DT2216

Protein target name:

BCL2L1

Mechanism of action:

DegraderPROTAC

Recommended Concentration for use in cells:

~~1 uM~~

Reviewer suggested Concentration for use in cells:

100-500 nM, maximum 1 uM

Probe Availability

MedChemExpress

Probe Information

Target and Probe Information

Target Details

Target class:

Other

Define other target class:

BCL2 family

Subclass:

Apoptosis Regulator

HUGO ID:

HGNC:992

Entrez gene ID:

598

UNIPROT ID:

Q07817

Target alias:

BCLXL, bcl-xl, BCL2 like 1, BCL-XL/S, BCL2L, BCLX, Bcl-X, PPP1R52, BCLL2L1

Probe Details

Chemical Probes Portal ID:

CPP2152

PubChem CID:

139331475

SMILES string:

CC1(C)CCC(C2=CC=C(Cl)C=C2)=C(C1)CN3CCN(C4=CC=C(C(NS(=O)(C5=CC(S(C(F)(F)F)(=O)=O)=C(N[C@@H](CSC6=CC=CC=C6)CCN7CCN(C(CCCCCC(N[C@@H](C(C)(C)C)C(N8[C@@H](C[C@@H](O)C8)C(N[C@H](C9=CC=C(C%10=C(N=CS%10)C)C=C9)C)=O)=O)=O)=O)CC7)C=C5)=O)=O)C=C4)CC3

InChi Key:

PXVFFBGSTYQHRO-REQIQPEASA-N

Control compound:

DT2216NC

Orthogonal probe:

A-1155463WEHI-539A-1331852

Physico-chemical properties

NMR:

1H NMR (400 MHz, CDCl3) δ 8.67 (s, 1H), 8.34 (d, J = 2.3 Hz, 1H), 8.08 (d, J = 9.1 Hz, 1H), 7.69 (d, J = 8.6 Hz, 2H), 7.46–7.27 (m, 12H), 7.14–6.93 (m, 3H), 6.76 (d, J = 8.6 Hz, 2H), 6.61 (d, J = 9.4 Hz, 1H), 6.34 (d, J = 8.7 Hz, 1H), 5.13–5.00 (m, 1H), 4.79–4.69 (m, 1H), 4.60 (d, J = 8.7 Hz, 1H), 4.49 (s, 1H), 4.10 (d, J = 11.4 Hz, 1H), 3.96–3.83 (m, 1H), 3.72–2.81 (m, 13H), 2.55–2.00 (m, 24H), 1.77–1.30 (m, 12H), 1.03 (s, 9H), 0.97 (s, 6H) ppm.

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

12.8 nM BCL-XL, 1.8 nM BCL-2, 300.9 nM BCL-W

Potency assay:

AlphaScreen, AlphaLITE

PDB ID for probe-target interaction (3D structure):

4QNQ

Structure-activity relationship:

Off target activity

Probe Selectivity in Vitro:

No changes in BCL-2 levels in all cells examined. In addition, DT2216 had no effect on the levels of BCL-W and MCL-1 in these cells.

In cell validation

On target activity in cells

Potency in cells:

DC50, Dmax, EC50

63 nM (DC50), 90.8% (Dmax), 52 nM (EC50)

Potency assay (cells):

HiBIT assay, immunoprecipitation of HA-ubiquitin and FLAG-BCL

Phenotypic Control Experiment:

Preincubation of MOLT-4 cells with an excess amount of ABT263 or VHL-L inhibited the formation of the BCL-XL–DT2216–VHL ternary complex. DT2216NC (control) fails to form ternary complex. DT2216 had no effect on the levels of BCL-XL in VHL-null 786-O renal cell carcinoma (RCC) cells.

Off target activity in cells

Probe Selectivity in Cell:

Proteomics shows that DT2216 does not degrade any other protein.

In vivo validation

Organism:

Mouse

Dose:

7.5-15 mg/Kg

Route of delivery:

Intravenous

Plasma half life:

6.3 (IP), 6.1 (IV) h

Systemic clearance:

0.42 ml/min/Kg

Primary Reference

Reference (doi):

10.1038/s41591-019-0668-z

Reference (PMID):

31792461

Endorsed probe

The Chemical Probes Portal endorses this compound as chemical probe for use in cells and in model organisms.

SAB Rating

Rating for use in Cells

3 review(s)

Rating for use in Model Organisms

3 review(s)

SAB Members

SAB Comments

It would be very useful to have information regarding the inhibition and degradation off-target profiling.

Jan 19 2021 - 12:30pm

DT2216 is a proteolysis-targeting chimera (PROTAC) based on the BH3-mimetic ABT263 which targets BCLXL to the Von Hippel-Lindau (VHL) E3 ligase for degradation. Unlike ABT263 (which functionally inhibits BCL2), DT2216 does not target BCL2 for degradation. Thus, relative to ABT263 it is a BCLXL selective compound. As BCLXL is an important determinant of platelet apoptosis, the clinical utility of ABT263 was limited by dose limiting yet on-target thrombocytopenia. As platelets have low VHL levels, dose limiting thrombocytopenia is not observed concurrent to clinically efficacious doses of DT2216. Thus, DT2216 represents a potent, cell active BCLXL-specific PROTAC which has anti-tumor activity in BCLXL-dependent tumors, without the toxicity of thrombocytopenia. This probe is highly specific for BCLXL and does not degrade other BCL2 family proteins. Much of the data presented in the initial description of this molecule was based on hematological cell lines (such as MOLT-4), but target degradation by this probe has also been verified in solid-organ cancer lines. Some caution needs to be exercised when determining dosage for in vitro cellular probing as the effective concentration range may vary and cells with low VHL (including platelets) will be resistant to the probe.

Jan 27 2021 - 12:05pm

DT2216 and its control substance DT2216NC represent a very good substance pair for investigating the role of BCL-XL degradation in both cells and animals. The authors present extensive evidence to support the specificity of DT2216 for BCL-XL degradation.

Feb 9 2021 - 7:01pm

DT2216

Probe Availability

Probe Information

Target Details

BCLXL, bcl-xl, BCL2 like 1, BCL-XL/S, BCL2L, BCLX, Bcl-X, PPP1R52, BCLL2L1

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

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DT2216

Probe Availability

Probe Information

Target Details

BCLXL, bcl-xl, BCL2 like 1, BCL-XL/S, BCL2L, BCLX, Bcl-X, PPP1R52, BCLL2L1

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?