| Targets |
Topoisomerase I. Dxd is a derivative of exatecan that acts as a topoisomerase I inhibitor, trapping the DNA-topoisomerase I complex and leading to irreversible DNA damage and apoptosis in rapidly dividing cells.
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|---|---|
| ln Vitro |
In cell-free assays, the GGFG linker is specifically cleaved by lysosomal proteases (e.g., cathepsin B) but remains stable in plasma. When incorporated into an ADC, the conjugate releases the Dxd payload intracellularly. The payload then diffuses to the nucleus to exert its cytotoxic effect by inhibiting topoisomerase I.
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| ln Vivo |
In vivo, when conjugated to a tumor-targeting antibody, the drug-linker conjugate significantly inhibits tumor growth in mouse xenograft models and can induce complete tumor regression. The hydrophilic PEG4 spacer increases the aqueous solubility of the ADC, allowing for higher drug-to-antibody ratios (DAR) without aggregation.
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| Enzyme Assay |
A general cell‑free protocol for a drug-linker conjugate: The compound is first conjugated to an antibody via maleimide-thiol chemistry. The resulting ADC is purified and analyzed to confirm its drug-to-antibody ratio (DAR). The ADC is then evaluated in a cell-free stability assay by incubation in human plasma at 37degC for 14 days, and intact ADC levels are measured by LC-MS.
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| Cell Assay |
A general cellular protocol for the ADC: Target-positive cancer cells (e.g., HER2-expressing cells) are plated in 96-well plates. The ADC is added in serial dilutions, and after 72-96 hours, cell viability is determined using a luminescent CellTiter-Glo assay. The half-maximal inhibitory concentration (IC50) is calculated.
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| Animal Protocol |
A general animal protocol for the ADC: Mice bearing subcutaneous xenografts of target-positive tumors are intravenously administered the ADC at doses of 1, 3, or 10 mg/kg. Tumor volume is measured with a caliper twice weekly. On study completion, tumors are collected for IHC and assessment of DNA damage (gamma-H2AX).
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| ADME/Pharmacokinetics |
General PK protocol: The ADC is administered intravenously to rats or cynomolgus monkeys. Blood is collected over 28 days. Plasma samples are analyzed using ligand-binding assays (e.g., ELISA) to measure total antibody concentrations and Dxd-conjugated antibody concentrations. The released payload is analyzed by LC-MS/MS.
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| Toxicity/Toxicokinetics |
General toxicity protocol: A 28-day repeat-dose IV toxicity study is performed in cynomolgus monkeys. Animals are administered the ADC at doses of 5, 20, or 60 mg/kg once weekly. Parameters include clinical signs, body weight, hematology, serum chemistry, and histopathology of the bone marrow, gastrointestinal tract, and liver.
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| References | |
| Additional Infomation |
Dxd is the cytotoxic component of the clinically approved ADC Enhertu (trastuzumab deruxtecan). The GGFG tetrapeptide linker is designed for cathepsin B-mediated cleavage, ensuring efficient payload release in the lysosomal compartment. Amino-PEG4-GGFG-Dxd has a molecular formula C53H66FN9O15 and a molecular weight of 1088.14 g/mol.
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| Exact Mass |
1087.466
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|---|---|
| CAS # |
2879227-88-8
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| PubChem CID |
171714382
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| Appearance |
Solid powder
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| Hydrogen Bond Donor Count |
8
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| Rotatable Bond Count |
30
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| Heavy Atom Count |
78
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| Complexity |
2200
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| Defined Atom Stereocenter Count |
3
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| InChi Key |
SCJCAOCGTRKMQY-KJDBIWOKSA-N
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| InChi Code |
InChI=1S/C53H66FN9O15/c1-3-53(72)36-22-41-49-34(27-63(41)51(70)35(36)28-78-52(53)71)48-38(10-9-33-31(2)37(54)23-39(62-49)47(33)48)60-46(68)29-77-30-59-44(66)25-58-50(69)40(21-32-7-5-4-6-8-32)61-45(67)26-57-43(65)24-56-42(64)11-13-73-15-17-75-19-20-76-18-16-74-14-12-55/h4-8,22-23,38,40,72H,3,9-21,24-30,55H2,1-2H3,(H,56,64)(H,57,65)(H,58,69)(H,59,66)(H,60,68)(H,61,67)/t38-,40-,53-/m0/s1
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| Chemical Name |
(2S)-2-[[2-[[2-[3-[2-[2-[2-(2-aminoethoxy)ethoxy]ethoxy]ethoxy]propanoylamino]acetyl]amino]acetyl]amino]-N-[2-[[2-[[(10S,23S)-10-ethyl-18-fluoro-10-hydroxy-19-methyl-5,9-dioxo-8-oxa-4,15-diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa-1,6(11),12,14,16,18,20(24)-heptaen-23-yl]amino]-2-oxoethoxy]methylamino]-2-oxoethyl]-3-phenylpropanamide
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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: 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)
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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.) |
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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