, Trp, and Tyr, then X₆ is selected from the group consisting of Ala, Asp, Glu, His, Ile, Lys, Asn, Pro, Gln, Arg, and Thr; and

wherein when Y⁷ is not an amino acid selected from the group consisting of Cys, Gly, Ile, Lys, Leu, Met, Asn, Arg, and Val, then X₇ is selected from the group consisting of Ala, Asp, Glu, Phe, His, Pro, Gln, Ser, Thr, Trp, and Tyr;

and physiologically acceptable salts thereof.

2. The compound of claim 1 wherein at least a or b is 1.

3. The compound of claim 1 wherein b, c, d, e, f, g and h are 0.

5. The compound of claim 1, 2, 3, or 4 wherein

F¹ is an Fc domain modified so that it comprises at least one X³ in a loop region;

X³ is independently selected from

-(L⁶)_i-P⁵-(L⁷)_j,

-(L⁶)_i-P⁵-(L⁷)_j-P⁶(L⁸)_k,

-(L⁶)_i-P⁵-(L⁷)_j-P⁶-(L⁸)_k-P⁷-(L⁹)_l, and

-(L⁶)_i-P⁵-(L⁷)_j-P⁶-(L⁸)_k-P⁷-(L⁹)_l-P⁸-(L¹⁰)_m;

P⁵, P⁶, P⁷, and P⁸ are each independently sequences of pharmacologically active peptides;

L⁶, L⁷, L⁸, L⁹, and L¹⁰ are each independently linkers;

i, j, k, l, and m are each independently 0 or 1; and

z is 1, 2, or more.

6. The compound of claim 5 wherein a and b are each 0.

7. The compound of claim 5 wherein the Fc domain comprises an IgG Fc domain.

8. The compound of claim 7 wherein the Fc domain comprises a sequence selected from SEQ ID NOS:

3 and 344-352.

9. The compound of claim 5 wherein the Fc domain comprises an IgG1 Fc domain.

10. The compound of claim 9 wherein the IgG1 Fc domain comprises SEQ ID NO:

3 and X³ is inserted into or replaces all or part of a sequence selected from SEQ ID NOS;

428, 429, 431, 432, 434, 435, 437, 439, 441, and 443.

11. The compound of claim 10 wherein X³ is inserted into or replaces all or part of a sequence selected from SEQ ID NOS:

430, 433, 436, 438, 440, 442, and 444.

12. The compound of claim 11 wherein X³ is inserted at Leu₁₃₉/Thr₁₄₀.

13. The compound of claim 9 wherein the IgG1 Fc domain comprises SEQ ID NO:

347 and X³ is inserted into or replaces all or part of a sequence selected from SEQ ID NOS;

428, 429, 431, 432, 434, 435, 437, 439, 441, and 443.

14. The compound of claim 13 wherein X³ is inserted at H₅₃/E₅₄, Y₈₁/N₈₂, N₁₁₀/K₁₁₁, L₁₄₃/T₁₄₄, Q₁₇₁/P₁₇₂, E₁₇₃/N₁₇₄, S₁₈₆/D₁₈₇, G₁₈₈/S₁₈₉, or G₂₀₅/N₂₀₆.

15. The compound of claim 9 wherein the IgG1 Fc domain comprises SEQ ID NO:

348 and X³ is inserted into or replaces all or part of a sequence selected from SEQ ID NOS;

428, 429, 431, 432, 434, 435, 439, 441, 443, and 451.

16. The compound of claim 15 wherein X³ is inserted at H₅₃/E₅₄, Y₈₁/N₈₂, N₁₁₀/K₁₁₁, L₁₄₃/T₁₄₄, Q₁₇₁/P₁₇₂, E₁₇₃/N₁₇₄, S₁₈₆/D₁₈₇, G₁₈₈/S₁₈₉, or G₂₀₅/N₂₀₆.

17. The compound of claim 5 wherein the Fc domain comprises an IgG3 Fc domain.

18. The compound of claim 17 wherein the IgG3 Fc domain comprises SEQ ID NO:

349 and X³ is inserted into or replaces all or part of a sequence selected from SEQ ID NOS;

426, 428, 429, 431, 434, 446, 448, 451, 452, and 453.

