Antibody-drug conjugates including antibody against human dlk1, and use thereof
A technology of antibody conjugates and antibodies, applied in medical preparations containing active ingredients, anti-animal/human immunoglobulins, anti-tumor drugs, etc.
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Embodiment 1
[0453] Production of Compounds 1, 2, 3 and 4
[0454]
[0455]
[0456] The compounds 1, 2, 3 and 4 were produced by the method described in International Publication Patent WO2017-089895.
[0457] Among the compounds 1, 2, 3 and 4, the structure of MMAE (Monomethyl Auristatin E) is as follows.
[0458]
Embodiment 2
[0459] Production of Compounds 5, 6, 7 and 8
[0460]
[0461]
[0462] The compounds 5, 6 and 7 were produced by the method described in Korean Laid-Open Patent No. 10-2018-0110645.
[0463]
[0464] The compound 8 was produced by the method described in Korean Application No. 10-2019-0000514.
Embodiment 3
[0465] Production of ADC
[0466] ADCs were produced in the following two steps, and LCB14-0511 and LCB14-0606, which are commonly used, were produced by the method described in Korean Laid-Open Patent No. 10-2014-0035393. The structural formulas of LCB14-0511 and LCB14-0606 are as follows:
[0467]
[0468] Step 1: Production of prenylated antibody
[0469] A prenylation reaction mixture of the antibody (18A5) produced by the method described in Korean Application No. 10-2018-0107639 was prepared and reacted at 30°C for 16 hours. The reaction mixture contained 24 μM antibody (18A5), and 200 nM FTase (Calbiochem, #344145) and 0.144 mM LCB14-0511 or LCB14-0606 buffer solution (50 mM Tris-HCl (pH 7.4), 5 mM MgCl 2 , 10μM ZnCl 2 , 0.144 mM DTT (Dithiothreitol, dithiothreitol)). After the reaction, the prenylated antibody was desalted using a G25 Sepharose column (AKTA purifier, GE healthcare) equilibrated with PBS (Phosphate Buffer Saline, phosphate buffer saline) buffer...
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