Journal: Nature Communications

Article Title: An orally available M pro /TMPRSS2 bispecific inhibitor with potent anti-coronavirus efficacy in vivo

doi: 10.1038/s41467-025-60832-z

Figure Lengend Snippet: a Schematic illustration of the screening workflow for the discovery of M pro /TMPRSS2 bispecific inhibitor. b Quantification of the subgenomic envelope (sgE) gene in VeroE6-TMPRSS2 cells ( n = 6) infected with wildtype SARS-CoV-2 and Alpha, Beta, Delta, Omicron (BA.1 and JN.1) variants in the presence or absence of TMP1. Lysates were harvested at 24 hours post infection (hpi). for one-step reverse transcription and quantitative polymerase chain reaction (RT-qPCR) analysis. c Infectious viral titres in the supernatants harvested at 24 hpi. from VeroE6-TMPRSS2 cells ( n = 4) infected with wildtype SARS-CoV-2 and Alpha, Beta, Delta, Omicron (BA.1 and JN.1) variants were determined by plaque assays. Number of plaques were normalized to those recovered from supernatants with mock treatment only. d Quantification of the viral Orf1b and N gene with RT-qPCR in primary human nasal epithelial cells (hNECs) ( n = 4), MRC-5 ( n = 6) and LLC-MK2 ( n = 6) cells infected with HCoV-HKU1, -OC43 and -NL63 at 48 hpi. e Infectious viral titres in the supernatants harvested at 24 hpi from Huh7 infected with HCoV-229E or VeroE6-TMPRSS2 cells ( n = 4) infected with SARS-CoV-1 and MERS-CoV were determined in Huh7 cells (for HCoV-229E) or VeroE6-TMPRSS2 (for SARS-CoV-1 and MERS-CoV) by plaque assays. Each data point represents one biological repeat. Data represent mean ± SEM from the indicated number of biological repeats. For box-and-whisker plot shown, whiskers bounded the min-max values of the data, the bounds of the box represented lower (Q1)/upper (Q3) quartiles, and the central value represented the median value. Data were obtained from three independent experiments. WT, wildtype SARS-CoV-2.

Article Snippet: The human nasal epithelial cells in air-liquid interface (ALI) culture were purchased from Epithelix (EP02MP, Epithelix, Switzerland) and maintained with MucilAir culture medium (EP04MM, Epithelix) until virus challenge.

Techniques: Infection, Reverse Transcription, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Whisker Assay

Journal: Nature Communications

Article Title: An orally available M pro /TMPRSS2 bispecific inhibitor with potent anti-coronavirus efficacy in vivo

doi: 10.1038/s41467-025-60832-z

Figure Lengend Snippet: a Pharmacokinetics of TMP1, Paxlovid and camostat oral delivery in mice ( n = 3). b Schematic illustration of SARS-CoV-2 infection in K18-hACE2 transgenic mice with therapeutic or prophylactic TMP1 treatment (Created in BioRender) . c Quantification of sgE gene of SARS-CoV-2 in the nasal turbinate and lung tissues of the infected mice ( n = 6) with prophylactic treatment. d Quantification of the infectious viral titres in the nasal turbinate and lung tissues of the infected mice ( n = 6) with prophylactic treatment. e Viral N protein expression in the nasal turbinate and lung tissues of infected mice ( n = 3) with prophylactic treatment was quantified with ImageJ. f Representative images of SARS-CoV-2 N protein expression (black arrow) in nasal turbinate and lung tissue of the infected mice. Scale bar represents 100 µm. g Histology analysis of the nasal turbinate and lung tissue of the infected mice by H&E staining. Scale bar represents 100 µm. Black arrowhead, nasal epithelial desquamation; open arrowhead, alveolar collapse; dashed circle, inflammation infiltrations in alveolar septa; asterisk, bronchiolar epithelium damage. h Body weight change of the female ( n = 6) and male ( n = 11 and 13 for vehicle and TMP1 group, respectively) infected mice with or without TMP1 prophylactic treatment. i Survival of the female ( n = 6) and male ( n = 11 and 13 for vehicle and TMP1 group, respectively) infected mice with or without TMP1 prophylactic treatment. j Quantification of sgE gene of SARS-CoV-2 in the nasal turbinate and lung tissues of the infected mice ( n = 10) with delayed therapeutic treatment. Each data point represents one biological repeat. Data represents mean ± SEM from the indicated number of biological repeats. Statistical significances were determined using one way-ANOVA with Dunnett’s multiple comparisons test ( c – e ), ( j ) and log-rank (Mantel-Cox) tests ( i ). Data were obtained from three independent experiments. * represented p < 0.05 and ** represented p < 0.01. *** represented p < 0.001, **** represented p < 0.0001. WT wildtype SARS-CoV-2, Veh vehicle, PAX Paxlovid.

Article Snippet: The human nasal epithelial cells in air-liquid interface (ALI) culture were purchased from Epithelix (EP02MP, Epithelix, Switzerland) and maintained with MucilAir culture medium (EP04MM, Epithelix) until virus challenge.

