RESUMEN
In an effort to discover novel, noncarbohydrate inhibitors of influenza virus neuraminidase we hypothesized that compounds which contain positively charged amino groups in an appropriate position to interact with the Asp 152 or Tyr 406 side chains might be bound tightly by the enzyme. Testing of 300 alpha- and beta-amino acids led to the discovery of two novel neuraminidase inhibitors, a phenylglycine and a pyrrolidine, which exhibited K(i) values in the 50 microM range versus influenza virus A/N2/Tokyo/3/67 neuraminidase but which exhibited weaker activity against influenza virus B/Memphis/3/89 neuraminidase. Limited optimization of the pyrrolidine series resulted in a compound which was about 24-fold more potent than 2-deoxy-2,3-dehydro-N-acetylneuraminic acid in an anti-influenza cell culture assay using A/N2/Victoria/3/75 virus. X-ray structural studies of A/N9 neuraminidase-inhibitor complexes revealed that both classes of inhibitors induced the Glu 278 side chain to undergo a small conformational change, but these compounds did not show time-dependent inhibition. Crystallography also established that the alpha-amino group of the phenylglycine formed hydrogen bonds to the Asp 152 carboxylate as expected. Likewise, the beta-amino group of the pyrrolidine forms an interaction with the Tyr 406 hydroxyl group and represents the first compound known to make an interaction with this absolutely conserved residue. Phenylglycine and pyrrolidine analogs in which the alpha- or beta-amino groups were replaced with hydroxyl groups were 365- and 2,600-fold weaker inhibitors, respectively. These results underscore the importance of the amino group interactions with the Asp 152 and Tyr 406 side chains and have implications for anti-influenza drug design.
Asunto(s)
Aminoácidos/farmacología , Antivirales/farmacología , Glicina/análogos & derivados , Neuraminidasa/antagonistas & inhibidores , Orthomyxoviridae/enzimología , Aminoácidos/química , Antivirales/química , Cristalografía por Rayos X , Glicina/farmacología , Hidroxilación , Modelos Moleculares , Neuraminidasa/química , Orthomyxoviridae/efectos de los fármacos , Conformación Proteica , Pirrolidinas/farmacologíaRESUMEN
The discovery of (+/-)-(2S,3R,4R)-2-(trifluoroacetamido)methyl-3-amino-1-(N'-ethyl-N'-isopropylcarbamyl)pyrrolidine-4-carboxylic acid (A-192558, 20e) as a potent inhibitor of influenza neuraminidase (NA) is described. Efficient syntheses of two core structures, cis-3-(allyloxycarbonyl)amino-1-(9'-fluorenylmethoxycarbonyl)pyrrolidine-4-carboxylic acid (7) and tert-butyl (+/-)-(2S,3R,4R)-2-aminomethyl-3-bis(tert-butyloxycarbonyl)amino-1-(N'-ethyl-N'-isopropylcarbamyl)pyrrolidine-4-carboxylate (18b), were developed. Starting with these core structures and using available structural information of the NA active site as the guide, analogues were synthesized in both the tri- and tetrasubstituted pyrrolidine series by means of high-throughput parallel synthesis in solid or solution phase for expeditious SAR. These studies accelerated the identification of (+/-)-(2S,3R,4R)-2-(trifluoroacetamido)methyl-3-amino-1-(N-ethyl-N-isopropylcarbamyl)pyrrolidine-4-carboxylate (20e, A-192558) as the most potent NA inhibitor in this series (IC50 = 0.2 microM against NA A and 8 microM against NA B). The X-ray crystallographic structure of A-192558 bound to NA revealed the predicted interaction of the carboxylic group with the positively charged pocket (Arg118, Arg292, Arg371) and interaction of the trifluoroacetamino residue with the hydrophobic pocket (Ile222, Trp178) of the enzyme active site. Surprisingly, the ethyl and isopropyl groups of the urea functionality induced a conformational change of Glu276, turning the Glu276/Glu277 hydrophilic pocket, which normally accommodates the triglycerol side chain of substrate sialic acid, into an induced hydrophobic pocket.
