| Size | Price | |
|---|---|---|
| 500mg | ||
| 1g | ||
| Other Sizes |
| Targets |
DGN549-L (1a) itself does not have a specific biological target. As part of an antibody-drug conjugate, it targets the antigen recognized by the antibody (e.g., FRα or CD123). Its mechanism of action is DNA alkylation after intracellular release. The catabolite identified from DGN549-L-based ADCs is the monoimine indolinobenzodiazepine aniline species (4). [1]
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| ln Vitro |
DGN549-L (1a) is not tested as a free drug; its activity is assessed as part of ADCs (e.g., ADC 2a and 3a). [1]
In FRα-positive cell lines (KB, T47D, NCI-H2110), the lysine-linked ADC 2a (DAR ~2.5) shows specific cytotoxicity with IC50 values (based on ADC concentration) of 5 × 10⁻¹² M (KB), 2 × 10⁻¹¹ M (T47D), and 4 × 10⁻¹¹ M (NCI-H2110). The corresponding IC50 values based on DGN549 payload concentration are 1 × 10⁻¹¹ M (KB), 6 × 10⁻¹¹ M (T47D), and 1 × 10⁻¹⁰ M (NCI-H2110). Cytotoxicity is blocked by excess unconjugated targeting antibody, confirming target specificity. [1] In CD123-positive cell lines (EOL-1, HNT-34, MV4-11), the lysine-linked ADC 3a (DAR ~2.5) shows specific cytotoxicity with IC50 values (based on ADC concentration) of 2 × 10⁻¹² M (EOL-1), 8 × 10⁻¹³ M (HNT-34), and 5 × 10⁻¹³ M (MV4-11). The corresponding IC50 values based on DGN549 payload concentration are 6 × 10⁻¹² M (EOL-1), 2 × 10⁻¹² M (HNT-34), and 2 × 10⁻¹² M (MV4-11). [1] Bystander activity: ADC 2a (lysine-linked) kills FRα-negative cells in mixed population assays at subnanomolar concentrations, consistent with release of a cell-permeable catabolite. [1] |
| ln Vivo |
DGN549-L (1a)-based ADCs show efficacy in mouse xenograft models. [1]
In NCI-H2110 (FRα-positive NSCLC) xenografts, ADC 2a (3 μg DGN549/kg) is minimally active (T/C = 32%); at 9 μg DGN549/kg, it induces durable complete regressions (6/6 complete). Activity is similar to site-specific ADC 2b. [1] In Molm-13 (CD123-positive AML) disseminated xenografts, ADC 3a at 0.3 μg DGN549/kg produces 59% increased life span; at 1 μg DGN549/kg, median survival exceeds 152 days (>443% ILS). The lysine-linked ADC 3a shows slightly lower activity compared to site-specific ADC 3b at the 0.3 μg DGN549/kg dose (59% vs 262% ILS). [1] |
| Cell Assay |
Cell binding assay (flow cytometry) : Cells (0.5–1 × 10⁵) are incubated with serial dilutions of test antibodies for 1 hour at 4°C, washed, and incubated with FITC-conjugated goat anti-human IgG secondary antibody for 1 hour at 4°C. After washing, cells are fixed and analyzed by flow cytometry. EC50 values for ADCs are comparable to parent antibodies. [1]
Cytotoxicity assay (CellTiter-Glo) : Cells (1500–3000/well) are seeded in 96-well plates and incubated overnight. Serial dilutions of ADCs are added with or without blocking antibody (20 μg/mL). After 4 days at 37°C, CellTiter-Glo reagent is added, and luminescence is measured. IC50 values are calculated. [1] Bystander activity assay : A mixed population of FRα-negative parental 300.19 cells and FRα-positive transfectants is treated with ADC. Cell viability is determined after 4 days using CellTiter-Glo. [1] Catabolite identification assay (LC-MS) : KB cells are treated with saturating concentration of ADC for 24 h. Catabolites in media are concentrated by binding to an anti-IGN antibody, extracted into acetone, dried, redissolved, and analyzed by UPLC coupled to high-resolution mass spectrometry. [1] |
| Animal Protocol |
Efficacy (NCI-H2110 xenograft) : Female CB-17 SCID mice (n=6/group) are inoculated subcutaneously with 1 × 10⁷ NCI-H2110 cells. When tumors reach ~100 mm³ (day 8), a single intravenous dose of ADC (3 or 9 μg DGN549/kg) is administered. Tumor size is measured twice weekly. [1]
