| Size | Price | Stock | Qty |
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| 25mg |
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| 50mg |
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| 100mg |
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| 250mg |
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| 500mg | |||
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| Targets |
Cleavable Linker
The molecular target of Boc-Dap-NE is not a biological target per se, as it is a chemical linker used in the synthesis of ADCs rather than a therapeutic agent. In the context of an ADC, the linker connects the antibody (which targets a cell surface antigen) to the cytotoxic drug payload. Boc-Dap-NE is a cleavable linker that is designed to release the drug payload upon enzymatic cleavage in the target cell. The dipeptide structure may be recognized and cleaved by lysosomal proteases such as cathepsin B, which are overexpressed in many cancer cells. The Boc-protected amino group provides stability during synthesis and can be deprotected when needed for conjugation. The ultimate biological target of the complete ADC would be the specific antigen recognized by the antibody, which could be HER2, CD30, CD33, or other tumor-associated antigens. The linker's role is to deliver the cytotoxic payload specifically to tumor cells, thereby maximizing efficacy and minimizing systemic toxicity. |
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| ln Vitro |
ADCs are made up of an antibody and an ADC cytotoxin that are joined together by an ADC linker.
In vitro activity of Boc-Dap-NE is not assessed as a standalone compound, as it is a linker molecule rather than a therapeutic agent. Its activity is evaluated in the context of the complete ADC, where the linker's role is to deliver the cytotoxic payload to target cells. In cell-based assays, ADCs constructed with Boc-Dap-NE would be tested for antigen-specific cytotoxicity against cancer cell lines expressing the target antigen. The linker's efficiency is assessed by measuring the potency of the ADC (IC50) compared to the free drug, as well as the selectivity for antigen-positive versus antigen-negative cells. The stability of the linker in serum is also evaluated to ensure that the drug is not released prematurely. The cleavage efficiency of the linker by cathepsin B or other lysosomal proteases is assessed in vitro using recombinant enzymes or cell lysates. Specific IC50 values would depend on the antibody, the drug payload, and the target antigen. |
| ln Vivo |
In vivo activity of Boc-Dap-NE is evaluated in the context of complete ADCs in animal models. ADCs incorporating this linker would be tested in xenograft models where human tumor cells expressing the target antigen are implanted in immunodeficient mice. The ADC would be administered intravenously at doses typically ranging from 1 to 30 mg/kg, often on a weekly or biweekly schedule for 2-4 weeks. Tumor growth inhibition would be measured, along with survival extension. The linker's in vivo stability, pharmacokinetics, and biodistribution would also be assessed. ADCs with cleavable linkers such as Boc-Dap-NE are designed to release the drug payload specifically within tumor cells, thereby maximizing efficacy and minimizing systemic toxicity. The in vivo efficacy of the ADC would depend on the antibody targeting, the potency of the drug payload, and the linker's cleavage efficiency in the tumor microenvironment.
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| Enzyme Assay |
For in vitro linker conjugation and stability assays with Boc-Dap-NE, the following protocol is used: The antibody (e.g., trastuzumab) is reduced with TCEP (tris(2-carboxyethyl)phosphine) at 37°C for 2 hours to generate free thiol groups on cysteine residues. The number of free thiols is determined using Ellman's reagent. Boc-Dap-NE is dissolved in DMSO and added to the reduced antibody in a buffer containing 20 mM histidine (pH 5.5) and 2 mM EDTA, at a molar ratio of 5-10:1 (linker:antibody). The conjugation reaction is carried out at room temperature for 1-2 hours. The excess linker is removed by dialysis or size-exclusion chromatography. The resulting ADC is characterized by HIC-HPLC, SEC-HPLC, and LC-MS to determine the drug-to-antibody ratio (DAR). For stability studies, the ADC is incubated in human plasma or serum at 37°C for up to 7 days, and samples are analyzed by SEC-HPLC to monitor aggregation and by LC-MS to assess drug release. For cathepsin B cleavage assays, the linker is incubated with recombinant cathepsin B at 37°C for 2-24 hours in acetate buffer (pH 5.5) containing DTT, and the released drug is quantified by HPLC.
