Zongertinib

Storage temp: -20℃

Solubility: DMSO: 45.91 mg/mL (85.72 mM)

Form: Solid

Color: Light Yellow to Green Yellow

1. Cell Experiment

DMSO : 30.77 mg/mL (57.45 mM; ultrasonic and warming and adjust pH to 7 with 1 M HCl and heat to 60°C; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

Preparing Stock Solutions：

Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.

Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

2. Animal Experiment

Select the appropriate dissolution method based on your experimental animal and administration route.

For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:

To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for in vivo experiments, it is recommended to prepare freshly and use it on the same day.

The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

Protocol 1

Add each solvent one by one: 10% DMSO, 40% PEG300, 5% Tween-80, 45% Saline

Solubility: ≥ 2.5 mg/mL (4.67 mM); Clear solution

This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).

Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.

Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.

Protocol 2

Add each solvent one by one: 10% DMSO, 90% (20% SBE-β-CD in Saline)

Solubility: ≥ 2.5 mg/mL (4.67 mM); Clear solution

This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).

Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.

Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.

In vivo, zongertinib demonstrated anti-tumor activity in mouse xenograft models of NSCLC with HER2 tyrosine kinase domain activating mutations. A key characteristic of zongertinib is its selectivity for HER2 - including exon 20 insertion mutations - while sparing wild-type epidermal growth factor receptor (EGFR), which is believed to limit EGFR-associated toxicities. In a preclinical proliferation assay, zongertinib showed 100-fold greater sparing activity for wild-type EGFR compared to its activity against the HER2 YVMA mutation. It is effective against various HER2 tyrosine kinase domain mutations, including A775-G776YVMA and P780-Y781insGSP. Zongertinib has also demonstrated intracranial activity, with an intracranial objective response rate of 37% and a disease control rate of 83% observed in patients with baseline brain metastases in a subanalysis of the Beamion LUNG-1 trial.

Human epidermal growth factor receptor 2 (HER2) mutations have emerged as important oncogenic drivers in non-small cell lung cancer (NSCLC), leading to uncontrolled cell growth and survival. Zongertinib is an irreversible tyrosine kinase inhibitor of HER2. Its acrylamide moiety forms a covalent bond with the cysteine 805 residue of the HER2 receptor, thereby inhibiting HER2 phosphorylation and the activation of downstream signaling pathways, including extracellular signal-regulated kinase (ERK), mitogen-activated protein kinases (MAPK), and PI3K/Akt. The inhibition of these pathways leads to a reduction in the proliferation of lung cancer cells harboring HER2 tyrosine kinase domain activating mutations.