Structure−Activity Relationships:  Analogues of the Dicaffeoylquinic and Dicaffeoyltartaric Acids as Potent Inhibitors of Human Immunodeficiency Virus Type 1 Integrase and Replication

Abstract

The dicaffeoylquinic acids (DCQAs) and dicaffeoyltartaric acids (DCTAs) are potent and selective inhibitors of human immunodeficiency virus type 1 (HIV-1) integrase. They also inhibit HIV-1 replication at nontoxic concentrations. Since integrase is an excellent target for anti-HIV therapy, structure-activity relationships were employed to synthesize compounds with: (1) improved potency against HIV-1 integrase, (2) improved anti-HIV effect in tissue culture, and (3) increased selectivity as indicated by low cellular toxicity. Thirty-four analogues of the DCTAs and DCQAs were synthesized and tested for cell toxicity, anti-HIV activity, and inhibition of HIV-1 integrase. Seventeen of the 34 analogues had potent activity against HIV-1 integrase ranging from 0. 07 to >10 microM. Seventeen analogues that were synthesized or purchased had no inhibitory activity against integrase at concentrations of 25 microM. Of the biologically active analogues, 7 of the 17 inhibited HIV replication at nontoxic concentrations. The most potent compounds were D-chicoric acid, meso-chicoric acid, bis(3,4-dihydroxydihydrocinnamoyl)-L-tartaric acid, digalloyl-L-tartaric acid, bis(3,4-dihydroxybenzoyl)-L-tartaric acid, dicaffeoylglyceric acid, and bis(3, 4-dihydroxyphenylacetyl)-L-tartaric acid. Anti-HIV activity of the active compounds in tissue culture ranged from 35 to 0.66 microM. Structure-activity relationships demonstrated that biscatechol moieties were absolutely required for inhibition of integrase, while at least one free carboxyl group was required for anti-HIV activity. These data demonstrate that analogues of the DCTAs and the DCQAs can be synthesized which have improved activity against HIV integrase.

Human Immunodeficiency Virus Type 1 (HIV-1) Integrase: Resistance to Diketo Acid Integrase Inhibitors Impairs HIV-1 Replication and Integration and Confers Cross-Resistance to l -Chicoric Acid

“…Suballelic reconstruction of reference HIV containing IN with the G140S substitution yielded HIV that was completely resistant to L-CA in tissue culture (37). HIV containing G140S also demonstrates resistance to L-CA analogs in tissue culture (36). Recombinant IN containing this substitution was impaired for catalysis and demonstrated eightfold resistance to L-CA and nearly fivefold resistance to L-731,988 in the strand transfer reaction (35).…”

“…Mutant IN genes were amplified with oligonucleotide primers containing NdeI and HindIII restriction sites (37) and then cloned into a pET-based protein expression vector (62). PCR conditions were as follows: initial denaturation for 2 min at 94°C, followed by 25 cycles at 94°C for 30 s, 55°C for 30 s, and 72°C for 75 s. Recombinant IN proteins were expressed in BL21 pLysS cells (Stratagene), as previously described (36). Cells were lysed by sonication and ultracentrifugation.…”

“…The lysate was dialyzed overnight against buffer A (20 mM HEPES, 1 M NaCl, 0.1% NP-40, 10% glycerol, and 5 mM ␤-mercaptoethanol). Five milliliters of each dialysate was purified via its N-terminal six-His tag with Ni 2ϩ affinity column chromatography (36). Protein purity was assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and protein concentration was determined using the Coomassie G-250 protein assay (Pierce).…”

“…The susceptibility of each mutant IN to L-CA or L-731,988 was determined essentially as previously described (36,37,49). Briefly, in triplicate, IN was incubated with V1-V2 as described above in the presence of 0.3 to 10.0 M inhibitor.…”

The economic potential of beach-cast seagrass – Cymodocea nodosa: a promising renewable source of chicoric acid

“…It has been shown to have immunostimulatory properties, promoting phagocyte activity in vitro and in vivo (Bauer et al 1989), and to inhibit hyaluronidase, a key enzyme involved in bacterial infection (Bauer 1998). In addition, CA has antiviral activity (Pellati et al 2004) and has been reported to inhibit HIV integrase and replication (King et al 1999, Lin et al 1999, Charvat et al 2006, Liu et al 2006. The activity of CA against herpes simplex virus has been demonstrated (Binns et al 2002).…”