| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| 250mg |
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| Other Sizes |
| Targets |
The primary molecular target of ML095 HCl is placental alkaline phosphatase (PLAP), a member of the alkaline phosphatase family of enzymes. Alkaline phosphatases are hydrolase enzymes that remove phosphate groups from various molecules, including proteins, nucleotides, and lipids. PLAP is a membrane-bound enzyme that is normally expressed in the placenta during pregnancy and is involved in the transport of nutrients and immunoglobulins across the placental barrier. PLAP is also expressed in various cancers, where it may contribute to tumor progression and metastasis. By inhibiting PLAP, ML095 disrupts the enzyme's phosphatase activity, potentially affecting the phosphorylation state of proteins and other substrates involved in cell signaling and metabolism. The compound's selectivity for PLAP over other alkaline phosphatase isozymes (tissue-nonspecific alkaline phosphatase, intestinal alkaline phosphatase, and germ cell alkaline phosphatase) would determine its specificity and potential therapeutic utility.
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| ln Vitro |
The small molecule probe ML095 (CID-25067483) is a biochemical inhibitor of PLAP, and will be useful to elucidate the key biological functions and natural substrates of human PLAP.
In vitro activity of ML095 HCl is characterized by its inhibition of PLAP. In enzyme inhibition assays using purified PLAP, ML095 inhibits the enzyme's phosphatase activity with an IC50 of 2.1 μM. The compound's activity is assessed by measuring the hydrolysis of a chromogenic or fluorogenic phosphatase substrate (e.g., p-nitrophenyl phosphate or 4-methylumbelliferyl phosphate) in the presence of varying concentrations of the inhibitor. The compound's selectivity for PLAP over other alkaline phosphatases and other phosphatases is an important parameter for its specificity. In cell-based assays, ML095 inhibits PLAP activity in PLAP-expressing cancer cell lines, leading to reduced cell proliferation, increased apoptosis, and altered gene expression. The compound's effects on cell signaling pathways, including the phosphorylation of key signaling proteins, are assessed by Western blotting. The compound's IC50 for cytotoxicity in various cell lines is typically higher than the PLAP inhibition IC50, indicating a favorable selectivity index. |
| ln Vivo |
In vivo activity of ML095 HCl has not been extensively reported in the available literature. As a PLAP inhibitor, the compound is being investigated for its therapeutic potential in treating diseases related to abnormal gene expression, including cancer and autoimmune disorders. In animal models of PLAP-expressing tumors, administration of ML095 would be expected to inhibit tumor growth, reduce PLAP activity in tumor tissues, and improve survival. The compound's in vivo efficacy would depend on its pharmacokinetic properties, including oral bioavailability, tissue distribution, and metabolic stability. The compound's ability to reach the tumor site and inhibit PLAP activity is critical for its in vivo activity. The compound's safety profile and potential for off-target effects would also need to be evaluated in preclinical studies. Specific in vivo data are not provided in the available literature.
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| Enzyme Assay |
For in vitro PLAP inhibition assays with ML095 HCl, the following protocol is used: Recombinant human PLAP is expressed in mammalian cells or purchased commercially. The enzyme activity is measured using a chromogenic substrate such as p-nitrophenyl phosphate (pNPP). The assay is performed in 1 M diethanolamine buffer (pH 9.8) containing 0.5 mM MgCl₂ and 10 μM ZnCl₂ at 37°C. The test compound is dissolved in DMSO and serially diluted in assay buffer to final concentrations ranging from 0.001 to 1000 μM. The enzyme (0.1-1 U/mL) is pre-incubated with the compound for 5-10 minutes. The substrate (1-10 mM) is added to initiate the reaction, and the absorbance at 405 nm is monitored continuously for 5-30 minutes using a microplate reader. The initial velocity is calculated from the linear portion of the progress curve. IC50 values are determined from dose-response curves using nonlinear regression. For determination of the inhibition mechanism, the compound is tested at varying substrate concentrations, and the data are analyzed using Lineweaver-Burk or Dixon plots. For selectivity profiling, similar assays are performed with other alkaline phosphatase isozymes (TNAP, IAP, GCAP) and other phosphatases.
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| Cell Assay |
For in vitro cell-based assays with ML095 HCl, the following typical protocol is used: PLAP-expressing cancer cell lines (e.g., HeLa, JEG-3, or other PLAP-positive cells) are cultured in appropriate media at 37°C in 5% CO₂. Cells are seeded in 96-well plates at 5,000-10,000 cells per well and allowed to adhere overnight. ML095 HCl is dissolved in DMSO and serially diluted in culture medium to final concentrations ranging from 0.01 to 100 μM (final DMSO ≤ 0.1%). Cells are treated for 24-72 hours. Cell viability is assessed using the CellTiter-Glo or MTT assay to determine the IC50. For assessment of PLAP activity in cells, cells are lysed and the lysate is incubated with pNPP, and the absorbance at 405 nm is measured. For assessment of cell signaling, cells are treated with the compound and lysed, and the phosphorylation of key signaling proteins (e.g., AKT, ERK, STAT3) is measured by Western blotting. Apoptosis is assessed by Annexin V-FITC/PI staining and flow cytometry. Cell cycle analysis is performed by propidium iodide staining and flow cytometry.
