Phosphoramidate Derivatives of Stavudine as Inhibitors of HIV: Unnatural Amino Acids May Substitute for Alanine

Some novel phosphoramidate derivatives of the nucleoside analogue stavudine have been prepared as membrane-soluble prodrugs of the bioactive free phosphate forms. Phenyl phosphates linked via nitrogen to methyl esterified amino acid analogues were studied, where the amino acid was an unnatural α-alkyl (or aryl) glycine or an α,α- dialkyl glycine. All compounds were characterized by a range of spectroscopic, spectrometric and analytical methods and were subjected to in vitro evaluation of their anti-human immunodeficiency virus efficacy. It is notable that certain unnatural amino acid derivatives could substitute for alanine with only a relatively small loss of activity and, moreover, that this activity did not fall-off with increasing alkyl chain length for the C2-C4 mono-alkyl series. These data are further probed by the application of our recently reported 31P-NMR-based carboxyl esterase assay, with informative results.

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Sterilization of drug-resistant influenza virus through genetic interference: use of unnatural amino acid-engineered virions

10.1101/2021.12.04.471209 ◽

2021 ◽

Author(s):

Xuesheng Wu ◽

Zhetao Zheng ◽

Hongmin Chen ◽

Haishuang Lin ◽

Yuelin Yang ◽

...

Keyword(s):

Influenza Virus ◽

Amino Acid ◽

Genetic Code ◽

Unnatural Amino Acids ◽

Influenza A ◽

Antiviral Agent ◽

Unnatural Amino Acid ◽

Drug Resistant ◽

Genetic Code Expansion

AbstractThe frequent emergence of drug resistance during the treatment of influenza A virus (IAV) infections highlights a need for effective antiviral countermeasures. Here, we present an antiviral method that utilizes unnatural amino acid-engineered drug-resistant (UAA-DR) virus. The engineered virus is generated through genetic code expansion to combat emerging drug-resistant viruses. The UAA-DR virus has unnatural amino acids incorporated into its drug-resistant protein and its polymerase complex for replication control. The engineered virus can undergo genomic segment reassortment with normal virus and produce sterilized progenies due to artificial amber codons in the viral genome. We validate in vitro that UAA-DR can provide a broad-spectrum antiviral strategy for several H1N1 strains, different DR-IAV strains, multidrug-resistant (MDR) strains, and even antigenically distant influenza strains (e.g., H3N2). Moreover, a minimum dose of neuraminidase (NA) inhibitors for influenza virus can further enhance the sterilizing effect when combating inhibitor-resistant strains, partly due to the promoted superinfection of unnatural amino acid-modified virus in cellular and animal models. We also exploited the engineered virus to achieve adjustable efficacy after external UAA administration, for mitigating DR virus infection on transgenic mice harboring the pair, and to have substantial elements of the genetic code expansion technology, which further demonstrated the safety and feasibility of the strategy. We anticipate that the use of the UAA-engineered DR virion, which is a novel antiviral agent, could be extended to combat emerging drug-resistant influenza virus and other segmented RNA viruses.

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Transferability of N-terminal mutations of pyrrolysyl-tRNA synthetase in one species to that in another species on unnatural amino acid incorporation efficiency

Amino Acids ◽

10.1007/s00726-020-02927-z ◽

2020 ◽

Author(s):

Thomas L. Williams ◽

Debra J. Iskandar ◽

Alexander R. Nödling ◽

Yurong Tan ◽

Louis Y. P. Luk ◽

...

