Protein kinases

• US 20060121475A1
• Filed: 09/05/2003
• Published: 06/08/2006
• Est. Priority Date: 09/06/2002
• Status: Abandoned Application

• Current Assignee: Ares Trading SA (Merck & Co., Inc.)
• Original Assignee: Ares Trading SA (Merck & Co., Inc.)
• Inventors: Yorke, Melanie, Fagan, Richard Joseph, Power, Christine, Davids, Andrew Robert, Phelps, Christopher Benjamin
• Application Number:
  US10/526,722
• Publication Number:
  US 20060121475A1
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A01K 2217/075 inducing loss of function, i.e. knock out
• CPC Class Codes: C12N 9/1205 Phosphotransferases with an alcohol group as acceptor (2.7.1), e.g. protein kinases

Abstract

The present invention relates to novel proteins, termed INSP081, INSP082, and INSP091, herein identified as members of the Germinal Center Kinase (GCK) subfamily of the STE20 family of protein kinases, preferably as NCK-interacting kinases (NIKs) and more preferably as NIK-like embryo specific kinases (NESKs), and to the use of these and nucleic acid sequences from the encoding genes in the diagnosis, prevention, and treatment of disease.

Claims

1-54. -54. (canceled)

55. A composition of matter:

(a) comprising an amino acid sequence as recited in SEQ ID NO;

2, SEQ ID NO;

4, SEQ ID NO;

6, SEQ ID NO;

8, SEQ ID NO;

10, SEQ ID NO;

12, SEQ ID NO;

14, SEQ ID NO;

16, SEQ ID NO;

18, SEQ ID NO;

20, SEQ ID NO;

22, SEQ ID NO;

24, SEQ ID NO;

26, SEQ ID NO;

28, SEQ ID NO;

30, SEQ ID NO;

32, SEQ ID NO;

34, SEQ ID NO;

36, SEQ ID NO;

38, SEQ ID NO;

40, SEQ ID NO;

42, SEQ ID NO;

44, SEQ ID NO;

46, SEQ ID NO;

48, SEQ ID NO;

50, SEQ ID NO;

52, SEQ ID NO;

54, SEQ ID NO;

56, SEQ ID NO;

58, SEQ ID NO;

60, SEQ ID NO;

62, SEQ ID NO;

64, SEQ ID NO;

66, SEQ ID NO;

68, SEQ ID NO;

70, SEQ ID NO;

72, SEQ ID NO;

74, SEQ ID NO;

76, SEQ ID NO;

78, SEQ ID NO;

80, SEQ ID NO;

82, SEQ ID NO;

84, SEQ ID NO;

86, SEQ ID NO;

88, SEQ ID NO;

90, SEQ ID NO;

92, SEQ ID NO;

94, SEQ ID NO;

96, SEQ ID NO;

98, SEQ ID NO;

100, SEQ ID NO;

102, SEQ ID NO;

104, SEQ ID NO;

106, SEQ ID NO;

108, SEQ ID NO;

110, SEQ ID NO;

112, SEQ ID NO;

114, SEQ ID NO;

116, SEQ ID NO;

118, SEQ ID NO;

120, SEQ ID NO;

122, SEQ ID NO;

124, SEQ ID NO;

126, SEQ ID NO;

128, SEQ ID NO;

130, SEQ ID NO;

132, SEQ ID NO;

134, SEQ ID NO;

136, SEQ ID NO;

138, SEQ ID NO;

140, SEQ ID NO;

142, SEQ ID NO;

144, SEQ ID NO;

146, SEQ ID NO;

148, SEQ ID NO;

150, SEQ ID NO;

152, SEQ ID NO;

154, SEQ ID NO;

156 or SEQ ID NO;

158;

(b) consisting of an amino acid sequence as recited in SEQ ID NO;

2, SEQ ID NO;

4, SEQ ID NO;

6, SEQ ID NO;

8, SEQ ID NO;

10, SEQ ID NO;

12, SEQ ID NO;

14, SEQ ID NO;

