Anti-herpes Simplex Research Articles

Stereoselective synthesis of 4'-selenonucleosides via seleno-Michael reaction as potent antiviral agents.

Based on the hypothesis that the bulky selenium atom, with 4p orbitals, can sterically hinder the approach of a cellular kinase to 5'-OH for phosphorylation, 4'-selenonucleosides with one-carbon homologation were designed and synthesized via a novel seleno-Michael reaction, with the stereoselectivity controlled by steric effects. 5'-Homo-4'-selenonucleosides (n = 2) demonstrated potent antiherpes simplex virus (HSV-1) activity, indicating that the bulky selenium atom might play a key role in preventing phosphorylation by cellular kinases, resulting in no antiviral activity.

Highly Oxygenated Limonoids and Lignans from Phyllanthus flexuosus.

Graphical Two new highly oxygenated limonoids, flexuosoids A (1) and B (2), and three new arylnaphthalene lignan glycosides, phyllanthusmins D–F (3–5), were isolated from the roots of Phyllanthus flexuosus, in addition to three known lignans, phyllanthusmin C, arabelline, and (+)-diasyringaresinol. Their structures were elucidated on the basis of detailed spectroscopic analysis and chemical methods. Compounds 1 and 2, two new decaoxygenated limonoids with a C-19/29 lactol bridge and heptaoxygenated substituents at C-1, C-2, C-3, C-7, C-11, C-17, and C-30, represent the second example of limonoids in the Euphorbiaceae family. Most of the isolates were tested for their antifeedant, anti-herpes simplex virus 1, and cytotoxic activities. The new limonoids 1 and 2 showed promising antifeedant activity against the beet army worm (Spodoptera exigua) with EC50 values of 25.1 and 17.3 μg/cm2, respectively. In addition, both of them displayed moderate cytotoxicity against the ECA109 human esophagus cancer cell line, along with the known lignan glycoside, phyllanthusmin C, with the IC50 values of 11.5 (1), 8.5 (2), and 7.8 (phyllanthusmin C) μM, respectively.Two new highly oxygenated limonoids, flexuosoids A (1) and B (2), and three new arylnaphthalene lignan glycosides, phyllanthusmins D–F (3–5), were isolated from the roots of Phyllanthus flexuosus, along with three known lignans. Flexuosoids A (1) and B (2) showed promising antifeedant activity against the beet army worm (Spodoptera exigua) with EC50 values of 25.1 and 17.3 μg/cm2, respectively. In addition, both of them and the known lignan glycoside, phyllanthusmin C, displayed moderate cytotoxicity against the ECA109 human esophagus cancer cell line, with the IC50 values of 11.5, 8.5, and 7.8 μM, respectively.Electronic supplementary materialThe online version of this article (doi:10.1007/s13659-014-0026-2) contains supplementary material, which is available to authorized users.

Autophagy Activation and Antiviral Activity by a Licorice Triterpene

The triterpene glycyrrhizic acid (GRA), the main product from the Glycyrrhiza glabra medicinal plant, is known for its antiinflammatory and antimicrobial activity. In this work, GRA was studied for its ability to induce the autophagic process activator Beclin 1 in epithelial cells and to observe how this property could influence its antiviral activity. After 24 h of treatment, GRA induced a Beclin 1 production that was more than twofold higher than that produced by rapamycin, used as a reference compound. When the compounds were added to HeLa cells together with the viruses, GRA demonstrated a strong antiherpes simplex virus type 1 (HSV1) activity, whereas rapamycin had no activity. However, if the compounds were added to the cells 24 h before the viruses, GRA induced the production of an even higher amount of Beclin 1 and showed an improved antiviral effect; under these conditions, rapamycin was also able to exert a significant anti-HSV1 activity. In conclusion, GRA is a strong inducer of the autophagy activator Beclin 1, which establishes a resistance state to HSV1 replication.

