BL-918, a racemic mixture, is a novel and potent small molecule activator of UNC-51-like kinase 1 (ULK1), inducing cytoprotective autophagy for Parkinson’s disease treatment. UNC-51-like kinase 1 (ULK1), the yeast Atg1 ortholog, is the sole serine-threonine kinase and initiating enzyme in autophagy, which may be regarded as a target in Parkinson's disease (PD). 33i (BL-918) as a potent activator of ULK1 by structure-based drug design. Subsequently, some key amino acid residues (Arg18, Lys50, Asn86, and Tyr89) were found to be crucial to the binding pocket between ULK1 and 33i by site-directed mutagenesis. Moreover, it was found that 33i induced autophagy via the ULK complex in SH-SY5Y cells. Intriguingly, this activator displayed a cytoprotective effect on MPP+-treated SH-SY5Y cells, as well as protected against MPTP-induced motor dysfunction and loss of dopaminergic neurons by targeting ULK1-modulated autophagy in mouse models of PD. Together, these results demonstrate the therapeutic potential to target ULK1, and 33i, the novel activator of ULK1, may serve as a candidate drug for future PD treatment.
Physicochemical Properties
| Molecular Formula | C23H15F8N3OS |
| Molecular Weight | 533.4369 |
| Exact Mass | 533.08 |
| CAS # | 2101517-69-3 |
| Related CAS # | (Rac)-BL-918;2435589-07-2 |
| PubChem CID | 146014432 |
| Appearance | White to off-white solid powder |
| Density | 1.5±0.1 g/cm3 |
| Index of Refraction | 1.592 |
| LogP | 6.85 |
| Hydrogen Bond Donor Count | 3 |
| Hydrogen Bond Acceptor Count | 10 |
| Rotatable Bond Count | 5 |
| Heavy Atom Count | 36 |
| Complexity | 737 |
| Defined Atom Stereocenter Count | 1 |
| SMILES | C1=CC=C(C=C1)[C@H](C(=O)NC2=C(C=C(C=C2)F)F)NC(=S)NC3=CC(=CC(=C3)C(F)(F)F)C(F)(F)F |
| InChi Key | BDBWQANRZRCMMD-LJQANCHMSA-N |
| InChi Code | InChI=1S/C23H15F8N3OS/c24-15-6-7-18(17(25)11-15)33-20(35)19(12-4-2-1-3-5-12)34-21(36)32-16-9-13(22(26,27)28)8-14(10-16)23(29,30)31/h1-11,19H,(H,33,35)(H2,32,34,36)/t19-/m1/s1 |
| Chemical Name | (2R)-2-[[3,5-bis(trifluoromethyl)phenyl]carbamothioylamino]-N-(2,4-difluorophenyl)-2-phenylacetamide |
| Synonyms | BL-918; BL 918; BL918 |
| HS Tariff Code | 2934.99.9001 |
| Storage |
Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition | Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs) |
Biological Activity
| ln Vitro | BL-918 (compound 33i) exhibits a high binding affinity for ULK1 (KD=0.719 μM) [1]. In neuronal-like SH-SY5Y cells, BL-918 (5 μM) induces autophagy over a 24-hour period [1]. -918 (0.5-50 μM; 24 hours) partially reverses MPP+-induced cell death, as determined by enhancing cell viability [1]. BL-918 (5 μM; duration 6-36 hours) time-drivenly increases the expression levels and phosphorylation status of LC3-II and Beclin-1, while reducing the levels of the autophagy substrate SQSTM1/p62. BL-918 increases the phosphorylation of ULK1 at Ser317 and Ser555, and decreases the phosphorylation of ULK1 at Ser757 [1]. Cell Autophagy Assay[1] |
| ln Vivo | Compound 33i, also known as BL-918 (compound 20, 40, or 80 mg/kg/day; border gavage; started two days prior to the first saline/MPTP injection and continued for five days following the last saline/MPTP injection), dramatically lowers dopamine (DA), homovanillic acid (HVA), and 3,4-diphenyl acetamide (DOPAC) levels [1]. |
| Cell Assay |
Cell Autophagy Assay[1] Cell Types: SH-SY5Y cells Tested Concentrations: 5 μM Incubation Duration: For 24 hrs (hours) Experimental Results: Induced autophagy. Cell viability assay [1] Cell Types: SH-SY5Y Cell Tested Concentrations: 0.5, 5, 50 μM Incubation Duration: 24 hrs (hours) Experimental Results: MPP+-induced cell death can be partially reversed, as determined by enhanced cell viability. Western Blot Analysis[1] Cell Types: SH-SY5Y Cell Tested Concentrations: 5 μM Incubation Duration: 6, 12, 24, 36 hrs (hours) Experimental Results: Time-dependent increase in the expression of LC3-II (key marker of autophagy), Beclin level-1 and its phosphorylation status, thereby reducing the levels of the selective autophagy substrate SQSTM1/p62. |
| Animal Protocol |
Animal/Disease Models: Male C57BL/6 mice (eight weeks old) weighing between 20 and 25 g [1] Doses: 20, 40 or 80 mg/kg Route of Administration: po (oral gavage); daily; first time Start 2 days before injecting saline/MPTP and continue for 5 days after the last injection of saline/MPTP. Experimental Results: Reduce the loss of DA and its metabolites. |
| References |
[1]. Small-Molecule Activator of UNC-51-Like Kinase 1 (ULK1) That Induces Cytoprotective Autophagy for Parkinson's Disease Treatment. J Med Chem. 2018 Apr 12;61(7):2776-2792. |
Solubility Data
| Solubility (In Vitro) | DMSO : ≥ 250 mg/mL (~468.66 mM) |
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (3.90 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 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.8746 mL | 9.3731 mL | 18.7463 mL | |
| 5 mM | 0.3749 mL | 1.8746 mL | 3.7493 mL | |
| 10 mM | 0.1875 mL | 0.9373 mL | 1.8746 mL |