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Conclusions: These data demonstrate that the majority ofMX001 treated patients had a > 99% decrease in viral load com-ared to baseline after 2 weeks of therapy. Antiviral activity ofMX001 was reduced in patients with high level resistance to CDV,ll of whom had received prior treatment with CDV. Finally, higherlasma exposure to CMX001 appeared to correlate with a moreapid response. CMX001 is a promising antiviral for severe aden-virus infection.

oi:10.1016/j.antiviral.2011.03.020

oster Session 1: Retroviruses, Respiratory Viruses, Emergingiruses and Antiviral Methods Chairs 4:00–6:00 pm Sofia 3 andyota

5

esign, Synthesis and West Nile Protease Inhibitory Activity ofovel Isatin Derivatives

eriyasamy Selvam 1,∗, Priya Srinivasan 2, Tanvi Khot 2, R.admanaban 2

Devaki Amma Memorial College of Pharmacy, Malapuram 673634,ndiaMicrobiology and Immunology, Georgetown School of Medicine,ashington, USA

Background: The mosquito-borne viral pathogens of global sig-ificance include the members of flavivirus genus of Flaviviridae

amily. Two important human pathogens are dengue and West Nileiruses which cause considerable morbidity and mortality through-ut tropical and subtropical regions of the world. No vaccines orntiviral therapeutics are available for these two pathogens. Theverall goal of our study is to develop potent inhibitors of Westile virus serine protease, which is a quintessential viral targets it is required for viral replication. In this study, we examinedhether derivatives of isatin (2,3-dioxoindole) could be versatile

ead compounds for structure–activity relationship (SAR) study.Methods: Novel isatin-sulphadimidine derivative synthesized

nd screened for their inhibitory activities of WNV protease in vitro.Results: The N-benzoyl derivative (SPIII-5H-BZ) and 5-chloro-

-acetyl derivative (SPIII-5Cl-AC) exhibited significant inhibitoryctivities against the WNV protease (IC50 values of 15 �M and.4 �M, respectively).

Conclusions: To our knowledge, this is the first report regardinghe inhibitory activities of isatin derivatives against the WNV serinerotease. Further work on SAR study for lead optimization is inrogress.

oi:10.1016/j.antiviral.2011.03.021

h 90 (2011) A21–A78

36

Effects of the Addition of Hiltonol® (Poly-ICLC) to a SARS-CoV SProtein Vaccine in Lethal SARS-CoV Mouse Model

Dale L. Barnard 1,∗, Miles Wandersee 1, Kevin Bailey 1, AaronSmith 1, Zach Vest 1, John D. Morrey 1, Andres M. Salazar 2

1 Utah State University, Logan, USA2 Oncovir, Inc., Washington, USA

The principal containment strategy for emerging diseases hasbeen rapid diagnosis/isolation followed by immunization, but thereis often a gap between isolation/identification and immuniza-tion. Host-targeted therapeutics that could provide immediate,broad-spectrum resistance to disease could fill this gap in pro-tection to allow time for agent specific immunization to takeeffect. Hiltonol® is a stabilized dsRNA therapeutic viral mimic orpathogen-associated molecular pattern (PAMP) that activates 2′5′

OAS, PKR, RIG-I, mda-5, DCs, natural killer cells interferons andvarious cytokines and chemokines, while at the same time accel-erating and enhancing the quality of adaptive cellular/humoralimmunity. It induces a broad-spectrum innate-immune antiviralstate lasting up to 3 weeks. The current study was done to seeif Hiltonol® could be used to protect during the time betweenthe isolation/identification of an agent and adaptive immunization.Mice immunized i.m. twice with S Protein vaccine in alum, 14 daysapart, were protected against death from viral challenge 14 daysafter the last immunization. A one time Immunization with vary-ing doses of vaccine (0.3, 1, 5 �g/mouse) along with simultaneousHiltonol® (10 �g/mouse) intranasally also significantly protectedmice against death (P < 0.001). Mice treated with 5 �g of vaccineand only 1 �g of Hiltonol®, showed diminished survival to only60%. These data suggest that the co-administration of Hiltonol® athigher doses of the S protein vaccine enhanced the protection ofmice against death. When mice were challenged with SARS-CoV3 days after one immunization with vaccine (0.3, 1, 5 �g/mouse)plus Hiltonol® at 10 �g, both given i.n., then all mice survivedthe infectious challenge. Mice receiving one course of vaccine plusalum and challenged on day 3 all died; mice receiving Hiltonol® at10 �g/mouse and challenged on day 3 all survived. Thus, it appearsthat Hiltonol® provided an immediate protection against diseasethat allowed time for the more specific vaccination strategy to takeeffect.