16, SEQ ID NO;

18, SEQ ID NO;

20, SEQ ID NO;

22, SEQ ID NO;

24, SEQ ID NO;

26, SEQ ID NO;

28, SEQ ID NO;

30, SEQ ID NO;

32, SEQ ID NO;

34, SEQ ID NO;

36, SEQ ID NO;

38, SEQ ID NO;

40, SEQ ID NO;

42, SEQ ID NO;

44, SEQ ID NO;

46, SEQ ID NO;

48, SEQ ID NO;

50, SEQ ID NO;

52, SEQ ID NO;

54, SEQ ID NO;

56, SEQ ID NO;

58, SEQ ID NO;

60, SEQ ID NO;

62, SEQ ID NO;

64, SEQ ID NO;

66, SEQ ID NO;

68, SEQ ID NO;

70, SEQ ID NO;

72, SEQ ID NO;

74, SEQ ID NO;

76, SEQ ID NO;

78, SEQ ID NO;

80, SEQ ID NO;

82, SEQ ID NO;

84, SEQ ID NO;

86, SEQ ID NO;

88, SEQ ID NO;

90, SEQ ID NO;

92, SEQ ID NO;

94, SEQ ID NO;

96, SEQ ID NO;

98, SEQ ID NO;

100, SEQ ID NO;

102, SEQ ID NO;

104, SEQ ID NO;

106, SEQ ID NO;

108, SEQ ID NO;

110, SEQ ID NO;

112, SEQ ID NO;

114, SEQ ID NO;

116, SEQ ID NO;

118, SEQ ID NO;

120, SEQ ID NO;

122, SEQ ID NO;

124, SEQ ID NO;

126, SEQ ID NO;

128, SEQ ID NO;

130, SEQ ID NO;

132, SEQ ID NO;

134, SEQ ID NO;

136, SEQ ID NO;

138, SEQ ID NO;

140, SEQ ID NO;

142, SEQ ID NO;

144, SEQ ID NO;

146, SEQ ID NO;

148, SEQ ID NO;

150, SEQ ID NO;

152, SEQ ID NO;

154, SEQ ID NO;

156 or SEQ ID NO;

158;

(c) that is a fragment of the amino acid sequence of (a) or (b) and which is a member of the Germinal Center Kinase (GCK) subfamily of the STE20 family of protein kinases, a NIK-like kinase, a NIK-like embryo specific kinase (NESK), or having an antigenic determinant in common with the polypeptide of (a) or (b);

(d) that is a functional equivalent of (a), (b) or (c);

(e) comprising an isolated polypeptide that is a functional equivalent of (a), (b) or (c) and is homologous to the amino acid sequence as recited in SEQ ID NO;

2, SEQ ID NO;

4, SEQ ID NO;

6, SEQ ID NO;

8, SEQ ID NO;

10, SEQ ID NO;

12, SEQ ID NO;

14, SEQ ID NO;

16, SEQ ID NO;

18, SEQ ID NO;

20, SEQ ID NO;

22, SEQ ID NO;

24, SEQ ID NO;

26, SEQ ID NO;

28, SEQ ID NO;

30, SEQ ID NO;

32, SEQ ID NO;

34, SEQ ID NO;

36, SEQ ID NO;

38, SEQ ID NO;

40, SEQ ID NO;

42, SEQ ID NO;

44, SEQ ID NO;

46, SEQ ID NO;

48, SEQ ID NO;

50, SEQ ID NO;

52, SEQ ID NO;

54, SEQ ID NO;

56, SEQ ID NO;

58, SEQ ID NO;

60, SEQ ID NO;

62, SEQ ID NO;

64, SEQ ID NO;

66, SEQ ID NO;

68, SEQ ID NO;

70, SEQ ID NO;

72, SEQ ID NO;

74, SEQ ID NO;

76, SEQ ID NO;

78, SEQ ID NO;

80, SEQ ID NO;

82, SEQ ID NO;

84, SEQ ID NO;

86, SEQ ID NO;

