NAV-2729

Biological Activity

NAV-2729 (Grassofermata) is a selective ARF6 inhibitor with IC50 value of 1.0 μM. NAV-2729 (Grassofermata) reduces uveal melanoma cell proliferation and tumorigenesis in a mouse model, confirming the functional relevance of this pathway and suggesting a therapeutic strategy for Gα-mediated diseases.

Mice experiments
The C57 mice expressing hSCARB2 (21‐day‐old) were infected with the viruses (2×10^6 PFU) via subcutaneously (s.c.) inoculation and were observed for clinical signs. Control‐infected mice were injected with the same volume of PBS. Control groups were subcutaneously (s.c.) injected with EV71 (2×10^6 PFU) and PBS, while experimental group were subcutaneously injected with EV71 (2 ×10^6 PFU) and NAV‐2729 (4.5mg/kg). NAV‐2729 was administered (s.c.) at the indicated dose and PBS was injected as a control. Survival rates changes of the mice were observed and recorded every day until 14 days postinfection (DPI). The organs and tissues were removed from euthanized mice at the indicated time points and fixed with 4% paraformaldehyde at 4°C for 24h. After fixation, the organs and tissues were subjected to histopathological and immuno histochemical (IHC) analysis.
https://pubmed.ncbi.nlm.nih.gov/37417384/

Burn sepsis model
Anaesthesia was performed by intramuscular injection of 100 mg/kg ketamine and 12.5 mg/kg xylazine hydrochloride. An adequate anesthetic depth was determined by toe pinch. Then the hair on the dorsal skin was removed using an electric clipper. All animals received a subcutaneous injection of 1 mL normal saline into the burn target area to protect from spinal cord damage during the burn injury. An approximately 30% total body surface area full-thickness scald burn was induced by exposing the shaved dorsal area to 98 °C water for 10 s, followed by intraperitoneal injection of 1 mL normal saline for fluid resuscitation. For analgesia, all animals received subcutaneously a 0.05 mg/kg buprenorphine 30 min prior to burn injury and two times a day for three days postoperatively. For burn wound infection, 5 mg/kg lipopolysaccharide (LPS) was intradermally injected at the burn eschar site immediately after the burn insult. The sham animals underwent a sham procedure that included all the interventions except for the thermal injury and LPS intradermally injection. For treatment intervention, immediately after the burn and sepsis stimuli were inflicted, mice in the treatment group received intraperitoneal injection of NAV-2729 dissolved in 5% dimethyl sulfoxide (DMSO), 30 mg/kg, once daily. In survival studies, NAV-2729 was given for 3 consecutive days. The dose and method of administration of NAV-2729 were chosen based on the results of our concentration gradient pilot study and by referring to relevant studies. The control mice received equivalent vehicle. The animals were then housed individually in sterile cages and were provided water and food freely. Mice were monitored for 7 days post wound infection in survival studies. At 24 h and 48 h after the burn sepsis injury was inflicted, 5–8 animals at each timepoint were euthanatized and the lung tissue and blood were sampled for further measurements.
https://pubmed.ncbi.nlm.nih.gov/38267288/

Protocol (for reference only)

Cell Experiment
Cell lines	CEK cells
Preparation method	To further investigate the effect of NAV-2729 on IBV infection, the cells were pre-treated with 10 µM NAV-2729 for 1 h prior to infection with IBV at an MOI of 0.01. Treatment with 10 µM inhibitor continued post-infection. After 48 h, cell supernatants were collected to determine viral titers and genomic copy numbers.
Concentrations	10 µM
Incubation time	48 h

Animal Experiment
Animal models	Male C57BL/6 mice
Formulation	5% dimethyl sulfoxide
Dosages	30 mg/kg
Administration	Intraperitoneal injection

Chemical Information

Molecular Weight	456.88
Formula	C25H17ClN4O3
CAS Number	419547-11-8
Solubility (25°C)	DMSO 46 mg/mL
Storage	Powder          -20°C   3 years ;  4°C   2 years In solvent       -80°C   6 months ;  -20°C   1 month

References

[1] Yoo JH, et al. Cancer Cell. ARF6 Is an Actionable Node that Orchestrates Oncogenic GNAQ Signaling in Uveal Melanoma.