DNA synthesis; antitumor antibiotic

Duocarmycin TM (60 μM; 4 d; BJAB and WSU-DLCL2 cells) is a cytotoxic drug that inhibits tumor cell proliferation [1].

Cell viability assay [1]
Cell Types: BJAB and WSU-DLCL2 Cell
Tested Concentrations: 60 μM
Incubation Duration: 4 days
Experimental Results: Inhibited tumor cell proliferation, the IC50 values of BJAB and WSU-DLCL2 cells were 0.153 μM and 0.079 μM respectively.

[1]. Immolation of p-Aminobenzyl Ether Linker and Payload Potency and Stability Determine the Cell-Killing Activity of Antibody-Drug Conjugates with Phenol-Containing Payloads. Bioconjug Chem. 2018 Feb 21;29(2):267-274.

Valine-citrulline (Val-Cit) dipeptides and p-aminobenzyl (PAB) spacers are commonly used as cleavable, self-cleaving linkers in antibody-drug conjugate (ADC) design, including the clinically approved ADC drug brentuximab vedotin (Adcetris). When the same linker is used to connect to the phenolic hydroxyl group of cyclopropylbenzoindolone (CBI) (P1), the resulting ADC1 exhibits reduced activity in cancer cell lines expressing the CD22 target (e.g., BJAB, WSU-DLCL2). In contrast, the conjugate (ADC2) of cyclopropylpyrroloindolone (CPI) (P2) shows higher activity, although both corresponding free drugs exhibit similar picomolar cytotoxic activity. Although the corresponding spirocyclized products of P1 and P2, responsible for DNA alkylation, are the main components in the buffer, the linker cleavage rate is slower when PAB is attached to the phenolic hydroxyl group of P1 in ether form (PABE) compared to P2. Further cleavage studies of two other PABE-linked substituted phenolic compounds showed that electron-withdrawing groups could accelerate cleavage, thereby releasing the acidic phenolic payload (delocalizing the negative charge on the expected anionic phenolic oxygen during cleavage). In contrast, efficient cleavage of LD4 did not produce active ADC4 because the cytotoxic efficacy of the payload (P4) was low. Furthermore, non-cleavage of LD5 did not affect the cytotoxic efficacy of its ADC5 because DNA alkylation of the centrally linked pyrrolobenzodiazepine did not require cleavage. Therefore, when using the Val-Cit-PAB linker to link antibodies to phenolic drugs, careful evaluation is required because linker cleavage, as well as the efficacy and stability of the payload, can affect the cytotoxic activity of antibody-drug conjugates (ADCs). [1]

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Solid-phase synthesis can rapidly generate peptide-drug conjugates. A peptide targeting the Thomsen-Friedenreich antigen (TFα) was conjugated to an alkylated subunit of the potent cytotoxic duocarmycin SA. This compound contains a linker that can be cleaved by cathepsin B and has been shown to be active and selective against tumor cell lines expressing TFα. https://pubs.acs.org/doi/10.1021/acs.bioconjchem.0c00282

C25H23CLN2O5

466.913525819778

466.129

157922-77-5

394851

Light yellow to yellow solid powder

4.6

2

5

5

33

712

1

ClC[C@@H]1CN(C(C2=CC3C=C(C(=C(C=3N2)OC)OC)OC)=O)C2C=C(C3C=CC=CC=3C1=2)O

NRHDGIYFJJUFKN-CQSZACIVSA-N

InChI=1S/C25H23ClN2O5/c1-31-20-9-13-8-17(27-22(13)24(33-3)23(20)32-2)25(30)28-12-14(11-26)21-16-7-5-4-6-15(16)19(29)10-18(21)28/h4-10,14,27,29H,11-12H2,1-3H3/t14-/m1/s1

[(1S)-1-(chloromethyl)-5-hydroxy-1,2-dihydrobenzo[e]indol-3-yl]-(5,6,7-trimethoxy-1H-indol-2-yl)methanone

Duocarmycin TM; 157922-77-5; [(1S)-1-(chloromethyl)-5-hydroxy-1,2-dihydrobenzo[e]indol-3-yl]-(5,6,7-trimethoxy-1H-indol-2-yl)methanone; CBI-TMI; CHEMBL66051; SCHEMBL12596987; (1S)-1-(chloromethyl)-3-(5,6,7-trimethoxy-1H-indole-2-carbonyl)-1H,2H,3H-benzo[e]indol-5-ol;

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : ≥ 50 mg/mL (~107.09 mM)

Solubility in Formulation 1: ≥ 2.5 mg/mL (5.35 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.

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Solubility in Formulation 2: ≥ 2.5 mg/mL (5.35 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 corn oil and mix evenly.

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 (Please use freshly prepared in vivo formulations for optimal results.)