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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
MCT4 1.3 nM (IC50)
AZD0095 specifically targets the monocarboxylate transporter 4 (MCT4), also known as SLC16A3. MCT4 is a transmembrane protein responsible for the export of lactate from cells, a process critical for regulating intracellular pH and supporting the Warburg effect (aerobic glycolysis) in cancer cells. AZD0095 is a potent and selective MCT4 inhibitor with an IC50 of 1.3 nM. It exhibits >1000-fold selectivity over MCT1 (the primary lactate importer), ensuring specificity for the lactate export pathway.. |
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| ln Vitro |
With an IC50 of 26 nM, AZD0095 (0-50 µM) suppresses the growth of NCI-H358 cells (high MCT4 expression)[1].
In vitro, AZD0095 is a selective and potent MCT4 inhibitor, demonstrating an IC50 of 1.3 nM.. In cell-based assays, AZD0095 (0-50 uM) inhibits the proliferation of NCI-H358 non-small cell lung cancer cells (which are MCT4-high) with an IC50 of 26 nM, indicating its ability to inhibit tumor cell growth effectively at low nanomolar concentrations.. |
| ln Vivo |
When combined with cediranib, AZD0095 (100 mg/kg, PO, twice day) reduces the growth of NCI-H358 xenograft tumors in mice[1].
In vivo, AZD0095 effectively inhibits tumor growth in a NCI-H358 xenograft model when used in combination with Cediranib, a VEGFR inhibitor.. While the single-agent activity might be modest, the combination therapy highlights the potential of AZD0095 to enhance the efficacy of anti-angiogenic agents. The mechanism is thought to be due to the reversal of the acidic tumor microenvironment (TME), which is immunosuppressive. By blocking lactate export, AZD0095 raises the pH inside the tumor, reducing immunosuppression and allowing for a more effective immune response. |
| Enzyme Assay |
The specific in vitro protocol for MCT4 inhibition is a radiolabeled lactate uptake assay using cells overexpressing MCT4. HEK-293 cells are stably transfected with human MCT4 and its ancillary protein CD147 (which is required for surface expression). Cells are seeded in 96-well plates. After reaching confluency, cells are washed and pre-incubated with varying concentrations of AZD0095 (0.001-1000 nM) in assay buffer (20 mM HEPES, pH 7.4, 5 mM KCl, 1 mM MgSO4, 1 mM CaCl2, 140 mM NaCl) for 15 minutes. The reaction is initiated by adding 100 uM of L-[14C]-lactic acid (1 uCi/mL) and incubated for 2-5 minutes. The uptake is terminated by washing the cells three times with ice-cold PBS containing 0.1 mM of the MCT inhibitor AR-C155858 to block further transport. The cells are lysed with 0.2 N NaOH, and the radioactivity is measured by liquid scintillation counting. The IC50 (1.3 nM) is calculated from the dose-response curve.
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| Cell Assay |
The in vitro cellular assay for AZD0095 involves the use of MCT4-high NCI-H358 lung cancer cells (or other cancer cell lines with high MCT4 expression). Cells are seeded in 96-well black-walled plates (1,000-2,000 cells/well) and cultured overnight. The medium is then replaced with fresh medium containing varying concentrations of AZD0095 (0.1-1000 nM). The cells are incubated for 72 hours (3 days) at 37degC. To measure cell viability, 10 uL of CCK-8 (Cell Counting Kit-8) solution is added to each well, and the plates are incubated for 2-4 hours. The absorbance at 450 nm is measured using a microplate reader. The IC50 for proliferation inhibition is calculated (reported as 26 nM for NCI-H358 cells). Additionally, to assess the mechanism, the extracellular pH of the culture medium can be measured using a pH meter or a pH-sensitive fluorescent probe, and the lactate concentration in the supernatant can be measured using a lactate assay kit..