Techniques: Drug discovery, Infection, Transgenic Assay, Expressing, Staining

Journal: Nature Communications

Article Title: An orally available M pro /TMPRSS2 bispecific inhibitor with potent anti-coronavirus efficacy in vivo

doi: 10.1038/s41467-025-60832-z

Figure Lengend Snippet: a Schematic illustration of SARS-CoV-2 infection in human nasal epithelial cells (hNECs). Differentiated hNECs maintained in air-liquid interface (ALI) culture were infected with SARS-CoV-2 Omicron JN.1 or KP.2 with vehicle only or TMP1 treatment until sample harvest at 48 hpi (Created in BioRender ). b Quantification of SARS-CoV-2 sgE gene in the infected cell lysates ( n = 5). c Quantification of the infectious viral titres in the apical supernatants ( n = 5). d Schematic illustration of the SARS-CoV-2 transmission study in golden Syrian hamsters (Created in BioRender ). e Quantification of sgE gene of SARS-CoV-2 in the nasal turbinate and lung tissues of the contact hamsters ( n = 6). f Quantification of the infectious viral titres in the nasal turbinate and lung tissues of the contact hamsters ( n = 6). g Quantification of viral N protein expression in nasal turbinate and lung tissues of the contact hamsters ( n = 3). Quantification was performed with ImageJ. h Representative images of SARS-CoV-2 N protein expression (black arrow) in nasal turbinate and lung tissue of the contact hamsters. Scale bar represents 100 µm. i Histology analysis of the nasal turbinate and lung tissue of the infected hamsters by H&E staining. Black arrowhead, nasal epithelial desquamation; dashed circle, necrotic cell debris in the nasal cavity; open arrowhead, hemorrhage in the alveolar septa; asterisk, alveoli collapse; cross, inflammatory infiltration in alveolar septa. Scale bar represents 100 µm. Each data point represents one biological repeat. Data represents mean ± SEM from the indicated number of biological repeats. Statistical significances were determined using two-tailed Student’s t -test ( b , c ) and ( e – g ). Data were obtained from three independent experiments. * represented p < 0.05 and ** represented p < 0.01. Veh vehicle.

Article Snippet: The human nasal epithelial cells in air-liquid interface (ALI) culture were purchased from Epithelix (EP02MP, Epithelix, Switzerland) and maintained with MucilAir culture medium (EP04MM, Epithelix) until virus challenge.

Techniques: Infection, Transmission Assay, Expressing, Staining, Two Tailed Test

Journal: Nature Communications

Article Title: An orally available M pro /TMPRSS2 bispecific inhibitor with potent anti-coronavirus efficacy in vivo

doi: 10.1038/s41467-025-60832-z

Figure Lengend Snippet: a Schematic illustration of SARS-CoV-1 infection in K18-hACE2 transgenic mice with vehicle or TMP1 treatment (Created in BioRender) . b Quantification of sgE gene of SARS-CoV-1 in the nasal turbinate and lung tissues of the infected mice with prophylactic treatment ( n = 8). c Quantification of the infectious viral titres in the nasal turbinate and lung tissues of the SARS-CoV-1-infected mice ( n = 8). d Viral N protein expression in the nasal turbinate and lung tissues of the SARS-CoV-1- infected mice ( n = 3) at 3 dpi. was quantified with ImageJ. e Representative images of N protein expression (black arrow) in nasal turbinate and lung tissues of the SARS-CoV-1-infected mice. Scale bar represents 100 µm. f Histology analysis of the nasal turbinate and lung tissues of the SARS-CoV-1-infected mice by H&E staining. Black arrowhead, nasal epithelial desquamation; asterisk, hemorrhage in nasal submucosal region; dashed circle, necrotic cell debris in nasal cavity; open arrowhead, alveolar collapse; cross, inflammatory infiltration. Scale bar represents 200 µm. g Schematic illustration of MERS-CoV MA infection in hDPP4-knockin (hDPP4-KI) transgenic mice with vehicle or TMP1 treatment (Created in BioRender107). h Quantification of N gene of MERS-CoV MA in the nasal turbinate and lung tissues of the infected mice ( n = 5). i Quantification of the infectious viral titres in the nasal turbinate and lung tissues of the MERS-CoV MA -infected mice ( n = 5). Each data point represents one biological repeat. Data represents mean ± SEM from the indicated number of biological repeats. Statistical significances were determined using two-tailed Student’s t -test ( b – d ) and ( h , i ). Data were obtained from three independent experiments. * represented p < 0.05 and ** represented p < 0.01. Veh vehicle.

Article Snippet: The human nasal epithelial cells in air-liquid interface (ALI) culture were purchased from Epithelix (EP02MP, Epithelix, Switzerland) and maintained with MucilAir culture medium (EP04MM, Epithelix) until virus challenge.

Techniques: Infection, Transgenic Assay, Expressing, Staining, Knock-In, Two Tailed Test