88, SEQ ID NO;

90, SEQ ID NO;

92, SEQ ID NO;

94, SEQ ID NO;

96, SEQ ID NO;

98, SEQ ID NO;

100, SEQ ID NO;

102, SEQ ID NO;

104, SEQ ID NO;

106, SEQ ID NO;

108, SEQ ID NO;

110, SEQ ID NO;

112, SEQ ID NO;

114, SEQ ID NO;

116, SEQ ID NO;

118, SEQ ID NO;

120, SEQ ID NO;

122, SEQ ID NO;

124, SEQ ID NO;

126, SEQ ID NO;

128, SEQ ID NO;

130, SEQ ID NO;

132, SEQ ID NO;

134, SEQ ID NO;

136, SEQ ID NO;

138, SEQ ID NO;

140, SEQ ID NO;

142, SEQ ID NO;

144, SEQ ID NO;

146, SEQ ID NO;

148, SEQ ID NO;

150, SEQ ID NO;

152, SEQ ID NO;

154, SEQ ID NO;

156 or SEQ ID NO;

158 and is a member of the Germinal Center Kinase (GCK) subfamily of the STE20 family of protein kinases;

(f) comprising an isolated polypeptide which is a fragment or functional equivalent of a polypeptide as set forth in (a), (b), (c), (d), or (e), and that has greater than 80% sequence identity with the amino acid sequence recited in SEQ ID NO;

2, SEQ ID NO;

4, SEQ ID NO;

6, SEQ ID NO;

8, SEQ ID NO;

10, SEQ ID NO;

12, SEQ ID NO;

14, SEQ ID NO;

16, SEQ ID NO;

18, SEQ ID NO;

20, SEQ ID NO;

22, SEQ ID NO;

24, SEQ ID NO;

26, SEQ ID NO;

28, SEQ ID NO;

30, SEQ ID NO;

32, SEQ ID NO;

34, SEQ ID NO;

36, SEQ ID NO;

38, SEQ ID NO;

40, SEQ ID NO;

42, SEQ ID NO;

44, SEQ ID NO;

46, SEQ ID NO;

48, SEQ ID NO;

50, SEQ ID NO;

52, SEQ ID NO;

54, SEQ ID NO;

56, SEQ ID NO;

58, SEQ ID NO;

60, SEQ ID NO;

62, SEQ ID NO;

64, SEQ ID NO;

66, SEQ ID NO;

68, SEQ ID NO;

70, SEQ ID NO;

72, SEQ ID NO;

74, SEQ ID NO;

76, SEQ ID NO;

78, SEQ ID NO;

80, SEQ ID NO;

82, SEQ ID NO;

84, SEQ ID NO;

86, SEQ ID NO;

88, SEQ ID NO;

90, SEQ ID NO;

92, SEQ ID NO;

94, SEQ ID NO;

96, SEQ ID NO;

98, SEQ ID NO;

100, SEQ ID NO;

102, SEQ ID NO;

104, SEQ ID NO;

106, SEQ ID NO;

108, SEQ ID NO;

110, SEQ ID NO;

112, SEQ ID NO;

114, SEQ ID NO;

116, SEQ ID NO;

118, SEQ ID NO;

120, SEQ ID NO;

122, SEQ ID NO;

124, SEQ ID NO;

126, SEQ ID NO;

128, SEQ ID NO;

130, SEQ ID NO;

132, SEQ ID NO;

134, SEQ ID NO;

136, SEQ ID NO;

138, SEQ ID NO;

140, SEQ ID NO;

142, SEQ ID NO;

144, SEQ ID NO;

146, SEQ ID NO;

148, SEQ ID NO;

150, SEQ ID NO;

152, SEQ ID NO;

154, SEQ ID NO;

156, SEQ ID NO;

158 or with an active fragment thereof;

(g) which is a polypeptide fragment, or functional equivalent thereof, and has an antigenic determinant in common with a polypeptide which consists of 7 or more amino acid residues from the amino acid sequence recited in SEQ ID NO;

