Kinase bypass: A new strategy for anti-HIV drug design

doi:10.1016/S0960-894X(00)80316-9

Bioorganic & Medicinal Chemistry Letters

Volume 3, Issue 6, June 1993, Pages 1207-1210

https://doi.org/10.1016/S0960-894X(00)80316-9 Get rights and content

Abstract

The 5′-bis(trichloroethyl) phosphate derivatives of several 3′-O-acyl thymidines were prepared from the thymidine phosphate. Even though the parent nucleosides are inactive as antiviral agents, the phosphates are selective inhibitors of the proliferation of HIV. This activity is attributed to a new mechanism of action, we herein describe as “kinase by-pass”.

The 5′-bis(trichloroethyl) phosphate derivatives of several 3′-O-acyl thymidines were prepared from the thymidine phosphate, which was prepared from thymidine in one step, using phosphorochloridate chemistry. Even though the parent nucleoside analogues are inactive as antivirals, the phosphate derivatives are selective inhibitors of the proliferation of HIV. This activity is attributed to a new mechanism of action we herein describe as “kinase by-pass”

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Aryl H-phosphonates. 19. New anti-HIV pronucleotide phosphoramidate diesters containing amino- and hydroxypyridine auxiliaries
2019, European Journal of Medicinal Chemistry
Citation Excerpt :
Apart from facilitating a diffusion through biological membranes, these type of compounds when inside the cell, undergo sequential chemical and enzymatic degradations to unleash phosphorylated antiviral metabolites [11–14]. In 1993 McGuigan et al. [15] reported on anti-HIV activity of AZT-phosphoramidate diesters bearing aryl and amino acid esters (alanine and its analogues) as phosphate masking groups. Bioactivation of these compounds commences with carboxylesterase-mediated hydrolysis of the amino acid ester part, followed by intramolecular displacement of the aryl group by the formed carboxylate [6,16–20].
We have designed a new type of AZT and ddU phosphoramidate diesters containing various combinations of 2-, 3-, 4-aminopyridine and 2-, 3-, 4-hydroxypyridine moieties attached to the phosphorus center, as potential anti-HIV pronucleotides. Depending on the pK_a values of the aminopyridines and the hydroxypyridines used, alternative synthetic strategies based on H-phosphonate chemistry were developed for their preparation. Synthetic aspects of these transformations and the biological activity of the synthesized compounds are discussed.
Norcantharimide analogues possessing terminal phosphate esters and their anti-cancer activity
2011, Bioorganic and Medicinal Chemistry
Citation Excerpt :
This in itself is not surprising. Simple alkyl esters are more resistant to hydrolysis than aromatic or trihaloalkyl phosphates and so it may be expected that the possibly bioactive ‘naked’ phosphate form of the molecule is not reached.44 In general both the phenyl and bis(trichloroethyl) analogues show far better bioactivity relative to the simple alkyl ester analogues which can be explained by this ease of hydrolysis.45,46
A family of norcantharidin analogues possessing a terminal alcohol (ethanol, propanol, butanol, pentanol, hexanol and cyclohexanol) moiety were treated with either chlorodiethyl, chlorodiphenyl or chloro-bis-trichloroethyl-phosphate to afford highly focused libraries of the corresponding phosphate esters. Subsequent biological screening against a panel of nine human cancer cell lines identified a trend between the ease of phosphate unmasking (phosphate ester hydrolysis) and cell death. The most potent analogues possessed either a diphenyl or a bis-trichloroethyl moiety. The effect of alkyl spacer was also examined with the hexyl analogues typically more potent. 4-Aza-4-(3-{bis(2,2,2-trichloroethyl)phosphate}propyl)-10-oxatricyclo[5.2.1.0]decane-3,5-dione (10b) was the most potent analogue synthesised with an average GI₅₀ of 11 μM across a panel of nine human carcinoma cell lines: colon carcinoma (HT29 and SW480); breast carcinoma (MCF-7); ovarian carcinoma (A2780); lung carcinoma (H460); skin carcinoma (A431); prostate carcinoma (DU145); neuronal carcinoma (BE2-C) and brain carcinoma (SJ-G2). This represents a fivefold improvement in anti-proliferative activity relative to the lead, norcantharidin.
<sup>31</sup>P NMR and genetic analysis establish hinT as the only Escherchia coli purine nucleoside phosphoramidase and as essential for growth under high salt conditions
2005, Journal of Biological Chemistry
Eukaryotic cells encode AMP-lysine (AMP-N-ϵ-(N-α-acetyl lysine methyl ester) 5′-phosphoramidate) hydrolases related to the rabbit histidine triad nucleotide-binding protein 1 (Hint1) sequence. Bacterial and archaeal cells have Hint homologs annotated in a variety of ways, but the enzymes have not been characterized, nor have phenotypes been described due to loss of enzymatic activity. We developed a quantitative ³¹P NMR assay to determine whether Escherichia coli possesses an adenosine phosphoramidase activity. Indeed, soluble lysates prepared from wild-type laboratory E. coli exhibited activity on the model substrate adenosine 5′-monophosphoramidate (AMP-NH₂). The E. coli Hint homolog, which had been comprehensively designated ycfF and is here named hinT, was cloned, overexpressed, purified, and characterized with respect to purine nucleoside phosphoramidate substrates. Bacterial hinT was several times more active than human or rabbit Hint1 on five model substrates. In addition, bacterial and mammalian enzymes preferred guanosine versus adenosine phosphoramidates as substrates. Analysis of the lysates from a constructed hinT knock-out strain of E. coli demonstrated that all of the cellular purine nucleoside phosphoramidase activity is due to hinT. Physiological analysis of this mutant revealed that the loss of hinT results in failure to grow in media containing 0.75 m KCl, 0.9 m NaCl, 0.5 m NaOAc, or 10 mm MnCl₂. Thus, cation-resistant bacterial cell growth may be dependent on the hydrolysis of adenylylated and/or guanylylated phosphoramidate substrates by hinT.
