| Size | Price | Stock | Qty |
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| 100mg |
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| 250mg |
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| 500mg |
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| 1g |
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| Other Sizes |
| Targets |
As a synthetic PROTAC linker, this compound has no direct biological target. It serves as a rigid, short spacer to connect an E3 ubiquitin ligase ligand and a target protein ligand. The cyclobutane ring imposes a fixed exit vector, which can pre-organize the PROTAC for optimal ternary complex formation. The primary amine can be acylated, and the primary alcohol can be activated (e.g., tosylate) for nucleophilic substitution.
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| ln Vitro |
PROTAC contains two distinct ligands linked by a single linker: one is the ligand for the E13 ubiquitin ligase, and the other is the ligand for the target protein. PROTAC utilizes the intracellular ubiquitin-proteasome system to selectively degrade the target protein.
No direct in vitro activity is reported for the isolated linker. In the final PROTAC, the rigid cyclobutyl linker influences degradation parameters (DC50 and Dmax). The constrained geometry can lead to improved degradation selectivity and lower DC50 values compared to flexible linkers. Degradation is measured in cell-based assays (Western blot or HTRF) using target-expressing cells after 24-48 hours of treatment. |
| ln Vivo |
No direct in vivo activity is reported for the linker alone. For the final PROTAC, in vivo efficacy is evaluated in animal models (e.g., xenografts) by measuring target protein knockdown in tissues via IHC or Western blot and assessing tumor growth inhibition. The cyclobutane ring is generally metabolically stable and can reduce off-target degradation compared to flexible linkers. The hydrochloride salt improves solubility for IV administration.
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| Enzyme Assay |
Not applicable for the isolated linker. For the final PROTAC, the primary amine is acylated with a carboxylic acid ligand using HATU or EDCI. The primary alcohol can be tosylated (TsCl, pyridine) or mesylated (MsCl, TEA) for nucleophilic substitution, or oxidized to an aldehyde/carboxylic acid for further conjugation. The cyclobutane ring structure is confirmed by NMR (characteristic upfield shifts). Binding affinity of the final PROTAC to its target is measured by SPR or TR-FRET.
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| Cell Assay |
For in vitro cell assays, the final PROTAC (containing this linker) is dissolved in DMSO to a 10-50 mM stock. Seed target-expressing cells (e.g., 5×10⁵ cells/well in 6-well plates) and allow to adhere overnight. Treat cells with increasing concentrations of the PROTAC (0.1 nM to 10 uM) for 48 hours. Harvest cells, wash with cold PBS, and lyse in RIPA buffer with protease inhibitors (30 min on ice). Centrifuge lysates at 16,000g for 15 min at 4degC. Quantify protein concentration by BCA assay. Perform Western blot using target-specific primary antibodies (1:1000) and HRP-conjugated secondary antibodies (1:5000). Develop with ECL and quantify band intensity by densitometry to determine DC50.
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| Animal Protocol |
For in vivo animal experiments, formulate the final PROTAC in a vehicle of 10% DMSO, 40% PEG300, 5% Tween 80, and 45% saline. Administer 10-30 mg/kg via intraperitoneal (IP) injection to BALB/c nude mice bearing subcutaneous xenografts (n=5 per group). Collect blood via tail vein at 0, 0.5, 1, 2, 4, 8, 12, and 24 hours post-dose. Separate plasma by centrifugation and store at -80degC. Add internal standard and acetonitrile to plasma (50 uL), vortex, centrifuge, and analyze supernatant by LC-MS/MS. Harvest tumors at termination for Western blot analysis of target protein levels.
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| ADME/Pharmacokinetics |
No PK data is available for the isolated linker. For the final PROTAC, the rigid cyclobutyl linker imparts good metabolic stability. Typical rodent PK parameters include a half-life (t1/2) of 3-8 hours, clearance (CL) of 15-35 mL/min/kg, volume of distribution (Vd) of 1-4 L/kg, and oral bioavailability (F%) of 10-25%. The polar hydroxymethyl and amine groups can increase solubility and reduce Vd. The primary amine may be acetylated in vivo.
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| Toxicity/Toxicokinetics |
No toxicity data is available for the isolated linker. For the final PROTAC, safety is evaluated in a 7-day exploratory toxicity study in rats (n=3/sex/group) at doses of 10, 30, and 100 mg/kg (IP). Endpoints include clinical observations, body weight, serum chemistry (ALT, AST, BUN, creatinine), and histopathology of major organs. Cyclobutane rings are generally non-toxic. The hydrochloride salt form is well-tolerated at typical doses.
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| Additional Infomation |
This PROTAC linker has the molecular formula C5H12ClNO and a molecular weight of 137.61 g/mol. It appears as a white to off-white crystalline solid. Purity is typically ≥95%. Storage: 2-8degC for 3 years (powder) or -80degC for 6 months (in solvent). Soluble in DMSO and water (due to hydrochloride salt). It is also known as cis-3-Amino-cyclobutanemethanol hydrochloride and 3-aminocyclobutane-1-methanol hydrochloride.
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| Molecular Formula |
C5H12CLNO
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|---|---|
| Molecular Weight |
137.61
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| CAS # |
130369-06-1
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| Appearance |
White to off-white solid powder
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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 | 7.2669 mL | 36.3346 mL | 72.6691 mL | |
| 5 mM | 1.4534 mL | 7.2669 mL | 14.5338 mL | |
| 10 mM | 0.7267 mL | 3.6335 mL | 7.2669 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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