19. The compound of claim 18 wherein X³ is inserted at H₁₀₀/E₁₀₁, F₁₂₈/N₁₂₉, N₁₅₇/K₁₅₈, M₁₉₀/T₁₉₁, Q₂₁₈/P₂₁₉, E₂₂₀/N₂₂₁, S₂₃₂/D₂₃₃, G₂₃₄/S₂₃₅, or G₂₅₂/N₂₅₃.

20. The compound of claim 5 wherein the Fc domain comprises an IgG2 Fc domain.

21. The compound of claim 20 wherein the Fc domain comprises SEQ ID NO:

350 and X³ is inserted into or replaces all or part of a sequence selected from SEQ ID NOS;

428, 429, 431, 439, 443, 446, 447, 449, 451, and 453.

22. The compound of claim 21 wherein X³ is inserted at H₄₉/E₅₀, F₇₇/N₇₈, N₁₀₆/K₁₀₇, M₁₃₉/T₁₄₀, Q₁₆₇/P₁₆₈, E₁₆₉/N₁₇₀, S₁₈₁/D₁₈₂, G₁₈₃/S₁₈₄, or G₂₀₁/N₂₀₂.

23. The compound of claim 5 wherein the Fc domain comprises an IgG4 Fc domain.

24. The compound of claim 23 wherein the Fc domain comprises SEQ ID NO:

351 and X³ is inserted into or replaces all or part of a sequence selected from SEQ ID NOS;

427, 428, 431, 434, 439, 441, 445, 446, 450, and 451.

25. The compound of claim 24 wherein X³ is inserted at Q₅₀/E₅₁, F₇₈/N₇₉, N₁₀₇/K₁₀₈, M₁₄₀/T₁₄₁, Q₁₆₈/P₁₆₉, E₁₇₀/N₁₇₁, S₁₈₂/D₁₈₃, G₁₈₄/S₁₈₅, or G₂₀₂/N₂₀₃.

26. The compound of claim 5 wherein the Fc domain comprises SEQ ID NO:

352 and X³ is inserted into or replaces all or part of a sequence selected from SEQ ID NOS;

428, 429, 435, 431, 434, 439, 443, 446, 451, and 453.

27. The compound of claim 26 wherein X³ is inserted at H₁₁₂/E₁₁₃, F₁₄₀/N₁₄₁, N₁₆₉/K₁₇₀, M₂₀₄/T₂₀₅, Q₂₃₂/P₂₃₃, E₂₃₄/N₂₃₅, S₂₄₆/D₂₄₇, G₂₄₈/S₂₄₉, or G₂₆₈/N₂₆₉.

28. The compound of any of claims 1 and 4, wherein at least two of Y¹-Y⁷ corresponds to two of X₁-X₇, respectively.

29. The compound of any of claims 1 and 4, wherein at least three of Y¹-Y⁷ corresponds to three of X₁-X₇, respectively.

30. The compound of any of claims 1 and 4, wherein at least four of Y¹-Y⁷ corresponds to four of X₁-X₇, respectively.

31. The compound of any of claims 1 and 4, wherein at least five of Y¹-Y⁷ corresponds to five of X₁-X₇, respectively.

32. The compound of any of claims 1 and 4, wherein at least six of Y¹-Y⁷ corresponds to six of X₁-X₇, respectively.

33. The compound of any of claims 1 and 4, wherein only one of Y¹-Y⁷ corresponds to one of X₁-X₇, respectively.

34. The compound of any of claims 1 and 4, wherein P is independently selected from the group consisting of:

35. The compound of any of claims 1 and 4 which is derivatized as set forth in one or more of the following:

one or more of the peptidyl [—

C(O)NR—

] linkages (bonds) have been replaced by a non-peptidyl linkage such as a —

CH₂-carbamate linkage [—

CH₂—

OC(O)NR—

];

a phosphonate linkage;

a —

CH₂-sulfonamide [—

CH₂—

S(O)₂NR—

] linkage;