2, SEQ ID NO;

4, SEQ ID NO;

6, SEQ ID NO;

8, SEQ ID NO;

10, SEQ ID NO;

12, SEQ ID NO;

14, SEQ ID NO;

16, SEQ ID NO;

18, SEQ ID NO;

20, SEQ ID NO;

22, SEQ ID NO;

24, SEQ ID NO;

26, SEQ ID NO;

28, SEQ ID NO;

30, SEQ ID NO;

32, SEQ ID NO;

34, SEQ ID NO;

36, SEQ ID NO;

38, SEQ ID NO;

40, SEQ ID NO;

42, SEQ ID NO;

44, SEQ ID NO;

46, SEQ ID NO;

48, SEQ ID NO;

50, SEQ ID NO;

52, SEQ ID NO;

54, SEQ ID NO;

56, SEQ ID NO;

58, SEQ ID NO;

60, SEQ ID NO;

62, SEQ ID NO;

64, SEQ ID NO;

66, SEQ ID NO;

68, SEQ ID NO;

70, SEQ ID NO;

72, SEQ ID NO;

74, SEQ ID NO;

76, SEQ ID NO;

78, SEQ ID NO;

80, SEQ ID NO;

82, SEQ ID NO;

84, SEQ ID NO;

86, SEQ ID NO;

88, SEQ ID NO;

90, SEQ ID NO;

92, SEQ ID NO;

94, SEQ ID NO;

96, SEQ ID NO;

98, SEQ ID NO;

100, SEQ ID NO;

102, SEQ ID NO;

104, SEQ ID NO;

106, SEQ ID NO;

108, SEQ ID NO;

110, SEQ ID NO;

112, SEQ ID NO;

114, SEQ ID NO;

116, SEQ ID NO;

118, SEQ ID NO;

120, SEQ ID NO;

122, SEQ ID NO;

124, SEQ ID NO;

126, SEQ ID NO;

128, SEQ ID NO;

130, SEQ ID NO;

132, SEQ ID NO;

134, SEQ ID NO;

136, SEQ ID NO;

138, SEQ ID NO;

140, SEQ ID NO;

142, SEQ ID NO;

144, SEQ ID NO;

146, SEQ ID NO;

148, SEQ ID NO;

150, SEQ ID NO;

152, SEQ ID NO;

154, SEQ ID NO;

156 or SEQ ID NO;

158;

(h) comprising a purified nucleic acid molecule which encodes a polypeptide according to (a), (b), (c), (d), (e), (f), or (g);

(i) comprising a purified nucleic acid sequence as recited in SEQ ID NO;

1, SEQ ID NO;

3, SEQ ID NO;

5, SEQ ID NO;

7, SEQ ID NO;

9, SEQ ID NO;

11, SEQ ID NO;

13, SEQ ID NO;

15, SEQ ID NO;

17, SEQ ID NO;

19, SEQ ID NO;

21, SEQ ID NO;

23, SEQ ID NO;

25, SEQ ID NO;

27, SEQ ID NO;

29, SEQ ID NO;

31, SEQ ID NO;

33, SEQ ID NO;

35, SEQ ID NO;

37, SEQ ID NO;

39, SEQ ID NO;

41, SEQ ID NO;

43, SEQ ID NO;

45, SEQ ID NO;

47 SEQ ID NO;

49, SEQ ID NO;

5 1, SEQ ID NO;

53, SEQ ID NO;

55, SEQ ID NO;

57, SEQ ID NO;

59, SEQ ID NO;

61, SEQ ID NO;

63, SEQ ID NO;

65, SEQ ID NO;

67, SEQ ID NO;

69, SEQ ID NO;

71, SEQ ID SEQ ID NO;

73, SEQ ID NO;

75, SEQ ID NO;

77, SEQ ID NO;

79, SEQ ID NO;

81, SEQ ID NO;

83, SEQ ID NO;

85, SEQ ID NO;

87, SEQ ID NO;

89, SEQ ID NO;

91, SEQ ID NO;