Asunto(s)
Antivirales/síntesis química , Inhibidores Enzimáticos/síntesis química , Neuraminidasa/antagonistas & inhibidores , Orthomyxoviridae/enzimología , Pirrolidinas/síntesis química , Antivirales/química , Cristalografía por Rayos X , Diseño de Fármacos , Inhibidores Enzimáticos/química , Modelos Moleculares , Estructura Molecular , Pirrolidinas/químicaAsunto(s)
Antivirales/química , Brotes de Enfermedades , Inhibidores Enzimáticos/química , Gripe Humana/tratamiento farmacológico , Neuraminidasa/antagonistas & inhibidores , Antivirales/farmacología , Brotes de Enfermedades/prevención & control , Diseño de Fármacos , Farmacorresistencia Microbiana , Inhibidores Enzimáticos/farmacología , Humanos , Gripe Humana/epidemiología , Gripe Humana/prevención & control , Gripe Humana/virologíaRESUMEN
Influenza A and B viruses belong to the Orthomyxoviridae family of viruses. These viruses are responsible for severe morbidity and significant excess mortality each year. Infection with influenza viruses usually leads to respiratory involvement and can result in pneumonia and secondary bacterial infections. Vaccine approaches to the prophylaxis of influenza virus infections have been problematic owing to the ability of these viruses to undergo antigenic shift by exchanging genomic segments or by undergoing antigenic drift, consisting of point mutations in the haemagglutinin (HA) and neuraminidase (NA) genes as a result of an error-prone viral polymerase. Historically, antiviral approaches for the therapy of both influenza A and B viruses have been largely unsuccessful until the elucidation of the X-ray crystallographic structure of the viral NA, which has permitted structure-based drug design of inhibitors of this enzyme. In addition, recent advances in the elucidation of the structure and complex function of influenza HA have resulted in the discovery of a number of diverse compounds that target this viral protein. This review article will focus largely on newer antiviral agents including those that inhibit the influenza virus NA and HA. Other novel approaches that have entered clinical trials or been considered for their clinical utility will be mentioned.
Asunto(s)
Gripe Humana/tratamiento farmacológico , Animales , Antivirales/farmacología , Antivirales/uso terapéutico , Humanos , Orthomyxoviridae/efectos de los fármacos , Orthomyxoviridae/fisiología , Replicación Viral/efectos de los fármacosAsunto(s)
Aminas/uso terapéutico , Antivirales/uso terapéutico , Inhibidores Enzimáticos , Neuraminidasa/antagonistas & inhibidores , Ácidos Siálicos/uso terapéutico , Virosis/tratamiento farmacológico , Animales , Brotes de Enfermedades , Diseño de Fármacos , Guanidinas , Hemaglutininas , Humanos , Vacunas contra la Influenza , Orthomyxoviridae/efectos de los fármacos , Orthomyxoviridae/enzimología , Orthomyxoviridae/fisiología , Oseltamivir , Piranos , Virosis/epidemiología , Virosis/prevención & control , Replicación Viral/efectos de los fármacos , ZanamivirRESUMEN
A series of influenza neuraminidase inhibitors with the cyclohexene scaffold containing lipophilic side chains have been synthesized and evaluated for influenza A and B neuraminidase inhibitory activity. The size and geometry of side chains have been modified systematically in order to investigate structure-activity relationships of this class of compounds. The X-ray crystal structures of several analogues complexed with neuraminidase revealed that the lipophilic side chains bound to the hydrophobic pocket consisted of Glu276, Ala246, Arg224, and Ile222 of the enzyme active site. The structure-activity relationship studies of this series have also demonstrated remarkably different inhibitory potency between influenza A and B neuraminidase. This indicated that the lipophilic side chains had quite different hydrophobic interactions with influenza A and B neuraminidase despite their complete homology in the active site. Influenza B neuraminidase appeared to be much more sensitive toward the increased steric bulkiness of inhibitors compared to influenza A neuraminidase. From the extensive structure-activity relationship investigation reported in this article, GS 4071 emerged as one of the most potent influenza neuraminidase inhibitors against both influenza A and B strains.
