Use of diazocine analogue in preparation of antineoplastic and antifungal medicine
An anti-tumor drug, anti-fungal drug technology, applied in the direction of anti-tumor drugs, anti-fungal agents, drug combinations, etc., can solve problems such as few researches
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Embodiment 1
[0056] How to prove that 2,6-bis-(6-mercapto-3-pyridine)-4,8-bis-(4-phenol)-[1,5]-diazapine inhibits fungi below by Example 1 Function (take Mucor as an example).
[0057] 1. Bacteria to be tested: Mucor biemalis 3.40
[0058] 2. TF medium (g / L) peptone 1%; glucose 1%; agar 1.6%; pH7.0
[0059] 3.2,6-bis-(6-mercapto-3-pyridine)-4,8-bis-(4-phenol)-[1,5]-diazocine concentration gradient acquisition: according to the distribution of the culture plate, Perform a ten-fold serial dilution of the stock solution, and the minimum dilution should reach 10 -6 , so that the determined MIC falls within the determined range
[0060] 4. Add 150 μl culture medium and 10 μl 2,6-bis-(6-mercapto-3-pyridine)-4,8-bis-(4-phenol)-[1,5] to each well of the 96-well plate -Diazocine solution and 50 μl mucormyces spore suspension, each drug concentration was repeated more than three times.
[0061] 5. Incubate at 30°C for 18 hours, subject to the growth visible to the naked eye in the control group...
Embodiment 2
[0065] How to prove that 2,6-bis-(6-mercapto-3-pyridine)-4,8-bis-(4-phenol)-[1,5]-diazocine is effective for tumor cells by Example 2 below Inhibitory effect (taking liver cancer cell Bel-7402 as an example).
[0066] 1. Cell line: Bel-7402 (adherent cells)
[0067] 2. Preparation of 2,6-bis-(6-mercapto-3-pyridine)-4,8-bis-(4-phenol)-[1,5]-diazocine solution: preparation concentration from 0.12mg / ml to 0.03 The mg / ml gradient decreases.
[0068] 3. PBS preparation: KCl 0.2g / l; KH 2 PO 4 0.2g / l; NaCl 8.0g / l; NaCl 2 HPO 4 .7H 2 O 1.56 g / l. D-Hanks solution: KCl 0.4g / l; KH 2 PO 4 0.06g / l; NaCl 8.0g / l; NaCl 2 HPO 4 .7H 2 O0.06g / l; NaHCO 3 0.35g / l; phenol red 0.02g / l.
[0069] MTT preparation: the concentration of the preservation solution is 5mg / ml, and the final concentration of the addition is 0.4mg / ml. Sterilize by filtration and store at -70°C. When in use, add MEM medium, make a 1:10 dilution, and filter to sterilize.
[0070] 4. Insert the cells into the ...
Embodiment 3
[0074]How to prove that 2,6-bis-(6-mercapto-3-pyridine)-4,8-bis-(4-phenol)-[1,5]diazocine can induce tumor cell growth by Example 3 Apoptosis occurs (taking cervical cancer cell Hela as an example).
[0075] 1. Add 2,6-bis-(6-mercapto-3-pyridine)-4,8-bis-(4-phenol)-[1,5]-diazoocine to the culture medium of Hela, the concentration 35ng / ml, incubated for 72 hours.
[0076] 2. In a 1.5ml microcentrifuge tube, centrifuge to pellet 1×10 6 -2×10 6 For Hela cells, wash the cells to be tested twice with PBS, remove the supernatant; add 400 μl cell lysate and 2 μl proteinase K, mix well, incubate in a 37°C water bath for 1 hour, add 2 μl RNase, and incubate for another 1 hour.
[0077] 3. In a desktop centrifuge, centrifuge at 14,000rpm at room temperature for 10 minutes, transfer the supernatant to another clean microcentrifuge tube; use phenol: chloroform = 1: 1, phenol: chloroform: isoamyl alcohol = 25: 24 : 1 solution and chloroform aliquots were extracted once for one hour eac...
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