Structure of 3-Bromopiperidine-2,6-dione
CAS No.: 62595-74-8
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The BI-3802 was designed by Boehringer Ingelheim and could be obtained free of charge through the Boehringer Ingelheim open innovation portal opnMe.com, associated with its negative control.
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PROTACs Targeting MLKL Protect Cells from Necroptosis
Rathje, Oliver H. ; Perryman, Lara ; Payne, Richard J. ; Hamprecht, Dieter W. ;
Abstract: Mixed Lineage Kinase domain-Like pseudokinase (MLKL) is implicated in a broad range of diseases due to its role as the ultimate effector of necroptosis and has therefore emerged as an attractive drug target. Here, we describe the development of PROteolysis TArgeting Chimeras (PROTACs) as a novel approach to knock down MLKL through chem. means. A series of candidate degraders were synthesized from a high-affinity pyrazole carboxamide-based MLKL ligand leading to the identification of a PROTAC mol. that effectively degraded MLKL and completely abrogated cell death in a TSZ model of necroptosis. By leveraging the innate ability of these PROTACs to degrade MLKL in a dose-dependent manner, the quant. relationship between MLKL levels and necroptosis was interrogated. This work demonstrates the feasibility of targeting MLKL using a PROTAC approach and provides a powerful tool to further our understanding of the role of MLKL within the necroptotic pathway.
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CAS No. : | 62595-74-8 |
Formula : | C5H6BrNO2 |
M.W : | 192.01 |
SMILES Code : | O=C(C(Br)CC1)NC1=O |
MDL No. : | MFCD11053059 |
InChI Key : | RYSICGXZRVMXDP-UHFFFAOYSA-N |
Pubchem ID : | 12570775 |
GHS Pictogram: |
![]() |
Signal Word: | Warning |
Hazard Statements: | H302-H315-H319-H332-H335 |
Precautionary Statements: | P280-P305+P351+P338-P310 |
Num. heavy atoms | 9 |
Num. arom. heavy atoms | 0 |
Fraction Csp3 | 0.6 |
Num. rotatable bonds | 0 |
Num. H-bond acceptors | 2.0 |
Num. H-bond donors | 1.0 |
Molar Refractivity | 39.02 |
TPSA ? Topological Polar Surface Area: Calculated from |
46.17 Ų |
Log Po/w (iLOGP)? iLOGP: in-house physics-based method implemented from |
0.94 |
Log Po/w (XLOGP3)? XLOGP3: Atomistic and knowledge-based method calculated by |
0.21 |
Log Po/w (WLOGP)? WLOGP: Atomistic method implemented from |
-0.19 |
Log Po/w (MLOGP)? MLOGP: Topological method implemented from |
0.48 |
Log Po/w (SILICOS-IT)? SILICOS-IT: Hybrid fragmental/topological method calculated by |
1.2 |
Consensus Log Po/w? Consensus Log Po/w: Average of all five predictions |
0.53 |
Log S (ESOL):? ESOL: Topological method implemented from |
-1.16 |
Solubility | 13.2 mg/ml ; 0.0687 mol/l |
Class? Solubility class: Log S scale |
Very soluble |
Log S (Ali)? Ali: Topological method implemented from |
-0.74 |
Solubility | 35.0 mg/ml ; 0.183 mol/l |
Class? Solubility class: Log S scale |
Very soluble |
Log S (SILICOS-IT)? SILICOS-IT: Fragmental method calculated by |
-1.72 |
Solubility | 3.64 mg/ml ; 0.019 mol/l |
Class? Solubility class: Log S scale |
Soluble |
GI absorption? Gatrointestinal absorption: according to the white of the BOILED-Egg |
High |
BBB permeant? BBB permeation: according to the yolk of the BOILED-Egg |
