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| 1mg |
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| Other Sizes |
| Targets |
diABZI-V/C-DBCO targets the STING (Stimulator of Interferon Genes) pathway. STING is an adaptor protein that plays a critical role in the innate immune response to cytosolic DNA, triggering the production of type I interferons (IFN-alpha and IFN-beta) and other pro-inflammatory cytokines. diABZI-V/C-DBCO is a prodrug-like STING agonist that is activated by cathepsin B, which is a protease highly expressed in the tumor microenvironment and within endosomal compartments. Upon cleavage by cathepsin B, the compound releases diABZI-amine, which is the active STING agonist. This activates STING, leading to downstream signaling via TBK1 and IRF3, and the production of interferons and other immune-stimulatory molecules.
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| ln Vitro |
diABZI-V/C-DBCO is a highly effective STING agonist that can release diABZI-amine under the action of cathepsin B to activate STING. By activating STING, it induces the production of interferons (e.g., IFN-beta) and other pro-inflammatory cytokines, thereby enhancing the immune system's response to tumors. The cathepsin B-dependent activation mechanism may provide enhanced tumor selectivity because cathepsin B is highly expressed in the tumor microenvironment and within endolysosomal compartments. In vitro, diABZI-V/C-DBCO is expected to potently activate STING in cells that express cathepsin B, leading to downstream signaling and cytokine production.
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| ln Vivo |
diABZI-V/C-DBCO is designed for use in targeted cancer immunotherapy, including ADC applications. When conjugated to a tumor-targeting antibody via the DBCO group (using click chemistry with azide-modified antibodies), the resulting ADC can deliver the STING agonist specifically to tumors. Upon internalization into cancer cells or tumor-associated immune cells, the conjugate is cleaved by cathepsin B to release the active diABZI-amine STING agonist. This leads to local activation of the STING pathway in the tumor microenvironment, promoting anti-tumor immunity. This strategy aims to activate innate immunity specifically within tumors while minimizing systemic immune activation and toxicity. In vivo studies are expected to show tumor growth inhibition, increased immune cell infiltration, and reduced systemic cytokine release compared to unconjugated STING agonists.
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| Enzyme Assay |
The activation of STING can be assessed using a STING binding or activation assay. Procedure: THP1-Blue ISG (interferon-stimulated gene) reporter cells, which are engineered to express an IRF-inducible secreted embryonic alkaline phosphatase (SEAP) reporter, are seeded in 96-well plates at 5×10^4 cells per well. diABZI-V/C-DBCO is added to cells at varying concentrations (e.g., 0.001, 0.01, 0.1, 1, 10, 100 nM) in the presence or absence of purified cathepsin B (e.g., 5 microg/mL) to simulate cleavage. After 18-24 hours of incubation at 37degC, the cell culture supernatant is harvested, and SEAP activity is measured by adding QUANTI-Blue reagent and reading absorbance at 620-655 nm. Alternatively, the binding of diABZI-amine to STING can be assessed by surface plasmon resonance (SPR) using recombinant human STING protein.
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| Cell Assay |
The STING-activating activity can also be assessed in human primary cells. Procedure: Human peripheral blood mononuclear cells (PBMCs) are isolated from healthy donor blood by Ficoll density gradient centrifugation. PBMCs are seeded in 96-well plates at 2×10^5 cells per well in RPMI-1640 medium with 10% FBS. Cells are treated with varying concentrations of diABZI-V/C-DBCO (e.g., 0.01, 0.1, 1, 10, 100 nM) for 6-24 hours. For cathepsin B-dependent activation, cells can be pre-treated with a cathepsin B inhibitor (e.g., CA-074Me, 10 microM) to confirm the mechanism. Cell culture supernatants are collected and assayed for IFN-beta, IL-6, TNF-alpha, and CXCL10 levels by ELISA. The expression of interferon-stimulated genes (ISGs) such as ISG15, MX1, and OAS1 can be measured by qRT-PCR from cell lysates. A strong induction of cytokines and ISGs indicates STING activation by diABZI-V/C-DBCO.
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| Animal Protocol |
The in vivo efficacy can be evaluated in a syngeneic mouse tumor model. Procedure: 6-8 week old female BALB/c mice are inoculated subcutaneously with 5×10^5 CT26 (colon carcinoma) or 4T1 (breast carcinoma) cells. When tumors reach an average size of 50-100 mm3, mice are randomized into groups (n=8-10 per group). diABZI-V/C-DBCO is administered intratumorally (direct injection into the tumor) at doses of 0.1, 0.3, 1 microg per tumor or as an ADC conjugate intravenously. Tumor volumes are measured every 2-3 days with digital calipers. Blood is collected at various time points to measure serum cytokine levels (IFN-beta, IL-6, TNF-alpha) by ELISA. At study endpoint (typically day 14-21), tumors and spleens are harvested. Tumor-infiltrating lymphocytes are analyzed by flow cytometry (CD8+ T cells, CD4+ T cells, NK cells, dendritic cells). Tumor tissue sections are analyzed for STING pathway activation (p-TBK1, p-IRF3) by immunohistochemistry. Tumor growth inhibition and increased immune cell infiltration indicate in vivo efficacy.
