| Size | Price | |
|---|---|---|
| Other Sizes |
| Targets |
Not applicable (synthetic linker, no pharmacological target). NH-bis(C1-Boc) is a non-cleavable linker that connects a monoclonal antibody to a cytotoxic payload in antibody-drug conjugates. The Boc protecting groups (tert-butoxycarbonyl) shield the amine functionalities during synthetic conjugation steps and are removed under acidic conditions to reveal free amines for further conjugation.
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|---|---|
| ln Vitro |
NH-bis(C1-Boc) itself has no direct biological activity. Its utility is defined by its chemical reactivity and stability. The linker is designed to be non-cleavable, meaning the antibody-payload conjugate remains intact until internalized into target cancer cells, where the entire conjugate is degraded in lysosomes, releasing the payload. This contrasts with cleavable linkers (e.g., hydrazone, disulfide, peptide-based) that release payloads extracellularly or in the endosome.
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| ln Vivo |
Since NH-bis(C1-Boc) is a synthetic chemical building block, in vivo activity is only relevant after incorporation into an ADC. ADCs utilizing non-cleavable linkers (e.g., Trastuzumab emtansine, T-DM1) have demonstrated potent in vivo antitumor activity, with linker stability contributing to a broader therapeutic window by reducing premature payload release and off-target toxicity. The DAR (drug-to-antibody ratio) and conjugation site affect ADC stability and efficacy.
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| Enzyme Assay |
Not applicable. NH-bis(C1-Boc) is a chemical reagent used in organic synthesis and ADC linker chemistry. Traditional pharmacological activity assays (receptor binding, enzyme inhibition) are not relevant for this compound. Instead, the following protocol describes typical Boc deprotection and conjugation steps. To remove Boc protecting groups, NH-bis(C1-Boc) is dissolved in anhydrous DCM (0.1-0.2 M) and treated with TFA (trifluoroacetic acid, 10-20 eq) at 0degC to 25degC for 30-60 minutes. Alternatively, HCl in dioxane (4 M, 5-10 eq) can be used. Reaction progress is monitored by TLC (silica gel, ethyl acetate/hexane 1:1, UV detection). After completion, solvents are removed under reduced pressure, and the deprotected diamine is precipitated or used directly in the next conjugation step without further purification.
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| Cell Assay |
Not applicable (no cellular target). NH-bis(C1-Boc) is typically used for chemical synthesis and not for direct addition to cell cultures. For ADC validation, the final ADC conjugate containing a linker derived from NH-bis(C1-Boc) can be tested in vitro. HER2-expressing SK-BR-3 breast cancer cells are seeded in 96-well plates (5,000 cells/well) and treated with varying concentrations of the ADC (0.001-100 nM) for 72-96 hours. Cell viability is measured using CellTiter-Glo. ADC internalization is confirmed by flow cytometry using anti-idiotype or anti-human Fc antibodies.
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| Animal Protocol |
Not applicable. NH-bis(C1-Boc) is not administered to animals. In vivo studies are performed after incorporation of the linker into an ADC. For ADC in vivo efficacy studies, female athymic nude mice bearing subcutaneous xenografts (e.g., NCI-N87 gastric or BT-474 breast tumors, 100-200 mm3) are randomized into treatment groups (n=8-10). ADC (1-10 mg/kg) or vehicle is administered intravenously once weekly for 2-4 weeks. Tumor volumes are measured twice weekly, and body weight is monitored for tolerability. Pharmacokinetic parameters (Cmax, AUC, clearance) are determined from plasma samples using ELISA (antibody-specific) or LC-MS/MS (payload-specific).
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| ADME/Pharmacokinetics |
NH-bis(C1-Boc) is a low-molecular-weight chemical linker (MW = 245.32) with a Boc-protected diamine structure. The linker is non-cleavable, providing enhanced plasma stability (half-life > 2 weeks for the final ADC) compared to cleavable linkers. It is designed for ADC construction through conjugation of the primary amine to the antibody (via amide or carbamate bonds) after Boc deprotection. The linker supports multiple payload or spacer attachments for flexible ADC design.
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| Toxicity/Toxicokinetics |
NH-bis(C1-Boc) is a chemical synthesis intermediate, not a therapeutic agent. Toxicity data are limited to standard laboratory safety: acute oral toxicity is not determined, but chemical hazard classification is not applicable for research use. As a diamine-based linker, it may cause skin and eye irritation. In the context of ADCs, the final conjugate‘s toxicity is primarily determined by the payload (e.g., maytansinoid, auristatin, calicheamicin) and the target antigen expression profile. Non-cleavable linkers generally reduce off-target toxicity.
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| Additional Infomation |
NH-bis(C1-Boc) is a building block for the development of antibody-drug conjugates, which are targeted cancer therapeutics. The bis-Boc-protected diamine structure allows selective deprotection and stepwise conjugation, enabling the synthesis of branched or dual-payload ADCs. It is widely used in academic and industrial ADC research for linker optimization, conjugation strategy development, and drug-to-antibody ratio (DAR) control. The non-cleavable nature of the linker provides a favorable pharmacokinetic profile for the resulting ADC, with reduced premature payload release in circulation. This compound is also used in the synthesis of PROTACs (proteolysis-targeting chimeras) and other bifunctional molecules where a stable diamine spacer is required.
|
| Molecular Formula |
C₁₂H₂₃NO₄
|
|---|---|
| Molecular Weight |
245.32
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| Exact Mass |
245.162
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| CAS # |
85916-13-8
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| PubChem CID |
4393691
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| Appearance |
Colorless to off-white solid powder
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| Density |
1.0±0.1 g/cm3
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| Boiling Point |
298.0±25.0 °C at 760 mmHg
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| Melting Point |
38-42ºC(lit.)
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| Flash Point |
134.0±23.2 °C
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| Vapour Pressure |
0.0±0.6 mmHg at 25°C
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| Index of Refraction |
1.446
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| LogP |
2.72
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
8
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| Heavy Atom Count |
17
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| Complexity |
245
|
| Defined Atom Stereocenter Count |
0
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| SMILES |
O=C(CNCC(OC(C)(C)C)=O)OC(C)(C)C
|
| InChi Key |
SMXMBXPLRFTROI-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C12H23NO4/c1-11(2,3)16-9(14)7-13-8-10(15)17-12(4,5)6/h13H,7-8H2,1-6H3
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| Chemical Name |
tert-butyl 2-[[2-[(2-methylpropan-2-yl)oxy]-2-oxoethyl]amino]acetate
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| Synonyms |
NHbis(C1Boc); NH bis(C1 Boc)
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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 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)
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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 | 4.0763 mL | 20.3815 mL | 40.7631 mL | |
| 5 mM | 0.8153 mL | 4.0763 mL | 8.1526 mL | |
| 10 mM | 0.4076 mL | 2.0382 mL | 4.0763 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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