Diagnostic or therapeutic somatostatin or bombesin analog conjugates and uses thereof
By linking bioactive peptides with connectors to maintain their activity and utilizing hydrophilic polymers to improve distribution, the problems of therapeutic toxicity and radiolabeling in tumor imaging are solved, enabling tumor-specific targeting and rapid excretion, thereby improving therapeutic efficacy and reducing toxicity.
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Embodiment 1
[0115] Preparation of the Dicesium Salt of (+)-Methotrexate (Methotrexate)
[0116] Methotrexate (0.75 mmol) was dissolved in 200 ml of water to which cesium bicarbonate (1.5 mmol) was added. The yellow solution was stirred for 4 hours, evaporated under reduced pressure, and the oily residue was dissolved in ethanol. Evaporate 3 times from absolute ethanol to remove residual water.
Embodiment 2
[0118] 2-Bromo-acetyl-D-tert-butyl-Ser-D-tert-butyl-Ser-D-tert-butyl-Ser-D-tert-butyl-Ser-D-tert-butyl-Ser-ε-tert-butyl Butoxycarbonyl-Lys-D-tert-Butyl-Tyr-D-tert-Butyl-Tyr-S-Trityl-Cys-Phe-D-Trp-ε-tert-Butoxycarbonyl-Lys-tert-Butyl Preparation of base-Thr-S-trityl-Cys-tert-butyl-Thr-Rink-amide-resin
[0119]Rink-amide MBHA polystyrene resin (0.25 mmol) [4-(2',4'-dimethoxyphenyl-Fmoc-(aminomethyl)phenoxyacetamido-norleucyl-methyl Diphenylmethylamine resin, 100-200 mesh, Novabiochem, San Diego, CA)] was added to the reaction vessel of a CS136 automated peptide synthesizer (CS Bio, Inc., San Carlos, CA) and swelled in DMF for approx. 1 hour. The resin was filtered and an excess of 20% piperidine in DMF was added and mixed for 2 minutes. The resin was filtered and again excess 20% piperidine was added and mixed for 20 minutes to ensure complete removal of the resin's Fmoc groups. After deprotection, the resin was washed 4 times with DMF, then the first protected amino acids, ...
Embodiment 3
[0122] Methotrexate-acetyl-D-tert-butyl-Ser-D-tert-butyl-Ser-D-tert-butyl-Ser-D-tert-butyl-Ser-D-tert-butyl-Ser-ε-tert-butyloxy ylcarbonyl-Lys-D-tert-butyl-Tyr-D-tert-butyl-Tyr-S-trityl-Cys-Phe-D-Trp-ε-tert-butoxycarbonyl-Lys-tert-butyl- Preparation of Thr-S-trityl-Cys-tert-butyl-Thr-Rink-amide-resin
[0123] The dicesium salt of methotrexate (0.75 mmol) prepared in Example 1 was dissolved in DMSO and added to the peptidyl resin prepared in Example 2 (0.25 mmol) in a round bottom flask. While heating in a water bath (40°C, 18 hours), the suspension was mixed gently, filtered and washed with copious DMF followed by methanol. After the final filtration, the derivatized resin was air dried overnight.
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