| Size | Price | Stock | Qty |
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| 50mg |
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| 100mg |
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| 250mg |
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| 1g |
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| Other Sizes |
| Targets |
The pharmacological target of the final product, MMAE, is tubulin, specifically the microtubule network within eukaryotic cells. MMAE is a potent antimitotic agent that binds to the vinca alkaloid binding site on tubulin, thereby inhibiting tubulin polymerization and disrupting the formation of microtubules. This leads to cell cycle arrest at the G2/M phase and the induction of apoptosis in rapidly dividing cancer cells. As an intermediate, this compound does not directly bind to tubulin; however, it is a structural fragment essential for the stereochemistry and activity of the final MMAE molecule.
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| ln Vitro |
MMAE, the final drug product, exhibits extremely potent in vitro cytotoxicity against a wide range of human cancer cell lines, with IC50 values typically in the picomolar to low nanomolar range. For example, MMAE has IC50 values of 0.07 nM against H3396 breast cancer cells and 0.5 nM against L540 lymphoma cells. The compound inhibits the proliferation of cancer cells by targeting the mitotic spindle and causing a block in the G2/M phase. The intermediate itself, however, is not biologically active; its activity is assessed based on its purity and ability to successfully synthesize functional MMAE. It is a building block and is not directly tested in cell viability assays.
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| ln Vivo |
The in vivo antitumor efficacy of MMAE, when used as the payload of an ADC, has been extensively validated in numerous mouse xenograft models. For instance, anti-CD30-MMAE ADC (brentuximab vedotin) has demonstrated profound and durable antitumor responses in models of Hodgkin's lymphoma and anaplastic large cell lymphoma at well-tolerated doses. Similarly, anti-Nectin-4-MMAE ADC has shown efficacy in bladder cancer models. The activity is mediated by the targeted delivery of the potent tubulin inhibitor directly to cancer cells. The intermediate-11 is not tested in vivo directly; instead, its value is in enabling the synthesis of the active pharmaceutical ingredient, MMAE.
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| Enzyme Assay |
Since Monomethyl auristatin E intermediate-11 is a synthetic precursor, no in vitro binding assays are performed on the intermediate itself. The activity of the final product, MMAE, is typically assessed using a tubulin polymerization assay. In this assay, purified tubulin (e.g., from bovine brain) is incubated with varying concentrations of MMAE in a buffer containing GTP. Polymerization is initiated by warming to 37degC, and the increase in absorbance at 340 nm (OD340) is measured over time using a spectrophotometer. The IC50 for inhibition of tubulin polymerization is calculated. Additionally, a competition binding assay with [3H]vinblastine can be used to confirm that MMAE binds to the vinca alkaloid site on tubulin. The intermediate is characterized by NMR and HPLC for structural authenticity and purity (≥98%).
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| Cell Assay |
The intermediate itself is not used in cell-based assays; instead, MMAE is evaluated in cell proliferation assays. Human cancer cell lines (e.g., HeLa, MCF-7, MDA-MB-468, or CD30+ Karpas 299) are seeded in 96-well plates and treated with serial dilutions of MMAE (or the ADC) for 72-96 hours. Cell viability is determined using a colorimetric assay such as MTT, CellTiter-Glo, or an ATP-based luminescence assay. IC50 values are derived by plotting the dose-response curves. To assess the mechanism, cells are stained with propidium iodide (PI) for cell cycle analysis by flow cytometry, and apoptosis is quantified using an Annexin V-FITC/PI double staining assay. The intermediate is used for synthesis and is not added directly to cells.
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| Animal Protocol |
In vivo experiments are performed with the complete ADC or the unconjugated MMAE payload. For xenograft models, immunodeficient mice (e.g., SCID or nude) are subcutaneously or orthotopically implanted with the relevant human cancer cell line (e.g., L540 lymphoma, MDA-MB-468 breast, or OVCAR-3 ovarian). Once tumors reach a certain volume (~100-200 mm3), mice are randomized and treated intravenously with the test ADC (or MMAE in a formulation) on a schedule such as q4d x3, q7d x3, or qd x5. Tumor volumes are measured twice weekly with digital calipers, and body weight is monitored for toxicity. At study endpoints, tumors are excised and analyzed for biomarkers such as phospho-histone H3 (pHH3) to confirm mitotic blockade, and TUNEL staining to measure apoptosis.