Keyword(s):

Amino Acids ◽

Amino Acid ◽

Genetic Code ◽

Unnatural Amino Acids ◽

Trna Synthetase ◽

Unnatural Amino Acid ◽

Amino Acid Incorporation ◽

Acid Incorporation ◽

Genetic Code Expansion ◽

Incorporation Efficiency

AbstractGenetic code expansion is a powerful technique for site-specific incorporation of an unnatural amino acid into a protein of interest. This technique relies on an orthogonal aminoacyl-tRNA synthetase/tRNA pair and has enabled incorporation of over 100 different unnatural amino acids into ribosomally synthesized proteins in cells. Pyrrolysyl-tRNA synthetase (PylRS) and its cognate tRNA from Methanosarcina species are arguably the most widely used orthogonal pair. Here, we investigated whether beneficial effect in unnatural amino acid incorporation caused by N-terminal mutations in PylRS of one species is transferable to PylRS of another species. It was shown that conserved mutations on the N-terminal domain of MmPylRS improved the unnatural amino acid incorporation efficiency up to five folds. As MbPylRS shares high sequence identity to MmPylRS, and the two homologs are often used interchangeably, we examined incorporation of five unnatural amino acids by four MbPylRS variants at two temperatures. Our results indicate that the beneficial N-terminal mutations in MmPylRS did not improve unnatural amino acid incorporation efficiency by MbPylRS. Knowledge from this work contributes to our understanding of PylRS homologs which are needed to improve the technique of genetic code expansion in the future.

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Novel Amino Acid Derivatives of Quinolines as Potential Antibacterial and Fluorophore Agents

Scientia Pharmaceutica ◽

10.3390/scipharm88040057 ◽

2020 ◽

Vol 88 (4) ◽

pp. 57

Author(s):

Oussama Moussaoui ◽

Rajendra Bhadane ◽

Riham Sghyar ◽

El Mestafa El Hadrami ◽

Soukaina El Amrani ◽

...

Keyword(s):

Amino Acid ◽

Methyl Esters ◽

Amino Acid Derivatives ◽

Bacterial Strains ◽

Fluorescent Properties ◽

Topoisomerase Iv ◽

Microdilution Method ◽

Hydrolysis Of ◽

Derivatives Of

A new series of amino acid derivatives of quinolines was synthesized through the hydrolysis of amino acid methyl esters of quinoline carboxamides with alkali hydroxide. The compounds were purified on silica gel by column chromatography and further characterized by TLC, NMR and ESI-TOF mass spectrometry. All compounds were screened for in vitro antimicrobial activity against different bacterial strains using the microdilution method. Most of the synthesized amino acid-quinolines show more potent or equipotent inhibitory action against the tested bacteria than their correspond esters. In addition, many of them exhibit fluorescent properties and could possibly be utilized as fluorophores. Molecular docking and simulation studies of the compounds at putative bacterial target enzymes suggest that the antimicrobial potency of these synthesized analogues could be due to enzyme inhibition via their favorable binding at the fluoroquinolone binding site at the GyrA subunit of DNA gyrase and/or the ParC subunit of topoisomerase-IV.

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Properties of phosphoenolpyruvate carboxykinase (guanosine triphosphate) synthesized in hepatoma cells in the presence of amino acid analogues

Biochemical Journal ◽

10.1042/bj1460585 ◽

1975 ◽

Vol 146 (3) ◽

pp. 585-593 ◽

Cited By ~ 10

Author(s):

S E Knowles ◽

J M Gunn ◽

L Reshef ◽

R W Hanson ◽

F J Ballard

Keyword(s):

Amino Acid ◽

Phosphoenolpyruvate Carboxykinase ◽

Hepatoma Cells ◽

Dibutyryl Cyclic Amp ◽

Enzyme Degradation ◽

Amino Acid Analogues ◽

Enzyme Molecules ◽

Natural Amino Acids

1. Phosphoenolpyruvate carboxykinase (GTP) was induced by a combination of dibutyryl cyclic AMP, theophyline and dexamethasone in Reuber H35 hepatoma cells under conditions where an amino acid in the medium was replaced by an appropriate analogue. 2. With canavanine replacing arginine or with 5-fluorotryptophan or 6-fluorotryptophan replacing tryptophan the induced enzyme had a lower catalytic activity-relative to antibody reactivity. 3. These aberrant enzyme molecules were heat-labile in vitro. 4. Measurements of enzyme degradation in vivo indicated that the canavanine-containing enzyme and the 6-fluorotryptophan-containing enzyme were degraded more rapidly than the enzyme containing all natural amino acids.

