Neublastin variants

• US 8,263,553 B2
• Filed: 08/18/2005
• Issued: 09/11/2012
• Est. Priority Date: 08/19/2004
• Status: Active Grant

• Current Assignee: Sinfonia Biotherapeutics AB
• Original Assignee: Biogen Idec MA Inc. (Biogen, Inc.)
• Inventors: Rossomando, Anthony, Silvian, Laura, Pepinsky, R. Blake
• Primary Examiner(s): Kolker, Daniel E
• Assistant Examiner(s): Emch, Gregory S
• Application Number:
  US11/573,773
• Publication Number:
  US 20080039385A1
• Time in Patent Office: 2,581 Days
• Field of Search: None
• US Class Current: 514/8.3
• 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: A61K 38/185 Nerve growth factor [NGF]; Brain derived neurotrophic factor [BDNF]; Ciliary neurotrophic factor [CNTF]; Glial derived neurotrophic factor [GDNF]; Neurotrophins, e.g. NT-3
• CPC Class Codes: A61K 47/60 the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/02 for peripheral neuropathies
• CPC Class Codes: A61P 25/04 Centrally acting analgesics, e.g. opioids
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 14/47 from mammals
• CPC Class Codes: C07K 14/4756 Neuregulins, i.e. p185erbB2 ligands, glial growth factor, heregulin, ARIA, neu differentiation factor
• CPC Class Codes: C07K 14/495 Transforming growth factor [TGF]
• CPC Class Codes: C07K 19/00 Hybrid peptides , i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

Compositions and methods for folding proteins belonging to the transforming growth factor beta superfamily are disclosed. The compositions and methods allow for the folding of such proteins when produced in an expression system that does not yield a properly folded, biologically active product.

Claims

1. A polypeptide comprising an amino acid sequence that is at least 80% identical to amino acids 15-113 of SEQ ID NO:

1, wherein the amino acid sequence comprises an amino acid other than arginine substituted at the positions corresponding to position 48 and position 49 of SEQ ID NO;

1, wherein the polypeptide, when dimerized, binds to a complex containing GFRalpha3 and RET.

2. The polypeptide of claim 1, wherein the arginine residue at the position corresponding to position 48 of SEQ ID NO:

1 is substituted with a non-conservative amino acid residue.

3. The polypeptide of claim 1, wherein the arginine residue at the position corresponding to position 48 of SEQ ID NO:

1 is substituted with glutamic acid.

4. The polypeptide of claim 1, wherein the arginine residue at the position corresponding to position 49 of SEQ ID NO:

1 is substituted with a non-conservative amino acid residue.

5. The polypeptide of claim 1, wherein the arginine residue at the position corresponding to position 49 of SEQ ID NO:

1 is substituted with glutamic acid.

6. The polypeptide of claim 1, wherein the arginine residue at the position corresponding to position 48 of SEQ ID NO:

1 and the arginine residue at the position corresponding to position 49 of SEQ ID NO;

1 are substituted with non-conservative amino acid residues.

7. The polypeptide of claim 1, wherein the arginine residue at the position corresponding to position 48 of SEQ ID NO:

1 and the arginine residue at the position corresponding to position 49 of SEQ ID NO;

1 are each substituted with glutamic acid.

8. The polypeptide of claim 1, wherein the amino acid sequence is at least 90% identical to amino acids 15-113 of SEQ ID NO:

1.

9. The polypeptide of claim 1, wherein the amino acid sequence is at least 95% identical to amino acids 15-113 of SEQ ID NO:

1.

13. The polypeptide of claim 6, wherein the amino acid sequence is at least 90% identical to amino acids 15-113 of SEQ ID NO:

1.

14. The polypeptide of claim 6, wherein the amino acid sequence is at least 95% identical to amino acids 15-113 of SEQ ID NO:

1.

15. The polypeptide of claim 7, wherein the amino acid sequence is at least 90% identical to amino acids 15-113 of SEQ ID NO:

1.

16. The polypeptide of claim 7, wherein the amino acid sequence is at least 95% identical to amino acids 15-113 of SEQ ID NO:

1.

20. A conjugate comprising the polypeptide of claim 1 conjugated to a non-naturally occurring polymer.

21. The conjugate of claim 20, wherein the non-naturally occurring polymer is a polyalkylene glycol.

22. The conjugate of claim 21, wherein the polyalkylene glycol is polyethylene glycol.

23. The conjugate of claim 22, wherein the polyethylene glycol is coupled to the polypeptide at the amino terminus.

24. The conjugate of claim 22, wherein the polyethylene glycol is coupled to the polypeptide at an internal polymer conjugation site.