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| Animal Protocol |
Animal/Disease Models: Murine NCI-H358 xenograft[1]
Doses: 100 mg/kg together with Cediranib (3 mg/kg) Route of Administration: Oral administration (po) Experimental Results: decreased tumor growth more efficiently than AZD0095 or AZD2171 alone. An in vivo protocol for AZD0095 involves a subcutaneous xenograft model using NCI-H358 non-small cell lung cancer cells. Female BALB/c nude mice (6-8 weeks old) are inoculated subcutaneously in the right flank with 5×10⁶ NCI-H358 cells. When tumors reach an average volume of 150-200 mm3, mice are randomized into groups (n=8-10). AZD0095 is formulated in a suitable vehicle (e.g., 10% DMSO/40% PEG300/5% Tween-80/45% saline) and administered orally by gavage at doses of 10, 30, or 100 mg/kg, twice daily (BID). The combination group receives AZD0095 (30 mg/kg, BID) plus Cediranib (1.5 mg/kg, once daily). Tumor volumes are measured twice weekly with calipers. Body weight is monitored as an indicator of toxicity. At study termination (21-28 days), tumors are harvested for Western blot analysis of MCT4 expression, immunohistochemistry (IHC) for CD31 (an angiogenesis marker), and for measurement of tumor pH using a microelectrode or a fluorescent pH probe (e.g., SNARF-4F). The combination group would show a significant reduction in tumor growth compared to monotherapies.. |
| ADME/Pharmacokinetics |
Detailed pharmacokinetic (PK) data for AZD0095 is not fully provided. It is described as an orally active inhibitor, indicating it has sufficient oral bioavailability. A standard PK study in mice would involve oral (PO) administration (10 mg/kg) and intravenous (IV) administration (1 mg/kg). Blood samples are collected at multiple time points (0.25, 0.5, 1, 2, 4, 6, 8, 12, and 24 hours post-dose). Plasma concentrations of AZD0095 are quantified by LC-MS/MS. Key parameters, including terminal half-life (T1/2), maximum plasma concentration (Cmax), area under the curve (AUC), and oral bioavailability (F%), would be calculated. The compound is likely metabolized by the liver, possibly by CYP450 enzymes. For research, it is typically dissolved in DMSO (100 mg/mL) for stock solutions and diluted in vehicle for in vivo administration.
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| Toxicity/Toxicokinetics |
Specific toxicological data for AZD0095 is not available. As an MCT4 inhibitor, the primary safety concern is the accumulation of lactate and hydrogen ions in tissues. This could potentially lead to muscle fatigue, lactic acidosis, and neurological disturbances. Standard safety assessment would include a 28-day repeat-dose oral toxicity study in rats and dogs to determine the No-Observed-Adverse-Effect Level (NOAEL). Key endpoints would include monitoring serum lactate and pH levels, liver function tests (ALT, AST), and histological examination of muscle, heart, and liver tissues. An in vitro hERG (human Ether-à-go-go-Related Gene) channel assay would be performed to assess the potential for cardiac QT prolongation.
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| References | |
| Additional Infomation |
AZD0095 is an investigational drug and is not approved for clinical use. It is a highly potent, selective, and orally active inhibitor of monocarboxylate transporter 4 (MCT4), with an IC50 of 1.3 nM and >1000-fold selectivity over MCT1.. MCT4 is upregulated in many cancers (e.g., pancreatic cancer, triple-negative breast cancer, NSCLC) to export the lactate produced by aerobic glycolysis, a hallmark of the Warburg effect. AZD0095 is a valuable tool for studying the role of MCT4 in tumor metabolism, the acidic tumor microenvironment (TME), and cancer immunotherapy.
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| Molecular Formula |
C27H32N8O2
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|---|---|
| Molecular Weight |
500.595384597778
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| Exact Mass |
500.264
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| CAS # |
2750001-23-9
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| PubChem CID |
146241312
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| Appearance |
Light yellow to yellow solid powder
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| LogP |
2.4
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
9
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| Rotatable Bond Count |
6
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| Heavy Atom Count |
37
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| Complexity |
733
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| Defined Atom Stereocenter Count |
1
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| SMILES |
CC1=C(C(C)=NC2=NC(C)=NN12)O[C@@H]1CCN(C2C=CC(C3N=NC(CN4CCOCC4)=CC=3)=CC=2)C1
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| InChi Key |
PHDAGSZQGNWGFE-XMMPIXPASA-N
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| InChi Code |
InChI=1S/C27H32N8O2/c1-18-26(19(2)35-27(28-18)29-20(3)32-35)37-24-10-11-34(17-24)23-7-4-21(5-8-23)25-9-6-22(30-31-25)16-33-12-14-36-15-13-33/h4-9,24H,10-17H2,1-3H3/t24-/m1/s1
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| Chemical Name |
4-[[6-[4-[(3R)-3-[(2,5,7-trimethyl-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)oxy]pyrrolidin-1-yl]phenyl]pyridazin-3-yl]methyl]morpholine
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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: 10 mg/mL (19.98 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: 1 mg/mL (2.00 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 10.0 mg/mL clear DMSO stock solution to 400 μL of PEG300 and mix evenly; then add 50 μL of Tween-80 to the above solution and mix evenly; then add 450 μL of normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 1 mg/mL (2.00 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 10.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 1 mg/mL (2.00 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.9976 mL | 9.9880 mL | 19.9760 mL | |
| 5 mM | 0.3995 mL | 1.9976 mL | 3.9952 mL | |
| 10 mM | 0.1998 mL | 0.9988 mL | 1.9976 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
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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