Cathelicidin LL-37 and HSV-1 Corneal Infection: Peptide Versus Gene Therapy.

To evaluate the potential utility of collagen-based corneal implants with anti-Herpes Simplex Virus (HSV)-1 activity achieved through sustained release of LL-37, from incorporated nanoparticles, as compared with cell-based delivery from model human corneal epithelial cells (HCECs) transfected to produce endogenous LL-37. We tested the ability of collagen-phosphorylcholine implants to tolerate the adverse microenvironment of herpetic murine corneas. Then, we investigated the efficacy of LL-37 peptides delivered through nanoparticles incorporated within the corneal implants to block HSV-1 viral activity. In addition, LL-37 complementary DNA (cDNA) was transferred into HCECs to confer viral resistance, and their response to HSV-1 infection was examined. Our implants remained in herpetic murine corneas 7 days longer than allografts. LL-37 released from the implants blocked HSV-1 infection of HCECs by interfering with viral binding. However, in pre-infected HCECs, LL-37 delayed but could not prevent viral spreading nor clear viruses from the infected cells. HCECs transfected with the LL-37 expressed and secreted the peptide. Secreted LL-37 inhibited viral binding in vitro but was insufficient to protect cells completely from HSV-1 infection. Nevertheless, secreted LL-37 reduced both the incidence of plaque formation and plaque size. LL-37 released from composite nanoparticle-hydrogel corneal implants and HCEC-produced peptide, both showed anti-HSV-1 activity by blocking binding. However, while both slowed down virus spread, neither was able on its own to completely inhibit the viruses. LL-37 releasing hydrogels may have potential utility as corneal substitutes for grafting in HSV-1 infected corneas, possibly in combination with LL-37 producing therapeutic cells.

Anti-Herpes Simplex Viruses activity of Camellia sinensis, member of the family Theaceae (green tea)

Background Medicinal plants were shown to possess several bioactivities of which Camellia sinensis, a popularly used refreshing beverage has been reported to have immune modulating and anti viral activities. From our lab we had already shown that C. sinensis extracts possessed anti-HIV and anti-HBV activities. In the current study we evaluated its anti-HSV activities. Methods Aqueous and methanolic extracts of C. sinensis were tested for their anti-HSV property by CPE reduction assay on Vero cell monolayers. Cytotoxicity of the extract was measured by MTT assay. Results Experiments showed that 12 µg/mL concentration of C. sinensis completely inhibited HSV-1 and 2 and the drug was found to be non toxic up to 10 mg/mL. Conclusion The study showed that C.sinesis extracts and EGCG compound are potent inhibitor of both HSV-1 and HSV-2 infection and it was found to be relatively non toxic.

Antiherpetic Mechanism of a Sulfated Derivative of Agaricus brasiliensis Fruiting Bodies Polysaccharide

Objective: To study the anti-herpes simplex virus (HSV) activity of a (1→6)-(1→3)-β-<smlcap>D</smlcap>-glucan isolated from Agaricus brasiliensis fruiting bodies (FR) as well as its chemically sulfated derivative (FR-S). Methods: The antiherpetic activity and mechanism of action was studied by viral plaque assay applying different methodological strategies. Results: Although FR presented no in vitro antiherpetic action at 1 mg/ml, FR-S displayed promising anti-HSV-1 and anti-HSV-2 activities in both simultaneous and postinfection treatments, resulting in selectivity indices (CC₅₀/EC₅₀) higher than 393. FR-S had no virucidal effect, but significantly suppressed HSV-1 (EC₅₀ = 0.32 µg/ml) and HSV-2 (EC₅₀ = 0.10 µg/ml) adsorption. FR-S was less effective on adsorption inhibition of mutant virus strains devoid of gC (HSV-1 gC^-39 and HSV-2 gCneg1), indicating a possible interaction with this glycoprotein. The reduction of viral adsorption upon cell pretreatment with FR-S also suggests its interaction with cellular components. FR-S inhibited HSV-1 (EC₅₀ = 8.39 µg/ml) and HSV-2 (EC₅₀ = 2.86 µg/ml) penetration more efficiently than heparin. FR-S reduced HSV-1 and HSV-2 cell-to-cell spread. A synergic effect between FR-S and acyclovir was also detected. Conclusions: FR-S displays an interesting mechanism of antiviral action and represents a promising candidate for the treatment and/or prevention of herpetic infections, to be used as a single therapeutic agent or in combination with acyclovir.