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| ADME/Pharmacokinetics |
The pharmacokinetic profile of MMAE (the final product) when administered as an ADC shows a relatively long circulation half-life (days) for the intact ADC, while the released MMAE has a short terminal half-life (approximately 24-48 hours). MMAE is a substrate for the efflux transporter P-glycoprotein (P-gp), which can affect its distribution and resistance profile. It is extensively metabolized by the cytochrome P450 enzyme CYP3A4. The intermediate-11, as a chemical building block, has no relevant PK properties itself. Its logP is 2.8, indicating moderate lipophilicity. It is a colorless to light yellow oil stored at -20degC.
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| Toxicity/Toxicokinetics |
The toxicity of the payload MMAE is primarily dose-limiting neutropenia and peripheral neuropathy due to its potent anti-tubulin activity, which affects rapidly dividing cells in the bone marrow and sensory neurons. MMAE alone is too toxic to be used as a systemic anticancer agent; it must be conjugated to an antibody for targeted delivery. The intermediate itself has not been characterized for specific toxicological endpoints; however, as a primary amine and an alcohol, it may cause skin and eye irritation and is harmful if ingested. Standard laboratory chemical safety practices, including the use of personal protective equipment (gloves, lab coat, eye protection) and proper ventilation, should be employed when handling this material. It is for research use only.
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| References |
[1]. Lisa Buckel, et al. Tumor radiosensitization by monomethyl auristatin E: mechanism of action and targeted delivery. Cancer Res. 2015 Apr 1;75(7):1376-87.
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| Additional Infomation |
Monomethyl auristatin E intermediate-11 is a specialized chemical building block used in the manufacture of the potent cytotoxic agent MMAE. MMAE is the "warhead" payload in several clinically approved and commercially successful antibody-drug conjugates (ADCs), including brentuximab vedotin (Adcetris®) for Hodgkin's lymphoma and systemic anaplastic large cell lymphoma, enfortumab vedotin (Padcev®) for urothelial cancer, and polatuzumab vedotin (Polivy®) for diffuse large B-cell lymphoma. The stereochemistry of this intermediate, specifically the (2S,3S) absolute configuration, is critical for the biological activity of the final drug, as it orients the benzyl and methyl groups in the correct spatial arrangement for binding to tubulin. This product is intended for research and pharmaceutical development purposes only.
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| Molecular Formula |
C14H23NO
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|---|---|
| Molecular Weight |
221.338524103165
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| Exact Mass |
221.177
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| CAS # |
870640-62-3
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| PubChem CID |
59021630
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| Appearance |
Colorless to light yellow oil
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| LogP |
2.8
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
2
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| Rotatable Bond Count |
6
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| Heavy Atom Count |
16
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| Complexity |
178
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| Defined Atom Stereocenter Count |
2
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| SMILES |
CC[C@H](C)[C@@H](CO)N(C)CC1=CC=CC=C1
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| InChi Key |
JPXSDNBZAYWWCL-GXTWGEPZSA-N
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| InChi Code |
InChI=1S/C14H23NO/c1-4-12(2)14(11-16)15(3)10-13-8-6-5-7-9-13/h5-9,12,14,16H,4,10-11H2,1-3H3/t12-,14+/m0/s1
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| Chemical Name |
(2S,3S)-2-[benzyl(methyl)amino]-3-methylpentan-1-ol
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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 | 4.5179 mL | 22.5897 mL | 45.1794 mL | |
| 5 mM | 0.9036 mL | 4.5179 mL | 9.0359 mL | |
| 10 mM | 0.4518 mL | 2.2590 mL | 4.5179 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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