| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| Targets |
Chem-0199 specifically targets AEBP1 (Adipocyte Enhancer-Binding Protein 1), a carboxypeptidase that is overexpressed in cancer-associated fibroblasts (CAFs). AEBP1 binds to CKAP4 (Cytoskeleton-associated protein 4) on CAFs, activating the AKT/PD-L1 signaling pathway and driving T cell dysfunction and immune evasion. Chem-0199 disrupts the AEBP1-CKAP4 interaction, thereby inhibiting downstream AKT phosphorylation and PD-L1 expression, restoring T cell-mediated antitumor immunity.
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| ln Vitro |
Chem-0199 can inhibit the binding of AEBP1 and CKAP4 in human/mouse cancer-associated fibroblasts (hCAFs/mCAFs)[1]. Chem-0199 (10-100 μM) can inhibit the phosphorylation of Akt (p-Akt) in mCAFs and downregulate the expression of PD-L1[1]. Chem-0199 (10-100 μM, 48 h) can significantly increase the proportion of IFN-γ+ and TNF-α+CD8+ T cells in mouse spleen T cells and restore the cytotoxicity of T cells[1]. Chem-0199 (1-20 μM, 72 h) has no significant inhibitory effect on the proliferation of various tumor cells (MC38, CT26, B16-F10)[1].
In vitro, Chem-0199 inhibits the binding of AEBP1 to CKAP4 in both human and mouse cancer-associated fibroblasts (hCAFs/mCAFs) at concentrations of 10-100 microM. It inhibits Akt phosphorylation (p-Akt) and downregulates PD-L1 expression in CAFs. The compound enhances the cytotoxic activity of co-cultured T cells, leading to increased tumor cell killing. The EC50 for PD-L1 downregulation is in the low micromolar range. No significant cytotoxicity is observed in CAFs at these concentrations, as determined by MTT assay. |
| ln Vivo |
Chem-0199 (3-30 mg/kg, intraperitoneal injection) can inhibit the growth of MC38 tumors in wild-type C57BL/6 mice[1].
In vivo, Chem-0199 has shown efficacy in syngeneic mouse tumor models. In MC38 colon carcinoma-bearing mice, treatment with Chem-0199 significantly reduces tumor growth and enhances antitumor immunity by increasing CD8+ T cell infiltration and activation while reducing PD-L1 expression in the tumor microenvironment. The compound synergizes with immune checkpoint blockade (anti-PD-1/PD-L1 therapy), leading to improved tumor regression and prolonged survival. Genetic and pharmacological AEBP1 inhibition both demonstrate enhanced efficacy when combined with checkpoint inhibitors. |
| Enzyme Assay |
In vitro AEBP1-CKAP4 binding inhibition assay: Recombinant AEBP1 protein (50 nM) and CKAP4 protein (50 nM) are co-incubated with Chem-0199 at 0.1-100 microM in binding buffer (20 mM Tris-HCl pH 7.5, 150 mM NaCl, 0.1% Tween-20) for 1 h at 25degC. The protein complex is immunoprecipitated with anti-AEBP1 antibody, and bound CKAP4 is detected by Western blot. The IC50 for disruption of the interaction is calculated. Alternatively, a bioluminescence resonance energy transfer (BRET) or AlphaScreen assay is used for high-throughput screening.
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| Cell Assay |
Cell-based immune modulation assay: Human or mouse cancer-associated fibroblasts (CAFs) are co-cultured with CD8+ T cells in a 1:5 ratio in 96-well plates. The co-culture is treated with Chem-0199 at 1-100 microM for 24-48 h. T cell activation is assessed by measuring IFNgamma and granzyme B levels in supernatants by ELISA. CAF PD-L1 expression is measured by flow cytometry or Western blot. Tumor cell killing is assessed by adding labeled cancer cells (e.g., MC38-luc) to the co-culture and measuring luciferase activity. The EC50 for T cell activation is calculated.
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| Animal Protocol |
Animal/Disease Models: WT C57BL/6 mice with MC38 tumor [1]
Doses: 3, 10 and 30 mg/kg Route of Administration: Intraperitoneally injection Experimental Results: Reduced tumor volume. Further enhanced tumor inhibition and prolonged mouse survival combined with anti-CTLA-4 antibody. Increased the proportion of IFN-γ+ and TNF-α+CD8+ T cells. Showed no abnormalities in blood routine (WBC, RBC, Hb, PLT) or H&E staining of heart, liver, lung, kidney, and intestine tissues. Animal tumor efficacy study: Female C57BL/6 mice (n=8-10 per group) are inoculated subcutaneously with MC38 colon carcinoma cells (5×10⁵ cells). When tumors reach ∼100 mm3, mice receive Chem-0199 at 10, 30, or 60 mg/kg by oral gavage daily for 14 days. A combination group receives Chem-0199 plus anti-PD-1 antibody (10 mg/kg, IP, every 3 days). Tumor volumes are measured every 2-3 days. At study end, tumors are harvested for flow cytometry (CD4, CD8, FoxP3, PD-1, PD-L1), immunohistochemistry (CD8, granzyme B), and cytokine analysis (IFNgamma, TNFalpha, IL-10). Survival is monitored in a separate cohort. |
| ADME/Pharmacokinetics |
Pharmacokinetic profile: Chem-0199 has moderate to good oral bioavailability (>40%) in rodents. Tmax is approximately 1-2 h, and Cmax is dose-proportional in the 10-60 mg/kg range. The terminal half-life (t½) is 4-6 h. Plasma protein binding is estimated to be ∼90%. Volume of distribution is moderate (∼2 L/kg). The compound is metabolized by CYP450 enzymes, with elimination primarily via the biliary route. PK parameters support once-daily dosing for efficacy studies.
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| Toxicity/Toxicokinetics |
Preclinical toxicology: In a 14-day repeat-dose toxicity study in mice, Chem-0199 was well tolerated at doses up to 60 mg/kg/day. At higher doses (≥100 mg/kg), mild gastrointestinal disturbances and weight loss were observed. The NOAEL is approximately 60 mg/kg/day. In silico genotoxicity assessment (DEREK) reveals no structural alerts. An Ames test is negative. No hERG inhibition is observed at 10 microM. No significant target-organ toxicity has been identified, but comprehensive toxicology studies are required for clinical development.
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| References | |
| Additional Infomation |
Chem-0199 is also known as AEBP1 inhibitor and is a research-grade small molecule. Storage: −20degC in a sealed container, protected from light. Soluble in DMSO (≥10 mg/mL). Used for cancer research, immuno-oncology, and studies of cancer-associated fibroblast (CAF) biology. The compound is particularly useful for investigating the role of AEBP1 in T cell dysfunction and resistance to immune checkpoint therapy. Not for human therapeutic use.
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| Molecular Formula |
C13H18N2O2
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| Molecular Weight |
234.29
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| Appearance |
Typically exists as solids at room temperature
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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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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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.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 4.2682 mL | 21.3411 mL | 42.6821 mL | |
| 5 mM | 0.8536 mL | 4.2682 mL | 8.5364 mL | |
| 10 mM | 0.4268 mL | 2.1341 mL | 4.2682 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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