No |
P-gp substrate? P-glycoprotein substrate: SVM model built on 1033 molecules (training set) |
No |
CYP1A2 inhibitor? Cytochrome P450 1A2 inhibitor: SVM model built on 9145 molecules (training set) |
No |
CYP2C19 inhibitor? Cytochrome P450 2C19 inhibitor: SVM model built on 9272 molecules (training set) |
No |
CYP2C9 inhibitor? Cytochrome P450 2C9 inhibitor: SVM model built on 5940 molecules (training set) |
No |
CYP2D6 inhibitor? Cytochrome P450 2D6 inhibitor: SVM model built on 3664 molecules (training set) |
No |
CYP3A4 inhibitor? Cytochrome P450 3A4 inhibitor: SVM model built on 7518 molecules (training set) |
No |
Log Kp (skin permeation)? Skin permeation: QSPR model implemented from |
-7.32 cm/s |
Lipinski? Lipinski (Pfizer) filter: implemented from |
0.0 |
Ghose? Ghose filter: implemented from |
None |
Veber? Veber (GSK) filter: implemented from |
0.0 |
Egan? Egan (Pharmacia) filter: implemented from |
0.0 |
Muegge? Muegge (Bayer) filter: implemented from |
1.0 |
Bioavailability Score? Abbott Bioavailability Score: Probability of F > 10% in rat |
0.55 |
PAINS? Pan Assay Interference Structures: implemented from |
0.0 alert |
Brenk? Structural Alert: implemented from |
2.0 alert: heavy_metal |
Leadlikeness? Leadlikeness: implemented from |
No; 1 violation:MW<1.0 |
Synthetic accessibility? Synthetic accessibility score: from 1 (very easy) to 10 (very difficult) |
2.34 |
* All experimental methods are cited from the reference, please refer to the original source for details. We do not guarantee the accuracy of the content in the reference.
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
62% | With N-ethyl-N,N-diisopropylamine; In N,N-dimethyl-formamide; for 16h; | 7- Chloro-1,2,3,4-tetrahydroisoquinoline (75.0 mg) was dissolved in DMF (0.7 mL). MA6-019 (86.0 mg) and DIPEA (a56.8 muL) were added into the mixture and stirred for 16 h. Purification by washing with aq. NaHCO3 (5 mL) and trituration from ethyl acetate/hexanes gave MA7-074 as a biege solid (77 mg, 62%). HPLC: >99% [tR = 11.6 min, 15% MeOH, 85 water (with 0.1% TFA), 20 min]. 1H NMR (400 MHz, DMSO) delta 10.71 (s, 1H), 7.17-7.11 (m , 3H), 3.94 (d, J = 16 Hz, 1H), 3.79 (d, J = 16 Hz, 1H), 3.65 (dd J = 8.0, J = 4.0, 1H), 2.90-2.95 (m, 1H), 2.90-2.81 (m, 1H), 2.70-2.80 (m, 2H), 2.65-2.50 (m, 2H), 2.20-2.08 (m, 1H), 1.95-1.85 (m, 1H). HRMS (ESI+): m/z calcd for C14H16ClN2O2 (M+H)+ 279.0894, found 279.0894, m/z calcd for C14H15ClN2O2Na (M+Na)+ 301.0714, found 301.0711. HPLC-MS: HPLC-MS (ESI+): m/z 279.2 [40%, (M+H)+], 277.1 [100%, (M-H)-]. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
9% | 3-(2-Oxo-4-phenylpyrrolidin-l-yl)piperidine-2,6-dione. To a stirred solution of 4- phenylpyrrolidin-2-one (21 1 mg, 1.31 mmol) in DMF (2 mL) was added NaH (57.6 mg, 1.44 mmol, 60% w/w dispersed into mineral oil) at 0 C under nitrogen atmosphere. The reaction mixture was stirred for 20 min at 0 C. To the above mixture was added dropwise a solution of 3- bromopiperidine-2,6-dione (125.7 mg, 0.65 mmol) in DMF (0.5 mL) at 0 C. The resulting mixture was stirred for additional 3 hours at room temperature. The resulting mixture was quenched with AcOH (0.5 mL) and concentrated under reduced pressure. The crude product was purified by prep-HPLC