| Size | Price | Stock | Qty |
|---|---|---|---|
| 10mg |
|
||
| 50mg | |||
| Other Sizes |
| Targets |
Antibody lysine residues (via NHS ester); azide-functionalized molecules (via DBCO for SPAAC); the disulfide bond is cleaved intracellularly by reducing agents (e.g., glutathione).
|
|---|---|
| ln Vitro |
ADC cytotoxins are connected to antibodies through an ADC connector to form ADCs [1].
DBCO-CONH-S-S-NHS ester is a cleavable ADC linker that integrates a DBCO click chemistry group, disulfide bonds, and an NHS ester. This combination enables site-specific conjugation and controlled payload release in antibody-drug conjugates. The DBCO moiety engages azide groups in a bioorthogonal manner, facilitating swift and selective conjugation while upholding native biological processes. This method is indispensable for constructing targeted drug delivery systems and diagnostic assays. The NHS ester group selectively reacts with primary amines on antibodies, enabling efficient labeling and tracking both in vitro and in vivo, essential for probing protein dynamics, interactions, and localization within cellular milieus. The disulfide bond provides a mechanism for controlled, intracellular drug release under reducing conditions (e.g., high glutathione concentrations in the cytoplasm). Proteins can be labeled with fluorescent dyes or other tags, ushering in diverse imaging applications. The linker has no inherent biological activity; its function is purely structural. |
| Enzyme Assay |
sulfo-SPDB is a chemical reagent, not a biologically active compound, and is characterized chemically rather than in enzyme/receptor assays. The purity (>95%) is confirmed by HPLC. The structure is verified by ¹H NMR spectroscopy, which shows characteristic signals for the NHS ester (delta ~2.8 ppm), the pyridyldithio group (delta ~7.2-8.5 ppm for pyridine aromatic protons), and the sulfonate group (delta ~3.0-4.0 ppm). The molecular weight (406.44) is confirmed by mass spectrometry (MS). The reactivity and specificity of the NHS ester group are tested by reaction with a model primary amine (e.g., benzylamine) in a buffer (e.g., PBS, pH 7.4) and monitoring the disappearance of the starting material by LC-MS over time; the half-life of the NHS ester in aqueous buffer is typically 1-2 hours. The pyridyldithio group content is quantified by a thiol release assay: the compound is treated with a reducing agent (e.g., dithiothreitol, DTT), which reduces the disulfide bond, releasing 2-thiopyridone that absorbs at 343 nm; the concentration is calculated using the extinction coefficient (ε343 = 8,080 M-¹cm-¹). For antibody conjugation, a model ADC is prepared by first reacting the NHS ester with antibody lysines at pH 8.0, followed by reduction of antibody interchain disulfides to generate free thiols, and then reaction with the pyridyldithio group. Conjugation efficiency is assessed by SEC-HPLC, and DAR is determined by hydrophobic interaction chromatography (HIC-HPLC) or LC-MS.
|
| Cell Assay |
DBCO-CONH-S-S-NHS ester is a chemical linker used to synthesize ADCs and other bioconjugates; it is not used directly in cell-based assays. The final ADC or bioconjugate is tested in cell-based assays. A typical cell-based assay for an ADC involves testing the ADC on antigen-positive and antigen-negative cells. Target cells are seeded in 96-well plates at 5×103 cells per well. The ADC is added at varying concentrations (typically 0.001-100 nM based on antibody concentration) and incubated for 72-120 hours. Cell viability is measured by CellTiter-Glo or MTT assays. The IC50 for antigen-positive cells is calculated. To confirm the role of the disulfide bond (cleavable mechanism), the assay may be performed in the presence of a reducing agent such as glutathione (10 mM) to enhance payload release, or an endocytosis inhibitor such as dynasore to block activity. For DBCO-based click chemistry applications, cells can be treated with an azide-functionalized probe, followed by addition of DBCO-CONH-S-S-NHS ester conjugated to a fluorophore, and labeling is visualized by confocal microscopy or flow cytometry. The disulfide bond can be cleaved by reducing agents to release the fluorophore.
|
| Animal Protocol |
DBCO-CONH-S-S-NHS ester is not used directly in animal studies. The final ADC or conjugate is tested in vivo. A typical in vivo protocol for an ADC built with this linker involves a murine xenograft model. Female athymic nude mice are injected subcutaneously with 5-10×10⁶ antigen-positive tumor cells in PBS mixed 1:1 with Matrigel. When tumors reach 100-200 mm3, mice are randomized into treatment groups (n=8-10). The ADC is formulated in PBS or a suitable vehicle and administered intravenously (i.v.) via the tail vein at doses of 1-10 mg/kg (based on antibody content). Control groups receive vehicle alone or non-targeting ADC. Tumor volumes are measured with calipers every 3-4 days, and body weights are recorded. Treatment is typically administered once weekly (QW) for 2-4 weeks. At study termination, tumors are excised, weighed, and analyzed for target protein expression and payload release. For imaging applications, DBCO-CONH-S-S-NHS ester conjugated to an imaging agent (e.g., near-infrared dye) may be administered to mice bearing tumors expressing an azide-tagged targeting moiety; the click reaction occurs in vivo, and the signal is visualized using an in vivo imaging system (IVIS).
