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HN C(NH2)NH SS HO 48 49 NHC(NH2) NH HO O S O S
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O H2N O 50 N N NH2
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CO2 Na N N O N 51
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Several other antiviral strategies have also been pursued for drug discovery. Because arenaviruses are enveloped viruses, modi cations of plasma membrane could affect viral replication. This approach was supported by the demonstration of inhibitory activity exhibited by fatty acids292 and myristic acids293 on Junin virus replication. The integrity of the actin micro laments might be required for optimal arenavirus multiplication as suggested by inhibition of Junin virus in
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OVERVIEW OF ANTIVIRAL DRUG DISCOVERY AND DEVELOPMENT
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N N F N S 52
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vitro by phenotiazines294 (exempli ed by tri uoroperazine 52). Lymphocytic choriomeningitis virus (LCMV) was shown to be sensitive to lethal mutagenesis induced by the mutagenic agent 5- uorouracil295and its genome could be cleaved by trans-acting ribozymes.296,297 Recently, a-dystroglycan has been identi ed as a cellular receptor for LCMV and Lassa virus (both belong to the Old World arenaviruses), suggesting a new target for drug discovery.298 300 However, New World arenaviruses (Junin, Machupo, and Guanarito) use an as yet unidenti ed receptor or coreceptor for binding. Screening in cell cultures identi ed a variety of classes of compounds with potential antiviral activity: brassinosteroids301 (e.g., 53), macrocyclic trichothecenes302 (e.g., 54), and 30 - uoro-30 -deoxyadenosine54 21. Another compound, 20 ,30 didehydro-30 -deoxythymidine (stampidine), improved survival in Lassa virusinfected mice.303
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CH3 CH3 H OH CH3 O O Br OH H3C 53 54 CH3O O CH3 OH H O O H
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OH H3C CH3 CH3
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INHIBITORS OF BUNYAVIRUSES
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The family of Bunyaviridae is composed of four genera: Bunyavirus (La Crosse virus), Hantavirus [Hantaan virus, Sin Nombre virus (SNV)], Nairovirus [CrimeanCongo hemorrhagic fever virus (CCHFV)], and Phlebovirus (RVFV, PTV, sand y fever virus). These viruses are distributed across the NIAID biodefense categories. As shown in Table 3.4, favorable outcomes were seen when ribavirin was used
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experimentally to treat CCHFV, La Crosse, and bunyavirus hemorrhagic fever with renal syndrome (HFRS), although the CCHFV and La Crosse trials were not blinded. No de nite ef cacy was observed in patients with hantavirus pulmonary syndrome (HPS).105,304 A very recent controlled trial further suggested that ribavirin might not be effective for treatment of hantavirus cardiopulmonary syndrome after onset of the cardiopulmonary phase.106 Since these viruses are BSL-3 or -4 agents, this presents a problem for routine antiviral testing. PTV, a phlebovirus related to RVFV, produces nonencephalitic lethal infections in mice. Because this virus presents a lower biohazard (BSL-2), it can serve as substitute in routine drug screening with the caveat that active compounds will have to be checked against the actual target virus eventually.79,278,305,306 Ribavirin is active against PTV in vitro and is protective when given either parenterally or orally, in single or multiple doses, to PTV-infected mice.44 In a report by Sidwell et al.,79 other active compounds included ribamidine, tiazofurin, pyrazofurin, and 3-deazaguanosine. Several immunomodulators, such as poly(ICLC), ampligen, 7-thia-8-oxoguanosine, and IFN-a, were also active.
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Ebola virus is classi ed as a select agent. Because of biosafety and biosecurity concerns, antiviral research has been conducted mainly by USAMRIID investigators. Huggins and co-workers recently established a lethal mouse model suitable for evaluation of prophylaxis and therapy of Ebola virus.307 Intraperitoneal administration (2.2 20 mg/kg), thrice daily, of C-c3Ado 10 signi cantly protected BALB/c mice from lethal infection with mouse-adapted Ebola Zaire virus, providing treatment was initiated on day 1, 0, or 1 relative to time of virus challenge.117,308 Treatment with 2.2 mg/kg initiated on day 3 postinfection still resulted in 40% survival. In another study, a single subcutaneous dose of 80 mg/kg or less of C-c3Ado 10, or of 1 mg/kg or less of c3-NPC A 9, provided equal or better protection, without causing toxicity.309 One dose of drug given on day 1 or 2 postinfection signi cantly reduced serum virus titers and resulted in survival of most or all animals. However, drug treatment given within 1 h after infection ( day 0 ) was less effective. In SCID mice, single or multiple drug treatment suppressed Ebola replication but did not prevent death.309 The prolonged ef cacy of these two SAH hydrolase inhibitors demonstrated a potential useful antiviral strategy in that drug treatment begins early in infection with high but nontoxic doses, in order to hold viral burden below the lethal threshold until the host immune system eliminates the infection.309 The remarkable antiviral activity observed with these compounds on day 1 or 2 postinfection, but not on the day of viral challenge, suggests that these compounds might have additional mechanisms of action, which contribute to the drug s effect after viral replication has begun, but before widespread dissemination of infection and extensive tissues damage have occurred. c3-NPC A was shown to induce a massive release of IFN-a when administered to Ebola-infected mice, but not uninfected mice, apparently reversing the virus-induced suppression of the type I
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