Evaluation of subchronic (13 weeks) and reproductive toxicity potential of intravaginal gel-microemulsion formulation of a dual-function phenyl phosphate derivative of bromo-methoxy zidovudine (compound WHI-05) in B<inf>6</inf>C<inf>3</inf>F<inf>1</inf> mice
2000, Contraception
Heterosexual transmission of human immunodeficiency virus (HIV) accounts for 90% of all new infections worldwide and significantly contributes to new acquired immunodeficiency syndrome (AIDS) cases in the United States. In a systematic effort to develop a microbicidal contraceptive capable of preventing HIV transmission as well as providing fertility control, we previously identified novel phenyl phosphate derivatives of 3′-azido-3′-deoxythymidine (zidovudine) which exhibit potent anti-HIV and spermicidal activities. This study reports the preliminary preclinical study of our lead compound WHI-05, 5-bromo-6-methoxy-5,6-dihydro-3′-azidothymidine-5′-(p-methoxyphenyl) methoxyalaninyl phosphate, for use as a dual-function topical microbicide. Acute toxicity studies have shown that WHI-05 has no detectable adverse effects on laboratory animals. The 13-week subchronic and reproductive toxicity potential of intravaginal gel-microemulsion formulation of WHI-05 were studied in mice to support its further development as a virucidal spermicide. Groups of 10 female B₆C₃F₁ mice were exposed intravaginally to a gel-microemulsion formulation containing 0%, 0.5%, 1.0%, or 2.0% WHI-05, 5 days/week for 13 consecutive weeks. On a molar basis, these concentrations represent 300 to 1200 times their in vitro spermicidal potency, and 1.5 × 10⁴ to 6.1 × 10⁴ times their in vitro anti-HIV activity. After 13 weeks of intravaginal treatment, one half of treated mice were evaluated for toxicity, and the other half were mated with untreated males to evaluate potential reproductive and developmental effects. Repetitive intravaginal application of WHI-05 to yield a local concentration 6.1 × 10⁴ times higher than its in vitro HIV IC₅₀ value and 1200 times higher than its spermicidal EC₅₀ value did not cause weight loss, morbidity, mortality, or specific tissue lesions detectable by histopathology. Furthermore, repeated intravaginal exposure of mice to WHI-05 for 13 weeks had no adverse effects on subsequent reproductive performance (100% fertile), neonatal survival (>96%), or pup development. These findings collectively show that the experimental dual-function anti-HIV and contraceptive agent, WHI-05, did not cause significant acute or subchronic and reproductive toxicity under the test conditions.
WHI-05, a novel bromo-methoxy substituted phenyl phosphate derivative of zidovudine, is a dual-action spermicide with potent anti-HIV activity
1999, Contraception
Heterosexual transmission of HIV to women is the fastest-growing mode of transmission. In a systematic effort to develop a microbicide capable of preventing HIV transmission as well as providing fertility control, novel phenyl phosphate derivatives of 3’-azido-3’-deoxythymidine (zidovudine, ZDV) have been identified that exhibit potent anti-HIV and spermicidal activities. This study reports the synthesis, characterization, and preclinical formulation of compound WHI-05, 5-bromo-6-methoxy-5,6-dihydro-3’-azidothymidine-5’-(p-methoxyphenyl) methoxyalaninyl phosphate. The anti-HIV activities of WHI-05 and ZDV were compared by measuring p24 antigen production and reverse transcriptase activity as markers of viral replication using human peripheral blood mononuclear cells (PBMC) infected with both ZDV-sensitive and ZDV-resistant strains of HIV. The sperm immobilizing activity (SIA) of WHI-05 was compared with that of ZDV and nonoxynol - 9 (N - 9) by computer-assisted sperm analysis (CASA). The effect of WHI-05 on sperm membrane integrity was examined by high resolution, low voltage scanning electron microscopy (HR-LVSEM). The in vitro cytotoxicity profile of WHI-05 versus N - 9 were compared using normal human vaginal, ectocervical, and endocervical epithelial cells. The in vivo vaginal tolerance, absorption, and toxicity of a 2% WHI-05 gel-microemulsion was tested in the rabbit. Whereas ZDV displayed potent anti-HIV activity but lacked SIA, WHI-05 elicited both potent anti-HIV activity and SIA. WHI-05 inhibited the replication of ZDV-sensitive as well as ZDV-resistant strains of HIV in PBMC. CASA combined with HR-LVSEM demonstrated that WHI-05-induced SIA was not associated with membrane damage. Unlike, N - 9, the spermicidal activity of WHI-05 was not associated with cytotoxicity to reproductive tract epithelial cells. Repetitive intravaginal application of a 2% WHI-05 gel-microemulsion did not damage the vaginal epithelium or cause local inflammation in the rabbit model. As a potent anti-HIV agent that has spermicidal activity and is devoid of mucosal toxicity, WHI-05 shows a unique clinical potential to become the active ingredient for a vaginal contraceptive for women who are at high risk for acquiring HIV by heterosexual vaginal transmission.
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Kinase bypass: A new strategy for anti-HIV drug design

Abstract

Antiviral Res.

Antiviral Res.

Tetrahedron Lett.

J. Biol. Chem.

J. Biol. Chem.

Proc. Natl. Acad. Sci. USA

Science

Antiviral Chem. Chemotter.

Antiviral Chem. Chemother.