|
| ADME/Pharmacokinetics |
DBCO-CONH-S-S-NHS ester is a cleavable linker, and its pharmacokinetics are studied as part of the complete ADC or conjugate. The linker itself is not administered as a standalone entity. For an ADC, the PK is characterized by the antibody portion, with the linker influencing stability and payload release. ADC PK is typically biphasic, with a distribution phase (alpha) and a terminal elimination phase (beta) with a half-life of 4-7 days for IgG-based ADCs. The stability of the disulfide bond in circulation is assessed by measuring the concentration of intact ADC versus released payload in plasma over time using LC-MS. In vitro plasma stability assays: the ADC is spiked into human or mouse plasma at 37degC for up to 14 days, and samples are taken at multiple time points for analysis. The DBCO group is stable in biological fluids. For small molecule conjugates, the DBCO group can react with endogenous azides or be reduced; however, the SPAAC reaction is highly selective. The NHS ester is hydrolyzed or reacted during the conjugation process and is not present in the final conjugate.
|
| Toxicity/Toxicokinetics |
DBCO-CONH-S-S-NHS ester is a chemical linker for research use; the toxicity profile is evaluated for the complete ADC or conjugate, not for the linker alone. For ADCs, the primary dose-limiting toxicities typically involve the cytotoxic payload (e.g., neutropenia, peripheral neuropathy, thrombocytopenia). The disulfide bond contributes to safety by enabling targeted payload release inside the tumor, reducing systemic toxicity. Premature cleavage in circulation would increase toxicity. The DBCO group is considered non-toxic and stable. For laboratory handling, DBCO-CONH-S-S-NHS ester should be handled with standard chemical safety precautions: use gloves, lab coat, eye protection. The NHS ester is reactive with amines (including those in skin and mucous membranes) and may cause irritation and sensitization. Avoid inhalation and contact with skin. The compound should be stored at -20degC under inert atmosphere (argon or nitrogen) to prevent hydrolysis of the NHS ester and oxidation of the disulfide bond. Solutions should be prepared fresh and used immediately. The compound is not intended for human use. Detailed toxicological data (LD50, genotoxicity, etc.) is not available for this specific linker.
|
| References | |
| Additional Infomation |
DBCO-CONH-S-S-NHS ester is a versatile cleavable linker widely used in the synthesis of antibody-drug conjugates (ADCs), bioconjugates, and imaging probes. It enables copper-free click chemistry via DBCO-SPAAC, which is highly selective and rapid, with second-order rate constants up to 1 M-¹s-¹. The disulfide bond allows controlled payload release in reducing environments (e.g., intracellular glutathione at ~1-10 mM vs. plasma at ~2-20 microM). The NHS ester is used for conjugation to primary amines on antibodies, proteins, or peptides. This linker is part of the growing class of "cleavable" ADC linkers, which also includes protease-cleavable (Val-Cit) and pH-sensitive (hydrazone) linkers. The product is intended for research and development use only, not for human therapeutic use. The compound should be stored at -20degC in a dry environment protected from light. The disulfide bond is susceptible to reduction by thiol-containing compounds; avoid co-storage with reducing agents such as DTT, TCEP, or beta-mercaptoethanol.
|
| Molecular Formula |
C28H27N3O6S2
|
|---|---|
| Molecular Weight |
565.660484552383
|
| Exact Mass |
565.134
|
| CAS # |
1435934-53-4
|
| PubChem CID |
98233194
|
| Appearance |
White to light yellow solid powder
|
| LogP |
1.7
|
| Hydrogen Bond Donor Count |
1
|
| Hydrogen Bond Acceptor Count |
8
|
| Rotatable Bond Count |
12
|
| Heavy Atom Count |
39
|
| Complexity |
992
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
S(CCNC(CCC(N1C2C=CC=CC=2C#CC2C=CC=CC=2C1)=O)=O)SCCC(=O)ON1C(CCC1=O)=O
|
| InChi Key |
RPNGUSKTYYIUDE-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C28H27N3O6S2/c32-24(29-16-18-39-38-17-15-28(36)37-31-26(34)13-14-27(31)35)11-12-25(33)30-19-22-7-2-1-5-20(22)9-10-21-6-3-4-8-23(21)30/h1-8H,11-19H2,(H,29,32)
|
| Chemical Name |
(2,5-dioxopyrrolidin-1-yl) 3-[2-[[4-(2-azatricyclo[10.4.0.04,9]hexadeca-1(16),4,6,8,12,14-hexaen-10-yn-2-yl)-4-oxobutanoyl]amino]ethyldisulfanyl]propanoate
|
| Synonyms |
DBCOCONHSSNHS ester; DBCO CONH S S NHS ester
|
| 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 : ~100 mg/mL (~176.78 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.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.7678 mL | 8.8392 mL | 17.6785 mL | |
| 5 mM | 0.3536 mL | 1.7678 mL | 3.5357 mL | |
| 10 mM | 0.1768 mL | 0.8839 mL | 1.7678 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
