| Targets |
(R)-DZD1516 targets the human epidermal growth factor receptor 2 (HER2), a member of the ErbB family of receptor tyrosine kinases. HER2 is known to be overexpressed or amplified in approximately 20-30% of breast cancers and is associated with aggressive tumor growth and poor clinical outcomes. This compound acts as a potent and selective inhibitor of HER2, blocking its kinase activity and preventing the subsequent activation of downstream signaling pathways, including the PI3K/AKT and MAPK/ERK cascades, which are critical for cell proliferation, survival, and metastasis. This selective HER2 inhibition makes it a valuable tool for studying the molecular mechanisms of HER2-driven cancers and for evaluating potential therapeutic interventions targeting this oncogenic driver.
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| ln Vitro |
In vitro studies have demonstrated that (R)-DZD1516 potently inhibits the phosphorylation of HER2 in various HER2-overexpressing cancer cell lines, leading to reduced downstream signaling and decreased cell proliferation. It effectively blocks the activation of the PI3K/AKT and MAPK pathways, resulting in cell cycle arrest and apoptosis in vitro. The compound shows high selectivity for HER2 over other kinases, minimizing off-target effects in cell-based assays. It has been evaluated in a panel of HER2-positive cancer cell lines, including BT-474, SK-BR-3, and NCI-N87, where it exhibits growth inhibitory effects with IC50 values in the low nanomolar range, consistent with its potent and selective mechanism of action.
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| ln Vivo |
In vivo studies have shown that (R)-DZD1516 exhibits significant anti-tumor efficacy in mouse xenograft models bearing HER2-positive human tumor cells. Oral administration of the compound leads to dose-dependent tumor growth inhibition, with tumor regression observed at optimal doses. In a xenograft model of HER2-driven gastric cancer, (R)-DZD1516 effectively reduces tumor volume and weight compared to vehicle controls. The compound's in vivo anti-tumor activity is correlated with reduced HER2 phosphorylation in the tumor tissues, confirming target engagement. It is well tolerated with minimal body weight changes at therapeutic doses, suggesting a favorable safety profile in these animal models.
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| Enzyme Assay |
A cell-free kinase assay protocol for (R)-DZD1516 involves measuring its ability to inhibit HER2 kinase activity. The protocol: (1) Prepare a reaction mixture containing 50 mM HEPES (pH 7.5), 10 mM MgCl2, 2 mM MnCl2, 1 mM DTT, 0.01% Tween-20, and purified recombinant HER2 kinase domain. (2) Add varying concentrations of (R)-DZD1516 (0.1-1000 nM) dissolved in DMSO (final DMSO ≤1%). (3) Add the substrate, a biotinylated peptide (e.g., poly-Glu-Tyr, 0.2 mg/mL), and 10 uM ATP (including 0.5 uCi [gamma-33P]-ATP per well). (4) Incubate at 30degC for 30 minutes. (5) Stop the reaction by adding 0.5 M EDTA to a final concentration of 30 mM. (6) Transfer the reaction mixture to streptavidin-coated plates, incubate for 15 minutes, and wash three times with PBS containing 0.05% Tween-20. (7) Add scintillation fluid and measure incorporated radioactivity using a scintillation counter. (8) Calculate the percent inhibition relative to control (DMSO only) and determine the IC50 by non-linear regression.
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| Cell Assay |
A standard cellular protocol for evaluating (R)-DZD1516 in HER2-positive cancer cells: (1) Culture HER2-overexpressing human breast cancer BT-474 cells in DMEM/F12 medium supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin at 37degC in 5% CO2. (2) Seed cells into 96-well plates at 5,000-10,000 cells/well and allow to attach overnight. (3) Prepare a 10 mM stock solution of (R)-DZD1516 in DMSO and serially dilute in culture medium (final DMSO concentration ≤0.5%). (4) Treat cells for 72 hours. (5) Add 20 microL of 5 mg/mL MTT solution to each well and incubate for 4 hours at 37degC. (6) Remove the medium and add 150 microL of DMSO to dissolve the formazan crystals. (7) Measure absorbance at 570 nm using a microplate reader. (8) Calculate the percentage of cell viability relative to the control. (9) For Western blotting, treat cells for 4-6 hours, lyse in RIPA buffer, and probe with antibodies against p-HER2, total HER2, p-AKT, and p-ERK.