In vitro evaluation of anti-herpes simplex-1 activity of three standardized medicinal plants from Lamiaceae.

Background:Rosmarinic acid (RA) is a phenolic acid with antioxidant and anti-viral effects. We have studied anti-herpes simplex virus type 1 (HSV-1) effect of three medicinal plants from Lamiaceae family which have been standardized on the basis of RA content.Materials and Methods:Methanolic extract of Teucrium polium, Ziziphora clinopoides, and Salvia rhytidea was prepared by maceration method and RA content of the plants was determined using a spectrophotometric method. Maximum nontoxic concentration (MNTC) of the extracts was determined using neutral red method. Serial dilutions of extracts up to MNTC were examined on Vero cells for anti-HSV-1 effect by plaque assay in comparison to acyclovir as a positive control.Results:Among the tested extracts, T. polium contained the highest percentage of RA (1.8%w/w) and exhibited the least toxicity (MNTC = 1000 μg/ml). The greatest anti-HSV-1 was shown by T. polium and Z. clinopoides extracts which exhibited both time and concentration-dependent plaque inhibition.Conclusion:Considering the low toxicity and significant anti-viral effect of T. polium extract, this plant would prove valuable as an active anti-viral drug.

Evaluation of cytotoxicity, anti-herpes simplex virus type 1 (HSV-1) and antibacterial activities of Ficus vasta and phytoconstituents

This study deals with the evaluation of in vitro cytotoxicity of the methanol 80% extract of Ficus vasta aerial parts in Vero cells, anti-HSV-1 and antibacterial activities and the determination of the phytochemical content of the F. vasta methanol extract. The extract was tested for its cytotoxicity in Vero cells and for anti-HSV-1 and against bacterial strains as S. aureus, S. epidermidis, E. coli and P. aeruginosa and also phytochemical constituents were detected. The results showed that this extract presented a CC50 value of 389 µg/mL, an intermediate value taking into account other researchers reported for some Moraceae species. Regarding the other biological activities, antiviral effect was not demonstrated by a HSV-1 challenge dilution of 100x tissue culture infective dose 50% (TCID50) and also by the disk-diffusion method at the higher non-toxic concentration of the extract (119 µg/mL). Phytochemical analysis of the methanol extract proved the presence of carbohydrates, tannins, flavonoids, coumarins, alkaloids and triterpenes. Chromatographic separation of the methanol extract resulted in the isolation and identification of ?-sitosterol, stigmasterol, lupeol, ursolic acid, naringenin, luteolin, quercetin, vitexin, quercetin 3-O-?-glucoside and rutin. The results indicated that F. vasta aerial parts methanol extract is potential candidate for experiments of biological activity screening that are not anti-HSV-1 or antibacterial against the strains evaluated in this study, to be carried on in the future.DOI: http://dx.doi.org/10.3329/icpj.v3i1.17295 International Current Pharmaceutical Journal, December 2013, 3(1): 211-218

Value of detection of anti-herpes simplex virus type 2 antibodies in the diagnosis of genital herpes before in vitro fertilization