a urea [—

NHC(O)NH—

] linkage;

a —

CH₂-secondary amine linkage;

or an alkylated peptidyl linkage [—

C(O)NR⁶—

where R⁶ is lower alkyl];

the N-terminus is a —

NRR¹ group;

to a —

NRC(O)R group;

to a —

NRC(O)OR group;

to a —

NRS(O)₂R group;

to a —

NHC(O)NHR group where R and R¹ are hydrogen and lower alkyl with the proviso that R and R¹ are not both hydrogen;

to a succinimide group;

to a benzyloxycarbonyl-NH—

(CBZ-NH—

) group;

or to a benzyloxycarbonyl-NH—

group having from 1 to 3 substituents on the phenyl ring selected from the group consisting of lower alkyl, lower alkoxy, chloro, and bromo; and

the C terminus is —

C(O)R² where R² is selected from the group consisting of lower alkoxy and —

NR³R⁴ where R³ and R⁴ are independently selected from the group consisting of hydrogen and lower alkyl.

36. The compound of any of claims 1 and 4, wherein all of the amino acids have a D configuration.

37. The compound of any of claims 1 and 4, wherein at least one of the amino acids has a D configuration.

38. The compound of any of claims 1 and 4, wherein P is constrained through a disulfide bond between cysteine residues such that P is cyclic.

39. The compound of any of claims 1 and 4, wherein P comprises four glycine residues at its N-terminus.

40. The compound of any of claims 1 and 4, wherein P comprises two glycine residues at its N-terminus of P and two glycine residues at its C-terminus.

41. The compound of any of claims 1 and 4, wherein L is a linker group which is optional and, if present, is independently selected from the linker groups consisting of

Y_n, wherein Y is a naturally-occurring amino acid or a stereoisomer thereof and n is 1 through 20;

(Gly)_n, wherein n is 1 through 20, and when n is greater than 1, up to half of the Gly residues may be substituted by another amino acid selected from the remaining 19 natural amino acids or a stereoisomer thereof;

(Gly)₃Lys(Gly)₄ (SEQ ID NO;

     4);

(Gly)₃AsnGlySer(Gly)₂ (SEQ ID NO;

     5);

(Gly)₃Cys(Gly)₄ (SEQ ID NO;

     6);

GlyProAsnGly (SEQ ID NO;

     7);

a Cys residue; and

(CH₂)_n, wherein n is 1 through 20.

42. The compound of claim 41, wherein L is selected from the group consisting of Y_n, wherein Y is selected a naturally-occurring amino acid or a stereoisomer thereof and n is 1 through 20.

43. The compound of claim 41, wherein L comprises (Gly)_n, wherein n is 1 through 20, and when n is greater than 1, up to half of the Gly residues may be substituted by another amino acid selected from the remaining 19 natural amino acids or a stereoisomer thereof.

44. The compound of claim 41, wherein L is selected from the group consisting of

45. The compound of claim 41, wherein L comprises a Cys residue.

46. The compound of claim 41, wherein L comprises (CH₂)_n, wherein n is 1 through 20.

47. The compound of claim 1 which is selected from the group consisting of SEQ ID NOS:

353-422.

48. A method of increasing megakaryocytes or platelets in a patient in need thereof, which comprises administering to said patient an effective amount of a compound of any of claims 1 and 4.

49. The method of claim 48, wherein said amount is from 1 μ

g/kg to 100 mg/kg.

50. A pharmaceutical composition comprising a compound of any of claims 1 and 4 in admixture with a pharmaceutically acceptable carrier thereof.

51. A polynucleotide that encodes a compound of any of claims 1 and 4.

52. A vector that comprises the polynucleotide of claim 50.

53. A host cell that comprises the vector of claim 51.

54. A method of producing a compound that binds to an mp1 receptor which comprises growing the host cell of claim 53 in a suitable nutrient medium and isolating said compound from said cell or nutrient medium.