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Engineering an acetyllysine reader with photocrosslinking amino acid for interactome profiling

Chemical Communications ◽

10.1039/d1cc04611j ◽

2021 ◽

Author(s):

Babu Sudhamalla ◽

Anirban Roy ◽

Soumen Barman ◽

Jyotirmayee Padhan

Keyword(s):

Amino Acids ◽

Amino Acid ◽

Protein Interactions ◽

Unnatural Amino Acids ◽

Protein Protein Interactions ◽

Site Specific ◽

Powerful Approach

The site-specific installation of light-activable crosslinker unnatural amino acids offers a powerful approach to trap transient protein-protein interactions both in vitro and in vivo. Herein, we engineer a bromodomain to...

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In vitro studies on the inhibition of colon cancer by amino acid derivatives of bromothiazole

Bioorganic & Medicinal Chemistry Letters ◽

10.1016/j.bmcl.2017.05.073 ◽

2017 ◽

Vol 27 (15) ◽

pp. 3507-3510 ◽

Cited By ~ 5

Author(s):

Nuno Vale ◽

Ana Correia-Branco ◽

Bárbara Patrício ◽

Diana Duarte ◽

Fátima Martel

Keyword(s):

Colon Cancer ◽

Amino Acid ◽

In Vitro Studies ◽

Amino Acid Derivatives ◽

Derivatives Of

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P-113d, an Antimicrobial Peptide Active againstPseudomonas aeruginosa, Retains Activity in the Presence of Sputum from Cystic Fibrosis Patients

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.45.12.3437-3444.2001 ◽

2001 ◽

Vol 45 (12) ◽

pp. 3437-3444 ◽

Cited By ~ 61

Author(s):

Umadevi S. Sajjan ◽

Linh T. Tran ◽

Nuria Sole ◽

Christopher Rovaldi ◽

Alan Akiyama ◽

...

Keyword(s):

Cystic Fibrosis ◽

Amino Acid ◽

Mirror Image ◽

Suppressive Therapy ◽

Significant Activity ◽

Amino Acid Residues ◽

Parent Molecule ◽

Histatin 5 ◽

Derivatives Of

ABSTRACT Antimicrobial peptides are a source of novel agents that could be useful for treatment of the chronic lung infections that afflict cystic fibrosis (CF) patients. Efficacy depends on antimicrobial activity against the major pathogens of CF patients, Pseudomonas aeruginosa, Staphylococcus aureus, and Haemophilus influenzae, in the environment of the CF patient's airway. We describe the in vitro efficacies of derivatives of histatins, which are histidine-rich peptides produced by the salivary glands of humans and higher primates. P-113, a peptide containing 12 of the 24 amino acid residues of the parent molecule, histatin 5, retained full antibacterial activity and had a good spectrum of activity in vitro against the prominent pathogens of CF patients. However, P-113 was not active in the presence of purulent sputum from CF patients. In contrast, P-113d, the mirror-image peptide with the amino acid residues in the d configuration, was stable in sputum, was as active as P-113 against pathogens of CF patients in the absence of sputum and retained significant activity in the presence of sputum from CF patients. Recombinant human DNase, which effectively liquefies sputum, enhanced the activity of P-113d in undiluted sputum against both exogenous (added) bacteria and endogenous bacteria. Because of its properties, P-113d shows potential as an inhalant in chronic suppressive therapy for CF patients.

