| Size | Price | Stock | Qty |
|---|---|---|---|
| 50mg |
|
||
| 100mg |
|
||
| 250mg | |||
| Other Sizes |
| Targets |
Amino-Tri-(carboxyethoxymethyl)-methane does not have a specific biological target, as it is a chemical linker rather than a pharmacologically active compound. Its function is purely structural, serving as a covalent connector between two or more molecular components. The "targets" of this linker are the specific functional groups on the molecules it is designed to conjugate. The three terminal carboxylic acid groups are the primary reactive sites, acting as targets for conjugation with molecules that contain primary amine groups. This reaction, typically facilitated by coupling reagents like EDC or HATU, forms stable amide bonds. The central amine group can also serve as a target for further functionalization, for example, by reacting with activated esters. The cleavable nature of the linker means it is designed to be a target for intracellular conditions, such as the acidic pH of lysosomes or the action of specific enzymes, which will break the linker and release the payload. Therefore, the biological "target" is the intracellular environment of the target cell. In the context of ADC design, the linker's role is to ensure that the potent cytotoxic drug is stably attached to the antibody in circulation but is efficiently released once the ADC is internalized by the cancer cell.
|
|---|---|
| ln Vitro |
ADC cytotoxins are connected to antibodies through an ADC connector to form ADCs [1]. Two distinct ligands, one for the E3 ubiquitin ligase and the other for the target protein, are present in PROTAC and are joined by a linker. PROTAC selectively degrades target proteins by means of the intracellular ubiquitin-proteasome system [2].
As a chemical linker, Amino-Tri-(carboxyethoxymethyl)-methane does not possess any direct in vitro biological activity, such as enzyme inhibition, receptor binding, or cytotoxicity. Its biological relevance is entirely indirect, stemming from its use in the synthesis of bioactive conjugates. The in vitro activity of this linker is assessed through the characterization of the final conjugates it helps to create. For instance, when used to synthesize an ADC, the activity of the conjugate is evaluated in cell-based assays for target binding, internalization, and cytotoxicity. The cleavable linker contributes to the conjugate's overall activity by ensuring the efficient release of the toxic payload inside the target cell, which is a critical factor for the ADC's potency. In PROTAC synthesis, the linker's role is to connect the target protein ligand and the E3 ligase ligand. Its length and flexibility are crucial for the formation of the ternary complex and the subsequent ubiquitination and degradation of the target protein. Thus, the "activity" of Amino-Tri-(carboxyethoxymethyl)-methane is a measure of its ability to function as a stable, cleavable bridge that enables the design of effective targeted therapeutics. |
| ln Vivo |
As a chemical linker, Amino-Tri-(carboxyethoxymethyl)-methane does not possess any direct in vivo biological activity. It is not administered as a therapeutic agent and does not exert pharmacological effects in animal models. Its in vivo relevance is strictly as a component of larger, biologically active conjugates. When incorporated into an ADC, the cleavable linker plays a critical role in the drug's in vivo efficacy. The linker must be stable in the bloodstream (to prevent premature release of the toxic payload and systemic toxicity) but readily cleavable once the ADC is internalized into the target cell (to ensure efficient drug release). This property is known as the "bystander effect" and is a key design consideration for ADCs. The specific in vivo activity—such as tumor regression—is determined by the conjugate's warhead and targeting ligand, but the linker's stability and cleavage kinetics are essential for the overall therapeutic outcome. Similarly, for a PROTAC, the linker's properties affect the compound's oral bioavailability, cellular permeability, and ability to form the ternary complex in vivo. Therefore, while Amino-Tri-(carboxyethoxymethyl)-methane has no direct in vivo activity, it is a critical component that influences the efficacy and safety of the therapeutics it is used to construct.