Asunto(s)
Acetamidas/farmacología , Antivirales/farmacología , Inhibidores Enzimáticos/farmacología , Virus de la Influenza A/enzimología , Virus de la Influenza B/enzimología , Neuraminidasa/antagonistas & inhibidores , Acetamidas/química , Antivirales/química , Cristalografía por Rayos X , Inhibidores Enzimáticos/química , Modelos Moleculares , Estructura Molecular , Oseltamivir , Relación Estructura-ActividadRESUMEN
The kinetics of inhibition of purified influenza neuraminidases from A/Tokyo/3/67 and B/Memphis/3/89 influenza viruses by (3R,4R,5S)-4-acetamido-5-amino-3-(1-ethylpropoxy)-1-cyclohexene- 1-carboxylic acid (GS4071) were investigated. Progress curve experiments established that GS4071 is a time dependent inhibitor of both A and B strains of influenza neuraminidase. The apparent association and dissociation rate constants, as well as the overall Ki values, were only modestly different for the two neuraminidase strains. The time dependent inhibition phenomenon, often referred to as slow-binding inhibition, appears to be a consequence of the very slow rate of dissociation of the compound from influenza neuraminidase.
Asunto(s)
Acetamidas/farmacología , Inhibidores Enzimáticos/farmacología , Virus de la Influenza A/efectos de los fármacos , Virus de la Influenza A/enzimología , Virus de la Influenza B/efectos de los fármacos , Virus de la Influenza B/enzimología , Neuraminidasa/antagonistas & inhibidores , Acetamidas/metabolismo , Sitios de Unión , Inhibidores Enzimáticos/metabolismo , Enlace de Hidrógeno , Cinética , Neuraminidasa/química , Oseltamivir , Conformación Proteica , Especificidad de la Especie , AguaRESUMEN
The synthesis and influenza neuraminidase inhibitory activity of a new series of C3-aza carbocyclic neuraminidase inhibitors are described. Analogues 3c and 3j, bearing a 3-pentyl group, exhibit influenza A inhibitory activities comparable to that of 1.
Asunto(s)
Antivirales/síntesis química , Compuestos Aza/síntesis química , Inhibidores Enzimáticos/síntesis química , Neuraminidasa/antagonistas & inhibidores , Orthomyxoviridae/efectos de los fármacos , Antivirales/farmacología , Compuestos Aza/farmacología , Inhibidores Enzimáticos/farmacología , Neuraminidasa/química , Orthomyxoviridae/enzimología , Relación Estructura-ActividadRESUMEN
A series of 94 benzoic acid derivatives was synthesized and tested for its ability to inhibit influenza neuraminidase. The enzyme-inhibitor complex structure was determined by X-ray crystallographic analysis for compounds which inhibited the enzyme. The most potent compound tested in vitro, 5 (4-acetylamino)-3-guanidinobenzoic acid), had an IC50 = 2.5 x 10(-6) M against N9 neuraminidase. Compound 5 was oriented in the active site of the neuraminidase in a manner that was not predicted from the reported active site binding of GANA (4) with neuraminidase. In a mouse model of influenza, 5 did not protect the mice from weight loss due to the influenza virus when dosed intranasally.
Asunto(s)
Antivirales/síntesis química , Benzoatos/síntesis química , Inhibidores Enzimáticos/síntesis química , Neuraminidasa/antagonistas & inhibidores , Orthomyxoviridae/efectos de los fármacos , Animales , Antivirales/farmacología , Benzoatos/farmacología , Ácido Benzoico , Diseño de Fármacos , Inhibidores Enzimáticos/farmacología , Femenino , Ratones , Ratones Endogámicos BALB C , Orthomyxoviridae/enzimología , Relación Estructura-ActividadRESUMEN
A single change (E119G) in the influenza A virus N9 neuraminidase (NA) results in resistance of the enzyme to the NA inhibitor 4-Guanidino-Neu5Ac2en (4-GuDANA). This change causes a salt link between Glu119, which sits in a pocket in the bottom of the active site of the enzyme, and the 4-guanidinium moiety of the inhibitor to be lost. NA "heads" of the resistant enzyme produced only a few small crystals under conditions in which the wild-type enzyme readily formed large crystals. These small crystals were of sufficient quality to yield X-ray crystallographic data which confirmed the E119G change and demonstrated the presence of electron density representing either a strong structural-water molecule or an anionic species in place of the glutamate carboxylate. NA heads of the resistant enzyme also have greatly reduced NA activity per milligram of total protein. We have now found that the mutant NA heads consist predominantly of monomers with a few dimers and tetramers, as determined by electron microscopic analysis of the protein. The low level of enzymatic activity as well as the small number of crystals obtained were probably from the few tetramers remaining intact in the preparation. The purified wild-type and 4-GuDANA-resistant enzymes were treated with the homobifunctional NHS-ester cross linker, DTSSP. SDS-PAGE analysis of the treated enzymes clearly revealed cross-linked dimers of the wild-type enzyme. In contrast, only a small proportion of the 4-GuDANA-resistant neuraminidase was cross-linked. An examination of the known X-ray crystallographic structure of the wild-type NA reveals a salt bridge between Glu119 and Arg156 of the same monomer. Arg156 is a conserved amino acid that is situated at the interface between monomers, and a salt link between this amino acid and Glu119 may contribute to the stability of enzyme tetramers. It is suggested that the E119G alteration in the 4-GuDANA-resistant NA leads to the abrogation of this interaction and thus to the instability of the NA tetramers.
