Thrombopoietic compounds

• US 20050080014A1
• Filed: 09/02/2004
• Published: 04/14/2005
• Est. Priority Date: 10/23/1998
• Status: Active Grant

• Current Assignee: Amgen Inc.
• Original Assignee: Amgen Inc.
• Inventors: Liu, Chuan-Fa, Feige, Ulrich, Cheetham, Janet C.
• Granted Patent: US 8,044,174 B2
• Time in Patent Office: Days
• US Class Current: 530/324
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 7/00 Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: A61P 7/04 Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents
• CPC Class Codes: A61P 7/06 Antianaemics
• CPC Class Codes: C07K 14/524 Thrombopoietin, i.e. C-MPL ligand
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• 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 2319/75 containing a fusion for activation of a cell surface receptor, e.g. thrombopoeitin, NPY and other peptide hormones
• 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

1-34. -34. (canceled)

35. A compound that binds to an mpl receptor comprising the structure

^TMP_1-(L₁)_n-TMP 2

wherein TMP₁ and TMP₂ are each independently selected from the group of core compounds comprising the structure selected from the group consisting of:

a.) X_2-X_3-X_4-X_5-X_6-X_7-X_8-X_9-X₁₀,

wherein,

X₂ is selected from the group consisting of Glu, Lys, and Val;

X₃ is selected from the group consisting of Gly and Ala;

X₄ is Pro;

X₅ is selected from the group consisting of Thr and Ser;

X₆ is selected from the group consisting of Leu, Ile, Val, Ala, and Phe;

X₇ is selected from the group consisting of Arg and Lys;

X₈ is selected from the group consisting of Gln, Asn, and Glu;

X₉ is selected from the group consisting of Trp, Tyr, Cys, Ala, and Phe;

X₁₀ is selected from the group consisting of Leu, Ile, Val, Ala, Phe, Met, and Lys;

b.) X_2-X_3-X_4-X_5-X_6-X_7-X_8-X_9-X₁₀,

wherein,

X₂ is selected from the group consisting of Glu, Asp, Lys, and Val;

X₃ is Ala;

X₄ is Pro;

X₅ is selected from the group consisting of Thr and Ser;

X₆ is selected from the group consisting of Leu, Ile, Val, Ala, and Phe;

X₇ is selected from the group consisting of Arg and Lys;

X₈ is selected from the group consisting of Gln, Asn, and Glu;

X₉ is selected from the group consisting of Trp, Tyr, Cys, Ala, and Phe;

X₁₀ is selected from the group consisting of Leu, Ile, Val, Ala, Phe, Met, and Lys;

c.) X_2-X_3-X_4-X_5-X_6-X_7-X_8-X_9-X₁₀,

wherein,

X₂ is selected from the group consisting of Glu, Asp, Lys, and Val;

X₃ is selected from the group consisting of Gly and Ala;

X₄ is Pro;

X₅ is Ser;

X₆ is selected from the group consisting of Leu, Ile, Val, Ala, and Phe;

X₇ is selected from the group consisting of Arg and Lys;

X₈ is selected from the group consisting of Gln, Asn, and Glu;

X₉ is selected from the group consisting of Trp, Tyr, Cys, Ala, and Phe;

X₁₀ is selected from the group consisting of Leu, Ile, Val, Ala, Phe, Met, and Lys;

d.) X_2-X_3-X_4-X_5-X_6-X_7-X_8-X_9-X₁₀,

wherein,

X₂ is selected from the group consisting of Glu, Asp, Lys, and Val;

X₃ is selected from the group consisting of Gly and Ala;

X₄ is Pro;

X₅ is selected from the group consisting of Thr and Ser;

X₆ is selected from the group consisting of Ile, Val, Ala, and Phe;

X₇ is selected from the group consisting of Arg and Lys;

X₈ is selected from the group consisting of Gln, Asn, and Glu;

X₉ is selected from the group consisting of Trp, Tyr, Cys, Ala, and Phe;

X₁₀ is selected from the group consisting of Leu, Ile, Val, Ala, Phe, Met, and Lys;

e.) X_2-X_3-X_4-X_5-X_6-X_7-X_8-X_9-X₁₀,

wherein,

X₂ is selected from the group consisting of Glu, Asp, Lys, and Val;

X₃ is selected from the group consisting of Gly and Ala;

X₄ is Pro;

X₅ is selected from the group consisting of Thr and Ser;

X₆ is selected from the group consisting of Leu, Ile, Val, Ala, and Phe;

X₇ is Lys;

X₈ is selected from the group consisting of Gln, Asn, and Glu;

X₉ is selected from the group consisting of Trp, Tyr, Cys, Ala, and Phe;