with the following conditions: Column: XBridge Shield RP18 EVO Column, 5 urn, 19 x 150 mm; Mobile Phase A: water (plus 0.05% FA), Mobile Phase B: ACN; Flow rate: 20 mL/min; Gradient: 15% B to 45% B in 7 min; Detector: UV 220 nm; Rt: 5.72 min). Desired fractions were collected and concentrated under reduced pressure. The residue was lyophilized to afford 3-(2-oxo-4-phenylpyrrolidin-l-yl)piperidine-2,6-dione, 29, as a white solid (29.4 mg, 9%):1H NMR (400 MHz, DMSO-d6) delta 10.94 (br s, 1H), 7.40 - 7.31 (m, 4H), 7.27 (dt, J = 5.9, 2.8 Hz, 1H), 4.93 - 4.86 (m, 1H), 3.79 - 3.56 (m, 2H), 3.33 - 3.15 (m, 1H), 2.90 - 2.69 (m, 2H), 2.60 - 2.50 (m, 1H), 2.48 - 2.36 (m, 1H), 2.35 - 2.16 (m, 1H), 1.92 - 1.80 (m, 1H); LC/MS (ESI, m/z): [(M + 1)]+= 273.2. | |
9% | To a stirred solution of 4- phenylpyrrolidin-2-one (21 1 mg, 1.31 mmol) in DMF (2 mL) was added NaH (57.6 mg, 1.44 mmol, 60% w/w dispersed into mineral oil) at 0 C under nitrogen atmosphere. The reaction mixture was stirred for 20 min at 0 C. To the above mixture was added dropwise a solution of 3- bromopiperidine-2,6-dione (125.7 mg, 0.65 mmol) in DMF (0.5 mL) at 0 C. The resulting mixture was stirred for additional 3 hours at room temperature. The resulting mixture was quenched with AcOH (0.5 mL) and concentrated under reduced pressure. The crude product was purified by prep-HPLC with the following conditions: Column: XBridge Shield RP18 EVO Column, 5 urn, 19 x 150 mm; Mobile Phase A: water (plus 0.05% FA), Mobile Phase B: ACN; Flow rate: 20 mL/min; Gradient: 15% B to 45% B in 7 min; Detector: UV 220 nm; Rt: 5.72 min). Desired fractions were collected and concentrated under reduced pressure. The residue was lyophilized to afford 3-(2-oxo-4-phenylpyrrolidin-l-yl)piperidine-2,6-dione, 1-29, as a white solid (29.4 mg, 9%): 1H NMR (400 MHz, OMSO-d6) delta 10.94 (br s, 1H), 7.40 - 7.31 (m, 4H), 7.27 (dt, J = 5.9, 2.8 Hz, 1H), 4.93 - 4.86 (m, 1H), 3.79 - 3.56 (m, 2H), 3.33 - 3.15 (m, 1H), 2.90 - 2.69 (m, 2H), 2.60 - 2.50 (m, 1H), 2.48 - 2.36 (m, 1H), 2.35 - 2.16 (m, 1H), 1.92 - 1.80 (m, 1H); LC/MS (ESI, m/z): [(M + 1)]+ = 273.2. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
31% | To a stirred solution of <strong>[19932-85-5]6-bromo-3H-1,3-benzoxazol-2-one</strong> (1, 6 g, 28.04 mmol) in THF (200 mL) was added sodium hydride (60% dispersion in mineral oil) (1.29 g, 56.07 mmol) portionwise and the mixture was heated at 60 C for 1 h. This mixture was added dropwise via cannula to a stirred solution of 3-bromopiperidine-2,6-dione (2, 8.07 g, 42.05 mmol) in THF (50 mL) at 60 C and stirred for 2 h. The reaction was quenched with saturated ammonium chloride solution and extracted with ethyl acetate. The organics were washed with water and brine, dried over anhydrous sodium sulphate and concentrated. The residue was purified by silica gel chromatography (50% Ethyl acetate:Hexanes) to obtain 3-(6-bromo-2-oxobenzo[d]oxazol-3(2H)-yl)piperidine-2,6-dione (3, 2.9 g, 8.71 mmol, 31% yield). LCMS (ES-): m/z 323.0 [M-H]- 1H NMR (400 MHz, DMSO- D6) ^ ^11.23 (s, 1H), 7.73 (s, 1H), 7.43 (d, J=8.4 Hz, 1H), 7.26 (d, J=8.36 Hz, 1H), 5.41-5.37 (m, 1H), 2.87-2.84 (m, 1H), 2.71-2.64 (m, 2H), 2.18-2.15 (m, 1H) |
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