Acknowledgement: Supported by Contract No. N01 AI-15435and HHSN272201000039I/HHSN27200002/A14 from NIAID, NIH.

doi:10.1016/j.antiviral.2011.03.022

37

Synthesis and Anti-HIV Activity of d-peptide Analogs as HIVFusion Inhibitors

Alice Baron 1,2,∗, Christine Kreuz 3, Gilles Gosselin 1, FredericLamaty 2, Jean Martinez 2, Dominique Surleraux 1, Claire Pierra 1,Pascal Clayette 3

1 Laboratoires Idenix, Montpellier, France2 Institut des Biomolécules Max Mousseron, Montpellier, France3 Bertin Pharma, Fontenay aux Roses, France

The identification of new anti-HIV molecules remains animportant challenge. To date, HIV viral entry becomes a promisingtarget for HIV drug development as illustrated for example by

the approval of Enfurvitide by regulatory agencies as HIV fusioninhibitor, and more recently, Maraviroc as HIV entry inhibitor.Although highly effective, Enfurvirtide, a C-peptide of 36 aminoacids mimicking the CHR region of gp41 has several serious

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imitations including protease degradation, high dosing require-ents, subcutaneous administration, and emergence of resistantIV strains. Based on these data, we present herein the synthesisnd the anti-HIV evaluation of new d-peptide analogs, expectedo be resistant to proteolytic degradation and endowed withmproved bioavailability, as specific inhibitors of the hydrophobeocket of the gp41N trimer. Using a similar cyclic d-peptidepproach, a consensual 8-mer sequence with 4 recurrent aminocids (d-Glu9, d-Trp10, d-Trp12, and d-Leu13) has been previouslyefined by other research groups. First, our efforts were devoted tonderstand the key elements responsible for their pharmacologicalctivity using molecular modelisation/dynamic approaches. Thisnitial study showed that d-Glu9, d-Trp10, d-Trp12, and d-Leu13nteract with residues in or on the edge (d-Glu9) of the hydrophobeocket and, that C- and N-terminal terminations govern struc-ural changes in cyclic peptides and probably their antiretroviralffects. Then, considering these data, more than 40 cyclic analogsere synthesized and evaluated against HIV-1-LAI by measuring

irus-induced cytopathogen effects (CPE) and the production ofhe major HIV nucleocapsid p24 protein in acutely infected MT-4ells. Retro and/or inverso peptides as well as 7-mer d-peptideshowed no antiviral effects. Only the substitution of d-Trp12 and-Leu13 residues by l-napthyl and/or cyclobutyl group(s) have

mproved antiretroviral efficacy.

oi:10.1016/j.antiviral.2011.03.023

8

ctivity of Novel Cyclophilin Inhibitors based on the Polyketide,anglifehrin A, against HIV

ichael Bobardt 1,∗, Steven Moss 2, Udayan Chatterji 1, Moham-ad Nur-E-Alam 2, Tony Warneck 2, Barrie Wilkinson 2, Philippeallay 1, Matthew Gregory 2

The Scripps Research Institute, La Jolla, USABiotica Technology Ltd, Cambridge, United Kingdom

Cyclophilin inhibitors, such as Cyclosporine A (CsA) and ana-ogues, such as DEBIO-025 (Alisporivir) have previously shownctivity in in vitro antiviral assays against HIV isolates (Ptak et al.,008), however, when tested clinically, DEBIO-025 only reducedIV-1 RNA levels by ≥0.5 and >1 log10 copies/mL in nine and twoatients respectively, whilst 27 of the treated patients showedo reduction in HIV-1 RNA levels (Steyn et al., 2006). DEBIO-025as trialled in HCV/HIV coinfected patients, showed better effi-

acy against HCV (Flisiak et al., 2008), and was followed by aubstantial focus in HCV treatment. Sanglifehrin A (SfA), a polyke-ide natural product, has been shown to bind Cyclophilins (CyPs),uch as CyPA, and to have antiviral activity in HCV replicon assays.ovel analogues with improved drug like properties were gen-rated through proprietary biosynthetic engineering technologynd semi-synthetic methods. These analogues were tested in vitrogainst HIV, and were seen to have significantly improved potency,oth in terms of EC50 and maximal antiviral effect. Novel SfAnalogues offer additional advantages over other virus-targetinggents under development for HIV and detailed profiling of selectedandidates is underway.