X₁₀ is selected from the group consisting of Leu, Ile, Val, Ala, Phe, Met, and Lys;

f.) X_2-X_3-X_4-X_5-X_6-X_7-X_8-X_9-X₁₀,

wherein,

X₂ is selected from the group consisting of Glu, Asp, Lys, and Val;

X₃ is selected from the group consisting of Gly and Ala;

X₄ is Pro;

X₅ is selected from the group consisting of Thr and Ser;

X₆ is selected from the group consisting of Leu, Ile, Val, Ala, and Phe;

X₇ is selected from the group consisting of Arg and Lys;

X₈ is selected from the group consisting of Gln and Asn;

X₉ is selected from the group consisting of Trp, Tyr, Cys, Ala, and Phe;

X₁₀ is selected from the group consisting of Leu, Ile, Val, Ala, Phe, Met, and Lys;

g.) X_2-X_3-X_4-X_5-X_6-X_7-X_8-X_9-X₁₀,

wherein,

X₂ is selected from the group consisting of Glu, Asp, Lys, and Val;

X₃ is selected from the group consisting of Gly and Ala;

X₄ is Pro;

X₅ is selected from the group consisting of Thr and Ser;

X₆ is selected from the group consisting of Leu, Ile, Val, Ala, and Phe;

X₇ is selected from the group consisting of Arg and Lys;

X₈ is selected from the group consisting of Gln, Asn, and Glu;

X₉ is selected from the group consisting of Tyr, Cys, Ala, and Phe;

X₁₀ is selected from the group consisting of Leu, Ile, Val, Ala, Phe, Met, and Lys; and

h.) X_2-X_3-X_4-X_5-X_6-X_7-X_8-X_9-X₁₀,

wherein,

X₂ is selected from the group consisting of Glu, Asp, Lys, and Val;

X₃ is selected from the group consisting of Gly and Ala;

X₄ is Pro;

X₅ is selected from the group consisting of Thr and Ser;

X₆ is selected from the group consisting of Leu, Ile, Val, Ala, and Phe;

X₇ is selected from the group consisting of Arg and Lys;

X₈ is selected from the group consisting of Gln, Asn, and Glu;

X₉ is selected from the group consisting of Trp, Tyr, Cys, Ala, and Phe;

X₁₀ is selected from the group consisting of Leu, Val, Ala, Phe, Met, and Lys; and

wherein L₁ is a linker; and

n is 0 or 1;

and physiologically acceptable salts thereof.

43. The compound according to claim 35 wherein said TMP₁ and TMP₂ are independently selected from the group consisting of:

X_2-X_4-X_5-X_6-X_7-X_8-X_9-X_10-X₁₁;

X_2-X_3-X_4-X_5-X_6-X_7-X_8-X_9-X_10-X_11-X₁₂;

X_2-X_3-X_4-X_5-X_6-X_7-X_8-X_9-X_10-X_11-X_12-X₁₃;

X_2-X_3-X_4-X_5-X_6-X_7-X_8-X_9-X_10-X_11-X_12-X_13-X₁₄;

X_1-X_2-X_3-X_4-X_5-X_6-X_7-X_8-X_9-X₁₀;

X_1-X_2-X_3-X_4-X_5-X_6-X_7-X_8-X_9-X_10-X₁₁;

X_1-X_2-X_3-X_4-X_5-X_6-X_7-X_8-X_9-X_10-X_11-X₁₂;

X_1-X_2-X_3-X_4-X_5-X_6-X_7-X_8-X_9-X_10-X_11-X_12-X₁₃; and

X_1-X_2-X_3-X_4-X_5-X_6-X_7-X_8-X_9-X_10-X_11-X_12-X_13-X₁₄,

wherein X₂-X₁₀ are as defined and X₁ and X₁₁-X₁₄ are selected from the group consisting of;

a.) X₁ is selected from the group consisting of Ile, Val, Leu, Ser, and Arg;

X₁₁ is selected from the group consisting of Ala, Ile, Val, Leu, Phe, Ser, Thr, Lys, His, and Glu;

X₁₂ is selected from the group consisting of Ala, Ile, Val, Leu, Phe, Gly, Ser, and Gln;

X₁₃ is selected from the group consisting of Arg, Lys, Thr, Val, Asn, Gln, and Gly; and

X₁₄ is selected from the group consisting of Ala, Ile, Val, Leu, Phe, Thr, Arg, Glu, and Gly;

b.) X₁ is selected from the group consisting of Ile, Ala, Val, Leu, Ser, and Arg;

X₁₁ is selected from the group consisting of Ala, Ile, Val, Leu, Phe, Thr, Lys, His, and Glu;

X₁₂ is selected from the group consisting of Ala, Ile, Val, Leu, Phe, Gly, Ser, and Gln;

X₁₃ is selected from the group consisting of Arg, Lys, Thr, Val, Asn, Gln, and Gly; and

X₁₄ is selected from the group consisting of Ala, Ile, Val, Leu, Phe, Thr, Arg, Glu, and Gly; and

c.) X₁ is selected from the group consisting of Ile, Ala, Val, Leu, Ser, and Arg;

X₁₁ is selected from the group consisting of Ala, Ile, Val, Leu, Phe, Ser, Thr, Lys, His, and Glu;

X₁₂ is selected from the group consisting of Ala, Ile, Val, Leu, Gly, Ser, and Gln;

X₁₃ is selected from the group consisting of Arg, Lys, Thr, Val, Asn, Gln, and Gly; and

X₁₄ is selected from the group consisting of Ala, Ile, Val, Leu, Phe, Thr, Arg, Glu, and Gly.