Objective To determine the prevalent herpes simplex virus (HSV) strain in patients with genital herpes (GH),and to evaluate the sensitivity,specificity,positive predictive value (PPV) and negative predictive value (NPV) of anti-herpes simplex virus type 2 (HSV2) IgG and IgM antibodies in the diagnosis of genital herpes (GH) before in vitro fertilization (IVF).Methods Totally,193 HSV2 clinical strains isolated in cell culture from the lesions of patients with GH in the Department of Dermatology,First Affiliated Hospital,Sun Yat-sen University between 2009 and 2011 were typed by using type-specific fluorescein isothiocyanate (FITC)-labelled anti-HSV monoclonal antibodies.Serum samples were obtained from 57 anti-HSV2 IgM/IgG antibody-positive females with suspected GH as well as their husbands (clinical observation group),68 HSV culture-positive patients diagnosed with GH (positive control group),and 120 children aged 8-12 years (negative control group).Enzyme-linked immunosorbent assay (ELISA) was performed to detect anti-HSV1/HSV2 IgG/IgM antibodies in these serum samples.Statistical analysis was carried out using chi-square test.Results There was a significant difference between the positive control group and negative control group in the positivity rate of anti-HSV1 IgG (89.71％ (61/68) vs.40.80％ (49/120),P ＜ 0.01) and anti-HSV2 IgG (91.18％ (62/68) vs.0,P ＜ 0.01),but not in that of anti-HSV1 IgM (20.59％ (14/68) vs.21.70％ (26/120),P ＞ 0.05) or anti-HSV2 lgM (13.24％ (9/68)vs.13.30％ (16/120),P ＞ 0.05).In the clinical observation group,the positivity rate of anti-HSV1 and anti-HSV2 IgM antibodies,anti-HSV1 and anti-HSV2 IgG antibodies was 80.70％ (46/57),91.23％ (52/57),84.21％ (48/57) and 14.04％ (8/57) respectively in the females,19.30％ (11/57),8.77％ (5/57),87.71％ (50/57),12.28％ (7/57)respectively in the males,with significant differences in the positivity rate of anti-HSV1 and-HSV2 IgM antibodies (both P ＜ 0.01),but not in that of anti-HSV 1 or-HSV2 IgG antibodies (both P ＞ 0.05).The sensitivity,specificity,PPV and NPV were 13.24％ (9/68),86.67％ (104/120),36.00％ (9/25) and 63.80％ (104/163) respectively for anti-HSV2 IgM antibody in the diagnosis of GH,91.18％ (62/68),100.00％ (120/120),100.00％ (62/62),and 95.24％ (120/126) respectively for anti-HSV2 IgG antibody.Conclusions HSV2 prevails in the patients with GH in this region,while HSV1 only amounts to 5.18％.The type-specific anti-HSV2 IgG antibody shows a higher specificity,sensitivity,PPV and NPV in the diagnosis of GH than anti-HSV2 IgM antibody,hence,the type-specific anti-HSV2 IgG antibody is superior to anti-HSV2 IgM antibody in diagnosing GH before assisted reproduction. Key words: Herpes genitalis; Simplex virus; Immunoglobulin G; Immunoglobulin M

Evaluation of PD 404,182 as an anti-HIV and anti-herpes simplex virus microbicide.

PD 404,182 (PD) is a synthetic compound that was found to compromise HIV integrity via interaction with a nonenvelope protein viral structural component (A. M. Chamoun et al., Antimicrob. Agents Chemother. 56:672-681, 2012). The present study evaluates the potential of PD as an anti-HIV microbicide and establishes PD's virucidal activity toward another pathogen, herpes simplex virus (HSV). We show that the anti-HIV-1 50% inhibitory concentration (IC50) of PD, when diluted in seminal plasma, is ∼1 μM, similar to the IC50 determined in cell culture growth medium, and that PD retains full anti-HIV-1 activity after incubation in cervical fluid at 37°C for at least 24 h. In addition, PD is nontoxic toward vaginal commensal Lactobacillus species (50% cytotoxic concentration [CC50], >300 μM), freshly activated human peripheral blood mononuclear cells (CC50, ∼200 μM), and primary CD4(+) T cells, macrophages, and dendritic cells (CC50, >300 μM). PD also exhibited high stability in pH-adjusted Dulbecco's phosphate-buffered saline with little to no activity loss after 8 weeks at pH 4 and 42°C, indicating suitability for formulation for transportation and storage in developing countries. Finally, for the first time, we show that PD inactivates herpes simplex virus 1 (HSV-1) and HSV-2 at submicromolar concentrations. Due to the prevalence of HSV infection, the ability of PD to inactivate HSV may provide an additional incentive for use as a microbicide. The ability of PD to inactivate both HIV-1 and HSV, combined with its low toxicity and high stability, warrants additional studies for the evaluation of PD's microbicidal candidacy in animals and humans.