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| Cell Assay |
For in vitro cell-based assays with ADCs containing Boc-Dap-NE, the following typical protocol is used: Target antigen-positive cancer cells (e.g., HER2-positive SK-BR-3 or BT-474 cells) and antigen-negative cells (e.g., MDA-MB-468) are cultured in appropriate media at 37°C in 5% CO₂. Cells are seeded in 96-well plates at 5,000-10,000 cells per well and allowed to adhere overnight. The ADC is serially diluted in culture medium to final concentrations ranging from 0.001 to 100 μg/mL (based on antibody concentration). Cells are treated with the ADC for 72-120 hours. Cell viability is assessed using the CellTiter-Glo or MTT assay. IC50 values are calculated from dose-response curves. For mechanistic studies, cells are treated with the ADC for 24-48 hours, and apoptosis is assessed by flow cytometry using Annexin V/PI staining. Cell cycle analysis is performed by propidium iodide staining. The specificity of the ADC is confirmed by the lack of activity against antigen-negative cells and by competition with excess unconjugated antibody. The intracellular trafficking of the ADC is assessed by confocal microscopy using fluorescently labeled antibodies.
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| Animal Protocol |
For in vivo animal studies with ADCs incorporating Boc-Dap-NE, the following general protocol is used: Female athymic nude mice (6-8 weeks old, 18-22 g) are subcutaneously injected with 5-10 × 10⁶ target antigen-positive tumor cells (e.g., BT-474 or NCI-N87 for HER2) in the flank. When tumors reach approximately 100-200 mm³, mice are randomized into treatment groups (n=8-10 per group). The ADC is administered intravenously via the tail vein at doses of 1, 3, 10, and 30 mg/kg on days 1, 8, 15, and 22 (weekly for 3-4 weeks). Tumor volumes are measured twice weekly with calipers and calculated as (length × width²)/2. Body weights are monitored for toxicity assessment. At the end of the study, tumors are excised, weighed, and processed for histopathology and immunohistochemistry. Blood samples are collected for pharmacokinetic analysis to measure ADC concentration, total antibody, and released drug.
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| Additional Infomation |
Boc-Dap-NE (CAS# 160800-65-7) is a cleavable ADC linker with a molecular formula of C23H36N2O5 and a molecular weight of 420.55 g/mol. It is used in the synthesis of antibody-drug conjugates (ADCs) and as an intermediate for MMAE synthesis. Future research could focus on optimizing the linker's stability and cleavage efficiency, developing new ADCs with improved therapeutic indices, and exploring its use in combination with different antibodies and payloads.
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| Molecular Formula |
C23H36N2O5
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|---|---|
| Molecular Weight |
420.542346954346
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| Exact Mass |
420.26
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| Elemental Analysis |
C, 65.69; H, 8.63; N, 6.66; O, 19.02
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| CAS # |
160800-65-7
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| Related CAS # |
Boc-Dap-NE;160800-65-7
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| PubChem CID |
20777964
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| Appearance |
White to off-white solid powder
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| LogP |
2.7
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
9
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| Heavy Atom Count |
30
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| Complexity |
570
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| Defined Atom Stereocenter Count |
5
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| SMILES |
C[C@H]([C@H]([C@@H]1CCCN1C(=O)OC(C)(C)C)OC)C(=O)N[C@H](C)[C@H](C2=CC=CC=C2)O
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| InChi Key |
XJDVGABQIFOWFC-SCQOQHIRSA-N
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| InChi Code |
InChI=1S/C23H36N2O5/c1-15(21(27)24-16(2)19(26)17-11-8-7-9-12-17)20(29-6)18-13-10-14-25(18)22(28)30-23(3,4)5/h7-9,11-12,15-16,18-20,26H,10,13-14H2,1-6H3,(H,24,27)/t15-,16-,18+,19-,20-/m1/s1
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| Chemical Name |
tert-butyl (S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidine-1-carboxylate
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| Synonyms |
Boc-Dap-NE;
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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) |
DMSO: ~100 mg/mL (~237.79 mM)
H2O: < 0.1 mg/mL (Insoluble) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (5.94 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (5.94 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 2.5 mg/mL (5.94 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. Solubility in Formulation 4: 10% DMSO+ 40% PEG300+ 5% Tween-80+ 45% saline: ≥ 2.5 mg/mL (5.94 mM) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.3779 mL | 11.8895 mL | 23.7790 mL | |
| 5 mM | 0.4756 mL | 2.3779 mL | 4.7558 mL | |
| 10 mM | 0.2378 mL | 1.1889 mL | 2.3779 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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