25. A conjugate comprising the polypeptide of claim 8 conjugated to a non-naturally occurring polymer.

26. The conjugate of claim 25, wherein the non-naturally occurring polymer is a polyalkylene glycol.

27. The conjugate of claim 26, wherein the polyalkylene glycol is polyethylene glycol.

28. The conjugate of claim 27, wherein the polyethylene glycol is coupled to the polypeptide at the amino terminus.

29. The conjugate of claim 27, wherein the polyethylene glycol is coupled to the polypeptide at an internal polymer conjugation site.

30. A conjugate comprising the polypeptide of claim 9 conjugated to a non-naturally occurring polymer.

31. The conjugate of claim 30, wherein the non-naturally occurring polymer is a polyalkylene glycol.

32. The conjugate of claim 31, wherein the polyalkylene glycol is polyethylene glycol.

33. The conjugate of claim 32, wherein the polyethylene glycol is coupled to the polypeptide at the amino terminus.

34. The conjugate of claim 32, wherein the polyethylene glycol is coupled to the polypeptide at an internal polymer conjugation site.

55. A fusion protein comprising the polypeptide of claim 1 and a heterologous amino acid sequence.

56. A fusion protein comprising the polypeptide of claim 8 and a heterologous amino acid sequence.

57. A fusion protein comprising the polypeptide of claim 9 and a heterologous amino acid sequence.

62. A dimer comprising two polypeptides according to claim 1.

63. A dimer comprising two polypeptides according to claim 8.

64. A dimer comprising two polypeptides according to claim 9.

73. A pharmaceutical composition comprising the polypeptide of claim 1 and a pharmaceutically acceptable carrier or excipient.

74. A pharmaceutical composition comprising the polypeptide of claim 8 and a pharmaceutically acceptable carrier or excipient.

75. A pharmaceutical composition comprising the polypeptide of claim 9 and a pharmaceutically acceptable carrier or excipient.

90. A method of treating a peripheral neuropathy, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 73.

91. A method of treating neuropathic pain, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 73.

92. The method of claim 91, wherein the neuropathic pain is associated with post-herpetic neuralgia.

93. The method of claim 91, wherein the neuropathic pain is associated with sciatica.

94. A method of treating painful diabetic neuropathy, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 73.

95. A method of activating the RET receptor in a human, the method comprising administering to the human an effective amount of the pharmaceutical composition of claim 73.

96. A method of treating a peripheral neuropathy, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 74.

97. A method of treating neuropathic pain, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 74.

98. The method of claim 97, wherein the neuropathic pain is associated with post-herpetic neuralgia.

99. The method of claim 97, wherein the neuropathic pain is associated with sciatica.

100. A method of treating painful diabetic neuropathy, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 74.

101. A method of activating the RET receptor in a human, the method comprising administering to the human an effective amount of the pharmaceutical composition of claim 74.

102. A method of treating a peripheral neuropathy, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 75.

103. A method of treating neuropathic pain, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 75.

104. The method of claim 103, wherein the neuropathic pain is associated with post-herpetic neuralgia.

105. The method of claim 103, wherein the neuropathic pain is associated with sciatica.

106. A method of treating painful diabetic neuropathy, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 75.

107. A method of activating the RET receptor in a human, the method comprising administering to the human an effective amount of the pharmaceutical composition of claim 75.

192. A nucleic acid comprising a sequence that encodes the polypeptide of claim 1.

193. An expression vector comprising the nucleic acid of claim 192.

194. A cell comprising the expression vector of claim 193.

195. A method of making a polypeptide, the method comprising:

providing the cell of claim 194, and

culturing the cell under conditions that permit expression of the nucleic acid.

196. A nucleic acid comprising a sequence that encodes the polypeptide of claim 8.

197. An expression vector comprising the nucleic acid of claim 196.

198. A cell comprising the expression vector of claim 197.

199. A method of making a polypeptide, the method comprising:

providing the cell of claim 198, and

culturing the cell under conditions that permit expression of the nucleic acid.

200. A nucleic acid comprising a sequence that encodes the polypeptide of claim 9.

201. An expression vector comprising the nucleic acid of claim 200.

202. A cell comprising the expression vector of claim 201.

203. A method of making a polypeptide, the method comprising:

providing the cell of claim 202, and

culturing the cell under conditions that permit expression of the nucleic acid.