Antiviral activity of the crude n-hexane extract from leaves of Piper lepturum var. angustifolium (C.DC.) Yunck. (Piperaceae)

Piper lepturum var. angustifolium (C.DC.) Yunck. belongs to the family Piperaceae which is widely found from North to South of Brazil. Crude n-hexane extract from leaves of P. lepturum var. angustifolium was assayed to anti-herpes simplex viruses (HSV) activity against HSV-1 and HSV-2 in cell cultures, incubated at 37°C for 48 h in a 5% CO2 atmosphere. Chemical profile of the extract was performed in a Shimadzu high performance liquid chromatography-diode array detector-ultraviolet (HPLC-DAD-UV), using a RP-18 column and mobile phase in gradient of acetonitrile and acid deionized water (pH = 3.0), at a flow rate of 1.0 ml/min. The HPLC analyses showed signals characteristics of non-polar compounds, mainly distributed between 50 and 70 min. UV spectra of the three main compounds (50.3% of the mixture) suggested isomers, with only one EŽmax at 235 nm. The crude n-hexanic extract showed 94.4% inhibition to HSV-1 with ED50 value of 5.2 µg/ml and selective index superior to 38.4. Considering HSV-2, the extract promoted 92.7% of inhibition with ED50 value of 1.1 µg/ml and selectivity index (SI) superior to 181.8. These results point out this species of Piperaceae as a source of active compounds for the treatment of infections caused by herpes simplex virus. Key words: Antiviral activity, herpes simplex virus type 1, herpes simplex virus type 2, Piper lepturum var. angustifolium, Piperaceae, high performance liquid chromatography-diode array detector-ultraviolet (HPLC-DAD-UV).

Turning an antiviral into an anticancer drug: Nanoparticle delivery of acyclovir monophosphate

Anti-herpes simplex virus (HSV) drug acyclovir (ACV) is phosphorylated by the viral thymidine kinase (TK), but not the cellular TK. Phosphorylated ACV inhibits cellular DNA synthesis and kills the infected cells. We hypothesize that ACV monophosphate (ACVP), which is an activated metabolite of ACV, should be efficient in killing cells independent of HSV-TK. If so, ACVP should be a cytotoxic agent if properly delivered to the cancer cells. The Lipid/Calcium/Phosphate (LCP) nanoparticles (NPs) with a membrane/core structure were used to encapsulate ACVP to facilitate the targeted delivery of ACVP to the tumor. The LCP NPs showed entrapment efficiency of ~70%, the nano-scaled particle size and positive zeta potential. Moreover, ACVP-loaded LCP NPs (A-LCP NPs) exhibited concentration-dependent cytotoxicity against H460 cells and increased S-phase arrest. More importantly, a significant reduction of the tumor volume over 4days following administration (p<0.05–0.005) of A-LCP NPs, suggests excellent in vivo efficacy. Whereas, two free drugs (ACV and ACVP) and blank LCP NPs showed little or no therapeutic effect. It was also found that the high efficacy of A-LCP NPs was associated with the ability to induce dramatic apoptosis of the tumor cells, as well as significantly inhibit tumor cell proliferation and cell cycle progression. In conclusion, with the help of LCP NPs, monophosphorylation modification of ACV can successfully modify an HSV-TK-dependent antiviral drug into an anti-tumor drug.

