Synthesis and Hybridization Studies of a New CPP-PNA Conjugate as a Potential Therapeutic Agent in Atherosclerosis Treatment

Abstract

The objective of this study was to design and synthesize a new CPP-PNA conjugate that would be able to penetrate endothelial cells, bind STAT1 mRNA and thereby block the activity of STAT1 (the Signal Transducer and Activator of Transcription 1), which is important in cases of vessel inflammation. In the course of the study, the TAMRA-PTD-4- Hal(traziole-Gly-PNA)-conjugate was successfully synthesized using a specific 1,3-dipolar Huisgen cycloaddition reaction known as a "click reaction". The hybridization properties of the conjugate to complementary hSTAT1 mRNA and hSTAT1 ssDNA fragments was verified by capillary electrophoresis (CE). Studies have shown that the attachment of a fluorescence-labeled peptide to a PNA sequence via a 1,2,3-triazole ring did not alter the binding properties of the PNA to the complementary hSTAT1 mRNA or hSTAT1 ssDNA fragments maintaining similar binding affinity. Furthermore, the data obtained suggest that the use of such a conjugate to modulate the activity and expression of STAT1 could provide a new therapeutic strategy for atherosclerosis treatment.

Vitamin B12 as a carrier of peptide nucleic acid (PNA) into bacterial cells

“…The azide derivative of this peptide ((KFF) 3 K-N 3 ) was attached to the alkyne-PEG5-PNA via 1,3-dipolar cycloaddition (Fig. 3d ) 43 . See Methods for detailed synthesis procedures.…”

“… 42 . Other conjugates were synthesized using copper-catalyzed azide-alkyne cycloaddition according to the procedures described in refs 39 , 43 . CuI (1.0 mg, 5 μmol) and TBTA (5.0 mg, 10 μmol) were dissolved in DMF/H 2 O (0.5 mL, 1:1 v/v) and stirred for 20 min.…”

Transportan 10 improves the pharmacokinetics and pharmacodynamics of vancomycin

“…TP10 and its analogues were synthesized according to the standard procedure of the solid phase peptide synthesis (SPPS) by using an automatic peptide synthesizer (Quartet, Protein Technologies Inc) with TentaGel S RAM resin (loading of amino groups − 0.25 mmol/g) 36,37 . Fmoc-protected amino acids were assembled as active derivatives in a 3-fold molar excess of O -(benzotriazole-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate (TBTU) with addition of N -hydroxybenzotriazole (HOBt) and N -methylmorpholine (NMM) (1:1:2) in the N , N -dimethylformoamide (DMF) solution for 2 × 30 min.…”

“…Conjugates of TP10 with Van were obtained by using “click chemistry” – the Cu(I)-catalyzed specific 1,3-dipolar Huisgen’s cycloaddition reaction 37 . The reactions of the alkyne functionalized TP10 analogues (0.8 μM each time) with 0.4 μM of azido functionalized Van-PEG 3 -N 3 (conjugate I ) or Van-PEG 4 -N 3 (conjugates II–IV ) were carried out in 1.5 ml of water/ tert -butanol medium (1:1 v/v) in the presence of 8 µ l of 0.1 M CuSO 4 × H 2 O and 4 µ l of freshly prepared solution of 0.5 M sodium ascorbate (2:1:2:5).…”

Preparation and cellular uptake of bicyclic‐peptide cargo clicked to cell penetrating peptides

“…It also facilitated the combinatorial preparation of one cargo and two CPPs. This strategy has been utilized for the attachment of proteins, oligonucleotides, and PNA to synthetic CPPs. To our knowledge, the conjugation of a bicyclic peptide cargo and a CPP via click chemistry has not been previously reported and can now be added to this list.…”