eferences

lisiak, et al., 2008. Hepatology 47 (3), 817–826.tak, et al., 2008. Antimicrob. Agents Chemother. 52 (4), 1302–1317.teyn, et al., 2006. Int. Conf. Retroviruses Opportunistic Infec, vol. 13 , Abs 516.

oi:10.1016/j.antiviral.2011.03.024

h 90 (2011) A21–A78 A29

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OBR-5-340—A Novel Pyrazolo-Pyrimidine Derivative withStrong Antiviral Activity Against Coxsackievirus B3 In Vitro andIn Vivo

Heike Braun 1,∗, Vadim A. Makarov 2, Olga B. Riabova 2, ElenaS. Komarova 2, Martina Richter 1, Peter Wutzler 1, MichaelaSchmidtke 1

1 Institute of Virology and Antiviral Therapy, Jena University Hospital,Jena, Germany2 Institute of Biochemistry, Russian Academy of Science, Moscow, Rus-sia

There is no specific medication to address diseases induced byCVB3, e.g. harmless respiratory infections, diarrhea, myocarditis,meningoenceophalitis, pneumonia, and pancreatitis until now.

In the present study, the toxicity and anti-CVB3 activity of OBR-5-340, a novel pyrazolo-pyrimidine derivative was determinedin vitro and in vivo. OBR-5-340 is well tolerated (CC50 > 500 �M)and exhibits strong dose-dependent antiviral activity against thepleconaril-resistant CVB3 31-1-93 in HeLa cells. The IC50 deter-mined for CVB3 31-1-93 was 0.03 �M. In contrast, pleconarilincluded as control was inactive. For the in vivo studies, adultmale NMRI mice were infected intraperitoneally with 5−4 pfu ofCVB3 31-1-93. OBR-340 and the control compound pleconaril wereadministered orally twice daily (100 mg/kg). In pleconaril- andplacebo-treated CVB3 31-1-93-infected mice disease onset wasobserved at day 3 or 4 p.i. Between day 5 and 10 p.i. severity ofdisease, lost of body weight were maximal (−15% related to day 0p.i.). OBR-5-340 treatment almost completely prevented CVB3 31-1-93-induced disease. No loss of body weight was observed andthe histopathological scores in heart and pancreas tissue as well asthe viral load in heart tissue were significantly reduced comparedto pleconaril- or placebo-treated mice. In conclusion, OBR-5-340exhibits a strong antiviral effect against a pleconaril-resistant CVB3variant in vitro and in vivo. It represents a promising new drugcandidate for the treatment of CVB3 infections.

doi:10.1016/j.antiviral.2011.03.025

40

Occurrence of Opportunistic Infections in People Living withHIV/AIDS Following Antiretroviral Therapy in West Bengal, India

Sayan Chakraborty ∗, Mehebubar Rahman, Bibhuti Saha

Calcutta School of Tropical Medicine, Kolkata, India

As per the data of 2008, approximately 33.4 million people areliving with HIV/AIDS worldwide and there were around 2.0 mil-lion AIDS related deaths in that year. Occurrence of opportunisticinfections (OIs) is the main cause of morbidity and mortality inHIV infected patients. OIs encompass a wide variety of microorgan-isms that produce fulminant infections in immunocompromisedHIV infected patients. HIV makes the infected person immunocom-promised by destroying his CD4 T-lymphocytes. CD4 T-lymphocytecount (or CD4 count) is a standard laboratory marker to monitordisease progression in HIV infected patients. When the CD4 countof a HIV infected person decreases he or she becomes susceptible toOIs due to the compromised immune system. Antiretroviral Ther-apy (ART) decreases multiplication HIV and thereby decreases CD4cell destruction. So there is increase in CD4 count and improvement

in immunity which leads to less occurrence of OIs in HIV patients.We followed 88 patients for 3 years, who were getting first line ARTat ART center, Calcutta School of Tropical Medicine. For analysis wedivided 88 patients into 5 groups based on their initial CD4 count i.e.