Artificially designed anti-HIV infection polypeptide, composition and use
A composition and application technology, applied in the fields of anti-HIV infection polypeptide, treatment or prevention of related diseases caused by HIV infection, non-physiologically toxic salt, acquired immunodeficiency syndrome, and can solve the problem of low bioavailability and drug resistance , limit the wide application of T20 and other issues
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Embodiment 1
[0141] Embodiment 1: the preparation of compound 6
[0142] A standard Fmoc solid-phase peptide synthesis method was used. All peptide sequences are amidated at the C-terminus and acetylated at the N-terminus. Rink Amide resin is selected, the amino acid is in L configuration, and the peptide chain is extended from the C-terminus to the N-terminus. The condensing agent is HBTU / HOBt / DIEA. The deprotecting agent is piperidine / DMF solution. The lysing agent is trifluoroacetic acid (TFA), and the crude peptide is dissolved in water and then freeze-dried for storage. Separation and purification are carried out by medium-pressure liquid chromatography Flash or high-pressure liquid chromatography (HPLC), and the pure peptide content is >95%. Matrix-assisted laser desorption time-of-flight mass spectrometry (MALDI-TOF-MS) confirmed the molecular weight of the peptide sequence.
[0143] Use CEM automatic microwave peptide synthesizer for peptide synthesis, the synthesis conditions...
Embodiment 2-68
[0154] Embodiment 2-68: Preparation of Compound 1-5, 7-68
[0155] Compounds No. 1-No. 5, No. 7-No. 68 were synthesized according to the method of Example (1), and the amino acids were all in L configuration, but the corresponding amino acid residues were replaced.
Embodiment 69
[0156] Example 69: Inhibition of HIV-1 Biological Activity Detection
[0157] Evaluation of compounds inhibiting HIV-1-mediated cell-cell fusion activity (IC 50 ):
[0158] Detection of HIV-1-Mediated Cell-Cell Fusion by Stain Transfer Assay: HIV-1 IIIB Infected H9 cells (H9 / HIV-1 IIIB ) was labeled with a fluorescent reagent Calcein-AM (Molecular Probes, Inc., Eugene, OR), and then, 50 μl of labeled H9 / HIV-1 was added to each well of a 96-well plate mB cells (2×10 5 / ml), and then add 50 μl of different concentrations of test samples (compounds prepared in the tested examples 1-34 and 44-53, two-fold serial dilution from a concentration of 250 μg / ml), and incubate at 37°C for 30min; then each well Add 100ul of MT-2 cells (1×10 6 / ml), incubated at 37°C for 2h. Confused and unfused Calcein-labeled HIV-1 infected MT-2 cells (MT-2 is a commonly used effector cell used in this experiment to simulate HIV-infected target cells, commercially available) with an inverted fluores...
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