ChemInform Abstract: Natural Products as a Source of Antiherpes Simplex Virus Agents

AbstractReview: 147 refs.

The identities and anti-herpes simplex virus activity of Clinacanthus nutans and Clinacanthus siamensis

ObjectiveTo distinguish the difference among the Clinacanthus nutans (Burm. f.) Lindau (C. nutans) and Clinacanthus siamensis Bremek (C. siamensis) by assessing pharmacognosy characteristics, molecular aspect and also to evaluate their anti-herpes simplex virus (HSV) type 1 and type 2 activities. MethodsMacroscopic and microscopic evaluation were performed according to WHO Geneva guideline. Stomatal number, stomatal index and palisade ratio of leaves were evaluated. Genomic DNA was extracted by modified CTAB method and ITS region was amplified using PCR and then sequenced. Dry leaves were subsequently extracted with n-hexane, dichloromethane and methanol and antiviral activity was performed using plaque reduction assay and the cytotoxicity of the extracts on Vero cells was determined by MTT assay. ResultsCross section of midrib and stem showed similar major components. Leaf measurement index of stomatal number, stomatal index and palisade ratio of C. nutans were 168.32±29.49, 13.83±0.86 and 6.84±0.66, respectively, while C. siamensis were 161.60±18.04, 11.93±0.81 and 3.37±0.31, respectively. The PCR amplification of ITS region generated the PCR product approximately 700 bp in size. There were 34 polymorphisms within the ITS region which consisted of 11 Indels and 23 nucleotide substitutions. The IC50 values of C. nutans extracted with n-hexane, dichloromethane and methanol against HSV-1 were (32.05±3.63) μg/mL, (44.50±2.66) μg/mL, (64.93±7.00) μg/mL, respectively where as those of C. siamensis were (60.00±11.61) μg/mL, (55.69±4.41) μg/mL, (37.39±5.85) μg/mL, respectively. Anti HSV-2 activity of n-hexane, dichloromethane and methanol C. nutans leaves extracts were (72.62±12.60) μg/mL, (65.19±21.45) μg/mL, (65.13±2.22) μg/mL, respectively where as those of C. siamensis were (46.52±4.08) μg/mL, (49.63±2.59) μg/mL, (72.64±6.52) μg/mL, respectively. ConclusionsThe combination of macroscopic, microscopic and biomolecular method are able to authenticate these closely related plants and both of them have a potency to be an anti-HSV agent.

Natural products as a source of anti-herpes simplex virus agents

Herpes simplex virus (HSV) infection is the major cause of neonatal and sporadic encephalitis as well as genital herpes globally. Although acyclovir (ACV) and related nucleoside analogs are valuable drugs for the treatment of infection and the suppression of dissemination, HSV remains highly prevalent worldwide. The emergence of resistance to ACV has also created an obstacle for the treatment of HSV-infected immunocompromised patients and highlights the need for new, safe and effective antiviral agents. Natural products, particularly traditional medicines, which have provided multiple novel drug leads, are known to be an important source of anti-HSV agents, even though their mechanisms of action and targets are unknown. In recent years, potential targets for anti-HSV agents have been identified, and these may be the targets of the effective natural products whose mechanisms are currently unknown. This review will focus on anti-HSV herbs, extracts, fractions and compounds from natural sources, in addition to potential anti-HSV targets. By creating a summary of the characteristics of active anti-HSV compounds and using that summary to mine natural product databases, a better drug discovery paradigm may be found.