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Synthesis and Evaluation of Some Symmetrical Phosphate Dimer Derivatives of 3′-Modified Nucleosides as Potential anti-HIV Agents

Antiviral Chemistry and Chemotherapy ◽

10.1177/095632029600700606 ◽

1996 ◽

Vol 7 (6) ◽

pp. 330-337 ◽

Cited By ~ 2

Author(s):

C. McGuigan ◽

H.-W. Tsang ◽

N. Mahmood ◽

A. J. Hay

Keyword(s):

Human Immunodeficiency Virus ◽

Analytical Data ◽

Modified Nucleosides ◽

Nucleoside Analogues ◽

Immunodeficiency Virus ◽

Anti Hiv Agents ◽

Derivatives Of ◽

Anti Hiv ◽

Hiv 1

Novel symmetrical nucIeotide-(5′,5′)-dimers of 3′-O-acetylthymidine, 3′-O-methylthymidine, 3′-O-ethylthymidine, 3′-O-n-propylthymidine and 3′-azido-3′-deoxythymidine (AZT) were synthesized as membrane soluble pro-drugs. These were prepared using phosphorodichloridate chemistry and were characterised by spectroscopic and analytical data. In-vitro evaluation of the derivatives in cells acutely infected with the human immunodeficiency virus (HIV-1) demonstrated a range of activities. These derivatives were generally found to display poor inhibition of HIV proliferation. Derivatives containing AZT moieties were found to be potent, but such compounds were less active than the parent nucleoside. The data indicated that the AZT-containing compounds act primarily via the release of the free nucleoside. However, in some cases, the dimers of certain inactive nucleoside analogues were found to be active. In these cases, release of the nucleoside alone cannot account for the activity.

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Highly water-soluble derivatives of the anesthetic agent propofol: in vitro and in vivo evaluation of cyclic amino acid esters

European Journal of Pharmaceutical Sciences ◽

10.1016/s0928-0987(03)00161-1 ◽

2003 ◽

Vol 20 (1) ◽

pp. 17-26 ◽

Cited By ~ 24

Author(s):

Cosimo Altomare ◽

Giuseppe Trapani ◽

Andrea Latrofa ◽

Mariangela Serra ◽

Enrico Sanna ◽

...

Keyword(s):

Amino Acid ◽

Water Soluble ◽

Anesthetic Agent ◽

Amino Acid Esters ◽

In Vivo Evaluation ◽

Derivatives Of ◽

Acid Esters

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Compensatory Substitutions Restore Normal Core Assembly in Simian Immunodeficiency Virus Isolates with Gag Epitope Cytotoxic T-Lymphocyte Escape Mutations

Journal of Virology ◽

10.1128/jvi.00068-06 ◽

2006 ◽

Vol 80 (16) ◽

pp. 8168-8177 ◽

Cited By ~ 27

Author(s):

Wendy W. Yeh ◽

Evan M. Cale ◽

Pimkwan Jaru-Ampornpan ◽

Carol I. Lord ◽

Fred W. Peyerl ◽

...

Keyword(s):

Human Immunodeficiency Virus ◽

Amino Acid ◽

Simian Immunodeficiency Virus ◽

Structural Integrity ◽

Viral Escape ◽

Amino Acid Substitutions ◽

Structural Constraints ◽

Immunodeficiency Virus ◽

Dominant Epitope

ABSTRACT The evolution of human immunodeficiency virus type 1 (HIV-1) and simian immunodeficiency virus (SIV) as they replicate in infected individuals reflects a balance between the pressure on the virus to mutate away from recognition by dominant epitope-specific cytotoxic T lymphocytes (CTL) and the structural constraints on the virus' ability to mutate. To gain a further understanding of the strategies employed by these viruses to maintain replication competency in the face of the intense selection pressure exerted by CTL, we have examined the replication fitness and morphological ramifications of a dominant epitope mutation and associated flanking amino acid substitutions on the capsid protein (CA) of SIV/simian-human immunodeficiency virus (SHIV). We show that a residue 2 mutation in the immunodominant p11C, C-M epitope (T47I) of SIV/SHIV not only decreased CA protein expression and viral replication, but it also blocked CA assembly in vitro and virion core condensation in vivo. However, these defects were restored by the introduction of upstream I26V and/or downstream I71V substitutions in CA. These findings demonstrate how flanking compensatory amino acid substitutions can facilitate viral escape from a dominant epitope-specific CTL response through the effects of these associated mutations on the structural integrity of SIV/SHIV.

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