Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2015 Nov 19;7(1):46-50.
doi: 10.1021/acsmedchemlett.5b00318. eCollection 2016 Jan 14.

iPSC Neuronal Assay Identifies Amaryllidaceae Pharmacophore with Multiple Effects against Herpesvirus Infections

James McNulty  1 , Leonardo D'Aiuto  2 , Yun Zhi  2 , Lora McClain  3 , Carlos Zepeda-Velázquez  1 , Spencer Ler  1 , Hilary A Jenkins  1 , Michael B Yee  4 , Paolo Piazza  5 , Robert H Yolken  6 , Paul R Kinchington  4 , Vishwajit L Nimgaonkar  3
Affiliations

iPSC Neuronal Assay Identifies Amaryllidaceae Pharmacophore with Multiple Effects against Herpesvirus Infections

James McNulty et al. ACS Med Chem Lett. .

Abstract

The Amaryllidaceae alkaloid trans-dihydrolycoricidine 7 and three analogues 8-10 were produced via asymmetric chemical synthesis. Alkaloid 7 proved superior to acyclovir, the current standard for herpes simplex virus, type 1 (HSV-1) infection. Compound 7 potently inhibited lytic HSV-1 infection, significantly reduced HSV-1 reactivation, and more potently inhibited varicella zoster virus (VZV) lytic infection. A configurationally defined (3R)-secondary alcohol at C3 proved crucial for efficacious inhibition of lytic HSV-1 infection.

Keywords: Amaryllidaceae; HSV-1; Herpesvirus; alkaloid; stem cells; varicella zoster.

PubMed Disclaimer

Conflict of interest statement

The authors declare no competing financial interest.

Figures

Figure 1
Figure 1
Structure of antiviral Amaryllidaceae alkaloids. Alkaloids 1–7 are natural products. Compounds 7–10 were accessed via total synthesis. Lycorine 1, pseudolycorine 2, pancratistatin 3, 7-deoxypancratistatin 4, narciclasine 5, 7-deoxynarciclasine 6, trans-dihydrolycoricidine 7, 3-epi-trans-dihydrolycoricidine 8, and 3-deoxy-trans-dihydrolycoricidine 9.
Scheme 1
Scheme 1
Asymmetric organocatalytic Michael-aldol [3 + 3]-cycloaddition reaction of cinnamaldehyde derivative 12 and azido-acetone derivatives 13a allows a one-step access to stereochemically defined cyclohexanol 14a permitting rapid asymmetric entry to natural alkaloid 7, while reaction of 13b leads to 14b, permitting access to C3-epimeric and deoxy analogues 8, 9, and 10.
Scheme 2
Scheme 2
Reagents and conditions: Yields of isolated products (%) are indicated. (a) 13b (1.00 equiv), 12 (1.05 equiv), 11 (10 mol %), quinidine (10 mol %), CH2Cl2, −10 °C → RT, 73%. (b) DMDC (3.0 equiv), H2, 10% Pd/C (0.075 equiv), EtOAc, 50 psi, RT, 79%. (c) Me4NHB(OAc)3 (4.0 equiv), AcOH (10.0 equiv), MeCN, RT, 90%. (d) AcCl (100 equiv). (e) Tf2O (5.0 equiv), DMAP (3.0 equiv), CH2Cl2, 0 °C → RT, 73%. (f) K2CO3 (0.10 equiv), H2O/MeOH 9:1, RT, 81%. DMDC = dimethyl dicarbonate, Tf = triflic, DMAP = 4-dimethylaminopyridine, Moc = methyloxycarbonyl.
Figure 2
Figure 2
Natural and synthetic Amaryllidaceae alkaloids selectively inhibit HSV-1 infection in Vero cells and iPSC-neurons. (a) Flow cytometry (FC) analysis of uninfected and HSV-1 infected Vero cells (MOI 1) 24 hpi. (b) Viral titer in Vero cell culture supernatants estimated with virus yield reduction assay using Vero cells 48 hpi. Plaques were not observed in cultures treated with compounds 3, 5, and 7. A very low titer was observed for ACV (33.3 pfu/mL). (c) FC analysis of uninfected and HSV-1 infected iPSC-neurons (MOI 0.3) 24 h postinfection (hpi). (d) Viral titer in iPSC-neuronal culture supernatants estimated with virus yield reduction assay using Vero cells 48 hpi. Plaques were observed only in cultures infected with supernatants deriving from untreated, infected neurons. Dimethyl sulfoxide (DMSO) is the vehicle. The data represent an average of three independent experiments. Error bars represent standard deviations.
Figure 3
Figure 3
Compound 7 significantly reduces HSV-1 reactivation and inhibits VZV lytic infection. (a–c) Sodium butyrate (NaB) induces HSV-1 reactivation in quiescent neuronal cultures (QCs) but not in infected cultures treated with 7 or 7 + IFN-α. Quiescent neuronal cultures were produced by incubating iPSC-neurons with HSV-1 in the presence of 5-bromodeoxyuridine (5BVdU) and interferon-α (IFN-α). Cells were also infected with HSV-1 in the presence of 7 or 7 + IFN-α. After 7 days, drugs were withdrawn (wd) and cells were either cultured in the presence of NaB to induce HSV-1 reactivation or vehicle for an additional 5 days. (a) Percentage of EGFP+ and EGFP-RFP+ cells. (b) Mean fluorescence intensity (MFI) of EGFP+ cells and EGFP-RFP+ cells. (c) Viral titer in culture supernatants estimated with virus yield reduction assay using Vero cells 48 hpi. Indicated p-values for comparisons in panel b involving EGFP and EGFP-RFP are depicted in green and red, respectively. The data represent an average of three independent experiments. Error bars represent standard deviations.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Kinchington P. R.; Leger A. J.; Guedon J. M.; Hendricks R. L. Herpes simplex virus and varicella zoster virus, the house guests who never leave. Herpesviridae 2012, 3, 5.10.1186/2042-4280-3-5. - DOI - PMC - PubMed
    1. Burrel S.; Aime C.; Hermet L.; Ait-Arkoub Z.; Agut H.; Boutolleau D. Surveillance of herpes simplex virus resistance to antivirals: a 4-year survey. Antiviral Res. 2013, 100, 365–72. 10.1016/j.antiviral.2013.09.012. - DOI - PubMed
    1. Jin Z. Amaryllidaceae and Sceletium alkaloids. Nat. Prod. Rep. 2013, 30, 849–68. 10.1039/c3np70005d. - DOI - PubMed
    1. Evidente A.; Kireev A. S.; Jenkins A. R.; Romero A. E.; Steelant W. F. A.; van Slambrouck S.; Kornienko A. Biological evaluation of structurally diverse amaryllidaceae alkaloids and their synthetic derivatives: discovery of novel leads for anticancer drug design. Planta Med. 2009, 75, 501–7. 10.1055/s-0029-1185340. - DOI - PMC - PubMed
    1. Gabrielsen B.; Monath T. P.; Huggins J. W.; Kefauver D. F.; Pettit G. R.; Groszek G.; Hollingshead M.; Kirsi J. J.; Shannon W. M.; Schubert E. M.; DaRe J.; Ugarkar B.; Ussery M.; Phelan M. J. Antiviral (RNA) activity of selected Amaryllidaceae isoquinoline constituents and synthesis of related substances. J. Nat. Prod. 1992, 55, 1569–81. 10.1021/np50089a003. - DOI - PubMed

LinkOut - more resources