VZV ischemic optic neuropathy and subclinical temporal artery infection without rash

A 75-year-old woman developed periorbital pain and blurred vision OS. Visual acuity (VA) was 20/40 OD, 20/400 OS with mild left relative afferent pupillary defect (APD). Left optic nerve was swollen and hyperemic with peripapillary flame hemorrhages (figure, A). Erythrocyte sedimentation rate (ESR) was 124 mm/h. She was treated with IV methylprednisolone, 250 mg every 6 h. On day 3, headache and vision improved. ESR was 98 mm/h and C-reactive protein was 1.40 mg/L. Rheumatoid factor, antinuclear antibodies, and antineutrophil cytoplasmic antibodies titers were negative. On day 4, left temporal artery biopsy revealed thickened intima and intact internal elastic lamina (figure, B) but no medial necrosis characteristic of giant cell arteritis (GCA). Sections of the temporal artery were deparaffinized and incubated with 10% normal sheep serum (NSS) in phosphate-buffered saline (PBS) for 1 hour at room temperature, rinsed 3 times in PBS, and incubated overnight at 4°C with polyclonal antibodies raised against the varicella-zoster virus (VZV) open reading frame 63 protein (1:1,000 dilution) or with normal rabbit serum (1:1,000 dilution). The next day, sections were washed 3 times in PBS, incubated with a 1:300 dilution of biotinylated goat antirabbit immunoglobulin G (IgG) in PBS containing 5% NSS, washed 3 times in PBS, incubated for 1 hour at room temperature with alkaline phosphatase–conjugated streptavidin (1:100 dilution), and washed 3 times with PBS. The color reaction was developed for 5–30 minutes with fresh fuchsin substrate system. Levamisole was added to the color reaction to block endogenous phosphatase. Uninfected and VZV-infected human fibroblast lung cells were used as controls (not shown). Steroids were changed to oral prednisone 60 mg daily. On day 7, brain MRI with gadolinium was negative. On day 9, pain and vision worsened. On day 11, orbital CT and head CT angiography were negative. On day 15, VA was 20/400 OS with relative left APD. On day 17, OS became blind without direct pupillary light reaction; fundus was obscured by vitreous hemorrhage. CSF contained 8 leukocytes/mm3, protein 72 mg/L, and glucose 54 mg/L. CSF cultures for bacteria, fungi, acid-fast bacilli, and cytology were negative. Because asymptomatic temporal artery biopsy was GCA-negative, VZV ischemic optic neuropathy (ION) was considered, and she was treated with IV acyclovir, 10 mg/kg every 8 hours for 7 days. On day 31, CSF contained anti-VZV IgG but not anti-herpes simplex virus IgG antibody, and serum-to-CSF ratio of anti-VZV IgG was reduced (14) compared to ratios for total IgG (121) and albumin (81). Immunohistochemistry and pathology revealed VZV antigen and neutrophils in the original left temporal artery specimen (figure, C). On day 31, she was treated with oral valacyclovir, 1 gram TID for 6 weeks; prednisone was reduced to 20 mg daily and tapered 5 mg/week. Six weeks later, pain resolved, and VA improved to finger counting. Left optic nerve was pale with clear margins and resolution of hemorrhage.

Immunomodulatory activity of an anti-HSV-1 synthetic stigmastane analog

Many viral infections are associated with the development of immunopathologies and autoimmune diseases, which are of difficult treatment and for which no vaccines are yet available. Obtaining compounds that conjugate both antiviral and immunomodulatory activities in the same molecule would be very useful for the prevention and/or treatment of these immunopathologies.The compound (22S,23S)-22,23-dihydroxystigmast-4-en-3-one (compound 1) displays anti-Herpes simplex virus type 1 activity in vitro and reduces the incidence of herpetic stromal keratitis (HSK) in mice, a chronic inflammatory syndrome induced by ocular HSV-1 infection.In the present study, compound 1 showed opposite immunomodulatory properties in vitro. It induced the release of pro-inflammatory cytokines in HSV-1-infected epithelial cells of ocular origin, and significantly reduced the production of these cytokines in LPS-activated macrophages. RNA microarrays revealed various overexpressed and repressed genes in compound 1 treated infected epithelial cells and activated macrophages, many of which are associated with innate immune responses and inflammatory processes. These immunomodulatory properties of compound 1, together with its previously reported antiviral activity, make it a potential drug for the treatment of HSK and many other immunopathologies of viral and non-viral origin.