46. The compound according to claim 35 wherein said TMP₁ and/or TMP₂ are 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 selected from the group consisting of;

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 —

CH_2—

secondary amine linkage; and

an alkylated peptidyl linkage [—

C(O)NR^6- here 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;

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.

47. The compound according to claim 35 wherein all of the amino acids have a D configuration.

48. The compound according to claim 35 wherein at least one of the amino acids has a D configuration.

49. The compound according to claim 35 which is cyclic.

50. The compound according to claim 35 wherein L₁ comprises a peptide.

51. The compound according to claim 50 wherein L₁ comprises Y_n, wherein Y is a naturally-occurring amino acid or a stereoisomer thereof and n is 1 through 20.

52. The compound according to claim 50 wherein L₁ 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.

53. The compound according to claim 50 wherein L₁ is selected from the group consisting of

54. The compound according to claim 50 wherein L₁ comprises a Cys residue.

55. A dimer of the compound according to claim 54.

56. The dimer according to claim 55 which is

57. The compound according to claim 35 wherein L₁ comprises (CH₂)_n, wherein n is 1 through 20.

58. The compound according to claim 35 or 43, which has the formula

(Fc)_m-(L₂)_q-TMP_1-(L₁)_n-TMP_2-(L₃)_r-(Fc)_p

wherein L₁, L₂ and L₃ are linker groups which are each 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;

     6) (Gly)₃AsnGlySer(Gly)₂;

(SEQ ID NO;

     7) (Gly)₃Cys(Gly)₄;

(SEQ ID NO;

     8) GlyProAsnGly;

(SEQ ID NO;

     9)

a Cys residue; and

(CH₂)_n, wherein n is 1 through 20

Fc is an Fc region of an immunoglobulin;

m, p, q and r are each independently selected from the group consisting of 0 and 1, wherein at least one of m or p is 1, and further wherein if m is 0 then q is 0, and if p is 0, then r is 0; and

physiologically acceptable salts thereof.

59. The compound according to claim 58 wherein L₁, L₂ and L₃ are each independently 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.

60. The compound according to claim 59 wherein L₁ 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.

61. The compound according to claim 59 wherein L₁, L₂ and L₃ are independently selected from the group consisting of

62. The compound according to claim 59 wherein L₁, L₂, or L₃ comprises a Cys residue.

63. A dimer of the compound according to claim 62.

64. The compound according to claim 59 wherein L₁, L₂ or L₃ comprises (CH₂)_n, wherein n is 1 through 20.

65. 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 according to claim 35.

66. The method according to claim 65, wherein said amount is from 1 μ

g/kg to 100 mg/kg.

67. A pharmaceutical composition comprising a compound according to claim 35 in admixture with a pharmaceutically acceptable carrier thereof.

68. A polynucleotide that encodes a compound according to claim 50.

69. A polynucleotide that encodes a compound according to claim 55.

70. A polynucleotide that encodes a compound according to claim 59.

71. A polynucleotide that encodes a compound according to claim 63.

72. A vector that comprises a polynucleotide according to claim 68.

75. A vector that comprises a polynucleotide according to claim 69.

76. A vector that comprises a polynucleotide according to claim 70.

77. A vector that comprises a polynucleotide according to claim 71.

78. A host cell that comprises a vector according to claim 72.

79. A host cell that comprises a vector according to claim 75.

80. A host cell that comprises a vector according to claim 76.

81. A host cell that comprises a vector according to claim 77.

82. A method of producing a compound which comprises growing a host cell according to claim 78 in a suitable nutrient medium and isolating said compound from said cell or nutrient medium.

83. A method of producing a compound which comprises growing a host cell according to claim 79 in a suitable nutrient medium and isolating said compound from said cell or nutrient medium.

84. A method of producing a compound which comprises growing a host cell according to claim 80 in a suitable nutrient medium and isolating said compound from said cell or nutrient medium.

85. A method of producing a compound which comprises growing a host cell according to claim 81 in a suitable nutrient medium and isolating said compound from said cell or nutrient medium.

36-42. -42. (canceled)

44-45. -45. (canceled)

73-74. -74. (canceled)