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. 2019 May;37(5):523-526.
doi: 10.1038/s41587-019-0073-7. Epub 2019 Apr 1.

Engineering a HER2-specific antibody-drug conjugate to increase lysosomal delivery and therapeutic efficacy

Jeffrey C Kang  1 , Wei Sun  1 , Priyanka Khare  1 , Mostafa Karimi  2 , Xiaoli Wang  1 , Yang Shen  2 , Raimund J Ober  3   4   5 , E Sally Ward  6   7   8
Affiliations

Engineering a HER2-specific antibody-drug conjugate to increase lysosomal delivery and therapeutic efficacy

Jeffrey C Kang et al. Nat Biotechnol. 2019 May.

Abstract

We improve the potency of antibody-drug conjugates (ADCs) containing the human epidermal growth factor receptor 2 (HER2)-specific antibody pertuzumab by substantially reducing their affinity for HER2 at acidic endosomal pH relative to near neutral pH. These engineered pertuzumab variants show increased lysosomal delivery and cytotoxicity towards tumor cells expressing intermediate HER2 levels. In HER2int xenograft tumor models in mice, the variants show higher therapeutic efficacy than the parent ADC and a clinically approved HER2-specific ADC.

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Figures

Figure 1.
Figure 1.
Acid-switching results in increased accumulation of ADCs in HER2-expressing tumor cells. Cells were incubated with 10 nM Alexa 488-labeled MMAE-conjugated antibody (WT, SG, YS or control hen egg lysozyme-specific antibody, C) for the indicated times at 37 °C, washed, incubated with 5 μg/ml Alexa 488-specific antibody and analyzed by flow cytometry. Mean fluorescence intensities (mean values of independent triplicate cell samples) for Alexa 488 fluorescence are shown. Error bars indicate SD. Statistically significant differences are indicated by * (unpaired two-tailed t-test). Two independent experiments were carried out with similar results.
Figure 2.
Figure 2.
ALTAs are more effective at reducing proliferation of HER2int-expressing cells and deliver increased levels of MMAE to target cells. (a) HER2-expressing cancer cells, or HER2-negative (MDA-MB-468) cells, were treated with MMAE-conjugated antibodies (WT, SG, YS or control hen egg lysozyme-specific antibody, C) for 4 days and cell viability determined. (b) MDA-MB-453 and JIMT-1 cancer cells were incubated with 10 nM MMAE-conjugated antibodies (WT, SG or YS) for 20 hours and cell-associated MMAE quantitated using LC-MS/MS. For (a) and (b), mean values of independent triplicate cell samples are shown and error bars indicate SD. Statistically significant differences between YS or SG and WT are indicated by * (one-way ANOVA with Tukey’s multiple comparison test; for panel (a), P values ranged from 0.001-0.0001 (MDA-MB-453), 0.003-0.008 (JIMT-1) and 0.001-0.05 (SK-OV-3)). (c) Female BALB/c SCID mice bearing MDA-MB-453 tumors were treated twice, with a 21 day interval (arrowheads), with 2 mg/kg ADC (WT-MMAE, SG-MMAE or YS-MMAE), T-DM1, unconjugated WT pertuzumab (WT) or vehicle (PBS) (n = 7 mice/group for YS-MMAE, T-DM1 or PBS; 8 mice/group for WT-MMAE, SG-MMAE or WT). (d) Female BALB/c SCID mice bearing JIMT-1 tumors were treated weekly (four times; arrowheads) with 2 mg/kg ADC (WT-MMAE or SG-MMAE), T-DM1, unconjugated WT or SG, or vehicle (PBS) (n = 6 mice/group for PBS; 7 mice/group for WT-MMAE, SG-MMAE, T-DM1, WT or SG). For (c) and (d), the mean tumor size for each treatment group is shown and error bars indicate SE. Statistically significant differences at the experimental endpoints are indicated by * (two-tailed Mann-Whitney U test and unpaired two-tailed t-test, respectively). For panels (a)-(d), at least two independent experiments were carried out with similar results.
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