Cationic Perylene Antivirals with Aqueous Solubility for Studies In Vivo

Abstract

Perylene-based compounds are attracting significant attention due to their high broad-spectrum antiviral activity against enveloped viruses. Despite unambiguous results of in vitro studies and high selectivity index, the poor water solubility of these compounds prevented in vivo evaluation of their antiviral properties. In this work, we synthesized a series of compounds with a perylene pharmacophore bearing positively charged substituents to improve the aqueous solubility of this unique type of antivirals. Three types of charged groups were introduced: (1) quaternary morpholinium salts (3a–b); (2) a 2′-O-l-valinyl-uridine hydrochloride residue (8), and (3) a 3-methylbenzothiazolium cation (10). The synthesized compounds were evaluated based both on antiviral properties in vitro (CHIKV, SARS-CoV-2, and IAV) and on solubility in aqueous media. Compound 10 has the greatest aqueous solubility, making it preferable for pre-evaluation by intragastrical administration in a mouse model of lethal influenza pneumonia. The results indicate that the introduction of a positively charged group is a viable strategy for the design of drug candidates with a perylene scaffold for in vivo studies.

Nucleoside Analogs and Perylene Derivatives Modulate Phase Separation of SARS-CoV-2 N Protein and Genomic RNA In Vitro

“…In addition to purine nucleoside derivatives, we considered a well-known cytidine analog NHC (β-D-N 4 -hydroxycytidine, Figure 4 a) [ 30 ] and several recently reported pyrimidine derivatives ( Figure 4 c,d) that inhibited SARS-CoV-2 in vitro. These derivatives included 5′-norcarbocyclic (NorC) analogs of 3H-pirrolo[2,3-d]-pyrimidine-2-one (NorC-24p) and 3H-furano[2,3-d]-pyrimidine-2-one (NorC-24f) nucleosides with a hydrophobic 4-pentylphenyl substituent ( Figure 4 c) [ 29 ] and perylene-harboring uridine analogs Peryl-8 and Peryl-5 (neutral and positively charged derivatives, respectively, Figure 4 d) [ 31 ]. To verify the importance of charged and hydrophobic fragments, we also tested the non-nucleoside neutral perylene derivatives Peryl-2a and Peryl-2b, their positively charged analogs Peryl-3a and Peryl-3b, and an additional positively charged compound with in vivo-confirmed antiviral activity Peryl-10 ( Figure 4 d) [ 31 ].…”

“…These derivatives included 5′-norcarbocyclic (NorC) analogs of 3H-pirrolo[2,3-d]-pyrimidine-2-one (NorC-24p) and 3H-furano[2,3-d]-pyrimidine-2-one (NorC-24f) nucleosides with a hydrophobic 4-pentylphenyl substituent ( Figure 4 c) [ 29 ] and perylene-harboring uridine analogs Peryl-8 and Peryl-5 (neutral and positively charged derivatives, respectively, Figure 4 d) [ 31 ]. To verify the importance of charged and hydrophobic fragments, we also tested the non-nucleoside neutral perylene derivatives Peryl-2a and Peryl-2b, their positively charged analogs Peryl-3a and Peryl-3b, and an additional positively charged compound with in vivo-confirmed antiviral activity Peryl-10 ( Figure 4 d) [ 31 ]. The codes of all small molecules were taken from the previous works [ 29 , 31 ] and the prefixes indicate small molecule types.…”

“…To verify the importance of charged and hydrophobic fragments, we also tested the non-nucleoside neutral perylene derivatives Peryl-2a and Peryl-2b, their positively charged analogs Peryl-3a and Peryl-3b, and an additional positively charged compound with in vivo-confirmed antiviral activity Peryl-10 ( Figure 4 d) [ 31 ]. The codes of all small molecules were taken from the previous works [ 29 , 31 ] and the prefixes indicate small molecule types.…”

“… Code Effects on the N-RNA Condensates a Antiviral Activity, IC50 ± SD, µM b S/S control ± SD Count/Count control ± SD ATP 0.4 ± 0.1 0.5 ± 0.2 - NHC 0.5 ± 0.3 0.4 ± 0.1 8 ± 5, refs. [ 29 , 31 ] c NorC-24p 11 ± 1 6.2 ± 0.8 21 ± 6, ref. [ 29 ] c NorC-24f 15 ± 1 9 ± 2 ≥50 c Flex-nt10 b 2 ± 1 4.7 ± 0.3 - Flex-ns10 2.9 ± 0.9 2.4 ± 0.4 >100 c Flex-ns12 0.4 ± 0.1 0.4 ± 0.1 >100 c Flex-dns12 3 ± 1 4 ± 1 >100 c Peryl-8 23 ± 7 1.4 ± 0.4 1.3 ± 0.4, ref.…”

“…[ 29 ] c NorC-24f 15 ± 1 9 ± 2 ≥50 c Flex-nt10 b 2 ± 1 4.7 ± 0.3 - Flex-ns10 2.9 ± 0.9 2.4 ± 0.4 >100 c Flex-ns12 0.4 ± 0.1 0.4 ± 0.1 >100 c Flex-dns12 3 ± 1 4 ± 1 >100 c Peryl-8 23 ± 7 1.4 ± 0.4 1.3 ± 0.4, ref. [ 31 ] c Peryl-5 50 ± 10 1.5 ± 0.4 >100, ref. [ 31 ] Peryl-2a 17 ± 6 1.8 ± 0.3 11 ± 2, ref.…”