Design, Synthesis, and Biological Evaluation of N-Alkylated Deoxynojirimycin (DNJ) Derivatives for the Treatment of Dengue Virus Infection

Abstract

We recently described the discovery of oxygenated N-alkyl deoxynojirimycin (DNJ) derivative 7 (CM-10-18) with antiviral activity against dengue virus (DENV) infection both in vitro and in vivo. This imino sugar was promising, but had an EC50 against DENV in BHK cells of 6.5 μM, which limited its use in in vivo. Compound 7 presented structural opportunities for activity relationship analysis, which we exploited and report here. These structure-activity relationship studies led to analogs 2h, 2l, 3j, 3l, 3v and 4b–4c with nanomolar antiviral activity (EC50 = 0.3–0.5 μM) against DENV infection, while maintaining low cytotoxicity (CC50 > 500 μM, SI > 1000). In male Sprague-Dawley rats, compound 3l was well tolerated at a dose up to 200 mg/kg and displayed desirable PK profiles, with significantly improved bioavailability (F = 92 ± 4%).

A Novel Dengue Virus Inhibitor, BP13944, Discovered by High-Throughput Screening with Dengue Virus Replicon Cells Selects for Resistance in the Viral NS2B/NS3 Protease

“…One strategy used for this purpose involves a specific target-based approach, e.g., an enzyme-based biochemical assay or structure-based virtual screen (38)(39)(40)(41)(42)(43)(44)(45)(46)(47)(48)(49). An alternative strategy is to use a replication-based approach, e.g., screening through virus infection assays or replicon cell lines (50)(51)(52). Although no drug is currently approved for the treatment of DENV infection, similar approaches have been successfully applied in the search for novel HCV inhibitors, suggesting the feasibility of these strategies.…”

Synthesis and Evaluation of Hybrid Structures Composed of Two Glucosylceramide Synthase Inhibitors

“…Yield 0.004 g (6 mmol, 25 % 1, 28.3, 28.5, 29.7, 31.2, 35.6 (CH 2 nonanoic amide), 50.5 (C-2'), 53.1 (C-1), 55.7 (C-6), 56.6 (C-1'), 63.6 (C-5), 64.7 (2 CH 2 O), 66.6 (C-2), 68.8 (C-3), 69.2 (C-4), 72.3 (C-3'), 114. 8,117.4,119.3 (CHarm),133.9,142.7,142.9 (Cq Carm), 178.3 ppm (C=O);I R( thin film): ñ = 3267, 2927,2856,1670,1508,1286,1199,1136,1066 propanoic acid), 50.7 (C-2'), 53.0 (C-1), 56.0 (OMe), 56.6 (C-1'), 57.3 (C-6), 63.5 (C-5), 64.5 (2 CH 2 O), 65.4 (CH 2 propanoic acid), 66.7 (C-2), 69.3 (C-4), 72.3 (C-3'), 114. 8,115.2,116.6,117.3,119.3 (CHarm),133.7,142.7,142.9,152.3,153.7 Compound 61:T he oxazolidine in compound 60 was hydrolyzed according to the saponification procedure described above.…”

“…8,117.4,119.3 (CHarm),133.9,142.7,142.9 (Cq Carm), 178.3 ppm (C=O);I R( thin film): ñ = 3267, 2927,2856,1670,1508,1286,1199,1136,1066 propanoic acid), 50.7 (C-2'), 53.0 (C-1), 56.0 (OMe), 56.6 (C-1'), 57.3 (C-6), 63.5 (C-5), 64.5 (2 CH 2 O), 65.4 (CH 2 propanoic acid), 66.7 (C-2), 69.3 (C-4), 72.3 (C-3'), 114. 8,115.2,116.6,117.3,119.3 (CHarm),133.7,142.7,142.9,152.3,153.7 Compound 61:T he oxazolidine in compound 60 was hydrolyzed according to the saponification procedure described above. The obtained imine 61 was immediately condensed with the hydroxysuccinimide esters 10, 42 and 55 as described in the general condensing procedure;L C-MS: t R = 3.65 min (linear gradient 0!50 % B);E SI-MS:…”

“…[6] This finding has been confirmedinr ecent years in several studies, and avariety of iminosugar-based GCS inhibitors that differ in the nature of the Nsubstituent buta lso in configurationo ft he piperidine iminosugar have been described. [7,8] Coinciding with these findings, studies were reported indicating that altered glycolipid levels, and in particulart he ganglioside GM3 (aNeu5Ac(2-3)-b-dGalp(1-4)-b-d-Glcp(1-1)Cer),w hichi st he result of ah igh-fat diet, negatively influences insulin signaling by interfering with the insulin receptor. [9] In 2007 we disclosed that AMP-DNM 2 decreases GM3 levels by partial inhibition of GCS in animal models of type 2d iabetes, and in this way restores insulin signaling.…”

“…[7b, 8,19] As econd strategy we recently investigated and that provedu nrewarding is testing am odel [20] proposed by Platt and Butters, who reasonedt hat N-butyldeoxynojirimycin acts on GCS not as ag lucose mimic, but rather as ac eramide mimic.I nt his model, which has not yet been substantiated due to al ack of structurald ata on GCS (the membrane-associatede nzyme has defied efforts to obtain crystallographic data), the hydroxy substituents at positions C2 and C3 and the hydroxymethyl group at C5 (glucopyranosen umbering) of the piperidinec ore mimic the two hydroxy groups of ceramide, with the N-alkyl chain as one of the ceramide alkyl chains. According to this model the C2 OH group would be redundant, and we thought to capitalize on this by testing N-alkylated derivatives of the natural product fagomine (2-deoxydeoxynojirimycin) in its eight configurational isomers.…”