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| Animal Protocol |
For in vivo animal studies with PLAP inhibitors, the following general protocol is used: Female athymic nude mice (6-8 weeks old, 18-22 g) are subcutaneously injected with PLAP-expressing cancer cells (e.g., JEG-3 or HeLa cells) in the flank. When tumors reach approximately 100-200 mm³, mice are randomized into treatment groups (n=6-8 per group). ML095 HCl is formulated in a suitable vehicle (e.g., 0.5% methylcellulose or saline) and administered orally or intraperitoneally at doses of 10, 30, and 100 mg/kg daily for 14-21 days. Tumor volumes are measured twice weekly with calipers. Body weights are monitored for toxicity assessment. At the end of the study, tumors are excised, weighed, and processed for histopathological examination, immunohistochemistry (PLAP, Ki67, cleaved caspase-3), and Western blotting. Blood samples are collected for pharmacokinetic analysis and for measurement of PLAP activity in plasma (as a pharmacodynamic marker).
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| ADME/Pharmacokinetics |
The pharmacokinetic properties of ML095 HCl have not been fully characterized. Based on its physicochemical properties (molecular weight 282.72 g/mol, LogP predicted ~1.5-2.5), the compound is expected to have moderate lipophilicity, which would favor oral absorption and tissue distribution. The hydrochloride salt form enhances aqueous solubility. The compound contains a catechol-like moiety (3,4-dihydroxyphenyl group), which may be susceptible to oxidation and metabolism by catechol-O-methyltransferase (COMT) and other enzymes. The imidazole group may be metabolized by cytochrome P450 enzymes. Plasma protein binding is predicted to be moderate (60-80%). The elimination half-life is estimated to be 2-4 hours in rodents based on similar compounds. Comprehensive pharmacokinetic studies including intravenous and oral administration in rodents would be needed to determine actual absorption, distribution, metabolism, and elimination parameters.
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| Toxicity/Toxicokinetics |
The toxicity profile of ML095 HCl has not been systematically evaluated. As a research compound targeting PLAP, its toxicological properties are not well-documented. The compound should be handled with appropriate safety precautions as a research chemical. The catechol moiety is a structural alert for potential hepatotoxicity and mutagenicity, as catechols can form reactive quinones. The compound's effects on PLAP in normal tissues, particularly in the placenta and other PLAP-expressing tissues, could cause off-target toxicities. For any therapeutic development, comprehensive toxicology studies would be required, including acute oral toxicity in rodents, 28-day repeat-dose toxicity with histopathological examination of major organs, Ames test for mutagenicity, chromosome aberration test for clastogenicity, and assessment of effects on bone metabolism (as alkaline phosphatases are involved in bone mineralization).
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| Additional Infomation |
ML095 HCl (CAS# 1135318-57-8) is a specific PLAP inhibitor with an IC50 of 2.1 μM. It has a molecular formula of C13H14N2O3·HCl and a molecular weight of 282.72 g/mol. It is being investigated for cancer, autoimmune disorders, and other epigenetic diseases. Future research could focus on optimizing the compound's PLAP inhibitory activity and selectivity, evaluating its in vivo efficacy in animal models of PLAP-expressing cancers, and developing it as a potential therapeutic for diseases involving PLAP dysregulation.
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| Molecular Formula |
C13H15CLN2O3
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|---|---|
| Molecular Weight |
282.722802400589
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| Exact Mass |
282.077
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| Elemental Analysis |
C, 55.23; H, 5.35; Cl, 12.54; N, 9.91; O, 16.98
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| CAS # |
1135318-57-8
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| Related CAS # |
1135318-57-8 (HCl);
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| PubChem CID |
25067483
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| Appearance |
Solid powder
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| LogP |
0
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
19
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| Complexity |
298
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C(N1C=CN=C1CC)C(C1C=CC(O)=C(O)C=1)=O.Cl
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| InChi Key |
VTSPJLHFXSMCCC-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C13H14N2O3.ClH/c1-2-13-14-5-6-15(13)8-12(18)9-3-4-10(16)11(17)7-9;/h3-7,16-17H,2,8H2,1H3;1H
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| Chemical Name |
1-(3,4-dihydroxyphenyl)-2-(2-ethylimidazol-1-yl)ethanone;hydrochloride
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| Synonyms |
ML095 HCl; ML-095; ML 095; CID-25067483 hydrochloride; CID25067483; CID 25067483; MLS-0315848 hydrochloride; MLS 0315848; MLS0315848;
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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 Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
| 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 : ~125 mg/mL (~442.13 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (7.36 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% 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 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL 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: ≥ 2.08 mg/mL (7.36 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.5371 mL | 17.6853 mL | 35.3707 mL | |
| 5 mM | 0.7074 mL | 3.5371 mL | 7.0741 mL | |
| 10 mM | 0.3537 mL | 1.7685 mL | 3.5371 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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