| Size | Price | |
|---|---|---|
| Other Sizes |
| Targets |
Non-cleavable Linker
The primary targets of (R)-Azetidine-2-carboxylic acid are the linker structures used in ADC and PROTAC technologies. As a non-cleavable ADC linker, it serves as a stable connector between the antibody and the cytotoxic payload in ADCs. As a PROTAC linker, it connects the ligand for the E3 ubiquitin ligase and the ligand for the target protein, enabling the selective degradation of target proteins via the intracellular ubiquitin-proteasome system. It does not have a traditional enzymatic or receptor target but is a key structural component in targeted protein degradation and antibody-drug conjugation strategies. |
|---|---|
| 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].
In vitro, (R)-Azetidine-2-carboxylic acid functions as a structural linker. ADC cytotoxins are connected to antibodies through this ADC linker to form stable ADCs. In the context of PROTACs, two distinct ligands—one for the E3 ubiquitin ligase and the other for the target protein—are joined by this linker. The resulting PROTAC molecule selectively degrades target proteins through the intracellular ubiquitin-proteasome system. The compound itself does not exhibit direct inhibitory or activating effects on cellular enzymes or receptors but enables the bioactivity of the conjugated molecule. |
| ln Vivo |
In vivo activity of (R)-Azetidine-2-carboxylic acid is derived from the ADC or PROTAC molecules in which it is incorporated. As a non-cleavable linker, it provides stability to the ADC in circulation, ensuring that the cytotoxic payload remains attached to the antibody until the ADC reaches the target cell. For PROTACs, the linker facilitates the formation of a ternary complex between the target protein and E3 ligase, leading to ubiquitination and subsequent proteasomal degradation of the target protein. The in vivo efficacy is dependent on the specific antibody or targeting ligand used in the conjugate.
|
| Enzyme Assay |
A typical in vitro enzyme/receptor binding assay for (R)-Azetidine-2-carboxylic acid involves testing the stability and integrity of the linker under physiological conditions. The compound is incubated in buffer solutions (e.g., PBS, pH 7.4) at 37°C for various time points (0, 24, 48, 72 hours) to assess its chemical stability and resistance to degradation. Samples are analyzed by HPLC or LC-MS to monitor the parent compound and detect any degradation products. The non-cleavable nature of the linker is confirmed by the absence of significant degradation over the tested period, indicating its suitability for stable conjugation in ADC applications.
|
| Cell Assay |
In vitro cellular assays for (R)-Azetidine-2-carboxylic acid typically involve evaluating the activity of the complete ADC or PROTAC molecule in which it is incorporated, rather than the linker alone. For ADC studies, cancer cell lines (e.g., HER2-positive SK-BR-3 cells) are treated with the ADC, and cell viability is measured using assays such as CCK-8 or MTT after 72 hours of incubation. For PROTAC studies, target protein degradation is assessed by Western blot analysis in treated cells. The linker itself does not directly affect cell viability but is essential for the function of the conjugated therapeutic molecule.
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| Animal Protocol |
In vivo animal studies for (R)-Azetidine-2-carboxylic acid are conducted using the final ADC or PROTAC construct. For ADC evaluation, tumor-bearing xenograft models (e.g., mice implanted with human cancer cells) are administered the ADC via intravenous injection at various doses. Tumor volume and body weight are monitored over 2-4 weeks to assess antitumor efficacy and tolerability. For PROTAC evaluation, pharmacokinetic and pharmacodynamic parameters are measured, including target protein degradation in tissues and assessment of downstream biological effects. The linker contributes to the overall stability and pharmacokinetic profile of the conjugate.
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| ADME/Pharmacokinetics |
The pharmacokinetic properties of (R)-Azetidine-2-carboxylic acid are determined by the ADC or PROTAC molecule in which it is incorporated. As a small molecule linker (MW 101.10), it has high aqueous solubility (≥175 mg/mL in water). The compound is stable as a powder at -20°C for 3 years and at 4°C for 2 years. In solvent, it is stable at -80°C for 6 months and at -20°C for 1 month. For in vivo formulations, the linker is typically conjugated to larger molecules, which determines the overall PK profile, including half-life, clearance, and tissue distribution.
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| Toxicity/Toxicokinetics |
The toxicity profile of (R)-Azetidine-2-carboxylic acid is primarily associated with the ADC or PROTAC molecules in which it is used. As a linker compound, it is considered to have low intrinsic toxicity. The compound is classified as a non-cleavable linker, which means it is designed to be stable and not release toxic metabolites. Standard toxicity studies for the final conjugate include assessment of body weight changes, clinical observations, hematological parameters, and histopathological examination of major organs in animal models. The linker itself does not exhibit significant cytotoxic effects in standard cell viability assays.
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| References | |
| Additional Infomation |
(R)-azacyclobutane-2-carboxylic acid is an azacyclobutane-2-carboxylic acid. It is the enantiomer of (S)-azacyclobutane-2-carboxylic acid.
(R)-Azetidine-2-carboxylic acid (CAS 7729-30-8) has a molecular formula of C₄H₇NO₂ and a molecular weight of 101.10. The compound appears as a white to off-white solid powder with a purity of ≥98%. It has a melting point of 209-211°C and a LogP of -0.83, indicating high hydrophilicity. The compound is not approved for clinical use and is intended for research purposes only. It is commonly used in the synthesis of ADCs and PROTACs for targeted cancer therapy and protein degradation research. The (R)-enantiomer is the specific stereoisomer used in these applications. |
| Molecular Formula |
C4H7NO2
|
|---|---|
| Molecular Weight |
101.10
|
| Exact Mass |
101.047
|
| CAS # |
7729-30-8
|
| PubChem CID |
637601
|
| Appearance |
White to off-white solid powder
|
| Density |
1.3±0.1 g/cm3
|
| Boiling Point |
242.0±33.0 °C at 760 mmHg
|
| Melting Point |
209-211ºC
|
| Flash Point |
100.1±25.4 °C
|
| Vapour Pressure |
0.0±1.0 mmHg at 25°C
|
| Index of Refraction |
1.499
|
| LogP |
-0.83
|
| Hydrogen Bond Donor Count |
2
|
| Hydrogen Bond Acceptor Count |
3
|
| Rotatable Bond Count |
1
|
| Heavy Atom Count |
7
|
| Complexity |
91.7
|
| Defined Atom Stereocenter Count |
1
|
| SMILES |
O([H])C(C1([H])C([H])([H])C([H])([H])N1[H])=O
|
| InChi Key |
IADUEWIQBXOCDZ-GSVOUGTGSA-N
|
| InChi Code |
InChI=1S/C4H7NO2/c6-4(7)3-1-2-5-3/h3,5H,1-2H2,(H,6,7)/t3-/m1/s1
|
| Chemical Name |
(2R)-azetidine-2-carboxylic acid
|
| 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 |
| 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
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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 | 9.8912 mL | 49.4560 mL | 98.9120 mL | |
| 5 mM | 1.9782 mL | 9.8912 mL | 19.7824 mL | |
| 10 mM | 0.9891 mL | 4.9456 mL | 9.8912 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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