10. A polypeptide comprising amino acids 15-113 of SEQ ID NO:

5.

11. The polypeptide of claim 10, wherein the polypeptide comprises amino acids 10-113 of SEQ ID NO:

5.

12. The polypeptide of claim 10, wherein the polypeptide comprises the amino acid sequence of SEQ ID NO:

5.

35. A conjugate comprising the polypeptide of claim 10 conjugated to a non-naturally occurring polymer.

36. The conjugate of claim 35, wherein the non-naturally occurring polymer is a polyalkylene glycol.

37. The conjugate of claim 36, wherein the polyalkylene glycol is polyethylene glycol.

38. The conjugate of claim 37, wherein the polyethylene glycol is coupled to the polypeptide at the amino terminus.

39. The conjugate of claim 37, wherein the polyethylene glycol is coupled to the polypeptide at an internal polymer conjugation site.

40. A conjugate comprising the polypeptide of claim 11 conjugated to a non-naturally occurring polymer.

41. The conjugate of claim 40, wherein the non-naturally occurring polymer is a polyalkylene glycol.

42. The conjugate of claim 41, wherein the polyalkylene glycol is polyethylene glycol.

43. The conjugate of claim 42, wherein the polyethylene glycol is coupled to the polypeptide at the amino terminus.

44. The conjugate of claim 42, wherein the polyethylene glycol is coupled to the polypeptide at an internal polymer conjugation site.

58. A fusion protein comprising the polypeptide of claim 10 and a heterologous amino acid sequence.

59. A fusion protein comprising the polypeptide of claim 11 and a heterologous amino acid sequence.

65. A dimer comprising two polypeptides according to claim 10.

66. A dimer comprising two polypeptides according to claim 11.

69. A dimer comprising two conjugates according to claim 42.

70. A dimer comprising two conjugates according to claim 43.

76. A pharmaceutical composition comprising the polypeptide of claim 10 and a pharmaceutically acceptable carrier or excipient.

77. A pharmaceutical composition comprising the polypeptide of claim 11 and a pharmaceutically acceptable carrier or excipient.

80. A pharmaceutical composition comprising the conjugate of claim 42 and a pharmaceutically acceptable carrier or excipient.

81. A pharmaceutical composition comprising the conjugate of claim 43 and a pharmaceutically acceptable carrier or excipient.

84. A pharmaceutical composition comprising the dimer of claim 66 and a pharmaceutically acceptable carrier or excipient.

86. A pharmaceutical composition comprising the dimer of claim 69 and a pharmaceutically acceptable carrier or excipient.

87. A pharmaceutical composition comprising the dimer of claim 70 and a pharmaceutically acceptable carrier or excipient.

108. A method of treating a peripheral neuropathy, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 76.

109. A method of treating neuropathic pain, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 76.

110. The method of claim 109, wherein the neuropathic pain is associated with post-herpetic neuralgia.

111. The method of claim 109, wherein the neuropathic pain is associated with sciatica.

112. A method of treating painful diabetic neuropathy, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 76.

113. A method of activating the RET receptor in a human, the method comprising administering to the human an effective amount of the pharmaceutical composition of claim 76.

114. A method of treating a peripheral neuropathy, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 77.

115. A method of treating neuropathic pain, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 77.

116. The method of claim 115, wherein the neuropathic pain is associated with post-herpetic neuralgia.

117. The method of claim 115, wherein the neuropathic pain is associated with sciatica.

118. A method of treating painful diabetic neuropathy, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 77.

119. A method of activating the RET receptor in a human, the method comprising administering to the human an effective amount of the pharmaceutical composition of claim 77.

132. A method of treating a peripheral neuropathy, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 80.

133. A method of treating neuropathic pain, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 80.

134. The method of claim 133, wherein the neuropathic pain is associated with post-herpetic neuralgia.

135. The method of claim 133, wherein the neuropathic pain is associated with sciatica.

136. A method of treating painful diabetic neuropathy, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 80.

137. A method of activating the RET receptor in a human, the method comprising administering to the human an effective amount of the pharmaceutical composition of claim 80.

138. A method of treating a peripheral neuropathy, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 81.

139. A method of treating neuropathic pain, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 81.

140. The method of claim 139, wherein the neuropathic pain is associated with post-herpetic neuralgia.

141. The method of claim 139, wherein the neuropathic pain is associated with sciatica.

142. A method of treating painful diabetic neuropathy, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 81.

143. A method of activating the RET receptor in a human, the method comprising administering to the human an effective amount of the pharmaceutical composition of claim 81.