Asunto(s)
Virus de la Influenza A/enzimología , Neuraminidasa/química , Mutación Puntual , Estructura Secundaria de Proteína , Animales , Embrión de Pollo , Simulación por Computador , Reactivos de Enlaces Cruzados , Cristalización , Cristalografía por Rayos X , Dimerización , Inhibidores Enzimáticos/farmacología , Guanidinas , Humanos , Cinética , Sustancias Macromoleculares , Microscopía Electrónica , Modelos Moleculares , Neuraminidasa/metabolismo , Neuraminidasa/ultraestructura , Piranos , Ácidos Siálicos/farmacología , Especificidad por Sustrato , ZanamivirRESUMEN
The reassortant influenza viruses, A/NWS-G70c with N9 neuraminidase (NA) and B/HK/8/73 (HG) with B/Lee/40 NA, were selected for resistance to 4-guanidino-Neu5Ac2en (4-GuDANA) by passaging the virus in stepwise increases in the concentration of 4-GuDANA. In the NA of resistant viruses, the absolutely conserved Glu 119, which lies in a pocket beneath the active site of the enzyme and interacts with the guanidinium moiety of 4-GuDANA, was changed to Gly. The mutant NA was >200-fold more resistant to 4-GuDANA than was the wild-type enzyme. During 72 h in cell culture, resistant A and B viruses displayed much less NA activity than did wild-type viruses but did undergo multicycle replication. While emergence of resistance to 4-GuDANA has not been observed in vivo, these results demonstrate that the development of resistance is possible and can be mediated by a single amino acid change in the active site of the viral NA.
Asunto(s)
Antivirales/farmacología , Inhibidores Enzimáticos/farmacología , Virus de la Influenza B/efectos de los fármacos , Neuraminidasa/antagonistas & inhibidores , Ácidos Siálicos/farmacología , Células Cultivadas , Resistencia a Medicamentos , Guanidinas , Virus de la Influenza A/efectos de los fármacos , Mutación , Neuraminidasa/genética , Piranos , ZanamivirRESUMEN
The active site of influenza virus neuraminidase (NA) is formed by 11 universally conserved residues. A guanidino group incorporated into two unrelated NA inhibitors was previously reported to occupy different negatively charged sites in the NA active site, A new inhibitor containing two guanidino groups was synthesized in order to utilize both sites in an attempt to acquire a combined increase in affinity. The X-ray crystal structures of the complexes show that the expected increase in affinity could not be achieved even though the added guanidino group binds to the negatively charged site as designed. This suggests that the ligand affinity to the target protein is contributed both from ligand-protein interactions and solvation/conformation energy of the ligand.
Asunto(s)
Guanidinas/farmacología , Hidroxibenzoatos/farmacología , Virus de la Influenza B/enzimología , Neuraminidasa/química , Sitios de Unión , Cristalografía por Rayos X , Inhibidores Enzimáticos/farmacología , Guanidinas/química , Humanos , Hidroxibenzoatos/química , Modelos Moleculares , Neuraminidasa/antagonistas & inhibidores , Agua/químicaRESUMEN
The design, synthesis, and in vitro evaluation of the novel carbocycles as transition-state-based inhibitors of influenza neuraminidase (NA) are described. The double bond position in the carbocyclic analogues plays an important role in NA inhibition as demonstrated by the antiviral activity of 8 (IC50 = 6.3 microM) vs 9 (IC50 > 200 microM). Structure-activity studies of a series of carbocyclic analogues 6a-i identified the 3-pentyloxy moiety as an apparent optimal group at the C3 position with an IC50 value of 1 nM for NA inhibition. The X-ray crystallographic structure of 6h bound to NA revealed the presence of a large hydrophobic pocket in the region corresponding to the glycerol subsite of sialic acid. The high antiviral potency observed for 6h appears to be attributed to a highly favorable hydrophobic interaction in this pocket. The practical synthesis of 6 starting from (-)-quinic acid is also described.