|
| Enzyme Assay |
There are no specific in vitro enzyme or receptor binding assays for Amino-Tri-(carboxyethoxymethyl)-methane, as it is not a biologically active compound that interacts with proteins. It is a chemical reagent, and its characterization is performed using analytical chemistry techniques. The quality and identity of the linker are typically confirmed by methods such as Nuclear Magnetic Resonance (NMR) spectroscopy, High-Performance Liquid Chromatography (HPLC), and Mass Spectrometry (MS) to verify its structure and purity (often >98%). If an assay were to be conducted to confirm its reactivity, it would be a chemical conjugation reaction rather than a biological binding assay. For example, the reactivity of the carboxylic acid groups could be tested by reacting the linker with a model amine-containing compound and monitoring the formation of the amide bond by HPLC or LC-MS. The cleavable nature of the linker could be tested in vitro by incubating it in simulated physiological conditions, such as a buffer at acidic pH (e.g., pH 5.0) or in the presence of specific enzymes, and monitoring the release of a model payload by HPLC. These are quality control and stability studies, not biological assays.
|
| Cell Assay |
There are no standard in vitro cell-based assays for Amino-Tri-(carboxyethoxymethyl)-methane, as it is not a bioactive molecule intended to affect cellular function. Its use in cell biology is indirect, as a reagent for constructing ADCs or PROTACs. In such cases, the cell-based assay would involve treating cells with the final conjugated therapeutic and then measuring a biological readout, such as cell viability (for an ADC's cytotoxicity) or target protein levels (for a PROTAC's degradation activity). The performance of the linker would be evaluated by the potency and selectivity of the conjugate, which depend on the linker's ability to release the payload or facilitate the ternary complex. However, there is no direct assay for the linker itself in cells, as it is not designed to interact with cells in its unconjugated form. The compound's solubility in DMSO (10 mM) and its stability under defined storage conditions ensure it can be used effectively in these applications without causing cellular artifacts. Any cell-based work involving Amino-Tri-(carboxyethoxymethyl)-methane is always part of a larger experimental design to evaluate a specific therapeutic conjugate.
|
| Animal Protocol |
Amino-Tri-(carboxyethoxymethyl)-methane is not used in in vivo animal experiments as a standalone compound, as it has no direct biological activity. It is a synthetic building block for creating ADCs and PROTACs. Animal studies involving this compound would only be conducted on the final, larger conjugates that incorporate it. In such studies, the linker's contribution to the overall pharmacokinetic and pharmacodynamic profile of the conjugate would be assessed. For example, researchers might compare the in vivo efficacy, stability, and toxicity of an ADC synthesized with a cleavable linker like this one to an ADC synthesized with a non-cleavable linker. The cleavable linker is expected to release the toxic payload inside the tumor, leading to better efficacy and a wider therapeutic index. However, there is no standard animal protocol for the linker itself. Its use is limited to the research and development phase, where it is employed in the chemical synthesis of test articles. The storage recommendations (-20°C for powder, -80°C for solution) are for maintaining the chemical integrity of the reagent. In summary, there are no in vivo animal experiments for Amino-Tri-(carboxyethoxymethyl)-methane as a single agent.
|
| ADME/Pharmacokinetics |
The pharmacokinetic (PK) properties of Amino-Tri-(carboxyethoxymethyl)-methane are not studied as it is not a therapeutic agent. However, its physicochemical properties provide insight into its behavior. It has a molecular weight of 337.32 g/mol and a molecular formula of C13H23NO9, indicating it is a relatively small, highly polar molecule with multiple ionizable carboxylic acid groups (pKa ~4-5). These properties suggest it would be highly water-soluble at physiological pH, but its logP is not reported. As a small, hydrophilic molecule, it would be expected to have a short half-life if introduced into the bloodstream, likely being rapidly cleared by renal filtration. However, when conjugated to larger molecules like antibodies, the overall PK is dominated by the larger component. The compound is a solid and is soluble in DMSO (10 mM). For storage, it is recommended to keep it at -20°C, where it is stable for up to three years, and for two years in solution at -80°C. The "PK" properties of this linker are thus not an independent consideration but are integral to the design and performance of the bioconjugates it helps to create, particularly its cleavable nature which is designed to release the payload specifically in the target tissue.
