Calcitonin receptor-binding peptides
A calcitonin receptor and calcitonin technology, applied in the direction of calcitonin, hormone peptide, isotope introduction into organic compounds, etc., can solve the technical difficulties of coupling while maintaining compound receptor specificity
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Embodiment 1
[0159] Synthesis of BAT chelating agent
[0160] BAT chelators, particularly S-cysteine-derived and ε-aminolysylamino-derived BAT chelators, were prepared according to the methods of commonly owned and co-pending U.S. Serial No. 08 / 414424, incorporated herein by reference .
Embodiment 2
[0162] Solid Phase Peptide Synthesis
[0163] Solid-phase peptide synthesis (SPPS) was performed on a 0.25 mmol scale using an Applied Biosystems Model 431A Peptide Synthesizer, using fluorenylmethoxycarbonyl (Fmoc) amino-terminal protection with dicyclohexylcarbodiimide / hydroxybenzo Triazole or 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate / hydroxybenzotriazole (HBTU / HOBT) coupling, and for carboxyl Use p-hydroxymethylphenoxymethyl-polystyrene (HMP) resin or Sasrin for terminal acid TM Either chlorotrityl free radical resin or Rink amide resin for carboxy-terminated amides.
[0164] Where appropriate, Fmoc-Cys(BAT) and Nα-Fmoc-Nε-(BAT)Lys were synthesized as described in commonly owned and co-pending US Serial No. 08 / 414424, incorporated herein by reference.
[0165] By using the appropriate 2-haloacid as the last residue of the coupling in SPPS, or by using 2-haloacid / diisopropylcarbodiimide / N-hydroxysuccinamide / NMP or 2-halogenated anhydride / diis...
Embodiment 3
[0181] General method for radiolabeling with technetium-99m
[0182] 0.1 mg of the peptide prepared in Example 2 was dissolved in 0.1 ml or 0.2 ml of water or 0.9% saline. Technetium-99m gluceptate was prepared by reconstituting Glucoscan vials (E.I. DuPont de Nemours, Inc., Wilmington, DE) with 0.25 ml technetium-99m sodium pertechnetate containing up to 200 mCi and left at room temperature 15 minutes. Then 25 μl technetium-99m gluconate was added to the peptide and the reaction was allowed to proceed for 15-60 minutes at room temperature or 10-30 minutes at 100° C. and then filtered through a 0.2 μm filter.
[0183] The purity of the technetium-99m-labeled peptides was determined by reverse-phase HPLC using the following conditions: Waters Delta Pak C18 column, 5 μ, 3.9 mm x 150 mm analytical column loaded with each labeled peptide at a solvent flow rate equal to 1 ml / min Eluting peptide (Delta-Pak). Gradient elution using a gradient of 20-50% solvent B / solvent A (solvent...
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