Asunto(s)
Acetamidas/síntesis química , Antivirales/síntesis química , Antivirales/farmacología , Ácidos Carboxílicos/síntesis química , Ciclohexanos/síntesis química , Virus de la Influenza A/efectos de los fármacos , Neuraminidasa/antagonistas & inhibidores , Ácidos Siálicos/síntesis química , Acetamidas/farmacología , Animales , Sitios de Unión/efectos de los fármacos , Ácidos Carboxílicos/farmacología , Línea Celular , Cristalografía por Rayos X , Ciclohexanos/farmacología , Ciclohexenos , Modelos Animales de Enfermedad , Diseño de Fármacos , Inhibidores Enzimáticos/síntesis química , Interacciones Hidrofóbicas e Hidrofílicas , Virus de la Influenza A/enzimología , Virus de la Influenza A/crecimiento & desarrollo , Ácido Quínico/química , Ácidos Siálicos/farmacología , Relación Estructura-Actividad , Ensayo de Placa ViralRESUMEN
The structure of Salmonella typhimurium LT2 neuraminidase (STNA) is reported here to a resolution of 1.6 angstroms together with the structures of three complexes of STNA with different inhibitors. The first is 2-deoxy-2,3-dehydro-N-acetyl-neuraminic acid (Neu5Ac2en or DANA), the second and third are phosphonate derivatives of N-acetyl-neuraminic acid (NANA) which have phosphonate groups at the C2 position equatorial (ePANA) and axial (aPANA) to the plane of the sugar ring. The complex structures are at resolutions of 1.6 angstroms, 1.6 angstroms and 1.9 angstroms, respectively. These analyses show the STNA active site to be topologically inflexible and the interactions to be dominated by the arginine triad, with the pyranose rings of the inhibitors undergoing distortion to occupy the space available. Solvent structure differs only around the third phosphonate oxygen, which attracts a potassium ion. The STNA structure is topologically identical to the previously reported influenza virus neuraminidase structures, although very different in detail; the root-mean-square (r.m.s) deviation for 210 C alpha positions considered equivalent is 2.28 angstroms (out of a total of 390 residues in influenza and 381 in STNA). The active site residues are more highly conserved, in that both the viral and bacterial structures contain an arginine triad, a hydrophobic pocket, a tyrosine and glutamic acid residue at the base of the site and a potential proton-donating aspartic acid. However, differences in binding to O4 and to the glycerol side-chain may reflect the different kinetics employed by the two enzymes.
Asunto(s)
Inhibidores Enzimáticos/química , Ácido N-Acetilneuramínico/análogos & derivados , Neuraminidasa/química , Salmonella typhimurium/enzimología , Ácidos Siálicos/química , Sitios de Unión , Catálisis , Modelos Moleculares , Neuraminidasa/antagonistas & inhibidores , Conformación Proteica , Ácidos Siálicos/farmacologíaRESUMEN
We report the selection and characterization of influenza A/NWS-G70c and B/HK/8/73 (HG) viruses which are resistant to the potent influenza neuraminidase inhibitor, 4-guanidino-Neu5Ac2en. Viruses were selected which replicated in MDCK cells in the presence of 20 micrograms/ml inhibitor. The neuraminidase of resistant viruses was > 200-fold more resistant to 4-guanidino-Neu5Ac2en than was the neuraminidase of the parent viruses. Although amounts of neuraminidase protein were similar in resistant and parent viruses, the enzyme activity of the resistant neuraminidase heads was reduced by > 95% for the substrates used. Relative to parent viruses, the resistant viruses replicated to equal or greater titers in tissue culture and in embryonated chicken eggs. Sequence analysis revealed a single nucleotide mutation in the neuraminidase gene of each virus resulting in the change of the conserved Glu 119 (which lies in a pocket beneath the active site of the enzyme) to Gly thus eliminating an electrostatic interaction with the C-4 guanidinium moiety of the inhibitor. Mutations (Asn-->Ser) at amino acids 145 and 150 were also found in the hemagglutinin gene of the B/HK/8/73 (HG) virus resistant to 4-guanidino-Neu5Ac2en. No changes were found in the hemagglutinin gene of the resistant A/NWS-G70c virus.