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| ADME/Pharmacokinetics |
Specific PK data for diABZI-V/C-DBCO is not provided. The compound has a high molecular weight of 1500.66 g/mol, which is typical for an ADC payload-linker. As a prodrug, it requires cathepsin B cleavage to become active. The DBCO group enables conjugation to antibodies via click chemistry (copper-free strain-promoted azide-alkyne cycloaddition). When conjugated to an antibody, the ADC will have PK properties typical of antibody therapeutics (long half-life, low volume of distribution). The released active metabolite, diABZI-amine, is a small molecule (MW ~500-600) that is expected to have good cell permeability.
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| Toxicity/Toxicokinetics |
Specific toxicity data for diABZI-V/C-DBCO is not available. As a STING agonist, systemic administration can cause cytokine release syndrome due to excessive production of interferons and pro-inflammatory cytokines. The cathepsin B-dependent activation mechanism of diABZI-V/C-DBCO is designed to restrict activation to the tumor microenvironment, potentially reducing systemic toxicity. Antibody conjugation further increases tumor targeting and reduces off-target activation. Comprehensive toxicological evaluation would be required for drug development.
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| References | |
| Additional Infomation |
diABZI-V/C-DBCO is an innovative STING agonist prodrug that is activated by cathepsin B, a protease highly expressed in the tumor microenvironment, and contains a DBCO group for conjugation to targeting ligands via click chemistry. STING agonists are a promising class of immunotherapies that activate the innate immune system to promote anti-tumor immunity. However, systemic administration of STING agonists is often associated with severe toxicity and cytokine storm. The cathepsin B-activatable prodrug strategy, combined with antibody targeting, aims to achieve tumor-localized STING activation while minimizing systemic exposure. The DBCO group enables copper-free click conjugation to azide-modified antibodies, allowing for the generation of stable ADCs. The compound has the molecular formula C78H89N19O13 and a molecular weight of 1500.66 g/mol. It is for research use only and is not an approved drug. The compound is typically stored at -20degC and protected from light to maintain stability.
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| Molecular Formula |
C78H89N19O13
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|---|---|
| Molecular Weight |
1500.66
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| Exact Mass |
1499.689
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| CAS # |
3035557-60-6
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| PubChem CID |
172872265
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| Appearance |
White to off-white solid powder
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| Hydrogen Bond Donor Count |
10
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| Rotatable Bond Count |
36
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| Heavy Atom Count |
110
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| Complexity |
3240
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| Defined Atom Stereocenter Count |
2
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| SMILES |
CCN1C(=CC(=N1)C)C(=O)NC2=NC3=C(N2C/C=C/CN4C5=C(C=C(C=C5OCCCNC(=O)OCC6=CC=C(C=C6)NC(=O)[C@H](CCCNC(=O)N)NC(=O)[C@H](C(C)C)NC(=O)CCCCC(=O)N7CC8=CC=CC=C8C#CC9=CC=CC=C97)C(=O)N)N=C4NC(=O)C1=CC(=NN1CC)C)C(=CC(=C3)C(=O)N)OC
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| InChi Key |
YQIYAYDEUDNEHB-UGNZTZBXSA-N
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| InChi Code |
InChI=1S/C78H89N19O13/c1-8-96-60(38-47(5)91-96)72(103)89-76-86-57-40-53(69(79)100)42-62(108-7)67(57)93(76)35-16-17-36-94-68-58(87-77(94)90-73(104)61-39-48(6)92-97(61)9-2)41-54(70(80)101)43-63(68)109-37-19-34-83-78(107)110-45-49-27-31-55(32-28-49)84-71(102)56(23-18-33-82-75(81)106)85-74(105)66(46(3)4)88-64(98)25-14-15-26-65(99)95-44-52-22-11-10-20-50(52)29-30-51-21-12-13-24-59(51)95/h10-13,16-17,20-22,24,27-28,31-32,38-43,46,56,66H,8-9,14-15,18-19,23,25-26,33-37,44-45H2,1-7H3,(H2,79,100)(H2,80,101)(H,83,107)(H,84,102)(H,85,105)(H,88,98)(H3,81,82,106)(H,86,89,103)(H,87,90,104)/b17-16+/t56-,66-/m0/s1
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| Chemical Name |
[4-[[(2S)-2-[[(2S)-2-[[6-(2-azatricyclo[10.4.0.04,9]hexadeca-1(16),4,6,8,12,14-hexaen-10-yn-2-yl)-6-oxohexanoyl]amino]-3-methylbutanoyl]amino]-5-(carbamoylamino)pentanoyl]amino]phenyl]methyl N-[3-[6-carbamoyl-3-[(E)-4-[5-carbamoyl-2-[(2-ethyl-5-methylpyrazole-3-carbonyl)amino]-7-methoxybenzimidazol-1-yl]but-2-enyl]-2-[(2-ethyl-5-methylpyrazole-3-carbonyl)amino]benzimidazol-4-yl]oxypropyl]carbamate
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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: (1). 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) |
DMSO : ~100 mg/mL (~66.64 mM; with ultrasonication)
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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 | 0.6664 mL | 3.3319 mL | 6.6637 mL | |
| 5 mM | 0.1333 mL | 0.6664 mL | 1.3327 mL | |
| 10 mM | 0.0666 mL | 0.3332 mL | 0.6664 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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