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A μ-opioid receptor modulator that works cooperatively with naloxone
Evan S. O& ; amp ; amp , et al. Nature,2024,631,686-693. DOI: 10.1038/s41586-024-07587-7 PubMed ID: 38961287
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Abstract: The μ-opioid receptor (μOR) is a well-established target for analgesia1, yet conventional opioid receptor agonists cause serious adverse efects, notably addiction and respiratory depression. These factors have contributed to the current opioid overdose epidemic driven by fentanyl2, a highly potent synthetic opioid. μOR negative allosteric modulators (NAMs) may serve as useful tools in preventing opioid overdose deaths, but promising chemical scafolds remain elusive. Here we screened a large DNA-encoded chemical library against inactive μOR, counter-screening with active, G-protein and agonist-bound receptor to ‘steer’ hits towards conformationally selective modulators. We discovered a NAM compound with high and selective enrichment to inactive μOR that enhances the afnity of the key opioid overdose reversal molecule, naloxone. The NAM works cooperatively with naloxone to potently block opioid agonist signalling. Using cryogenic electron microscopy, we demonstrate that the NAM accomplishes this efect by binding a site on the extracellular vestibule in direct contact with naloxone while stabilizing a distinct inactive conformation of the extracellular portions of the second and seventh transmembrane helices. The NAM alters orthosteric ligand kinetics in therapeutically desirable ways and works cooperatively with low doses of naloxone to efectively inhibit various morphine-induced and fentanyl-induced behavioural efects in vivo while minimizing withdrawal behaviours. Our results provide detailed structural insights into the mechanism of negative allosteric modulation of the μOR and demonstrate how this can be exploited in vivo.
Purchased from AmBeed: 52409-22-0 ; 93267-04-0 ; 220728-62-1
CAS No. : | 93267-04-0 | MDL No. : | MFCD00216579 |
Formula : | C9H16INO4 | Boiling Point : | - |
Linear Structure Formula : | ICH2CH(NHCO2C(CH3)3)CO2CH3 | InChI Key : | UGZBFCCHLUWCQI-LURJTMIESA-N |
M.W : | 329.13 | Pubchem ID : | 10903591 |
Synonyms : |
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Signal Word: | Warning | Class: | N/A |
Precautionary Statements: | P261-P305+P351+P338 | UN#: | N/A |
Hazard Statements: | H302-H315-H319-H335 | Packing Group: | N/A |
GHS Pictogram: |
* 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 |
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63% | Degassed anhydrous DMF (42 mL) was added to zinc (13.72 g, 209.8 mmol) under a flow of argon. Chlorotrimethylsilane (5.30 mL, 42.0 mmol) was added and the mixture stirred vigorously for 30 min. Stirring was stopped and the zinc was allowed to settle. The supernatant was decanted under a flow of argon and the zinc washed with degassed DMF (2 × 20 mL). A solution of Boc-3-iodoalanine methyl ester (29.9 g, 90.9 mmol) in degassed DMF (75 mL) was added to the zinc. Upon addition an exotherm was observed. The cloudy gray solution was stirred for 30 min at room temperature and then the zinc was allowed to settle. This was repeated for a second batch of zincate. The two batches of zincate were combined by decanting the supernatant into a clean flask under a flow of argon. A solution of bromide 8 (40.0 g, 140 mmol) in degassed DMF (120 mL), palladium acetate (1.57 g, 6.99 mmol), and X-Phos (6.67 g, 14.0 mmol) were added sequentially. The reaction was heated at 40 C. After 16 h the reaction mixture was poured into water (400 mL) and ethyl acetate (250 mL) was added. The mixture was filtered through a pad of celite, washing the filter cake with ethyl acetate (2 × 50 mL). The layers were separated, the aqueous extracted with ethyl acetate (100 mL) and the organics combined. The organics were then washed with brine (5 × 400 mL), dried (MgSO4), and concentrated in vacuo. The resulting residue was purified by dry flash chromatography (2% ethyl acetate in heptane to 20%) to give compound 9 as a brown solid (35.95 g, 63%).1H NMR (400 MHz, CDCl3): 7.84 (s, 1H), 7.47-7.36 (m, 2H), 5.22-5.14 (m, 1H), 4.72-4.63 (m, 1H), 3.74 (s, 3H), 3.57 (dd, 1H), 3.25-3.15 (m, 1H), 1.33 (s, 9H). |
Yield | Reaction Conditions | Operation in experiment |
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0.16 g | With dicyclohexyl-(2',6'-dimethoxybiphenyl-2-yl)-phosphane; tris-(dibenzylideneacetone)dipalladium(0); iodine; zinc; In N,N-dimethyl-formamide; at 20℃; for 16h;Inert atmosphere; Sealed tube; | To an oven dried 8 mL vial with teflon cap purged with N2 was added Zinc dust (298 mg, 4.56 mmol), DMF(1.5 mL), and iodine (57.8 mg, 0.228 mmol). To this mixture was added (R)-methyl 2-((tert-butoxycarbonyl)amino)-3-iodopropanoate (500 mg, 1.519 mmol), immediately followed by iodine (57.8 mg, 0.228 mmol).Pd2(dba)3 (69.6 mg, 0.076 mmol), 2-DICYCLOHEXYLPHOSPHINO-2?,6?-DIMETHOXYBIPHENYL(62.4 mg, 0.152 mmol), <strong>[3032-81-3]1,3-dichloro-5-iodobenzene</strong> (622 mg, 2.279 mmol) and the reaction mixture wasallowed to stir at rt for 16 h. The crude mixture was diluted in EtOAc (30 mL) and DMF was removed withfour aqueous washes. The organic phase was dried over anhydrous sodium sulfate. The solution was filteredand concentrated, and the crude was purified by silica gel chromatography using 100percent hexanes to 30percentEtOAc/Hexanes. The desired product was obtained as a pale yellow oil, (S)-methyl 2-((tertbutoxycarbonyl)amino)-3-(3,5-dichlorophenyl)propanoate, 0.160 g yield in 77percent purity by LCMS analysis.Analysis LCMS condition F: Retention time=2.84 min; ESI-MS(+) m/z 348.1 (M+H). |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
Step A: Methyl N-(tert-butoxycarbonyl)-4-(2-fluorophenyl)-5-nitrilonorvalinate (0251) To a solution of methyl N-(tert-butoxycarbonyl)-3-iodo-D-alaninate (5.00 g, 15.2 mmol) in DMF (20 mL) was added cesium carbonate (5.44 g, 16.7 mmol) and the mixture was stirred at 23 C. for 2 hours. (2-Fluorophenyl)acetonitrile (5.87 mL, 45.6 mmol) and cesium carbonate (7.42 g, 22.8 mmol) were added and the resulting mixture was stirred for 1 hour. The mixture was filtered and water was added to the filtrate. The mixture was extracted with ethyl acetate (3×). The combined organic extracts were washed with water (3×), brine, dried over sodium sulfate, filtered and concentrated. Purification by silica gel chromatography (0% ethyl acetate?50% ethyl acetate/hexane) gave the title compound as a racemic mixture of cis and trans diastereomers. MS: m/z=378.1 (M+CH3CN+1). |
[ 170848-34-7 ]
(S)-Methyl 2-((tert-butoxycarbonyl)amino)-3-iodopropanoate
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