93, SEQ ID NO;

95, SEQ ID NO;

97, SEQ ID NO;

99, SEQ ID NO;

101, SEQ ID NO;

103, SEQ ID NO;

105, SEQ ID NO;

107, SEQ ID NO;

109, SEQ ID NO;

111, SEQ ID NO;

113, SEQ ID NO;

115, SEQ ID NO;

117, SEQ ID NO;

119, SEQ ID NO;

121, SEQ ID NO;

123, SEQ ID NO;

125, SEQ ID NO;

127, SEQ ID NO;

129, SEQ ID NO;

131, SEQ ID NO;

133, SEQ ID NO;

135, SEQ ID NO;

137, SEQ ID NO;

139, SEQ ID NO;

141, SEQ ID NO;

143, SEQ ID NO;

145, SEQ ID NO;

147, SEQ ID NO;

149, SEQ ID NO;

151, SEQ ID NO;

153, SEQ ID NO;

155, SEQ ID NO;

157 or is a redundant equivalent or fragment thereof;

(j) consisting of a purified nucleic acid sequence as recited in SEQ ID NO;

1, SEQ ID NO;

3, SEQ ID NO;

5, SEQ ID NO;

7, SEQ ID NO;

9, SEQ ID NO;

11, SEQ ID NO;

13, SEQ ID NO;

15, SEQ ID NO;

17, SEQ ID NO;

19, SEQ ID NO;

21, SEQ ID NO;

23, SEQ ID NO;

25, SEQ ID NO;

27, SEQ ID NO;

29, SEQ ID NO;

31, SEQ ID NO;

33, SEQ ID NO;

35, SEQ ID NO;

37, SEQ ID NO;

39, SEQ ID NO;

41, SEQ ID NO;

43, SEQ ID NO;

45, SEQ ID NO;

47 SEQ ID NO;

49, SEQ ID NO;

51, SEQ ID NO;

53, SEQ ID NO;

55, SEQ ID NO;

57, SEQ ID NO;

59, SEQ ID NO;

61, SEQ ID NO;

63, SEQ ID NO;

65, SEQ ID NO;

67, SEQ ID NO;

69, SEQ ID NO;

71, SEQ ID SEQ ID NO;

73, SEQ ID NO;

75, SEQ ID NO;

77, SEQ ID NO;

79, SEQ ID NO;

81, SEQ ID NO;

83, SEQ ID NO;

85, SEQ ID NO;

87, SEQ ID NO;

89, SEQ ID NO;

91, SEQ ID NO;

93, SEQ ID NO;

95, SEQ ID NO;

97, SEQ ID NO;

99, SEQ ID NO;

101, SEQ ID NO;

103, SEQ ID NO;

105, SEQ ID NO;

107, SEQ ID NO;

109, SEQ ID NO;

111, SEQ ID NO;

113, SEQ ID NO;

115, SEQ ID NO;

117, SEQ ID NO;

119, SEQ ID NO;

121, SEQ ID NO;

123, SEQ ID NO;

125, SEQ ID NO;

127, SEQ ID NO;

129, SEQ ID NO;

131, SEQ ID NO;

133, SEQ ID NO;

135, SEQ ID NO;

137, SEQ ID NO;

139, SEQ ID NO;

141, SEQ ID NO;

143, SEQ ID NO;

145, SEQ ID NO;

147, SEQ ID NO;

149, SEQ ID NO;

151, SEQ ID NO;

153, SEQ ID NO;

155, SEQ ID NO;

157 or is a redundant equivalent or fragment thereof;

(k) comprising a purified nucleic acid molecule which hybridizes under high stringency conditions with a nucleic acid molecule according to any one of (h), (i) or

     0);

(l) comprising a vector that comprises a nucleic acid molecule according to (h), (i),

     0) or (k);

(m) comprising a host cell transformed with a vector according to (1);

(n) comprising a ligand which binds specifically to the Germinal Center Kinase (GCK) subfamily of the STE20 family of protein kinases;

