Basic Information
Remarks
Chemical Identifiers
Basic Properties
Lipinski RO5
Hydrophobic Tag Properties
Mechanism of Action
POI (Protein of Interest)
Amino Acid Sequence
3D Structure (/structures/AF-J7MA22-F1-model_v6.cif)
Function
Essential for the formation and stability of microtubules . Required for the organization of the mitotic spindle and for the proper attachment of kinetochores to MTs. Promotes the recruitment of NUDC to the mitotic spindle for mitotic progression.
Degradation Pathway
Mechanism
The ubiquitin-proteasome pathway (UPP) constitutes the principal pathway for protein degradation in eukaryotic cells, mediating the breakdown of target proteins through ubiquitin tagging and subsequent proteasomal degradation. In an ATP-dependent manner, the ubiquitin-activating enzyme (E1) activates ubiquitin (Ub) and conjugates it to a ubiquitin-conjugating enzyme (E2). E3 ligase subsequently catalyzes the transfer of Ub to the target protein, leading to the formation of a polyubiquitin chain that completes the ubiquitination mark. The 19S regulatory particle of the 26S proteasome recognizes this ubiquitination signal and, after deubiquitination, mediates the ATP-dependent delivery of the substrate into the 20S proteasome for degradation.
Description
To investigate the mechanism of the degradation of Hyt-9 in H3122 cells, we performed rescue assays. Pretreatment with the proteasome inhibitor MG-132 (2μM) blocked ALK degradation induced by Hyt-9 in H3122 cells (Figure 4A), suggesting that engagement of the proteasome pathway is essential for the observed ALK degradation. Pretreatment with the lysosomal inhibitor chloroquine did not affect ALK degradation (Figure 4A), suggesting that norbornene-induced degradation does not involve autophagy/lysosome pathways. These results suggested that ALK degradation caused by norbornene-based HyT was mediated by inducing the destabilization of ALK, thereby recruiting Hsp70 without directly binding and then degrading the protein by the proteasome (Figure 4F).
Activity Data
Description
compound Hyt-9 potently induced EML4-ALK protein degradation in a dose-dependent manner with a DC50 of 134 nM after 24 h of treatment (Figure 2A, B).
Description
More over, compound Hyt-9 exhibited more potent antiproliferative activity in H3122 cells, with an IC50 value of 0.07±0.01μM (Figure 1C). (C) Cell growth inhibition in H3122 cells. Cells were treated with the indicated concentrations of compounds Hyt-7‒10 for 72 h (0.032, 0.063, 0.125, 0.250, 0.500, 1.000, 2.000, and 4.000μM from left to right), and IC50 values were determined using MTT assays.
Description
Furthermore, we investigated whether Hyt-9 could exhibit advantages over ALK inhibitor A1, A1-based PROTAC Q2, and adamantane-based HyT Hyt-6 administered P.O. every other day for 21 days(Supporting Information, Figure S9).Hyt-9(20 mg/kg) potently inhibited tumor growth with a tumor inhibition rate of 58.03 %, which was superior to that of A1(42.20 %), Q2(44.61 %), and Hyt-6 (43.12 %) at the same dosing schedule (Supporting Information, Figure S9A, B, and C)
PK (Pharmacokinetics)
| Route | Dose | T1/2 | Tmax | Cmax | AUC | MRT | CL | F | Vd | Vz |
|---|---|---|---|---|---|---|---|---|---|---|
| I.V. | 1 mg/kg | 4.29±0.65 h | 0.08 h | 200.4±33.0 ng/mL | (0-t)316.0±62.3 h*ng/mL (0-∞)418.8±105.2 h*ng/mL | (last)5.37±0.81 h | (F_obs)41.4±9.4 Ml/min/kg | 13.1±1.8 L/kg | ||
| P.O. | 10mg/kg | 5.45±1.81 h | 1.33±0.58 h | 53.1±12.5 ng/mL | (0-t)270.5±100.8 h*ng/mL （0-∞）270.5±100.8 h*ng/mL | (last)8.6±2.8 h | 8.6±3.2 % | 292.4±102.1 L/kg |
Disease Information
| Disease | ICD-11 | Source |
|---|---|---|
| lung adenocarcinoma | 2C25.0 | Cell Line Origin |
| Hemimegalencephaly | LA05.1 | Gene-Disease Association |
| Familial benign macrocephaly | LB70.3 | Gene-Disease Association |
| Overgrowth Syndrome | LD2C | Gene-Disease Association |
| Hydrocephalus | 8D64.Z | Gene-Disease Association |