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| Animal Protocol |
An in vivo protocol for evaluating (R)-DZD1516 in a xenograft mouse model: (1) Culture BT-474 or NCI-N87 cancer cells and harvest. (2) Inoculate 5×10⁶ cells subcutaneously into the right flank of 6-8 week old female BALB/c nude mice. (3) When tumors reach an average volume of 150-200 mm3 (approximately 14-21 days), randomize mice into groups (n=6-8 per group). (4) Formulate (R)-DZD1516 in a vehicle of 10% DMSO, 40% PEG300, 5% Tween 80, and 45% saline. (5) Administer the compound orally (PO) daily at doses of 1, 3, and 10 mg/kg. (6) Measure tumor size using calipers twice weekly and calculate tumor volume using the formula (width2 × length)/2. (7) Record body weights to monitor toxicity. (8) After 14-21 days of treatment, euthanize mice and excise tumors for weighing. (9) Collect tumors and snap-freeze for Western blotting to assess p-HER2 levels. (10) Analyze tumor growth inhibition (TGI) compared to vehicle controls.
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| ADME/Pharmacokinetics |
Pharmacokinetic properties of (R)-DZD1516 have been characterized in preclinical species. Following oral administration, the compound exhibits moderate to good oral bioavailability. The compound has a molecular weight of 547.56, and it is highly soluble in DMSO (≥100 mg/mL) but has limited aqueous solubility. In rodent PK studies, (R)-DZD1516 demonstrates moderate clearance and a half-life (t½) of approximately 2-4 hours. The compound shows good plasma exposure (AUC) that increases dose-proportionally. Peak plasma concentrations (Cmax) are achieved within 1-2 hours post-administration (Tmax). Tissue distribution studies indicate good tumor penetration, consistent with its in vivo efficacy. For storage, the powder is stable at -20degC for up to 3 years, and stock solutions in DMSO can be stored at -80degC for up to 6 months or at -20degC for 1 month, protected from moisture and light.
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| Toxicity/Toxicokinetics |
No formal toxicity studies have been published for (R)-DZD1516, as it is a research compound and not an approved drug. Based on its mechanism as a HER2 inhibitor, potential toxicities may include gastrointestinal effects (diarrhea, nausea), skin rash, and fatigue, which are common among tyrosine kinase inhibitors. However, its high selectivity for HER2 may reduce off-target toxicities compared to other less selective kinase inhibitors. In preclinical xenograft models, (R)-DZD1516 is well tolerated at therapeutic doses with no significant body weight loss or behavioral changes observed. No data are available regarding acute toxicity (LD50), genotoxicity, or reproductive toxicity. Standard safety precautions for handling HER2 inhibitors should be followed.
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| References | |
| Additional Infomation |
(R)-DZD1516 is a research compound and is not an FDA-approved drug. It is the R enantiomer of DZD1516, a potent and selective HER2 inhibitor. HER2 (human epidermal growth factor receptor 2) is a receptor tyrosine kinase whose overexpression drives multiple cancers, including breast and gastric cancers. The compound is used in preclinical research to study HER2-driven carcinogenesis and to evaluate HER2-targeted therapeutic strategies. No clinical trials have been reported for (R)-DZD1516. The compound is for research use only and not intended for human therapeutic use. It is also known by its catalog number, and its IUPAC name is 4-quinazolinamine, 5-[(4R)-3,3-difluoro-1-methyl-4-piperidinyl]oxy-7-methoxy-N-[3-methyl-4-([1,2,4]triazolo[1,5-a]pyridin-7-yloxy)phenyl].
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| CAS # |
2387570-03-6
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| Appearance |
Solid powder ; White to off-white
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : ~100 mg/mL (~182.63 mM; with ultrasonication)
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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