Asunto(s)
Antivirales/farmacología , Inhibidores Enzimáticos/farmacología , Virus de la Influenza A/efectos de los fármacos , Virus de la Influenza B/efectos de los fármacos , Neuraminidasa/antagonistas & inhibidores , Ácidos Siálicos/farmacología , Secuencia de Aminoácidos , Animales , Línea Celular , Embrión de Pollo , Farmacorresistencia Microbiana , Guanidinas , Glicoproteínas Hemaglutininas del Virus de la Influenza , Hemaglutininas Virales/genética , Humanos , Virus de la Influenza A/enzimología , Virus de la Influenza B/enzimología , Datos de Secuencia Molecular , Mutación , Neuraminidasa/genética , Piranos , Homología de Secuencia de Aminoácido , ZanamivirRESUMEN
Influenza virus sialidase is a surface enzyme that is essential for infection of the virus. The catalytic site is highly conserved among all known influenza variants, suggesting that this protein is a suitable target for drug intervention. The most potent known inhibitors are analogs of 2-deoxy-2,3-didehydro-N-acetylneuraminic acid (Neu5Ac2en), particularly the 4-guanidino derivative (4-guanidino-Neu5Ac2en). We utilized the benzene ring of 4-(N-acetylamino)benzoic acids as a cyclic template to substitute for the dihydropyran ring of Neu5Ac2en. In this study several 3-(N-acylamino) derivatives were prepared as potential replacements for the glycerol side chain of Neu5Ac2en, and some were found to interact with the same binding subsite of sialidase. Of greater significance was the observation that the 3-guanidinobenzoic acid derivative (equivalent to the 4-guanidino grouping of 4-guanidino-Neu5Ac2en), the most potent benzoic acid inhibitor of influenza sialidase thus far identified (IC50 = 10 microM), occupied the glycerol-binding subsite on sialidase as opposed to the guanidino-binding subsite. This benzoic acid derivative thus provides a new compound that interacts in a novel manner with the catalytic site of influenza sialidase.
Asunto(s)
Antiinfecciosos/farmacología , Benzoatos/farmacología , Virus de la Influenza A/enzimología , Virus de la Influenza B/enzimología , Ácido N-Acetilneuramínico/análogos & derivados , Neuraminidasa/antagonistas & inhibidores , Antiinfecciosos/química , Benzoatos/química , Ácido Benzoico , Cristalografía por Rayos X , Espectroscopía de Resonancia Magnética , Neuraminidasa/química , Ácidos Siálicos/química , Ácidos Siálicos/farmacología , Relación Estructura-ActividadRESUMEN
Influenza virus neuraminidase (NA) of the N9 subtype also possesses hemagglutinin activity and the hemagglutinating, or hemabsorbing (HB), site is distinct from the catalytic site. Previous results suggested that the NA was binding to sialic acid on the red cell surface, but we now report that the HB receptor is not sensitive to N9 influenza neuraminidase activity. Cell lines that constitutively express N9 or N2 neuraminidase have been used to further investigate the specificity of red blood cell binding to the HB site. The results suggest that the ligand is N-acetylneuraminic acid in a linkage or environment that is not sensitive to influenza virus neuraminidase, but which is released by the broadly specific bacterial sialidases from Micromonospora viridifaciens or Arthrobacter ureafaciens.
Asunto(s)
Eritrocitos/inmunología , Hemaglutininas Virales/inmunología , Neuraminidasa/inmunología , Orthomyxoviridae/inmunología , Animales , Células CHO , Línea Celular , Pollos , Cricetinae , Eritrocitos/virología , Humanos , Neuraminidasa/clasificación , Orthomyxoviridae/enzimologíaRESUMEN
A strategy was developed to design non-carbohydrate inhibitors of influenza virus neuraminidase. Using an iterative cycle of modeling, synthesis, biological testing and X-ray crystallography structure determination, a series of inhibitors based on benzoic acid were produced. The refined structures of three compounds complexed with neuraminidase are reported. The results demonstrate the success of this structure-based drug-design strategy.