(o) comprising a ligand that binds specifically to a polypeptide according to (a), (b), (c), (d), (e), (f) or (g);

(p) comprising a compound that increases or decreases the level of expression or activity of a polypeptide according to (a), (b), (c), (d), (e), (f) or (g);

(q) comprising a compound that binds to a polypeptide according to (a), (b), (c), (d), (e), (f) or (g) without inducing any of the biological effects of the polypeptide;

(r) comprising a pharmaceutical composition comprising a pharmaceutically acceptable carrier and (i) a polypeptide according to (a), (b), (c), (d), (e), (f) or (g);

or (ii) a polynucleotide according to (h), (i), (j), (k) or (l);

(s) comprising a vaccine composition comprising (i) a polypeptide according to (a), (b), (c), (d), (e), (f) or (g);

or (ii) a polynucleotide according to (h), (i), (j), (k) or (l);

(t) comprising a kit comprising a first container containing a nucleic acid probe that hybridizes under stringent conditions with a nucleic acid molecule according to (h), (i), (j), (k);

a second container containing primers useful for amplifying said nucleic acid molecule; and

instructions for using the probe and primers for facilitating the diagnosis of disease;

(u) comprising the kit of (t), further comprising a third container holding an agent for digesting un-hybridized RNA;

(v) comprising a kit comprising an array of nucleic acid molecules, at least one of which is a nucleic acid molecule according (h), (i),

     0), (k);

(w) comprising a kit comprising one or more antibodies that bind to a polypeptide as recited in any one of (a), (b), (c), (d), (e), (f) or (g); and

a reagent useful for the detection of a binding reaction between said antibody and said polypeptide;

or

(x) comprising a transgenic or knockout non-human animal that has been transformed to express higher, lower or absent levels of a polypeptide according (a), (b), (c), (d), (e), (f) or (g).

56. The composition of matter according to claim 55(o), wherein said ligand is an antibody.

57. The composition of matter according to claim 55(q), wherein said compound is a natural or modified substrate, ligand, enzyme, receptor or structural or functional mimetic.

58. A method of diagnosing a disease in a patient comprising assessing the level of expression of a natural gene encoding a polypeptide or assessing the activity of a polypeptide according to claim 55 in tissue from said patient and comparing said level of expression or activity to a control level, wherein a level that is different to said control level is indicative of disease.

59. A method according to claim 58 that is carried out in vitro.

60. A method of using a composition of matter according to claim 55 for:

a) screening for a compound effective to treat disease, by contacting a non-human transgenic animal transformed to express higher, lower or absent levels of a polypeptide according to claim 55 with a candidate compound and determining the effect of the compound on the disease of the animal;

b) the identification of a compound that is effective in the treatment, diagnosis or both treatment and diagnosis of disease, comprising contacting a polypeptide or a polynucleotide according to claim 55 with one or more compounds suspected of possessing binding affinity for said polypeptide or nucleic acid molecule, and selecting a compound that binds specifically to said nucleic acid molecule or polypeptide;

c) for treating a disease in a patient comprising administering to the patient a polypeptide, a nucleic acid molecule, a vector, a host cell, a ligand, a compound or a pharmaceutical composition according to claim 55;

or

d) for monitoring the therapeutic treatment of disease in a patient, comprising monitoring over a period of time the level of expression or activity of a polypeptide or a polynucleotide according to claim 55 in tissue from said patient, wherein altering said level of expression or activity over the period of time towards a control level is indicative of regression of said disease.

61. The method according to claim 60, wherein, for diseases in which the expression of the natural gene or the activity of the polypeptide is lower in a diseased patient when compared to the level of expression or activity in a healthy patient, the polypeptide, nucleic acid molecule, vector, ligand, compound or composition administered to the patient is an agonist.

62. The method according to claim 60, wherein, for diseases in which the expression of the natural gene or activity of the polypeptide is higher in a diseased patient when compared to the level of expression or activity in a healthy patient, the polypeptide, nucleic acid molecule, vector, ligand, compound or composition administered to the patient is an antagonist.