156. A method of treating a peripheral neuropathy, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 84.

157. A method of treating neuropathic pain, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 84.

158. The method of claim 157, wherein the neuropathic pain is associated with post-herpetic neuralgia.

159. The method of claim 157, wherein the neuropathic pain is associated with sciatica.

160. A method of treating painful diabetic neuropathy, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 84.

161. A method of activating the RET receptor in a human, the method comprising administering to the human an effective amount of the pharmaceutical composition of claim 84.

168. A method of treating a peripheral neuropathy, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 86.

169. A method of treating neuropathic pain, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 86.

170. The method of claim 169, wherein the neuropathic pain is associated with post-herpetic neuralgia.

171. The method of claim 169, wherein the neuropathic pain is associated with sciatica.

172. A method of treating painful diabetic neuropathy, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 86.

173. A method of activating the RET receptor in a human, the method comprising administering to the human an effective amount of the pharmaceutical composition of claim 86.

174. A method of treating a peripheral neuropathy, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 87.

175. A method of treating neuropathic pain, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 87.

176. The method of claim 175, wherein the neuropathic pain is associated with post-herpetic neuralgia.

177. The method of claim 175, wherein the neuropathic pain is associated with sciatica.

178. A method of treating painful diabetic neuropathy, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 87.

179. A method of activating the RET receptor in a human, the method comprising administering to the human an effective amount of the pharmaceutical composition of claim 87.

204. A nucleic acid comprising a sequence that encodes the polypeptide of claim 10.

205. An expression vector comprising the nucleic acid of claim 204.

206. A cell comprising the expression vector of claim 205.

207. A method of making a polypeptide, the method comprising:

providing the cell of claim 206, and

culturing the cell under conditions that permit expression of the nucleic acid.

208. A nucleic acid comprising a sequence that encodes the polypeptide of claim 11.

209. An expression vector comprising the nucleic acid of claim 208.

210. A cell comprising the expression vector of claim 209.

211. A method of making a polypeptide, the method comprising:

providing the cell of claim 210, and

culturing the cell under conditions that permit expression of the nucleic acid.

17. A polypeptide consisting of amino acids 15-113 of SEQ ID NO:

5.

18. A polypeptide consisting of amino acids 10-113 of SEQ ID NO:

5.

45. A conjugate comprising the polypeptide of claim 18 conjugated to a non-naturally occurring polymer.

46. The conjugate of claim 45, wherein the non-naturally occurring polymer is a polyalkylene glycol.

47. The conjugate of claim 46, wherein the polyalkylene glycol is polyethylene glycol.

48. The conjugate of claim 47, wherein the polyethylene glycol is coupled to the polypeptide at the amino terminus.

49. The conjugate of claim 47, wherein the polyethylene glycol is coupled to the polypeptide at an internal polymer conjugation site.

60. A fusion protein comprising the polypeptide of claim 18 and a heterologous amino acid sequence.

67. A dimer comprising two polypeptides according to claim 18.

78. A pharmaceutical composition comprising the polypeptide of claim 18 and a pharmaceutically acceptable carrier or excipient.

120. A method of treating a peripheral neuropathy, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 78.

121. A method of treating neuropathic pain, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 78.

122. The method of claim 121, wherein the neuropathic pain is associated with post-herpetic neuralgia.

123. The method of claim 121, wherein the neuropathic pain is associated with sciatica.

124. A method of treating painful diabetic neuropathy, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 78.

125. A method of activating the RET receptor in a human, the method comprising administering to the human an effective amount of the pharmaceutical composition of claim 78.

212. A nucleic acid comprising a sequence that encodes the polypeptide of claim 18.

213. An expression vector comprising the nucleic acid of claim 212.

214. A cell comprising the expression vector of claim 213.

215. A method of making a polypeptide, the method comprising:

providing the cell of claim 214, and

culturing the cell under conditions that permit expression of the nucleic acid.

19. A polypeptide consisting of the amino acid sequence of SEQ ID NO:

5.

