Multi-modal leucocyte molecule probe compound and its preparation method and use
A molecular probe and compound technology, applied in the field of multifunctional probe compounds, can solve the problems of low sensitivity, poor specificity of inflammation, high cost, etc.
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Embodiment 1
[0069] Embodiment 1: Use solid-phase chemical method to synthesize double label probe precursor compound cFLFLF-Fl-HYNIC (formula III) and cFLFLF-Fl-NOTA (formula V), as scheme 1:
[0070] Using a peptide synthesizer, through standard Fmoc chemistry, compound 1 is gradually coupled on wangresin, and after removing the protective group MTT of the side chain, it is coupled with Fmoc-N-amido-PEG24-Acid to generate compound 2, and further removed Remove Fmoc protection and couple with Fmoc-Cys(Trt)-OH to generate compound 3. After removing Fmoc protection on the structure of compound 3, couple with NHS-HYNIC-Boc or p-SCN-Bn-NOTA respectively, and then in the mass In 1% trifluoroacetic acid-dichloromethane, wangresin was cleaved and the protecting group Trt on the cysteine side chain thiol was removed to generate compound 4 (cFLFLFK-PEG24(SH)-HYNIC) and compound 5, respectively. (cFLFLFK-PEG24(SH)-NOTA). After HPLC purification, the sulfhydryl groups on compound 4 and compound 5...
Embodiment 2
[0072] Example 2, the ECT probe precursor compound cFLFLF-PEG24-HYNIC (formula VIII) was synthesized using solid phase chemistry, and the PET probe precursor compound cFLFLF-PEG24-NOTA (formula X) was shown in Scheme 2:
[0073] Using a peptide synthesizer, through standard Fmoc chemistry, compound 2 was gradually coupled on wangresin, and after further removal of Fmoc protection, compound 6 and Compound 7, then cut wangresin in 1% trifluoroacetic acid-dichloromethane to generate ECT probe precursor compound cFLFLF-PEG24-HYNIC (formula VIII) and PET probe precursor compound cFLFLF-PEG24 -NOTA (Formula X) (as shown in Scheme 2), purified by HPLC, and freeze-dried to obtain the target compound. The molecular weight of this compound was identified by using a MULDI-TOF mass spectrometer. The calculated and experimental molecular weights of cFLFLF-PEG24-HYNIC (Formula VIII) were 2,405 and 2,404, respectively; the calculated and experimental molecular weights of cFLFLF-PEG24-NOTA (...
Embodiment 3
[0076] Embodiment 3, synthetic multimodal leukocyte molecular probe compound cFLFLF-Fl- 99m Tc (formula IV), such as scheme 3:
[0077] Take 50 μg of cFLFLF-Fl-HYNIC (formula III), weigh 6.7 mg of tricine into an eppendorf vial, add 200 μL of normal saline to dissolve, then add 200 μL of TPPTS (5 mg / mL aqueous solution), and finally add 20 μL of 1 mg / mL stannous chloride hydrochloric acid solution (1M hydrochloric acid solution), then immediately add 0.4mL[ 99m Tc] pertechnetate (20mCi), react at 60°C for 10 minutes, let stand at room temperature for 10 minutes, use a C18 separation column (Sep-Paklightcartridge) to separate the salt in the reaction mixture, and finally use 1mL ethanol to elute the labeled compound, then add 1mL normal saline was mixed to obtain the cFLFLF-Fl- to be injected 99m Tc (Formula IV).
[0078] Scheme 3 chemical synthesis route:
[0079]
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