Efficacy (Molm-13 disseminated xenograft) : Female thymic nude mice (n=10/group) are inoculated intravenously with 1 × 10⁷ Molm-13 cells. On day 7, a single intravenous dose of ADC (0.1, 0.3, or 1 μg DGN549/kg) is administered. To block Fc receptors, mice receive 400 mg/kg of a non-targeting IgG1 antibody intraperitoneally 24 h before ADC, followed by 100 mg/kg on days 4 and 9. [1] Pharmacokinetics : Non-tumor-bearing CD-1 mice receive a single intravenous dose of ADC (2.5 mg antibody/kg). Plasma samples are collected at various time points (2 min to 28 days). ADC concentrations are measured by two ELISA methods: one for total antibody (anti-human IgG capture) and one for conjugated antibody (anti-IGN capture). [1] |
| ADME/Pharmacokinetics |
For lysine-linked ADC 2a (DGN549-L conjugated to mAb1) in CD-1 mice at 2.5 mg antibody/kg: total antibody Cmax = 40.3 μg/mL, AUC0-∞ = 6,550 h•μg/mL, clearance = 0.45 mL/h/kg, t½ = 4.8 days, Vss = 29.7 mL/kg. Conjugated antibody Cmax = 45.3 μg/mL, AUC0-∞ = 5,578 h•μg/mL, clearance = 0.45 mL/h/kg, t½ = 4.5 days, Vss = 29.7 mL/kg. Compared to site-specific ADC 2b, lysine-linked ADC 2a shows 1.6-fold lower exposure and 1.7-fold faster clearance. [1]
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| Toxicity/Toxicokinetics |
No direct toxicity data (e.g., MTD) for DGN549-L (1a) as a free drug are reported. However, mouse tolerability studies of ADC 2a (lysine-linked DGN549 ADC) show that the maximum tolerated dose (MTD) is approximately 50–60 μg DGN549/kg (approx 2.5–3 mg antibody/kg). This is approximately 2-fold lower than the MTD of the corresponding site-specific ADC 2b (100–125 μg DGN549/kg), indicating that the lysine-linked ADC is less well tolerated. [1]
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| References | |
| Additional Infomation |
Synthesis: DGN549-L (1a) is an NHS ester-bearing IGN payload used for lysine conjugation. [1]
Conjugation method (lysine) : DGN549-L is conjugated to antibodies via NHS ester chemistry targeting lysine residues, yielding heterogeneous ADCs with DAR between 2.5 and 3.0. [1] Catabolite: The only major catabolite species identified from both lysine-linked and CYSAMB DGN549 ADCs is the monoimine indolinobenzodiazepine aniline species (4), confirming that the conjugation chemistry does not alter the mechanism of action. [1] Hydrophobicity: By HIC analysis, lysine-linked DGN549 ADCs elute as a broad unresolved peak over approximately 45 min of the HIC gradient, reflecting heterogeneity in both DAR and conjugation sites. [1] |
| Molecular Formula |
C58H58N8O13
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|---|---|
| Molecular Weight |
1075.13
|
| Exact Mass |
1074.412
|
| CAS # |
1884276-68-9
|
| PubChem CID |
121265250
|
| Appearance |
Typically exists as solid at room temperature
|
| LogP |
4.3
|
| Hydrogen Bond Donor Count |
4
|
| Hydrogen Bond Acceptor Count |
15
|
| Rotatable Bond Count |
20
|
| Heavy Atom Count |
79
|
| Complexity |
2270
|
| Defined Atom Stereocenter Count |
4
|
| SMILES |
C[C@@H](C(=O)N[C@@H](C)C(=O)NC1=CC(=CC(=C1)COC2=C(C=C3C(=C2)N=C[C@@H]4CC5=CC=CC=C5N4C3=O)OC)COC6=C(C=C7C(=C6)NC[C@@H]8CC9=CC=CC=C9N8C7=O)OC)NC(=O)CCCCC(=O)ON1C(=O)CCC1=O
|
| InChi Key |
ZJAVYEURWFEFIB-RUUJEGCPSA-N
|
| InChi Code |
InChI=1S/C58H58N8O13/c1-32(61-51(67)15-9-10-16-54(70)79-66-52(68)17-18-53(66)69)55(71)62-33(2)56(72)63-38-20-34(30-77-49-26-43-41(24-47(49)75-3)57(73)64-39(28-59-43)22-36-11-5-7-13-45(36)64)19-35(21-38)31-78-50-27-44-42(25-48(50)76-4)58(74)65-40(29-60-44)23-37-12-6-8-14-46(37)65/h5-8,11-14,19-21,24-28,32-33,39-40,60H,9-10,15-18,22-23,29-31H2,1-4H3,(H,61,67)(H,62,71)(H,63,72)/t32-,33-,39-,40-/m0/s1
|
| Chemical Name |
(2,5-dioxopyrrolidin-1-yl) 6-[[(2S)-1-[[(2S)-1-[3-[[(12aS)-8-methoxy-6-oxo-12a,13-dihydroindolo[2,1-c][1,4]benzodiazepin-9-yl]oxymethyl]-5-[[(12aS)-8-methoxy-6-oxo-11,12,12a,13-tetrahydroindolo[2,1-c][1,4]benzodiazepin-9-yl]oxymethyl]anilino]-1-oxopropan-2-yl]amino]-1-oxopropan-2-yl]amino]-6-oxohexanoate
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| Synonyms |
DGN549-L; 1884276-68-9; orb1691407; SCHEMBL17665043;
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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 | 0.9301 mL | 4.6506 mL | 9.3012 mL | |
| 5 mM | 0.1860 mL | 0.9301 mL | 1.8602 mL | |
| 10 mM | 0.0930 mL | 0.4651 mL | 0.9301 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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