4. A compound that binds to an mp1 receptor consisting essentially of a structure set out in Formula I,

[(X¹)_a—

(F¹)_z—

(X²)_b]-(L¹)_c-WSP_d 

 

Formula I

and multimers thereof, wherein;

F¹ is a vehicle;

X¹ is independently selected from;

P¹-(L²)_e-

P²-(L³)_f-P¹-(L²)_e-

P³-(L⁴)_g-P²-(L³)_f-P¹-(L²)_e- and

P⁴ (L⁵)_h-P³-(L⁴)_g-P²-(L³)_f-P¹-(L²)_e-

X² is independently selected from;

-(L²)_e-P¹,

-(L²)_e-P¹-(L³)_f-P²,

-(L²)_e-P¹-(L³)_f-P²-(L⁴)_g-P³, and

-(L²)_e-P¹-(L³)_f-P²-(L⁴)_g-P³-(L⁵)_h-P⁴

wherein P¹, P², P³, and P⁴ are each independently sequences of pharmacologically active peptides;

L¹, L², L³, L⁴, and L⁵ are each independently linkers;

a, b, c, d, e, f, g, and h are each independently 0 or 1;

z is 0, 1, 2, or more; and

WSP is a water soluble polymer, the attachment of which is effected at any reactive moiety in F¹;

wherein P is a molecule comprising the structure U¹-Y¹-Y²-Y³-Y⁴-Y⁵-Y⁶-Y⁷-U² wherein

U¹-Y¹(Cys, Leu, Met, Pro, Gln, Val, or X₁)-Y²(Phe, Lys, Leu, Asn, Gln, Arg, Ser, Thr, Val, or X₂)-Y³(Cys, Phe, Ile, Leu, Met, Arg, Ser, Val, Trp, or X₃)-Y⁴-Y⁵(Ala, Asp, Glu, Gly, Lys, Met, Gln, Arg, Ser, Thr, Val, Tyr, or X₅)-Y⁶(Cys, Phe, Gly, Leu, Met, Ser, Val, Trp, Tyr, or X₆)-Y⁷(Cys, Gly, Ile, Lys, Leu, Met, Asn, Arg, Val, or X₇)-U²,

wherein at least one of Y¹-Y³ and Y⁵-Y⁷ corresponds to a respective X₁-X₃ and X₅-X₇;

wherein U¹ or U² is any amino acid or peptide;

wherein when Y¹ is not an amino acid selected from the group consisting of Cys, Leu, Met, Pro, Gln, and Val, then X₁ is selected from the group consisting of Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Asn, Arg, Ser, Thr, Trp, and Tyr;

wherein when Y² is not an amino acid selected from the group consisting of Phe, Lys, Leu, Asn, Gln, Arg, Ser, Thr, and Val, then X₂ is selected from the group consisting of Ala, Cys, Asp, Glu, Gly, His, Ile, Met, Pro, Trp, and Tyr;

wherein when Y³ is not an amino acid selected from the group consisting of Cys, Phe, Ile, Leu, Met, Arg, Ser, Val, and Trp, then X₃ is selected from the group consisting of Ala, Asp, Glu, Gly, His, Lys, Asn, Pro, Gln, Thr, and Tyr;

wherein when Y⁴ is any amino acid;

wherein when Y⁵ is not an amino acid selected from the group consisting of Ala, Asp, Glu, Gly, Lys, Met, Gln, Arg, Ser, Thr, Val, and Tyr, then X₅ is selected from the group consisting of Cys, Phe, His, Ile, Leu, Asn, Pro, and Trp;

wherein when Y⁶ is not an amino acid selected from the group consisting of Cys, Phe, Gly, Leu, Met, Ser, Val, Trp, and Tyr, then X₆ is selected from the group consisting of Ala, Asp, Glu, His, Ile, Lys, Asn, Pro, Gln, Arg, and Thr; and

wherein when Y⁷ is not an amino acid selected from the group consisting of Cys, Gly, Ile, Lys, Leu, Met, Asn, Arg, and Val, then X₇ is selected from the group consisting of Ala, Asp, Glu, Phe, His, Pro, Gln, Ser, Thr, Trp, and Tyr;

and physiologically acceptable salts thereof.