| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| 50mg |
|
||
| 100mg |
|
||
| Other Sizes |
| Targets |
Fmoc-Val-Ala-aminomethyl acetate itself does not have a direct biological target. Its role is as a linker component in ADCs, where it covalently attaches the antibody to a cytotoxic drug. Upon incorporation into an ADC, the Val-Ala dipeptide sequence serves as a cleavable moiety that is recognized by specific enzymes, such as cathepsin B, which are overexpressed in the tumor microenvironment. This enzymatic cleavage enables the targeted release of the cytotoxic payload inside cancer cells. Therefore, the relevant target is the intracellular protease that cleaves the Val-Ala linker, with the ultimate pharmacological target being the cancer cell's proliferative machinery affected by the released drug.
|
|---|---|
| ln Vitro |
There is no reported direct biological activity or in vitro efficacy for Fmoc-Val-Ala-aminomethyl acetate itself, as it is a synthetic chemical intermediate, not a biologically active compound. Its activity is defined by its utility as a research tool. In the context of ADC development, the linker is assessed in vitro for its stability in plasma and its cleavability by target proteases. The efficacy of an ADC containing this linker is evaluated in cell viability assays against specific cancer cell lines. The linker-payload conjugate's ability to be internalized and release the drug is confirmed through LC-MS/MS detection of the released payload in cell lysates.
|
| ln Vivo |
No in vivo activity is reported for Fmoc-Val-Ala-aminomethyl acetate as a standalone compound. Its in vivo evaluation is conducted as part of a complete ADC molecule that includes an antibody and a payload. In such constructs, the ADC's anti-tumor efficacy is assessed in mouse xenograft models, where it can lead to significant tumor growth inhibition. The safety and pharmacokinetics of the ADC are also evaluated in animal models to determine its therapeutic window. The linker's stability in circulation and efficient cleavage in the tumor microenvironment are critical for the ADC's in vivo performance.
|
| Enzyme Assay |
A typical non-cellular assay for this type of linker involves a stability test in human or mouse plasma. The linker-payload conjugate is incubated in plasma at 37degC for up to 24-48 hours. At predetermined time points, aliquots are taken, and proteins are precipitated. The supernatant is analyzed by LC-MS/MS to quantify the amount of intact linker-payload and released payload. This helps determine the linker's susceptibility to premature cleavage. Another assay uses purified cathepsin B to confirm enzyme-specific cleavage, where the linker is incubated with the enzyme in an appropriate buffer, and the release of the payload is monitored via HPLC or LC-MS.
|
| Cell Assay |
There is no cell-based assay for the linker alone. However, a standard in vitro protocol for evaluating an ADC containing this linker uses a panel of cancer cell lines. Cells are seeded in 96-well plates and treated with serial dilutions of the ADC. After 72-96 hours, cell viability is measured using a luminescent CellTiter-Glo assay to calculate the half-maximal inhibitory concentration (IC₅0). To confirm the mechanism, an excess of unconjugated antibody is used in a competition assay, and cytotoxicity is compared in protease-positive vs. protease-negative cell lines. Uptake and payload release are also visualized via confocal microscopy using fluorescently labeled ADCs.
|
| Animal Protocol |
Animal studies for the linker are conducted as part of an ADC. In a typical efficacy study, 5-6-week-old female BALB/c nude mice are implanted subcutaneously with 5 × 10⁶ cancer cells. When tumors reach an average size of 100-150 mm3, mice are randomized and treated intravenously with the ADC every 4 days for a total of 3-4 doses. Tumor volumes are measured twice weekly with calipers, and body weight is monitored as a safety indicator. At study termination, tumors are excised, weighed, and analyzed for proliferation markers (Ki-67). Tissues are also collected for biodistribution analysis of the payload.
|
| ADME/Pharmacokinetics |
Pharmacokinetic properties are not determined for the isolated linker but for the final ADC. A typical ADC has a long terminal half-life (t1/2) of several days in humans due to the antibody component. The clearance is slow, and volume of distribution is limited. The linker-payload conjugate is designed to be stable in circulation, with minimal release of the free payload in the bloodstream. The primary route of elimination for the ADC is proteolytic degradation, and the released payload is metabolized, primarily in the liver, and excreted in the feces.
|
| Toxicity/Toxicokinetics |
No toxicity data is available for Fmoc-Val-Ala-aminomethyl acetate alone. As a research reagent, standard safety precautions should be taken, assuming it is toxic. The toxicity of an ADC containing this linker is evaluated in preclinical models. Typical findings include bone marrow suppression and hepatotoxicity, which are often attributed to the payload and are dose-limiting. In animal models, a maximum tolerated dose (MTD) is established, and toxicities are monitored through body weight changes, complete blood counts (CBC), and serum chemistry analysis for liver and kidney function, which are correlated with histopathological findings in tissues.
|
| References |
[1]. Charng-Sheng TSAI, et al. Antibody drug conjugates. Patent. WO2023125530
|
| Additional Infomation |
Fmoc-Val-Ala-aminomethyl acetate is not a drug and is not approved for clinical use. It is exclusively a research-grade chemical intermediate for the synthesis of antibody-drug conjugates (ADCs) in oncology. Its mechanism of action is predicated on its use as a cleavable linker that is stable in systemic circulation but is specifically cleaved by intracellular proteases, such as cathepsins, within target cancer cells, enabling the targeted delivery and release of a cytotoxic payload. No clinical trials have been registered for this chemical. For research use only; not for human therapeutic or diagnostic use.
|
| Exact Mass |
481.221
|
|---|---|
| CAS # |
2505045-86-1
|
| PubChem CID |
159391478
|
| Appearance |
White to off-white solid powder
|
| Hydrogen Bond Donor Count |
3
|
| Hydrogen Bond Acceptor Count |
6
|
| Rotatable Bond Count |
11
|
| Heavy Atom Count |
35
|
| Complexity |
750
|
| Defined Atom Stereocenter Count |
2
|
| SMILES |
C[C@@H](C(=O)NCOC(=O)C)NC(=O)[C@H](C(C)C)NC(=O)OCC1C2=CC=CC=C2C3=CC=CC=C13
|
| InChi Key |
MZBRJYLMNFPXHD-HJPURHCSSA-N
|
| InChi Code |
InChI=1S/C26H31N3O6/c1-15(2)23(25(32)28-16(3)24(31)27-14-35-17(4)30)29-26(33)34-13-22-20-11-7-5-9-18(20)19-10-6-8-12-21(19)22/h5-12,15-16,22-23H,13-14H2,1-4H3,(H,27,31)(H,28,32)(H,29,33)/t16-,23-/m0/s1
|
| Chemical Name |
[[(2S)-2-[[(2S)-2-(9H-fluoren-9-ylmethoxycarbonylamino)-3-methylbutanoyl]amino]propanoyl]amino]methyl acetate
|
| HS Tariff Code |
2934.99.9001
|
| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: (1). This product requires protection from light (avoid light exposure) during transportation and storage. (2). Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
|
| Solubility (In Vitro) |
DMSO: 50 mg/mL (103.83 mM)
|
|---|---|
| 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