Association of Anti–Herpes Simplex Virus IgG in Tears and Serum With Clinical Presentation in Patients With Presumed Herpetic Simplex Keratitis

To assess the clinical relevance of tear anti-herpes simplex virus (HSV) antibody measurement for the diagnosis of herpes simplex keratitis. Records of 364 patients clinically suspect of HSV-related keratitis who had tear anti-HSV IgG assessment (tear-quantified anti-HSV IgG/filtrated IgG ratio) in our institution between January 2000 and August 2008 were retrospectively analyzed. Patients were classified into 4 groups as follows: group 1, anti-HSV IgG negative in serum and tears; group 2, anti-HSV IgG negative in tears and positive in serum; group 3, anti-HSV IgG nonsignificantly positive in tears and positive in serum; and group 4, anti-HSV IgG significantly positive in serum and tears. Randomly selected patient charts from each group were reviewed for clinical data. The prevalence of anti-HSV IgG in blood increased with age from >70% before 20 years to 95% after 70 years. The prevalence of anti-HSV IgG in tears increased with age from 20% before 20 years to >50% after 70 years. The presence (either significant or not) of anti-HSV IgG in tears was significantly associated with decreased corneal sensation, presence of stromal opacities, and with neurotrophic keratitis. Logistic regression showed no significant association between age and clinical signs except for herpetic ulcers and herpetic necrotizing keratitis. Tear production of anti-HSV IgG increases with age, and it is associated with sequelae of herpes simplex keratitis. Conversely, it is poorly associated with clinical signs of acute herpes simplex keratitis.

Inhibition of Herpes Simplex Virus Type 1 by Thymol-Related Monoterpenoids

This study examined the anti-herpes simplex virus type I activity of the major constituents of several essential oils. Plaque reduction assays were performed to evaluate anti-herpes simplex virus type I activity. Thymol and carvacrol both possessed significant antiviral activity with an IC₅₀ of 7 µM, and herpes simplex virus type I was 90 % inactivated within 1 hr. The mode of antiviral action was shown to affect the virion directly. Evidence was also observed by electron microscopy. Evaluation of the structural requirements for antiviral activity of thymol-related monoterpenoids revealed that aliphatic side chains had a minor effect, while a hydrophilic group on the benzene ring was sufficient for activity. Our results suggest that thymol and carvacrol are potential candidates for topical therapeutic application to reduce herpes simplex virus transmission.

Anti-herpes simplex virus efficacies of 2-aminobenzamide derivatives as novel HSP90 inhibitors

After the widespread use of the acyclic purine nucleoside analogues for therapy of herpes simplex virus (HSV) infection since the 1980s, new antiviral strategies are urgently needed to counter the emergence of drug-resistant clinical isolates. In this report, we define the anti-HSV efficacies of three optimized 2-aminobenzamide derivatives in vitro and in vivo. The synthetic analogues SNX-25a, SNX-2112 and SNX-7081, which selectively bind to the N-terminal ATP pocket of heat shock protein 90 (HSP90), exhibited significant anti-HSV-1 and HSV-2 activities at non-cytotoxic concentrations in Vero cells, with EC50 values close to that of acyclovir (ACV). The in vivo antiviral potentials were then confirmed using a herpes simplex keratitis (HSK) rabbit model, where eye gels containing 0.1% or 0.025% SNX-25a displayed the highest efficacies against HSV-1 infection, which were better than that obtained with 0.1% ACV. SNX-2112 and SNX-7081 gels were also effective against HSV-1 with different magnitude of activities. Our results for the first time confirmed the anti-HSV efficacies of these 2-aminobenzamide derivatives and suggest that with alternative mechanisms of action these novel HSP90 inhibitors, especially SNX-25a, could be potent as new anti-HSV clinical trial candidates.