|
| Toxicity/Toxicokinetics |
As a PEG-based linker, Amino-Tri-(carboxyethoxymethyl)-methane is considered to have low toxicity. PEG polymers are widely regarded as biocompatible, non-toxic, and non-immunogenic, which is why they are frequently used in pharmaceuticals and biomedical research. The compound is classified as a research reagent, and standard laboratory safety precautions should be followed when handling it. The safety of this linker is primarily considered in the context of its use in ADCs. A cleavable linker is designed to be stable in circulation but to release its payload inside the target cell. If the linker were to be cleaved prematurely, it could lead to the release of the toxic payload into the bloodstream, causing off-target toxicity. Therefore, the toxicity of the linker itself is not the primary concern, but rather the toxicity of the payload it is designed to carry if the linker fails to perform its function correctly. Its toxicity profile has not been extensively studied independently, but due to its intended use and chemical class, it is not expected to be acutely toxic. All handling should be done with appropriate personal protective equipment (PPE) in a well-ventilated area.
|
| References |
[1]. Markus Ribbert, et al. Self coupling recombinant antibody fusion proteins. WO2009013359A2.
[2]. David Margulies, et al. Fluorescent molecular sensor for targeting changes in protein surfaces, and methods of use thereof. WO2015166491A2. |
| Additional Infomation |
Amino-Tri-(carboxyethoxymethyl)-methane is a research-use-only reagent and is not a drug, nor is it approved for any clinical or therapeutic use. It is a highly versatile chemical tool with key applications in biomedical research and industry. Its primary application is as a cleavable PEG linker in the synthesis of antibody-drug conjugates (ADCs) and PROTACs. In ADCs, it serves as a bridge to connect a potent cytotoxic drug to a targeting antibody, ensuring the drug is delivered specifically to cancer cells and released intracellularly. In PROTACs, it links a target protein ligand to an E3 ligase ligand, facilitating targeted protein degradation. Beyond these cutting-edge therapeutic applications, Amino-Tri-(carboxyethoxymethyl)-methane also functions as a chelating agent in various industries. It is used in water treatment to bind and sequester metal ions, preventing scale formation and removing heavy metals. It is also a key ingredient in biodegradable detergents and cleaning agents, where its chelating properties enhance cleaning efficacy while being environmentally sustainable. Its trifunctional structure and cleavable nature make it a unique and valuable compound for both advanced drug delivery and industrial applications.
|
| Molecular Formula |
C13H23NO9
|
|---|---|
| Molecular Weight |
337.323024988174
|
| Exact Mass |
337.137
|
| CAS # |
174362-95-9
|
| Related CAS # |
Amino-Tri-(carboxyethoxymethyl)-methane hydrochloride;1416771-72-6
|
| PubChem CID |
58107553
|
| Appearance |
Colorless to off-white Solid-Liquid Mixture
|
| LogP |
-5.1
|
| Hydrogen Bond Donor Count |
4
|
| Hydrogen Bond Acceptor Count |
10
|
| Rotatable Bond Count |
15
|
| Heavy Atom Count |
23
|
| Complexity |
331
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
C(COCC(COCCC(=O)O)(COCCC(=O)O)N)C(=O)O
|
| InChi Key |
PZOSHMSOMAHCEX-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C13H23NO9/c14-13(7-21-4-1-10(15)16,8-22-5-2-11(17)18)9-23-6-3-12(19)20/h1-9,14H2,(H,15,16)(H,17,18)(H,19,20)
|
| Chemical Name |
3-[2-amino-3-(2-carboxyethoxy)-2-(2-carboxyethoxymethyl)propoxy]propanoic acid
|
| Synonyms |
PUN62959 PUN 62959 PUN-62959Amino-Tri-(carboxyethoxymethyl)-methane
|
| 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: This product requires protection from light (avoid light exposure) during transportation and storage. |
| 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) |
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
|
|---|---|
| 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 | 2.9645 mL | 14.8227 mL | 29.6454 mL | |
| 5 mM | 0.5929 mL | 2.9645 mL | 5.9291 mL | |
| 10 mM | 0.2965 mL | 1.4823 mL | 2.9645 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
