| Size | Price | Stock | Qty |
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| 50mg |
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| 100mg |
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| 250mg |
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| Targets |
Boc-Val-Ala-PAB-PNP is an ADC linker that enables targeted delivery of cytotoxic drugs to cancer cells. The compound’s Val-Ala dipeptide sequence is recognized and cleaved by cathepsin B, a lysosomal protease that is upregulated in many cancer types. The PAB (p-aminobenzyl) group serves as a self-immolative spacer that, upon enzymatic cleavage, undergoes spontaneous decomposition to release the drug payload. The PNP ester allows conjugation to amine groups on drugs or antibodies. The Boc protecting group is removed during synthesis to allow further conjugation. This design enables tumor-specific drug release, reducing systemic toxicity.
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| ln Vitro |
ADC cytotoxins are connected to antibodies through an ADC connector to form ADCs [1].
In vitro activity of Boc-Val-Ala-PAB-PNP is demonstrated in ADC synthesis and drug release assays. The compound is used to conjugate cytotoxic drugs to antibodies, creating ADCs that can selectively kill cancer cells. In vitro cytotoxicity assays with ADC-treated cancer cells demonstrate the effectiveness of the drug release mechanism. The Val-Ala linker is cleaved by cathepsin B, releasing the drug payload in a cell-specific manner. The PNP ester facilitates efficient conjugation, and the Boc protecting group allows for controlled synthesis. These in vitro assays confirm the linker’s functionality and its utility in ADC development. |
| ln Vivo |
In vivo activity of Boc-Val-Ala-PAB-PNP is evaluated in animal models of cancer using ADCs synthesized with this linker. Tumor xenograft models are used to assess the efficacy of ADCs containing the Boc-Val-Ala-PAB-PNP linker. The ADCs are administered intravenously, and tumor growth inhibition and survival are monitored. The tumor-specific cleavage of the Val-Ala linker by cathepsin B enables selective drug release within the tumor, enhancing efficacy and reducing off-target toxicity. These in vivo studies are critical for evaluating the therapeutic potential of ADCs incorporating this linker.
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| Enzyme Assay |
In vitro assays for Boc-Val-Ala-PAB-PNP involve testing the linker’s cleavage by cathepsin B. Recombinant cathepsin B or tumor cell lysates are incubated with the linker or the ADC containing the linker. The release of the drug payload is measured by HPLC or LC-MS. The efficiency of conjugation to antibodies is assessed by SDS-PAGE or mass spectrometry. The PNP ester activation allows for conjugation to amine groups, and the reaction can be monitored by UV spectroscopy. These assays confirm the linker’s functionality and its ability to release drugs in a cathepsin B-dependent manner.
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| Cell Assay |
In vitro cellular assays for Boc-Val-Ala-PAB-PNP are conducted using ADCs synthesized with this linker. Cancer cell lines that overexpress cathepsin B (e.g., MDA-MB-231, HT-1080) are treated with the ADC. Cell viability is measured using MTT or CellTiter-Glo assays to assess cytotoxicity. Cathepsin B inhibitors can be used to confirm that the cytotoxicity is dependent on cathepsin B-mediated cleavage. Drug release can be measured in cell culture supernatants by LC-MS. These assays demonstrate the tumor-specific cytotoxicity of ADCs containing the Boc-Val-Ala-PAB-PNP linker.
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| Animal Protocol |
In vivo animal experiments with Boc-Val-Ala-PAB-PNP are conducted in mouse xenograft models of cancer. Immunodeficient mice are engrafted with human cancer cell lines that overexpress cathepsin B. ADCs synthesized with the linker are administered intravenously at varying doses. Tumor growth is measured by caliper, and survival is monitored. Pharmacodynamic endpoints include tumor drug levels, cathepsin B activity, and apoptosis markers. These studies evaluate the in vivo efficacy and safety of ADCs containing the Boc-Val-Ala-PAB-PNP linker.
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| ADME/Pharmacokinetics |
Pharmacokinetic data for Boc-Val-Ala-PAB-PNP are relevant in the context of ADCs synthesized with this linker. The linker itself is not administered as a free drug but as part of an ADC. The pharmacokinetics of the ADC are influenced by the properties of the antibody, the linker, and the drug payload. The linker is designed to be stable in circulation and cleaved intracellularly by cathepsin B. The Boc protecting group is removed during synthesis, and the final ADC’s PK is determined by the intact conjugate. Detailed PK studies are conducted for each ADC individually.
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| Toxicity/Toxicokinetics |
Toxicological data for Boc-Val-Ala-PAB-PNP are evaluated as part of the safety assessment of ADCs incorporating this linker. The linker itself is not typically tested alone, but the toxicity of the ADC is assessed in preclinical studies. The tumor-specific cleavage of the Val-Ala linker by cathepsin B is intended to reduce systemic toxicity. However, off-target cleavage or payload release can lead to toxicity. Standard toxicology studies in rodents and non-rodents are conducted to assess the safety of ADCs containing this linker.
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| References |
[1]. Beck A, et al. Strategies and challenges for the next generation of antibody-drug conjugates. Nat Rev Drug Discov. 2017 May;16(5):315-337.
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| Additional Infomation |
Boc-Val-Ala-PAB-PNP is a cleavable peptide linker used in the synthesis of antibody-drug conjugates (ADCs). It contains a Val-Ala dipeptide sequence that is cleaved by cathepsin B, a tumor-associated protease. The PAB group provides a self-immolative spacer, and the PNP ester enables conjugation to drugs or antibodies. The Boc group protects the amine during synthesis. This linker enables tumor-specific drug release, improving the therapeutic index of ADCs. It is a key building block in the development of next-generation ADCs for cancer therapy.
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| Molecular Formula |
C27H34N4O9
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|---|---|
| Molecular Weight |
558.5803
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| Exact Mass |
558.232
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| CAS # |
1884578-00-0
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| PubChem CID |
100029284
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| Appearance |
White to off-white solid powder
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| LogP |
4.4
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
9
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| Rotatable Bond Count |
13
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| Heavy Atom Count |
40
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| Complexity |
883
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| Defined Atom Stereocenter Count |
2
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| SMILES |
C[C@@H](C(=O)NC1=CC=C(C=C1)COC(=O)OC2=CC=C(C=C2)[N+](=O)[O-])NC(=O)[C@H](C(C)C)NC(=O)OC(C)(C)C
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| InChi Key |
PGJUTVVCCBCEIY-JTSKRJEESA-N
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| InChi Code |
InChI=1S/C27H34N4O9/c1-16(2)22(30-25(34)40-27(4,5)6)24(33)28-17(3)23(32)29-19-9-7-18(8-10-19)15-38-26(35)39-21-13-11-20(12-14-21)31(36)37/h7-14,16-17,22H,15H2,1-6H3,(H,28,33)(H,29,32)(H,30,34)/t17-,22-/m0/s1
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| Chemical Name |
[4-[[(2S)-2-[[(2S)-3-methyl-2-[(2-methylpropan-2-yl)oxycarbonylamino]butanoyl]amino]propanoyl]amino]phenyl]methyl (4-nitrophenyl) carbonate
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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 | 1.7903 mL | 8.9513 mL | 17.9025 mL | |
| 5 mM | 0.3581 mL | 1.7903 mL | 3.5805 mL | |
| 10 mM | 0.1790 mL | 0.8951 mL | 1.7903 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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