50. A conjugate comprising the polypeptide of claim 19 conjugated to a non-naturally occurring polymer.

51. The conjugate of claim 50, wherein the non-naturally occurring polymer is a polyalkylene glycol.

52. The conjugate of claim 51, wherein the polyalkylene glycol is polyethylene glycol.

53. The conjugate of claim 52, wherein the polyethylene glycol is coupled to the polypeptide at the amino terminus.

54. The conjugate of claim 52, wherein the polyethylene glycol is coupled to the polypeptide at an internal polymer conjugation site.

61. A fusion protein comprising the polypeptide of claim 19 and a heterologous amino acid sequence.

68. A dimer comprising two polypeptides according to claim 19.

71. A dimer comprising two conjugates according to claim 52.

72. A dimer comprising two conjugates according to claim 53.

79. A pharmaceutical composition comprising the polypeptide of claim 19 and a pharmaceutically acceptable carrier or excipient.

82. A pharmaceutical composition comprising the conjugate of claim 52 and a pharmaceutically acceptable carrier or excipient.

83. A pharmaceutical composition comprising the conjugate of claim 53 and a pharmaceutically acceptable carrier or excipient.

85. A pharmaceutical composition comprising the dimer of claim 68 and a pharmaceutically acceptable carrier or excipient.

88. A pharmaceutical composition comprising the dimer of claim 71 and a pharmaceutically acceptable carrier or excipient.

89. A pharmaceutical composition comprising the dimer of claim 72 and a pharmaceutically acceptable carrier or excipient.

126. A method of treating a peripheral neuropathy, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 79.

127. A method of treating neuropathic pain, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 79.

128. The method of claim 127, wherein the neuropathic pain is associated with post-herpetic neuralgia.

129. The method of claim 127, wherein the neuropathic pain is associated with sciatica.

130. A method of treating painful diabetic neuropathy, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 79.

131. A method of activating the RET receptor in a human, the method comprising administering to the human an effective amount of the pharmaceutical composition of claim 79.

144. A method of treating a peripheral neuropathy, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 82.

145. A method of treating neuropathic pain, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 82.

146. The method of claim 145, wherein the neuropathic pain is associated with post-herpetic neuralgia.

147. The method of claim 145, wherein the neuropathic pain is associated with sciatica.

148. A method of treating painful diabetic neuropathy, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 82.

149. A method of activating the RET receptor in a human, the method comprising administering to the human an effective amount of the pharmaceutical composition of claim 82.

150. A method of treating a peripheral neuropathy, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 83.

151. A method of treating neuropathic pain, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 83.

152. The method of claim 151, wherein the neuropathic pain is associated with post-herpetic neuralgia.

153. The method of claim 151, wherein the neuropathic pain is associated with sciatica.

154. A method of treating painful diabetic neuropathy, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 83.

155. A method of activating the RET receptor in a human, the method comprising administering to the human an effective amount of the pharmaceutical composition of claim 83.

162. A method of treating a peripheral neuropathy, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 85.

163. A method of treating neuropathic pain, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 85.

164. The method of claim 163, wherein the neuropathic pain is associated with post-herpetic neuralgia.

165. The method of claim 163, wherein the neuropathic pain is associated with sciatica.

166. A method of treating painful diabetic neuropathy, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 85.

167. A method of activating the RET receptor in a human, the method comprising administering to the human an effective amount of the pharmaceutical composition of claim 85.

180. A method of treating a peripheral neuropathy, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 88.

181. A method of treating neuropathic pain, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 88.

182. The method of claim 181, wherein the neuropathic pain is associated with post-herpetic neuralgia.

183. The method of claim 181, wherein the neuropathic pain is associated with sciatica.

184. A method of treating painful diabetic neuropathy, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 88.

185. A method of activating the RET receptor in a human, the method comprising administering to the human an effective amount of the pharmaceutical composition of claim 88.

186. A method of treating a peripheral neuropathy, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 89.

187. A method of treating neuropathic pain, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 89.

188. The method of claim 187, wherein the neuropathic pain is associated with post-herpetic neuralgia.

189. The method of claim 187, wherein the neuropathic pain is associated with sciatica.

190. A method of treating painful diabetic neuropathy, the method comprising administering to a human in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 89.

191. A method of activating the RET receptor in a human, the method comprising administering to the human an effective amount of the pharmaceutical composition of claim 89.

216. A nucleic acid comprising a sequence that encodes the polypeptide of claim 19.

217. An expression vector comprising the nucleic acid of claim 216.

218. A cell comprising the expression vector of claim 217.

219. A method of making a polypeptide, the method comprising:

providing the cell of claim 218, and

culturing the cell under conditions that permit expression of the nucleic acid.