Filterable duocarmycin-containing antibody-drug conjugate compositions and related methods
A technology of duocarmycin and composition, applied in the field of approving batches of ADCs containing duocarmycin, can solve the problems of inaccurate determination of free linker-drug content, insufficient free linker-drug amount, etc.
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Embodiment 1
[0131] Example 1 - Outer Standard Sample
[0132] The non-conjugated form of methalin joints - drug VC-SECO-DUBA (SYD980) (0.25 μg / ml) was analyzed by the RP-UPLC method discussed above. figure 1 It is a representative of the chromatogram obtained. figure 1 A clear peak corresponding to unconfixed multiplial canceam joints - drug (Vc-SECO-DUBA) is shown at 6.205 minutes.
Embodiment 2-A
[0133] Example 2-ADC analysis
[0134] Samples from Tru-Uplc monurazine batch were treated in accordance with the above sample preparation procedure, and analyzed using the above RP-UPLC method. The columns obtained are displayed in disadvantages - drug VC-SECO-DUBA. figure 2 It is a representative of the chromatogram obtained, and peaks were detected in 6.2 minutes or near it. This analysis determined that the quantity of the drug in the sample - the amount of drug was lower than the detection limit, and thus below the predetermined level (ie, pass / fail detection test). Therefore, the release / approval of the batch is appropriate.
Embodiment 3
[0135] Comparison of Example 3 - Composition
[0136] 100 μL of the sample containing Samples containing trastum monoclonarazine (10 mg / ml) and Vc-SECO-DUBA (0.25 μg / ml) was introduced into a 10K filter ( Omega centrifugal device). Dilution media (300 μL) is added to the filter, the dilution medium comprises water, acetonitrile (ACN) or methanol (MeOH) used at different percent, and different percent acid (formic acid, trifluoroacetic acid (TFA) or hydrochloric acid) . The samples and dilution media on the filter were homogenized and centrifuged at 14,000 g for 15 minutes. The filtrate was homogenized and transferred to the HPLC vial. The amount of VC-SECO-DUBA (SYD980, ie, unconfixed multiplied metamycline-drug) is used in the RP-UPLC-UV detection filtrate, and the formula (Vi) is used to quantify . The results are summarized in Table 2.
[0137] Table 2. Recycling results of VC-SECO-DUBA
[0138]
[0139] A: Relative standard deviation of three samples
[0140] B: Only tw...
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