Preclinical Efficacy and Safety Studies of Formulation SSV-003, a Potent Anti-Viral Herbal Formulation

Figures & data

Table 1 IC₅₀, Percent Cytotoxicity, and Percent CPE of Test Compounds (Influenza A-H1N1-VR219)

S. No	Compound	IC50 (µg/mL) against H1N1	% Cytotoxicity against MDCK Cells at 50 µg/mL	% CPE against H1N1) at 50 µg/mL
1	SSV-003	<0.39*	0	0
2	Vitamin C (Commercial preparation)	>50**	0	91.1
3	Ribavirin (Control)	1.38	0	0

Notes: *IC₅₀ is lower than the lowest concentration tested, **IC₅₀ is higher than the highest concentration tested.

Table 2 Percent CPE of Test Compounds at Different Tested Concentrations (Influenza A-H1N1-VR219)

S. No	Compound	% CPE at Tested Concentrations (µg/mL)
		50	25	12.5	6.25	3.125	1.5625	0.7813	0.39
1	SSV-003	0	0	0	0	0	0	0	0
2	Vitamin C (commercial preparation)	91.1	91.3	93	92.9	92.5	91.8	92.8	93.7
3	Ribavirin (Control)	0	0	0	0	14.7	49.3	69.9	95.9

Table 3 IC₅₀, Percent Cytotoxicity, and Percent CPE of Test Compounds (Beta Corona-OC43)

S. No	Compound	IC50 (µg/mL) against Betacorona (OC43)	% Cytotoxicity against HCT-8 Cells at 50 µg/mL	% CPE against Betacorona (OC43) at 50 µg/mL
1	SSV-003	2.26	0	0
2	Vitamin C (commercial preparation)	>50**	0	93
3	Ribavirin (Control)	2.25	0	0
4	Remdisivir (Control)	6.23	0	0

Note: **IC50 is higher than the highest concentration tested.

Table 4 Percent CPE of Test Compounds at Different Tested Concentrations (Beta Corona-OC43)

S. No	Compound	% CPE at Tested Concentrations (µg/mL)
		50	25	12.5	6.25	3.125	1.5625	0.7813	0.39
1	SSV-003	0	0	0	0	53.83	71.56	69.9	83.70
2	Vitamin C (commercial preparation)	93	94	93.34	93.42	94.29	94	93.65	94.56
3	Ribavirin	0	0	0	0	14.7	49.3	69.9	95.9
4	Remdisivir	0	0	31	51	79	86	92	96.0

Table 5 Median Tissue Culture Infectious Dose in Mice Lungs following Treatment with Test Compounds

Group	Treatment	Dose/Route*	TCID50 Titer (Mean±SD)	% Drop in Infection Compared to Infection CTRL
G1	Infected control	Vehicle (CMC)/Oral	4.61±0.20	0%
G2	Remdisivir	25 mg/kg SC	2.21±0.21*	99.6%
G3	SSV-003	500 mg/kg Oral	3.76±0.50*^$	85.9%
G4	SSV-003	1,000 mg/kg Oral	3.34±0.26*^$	94.6%
G5	SSV-003	1,500 mg/kg Oral	3.30±0.55*^$	95.1%

Notes: All values are expressed as the mean±SD, Significant differences between the treated group and the Virus control were analyzed by One-way ANOVA, followed by a Dunnett’s multiple comparison test, using GraphPad Prism at 95% confidence levels. *P<0.05 vs Virus control and ^$P<0.05 vs Remdesivir.

Figure 1 The effect of SSV-003 on cellular biomarker CD4+ after treatment. All values are expressed as the mean±SD (n=3, pooled samples of three animals in triplicate). Significant differences between the treated group and Virus control were analyzed by One-way ANOVA, followed by a Dunnett’s multiple comparison test, using GraphPad Prism at 95% confidence levels. *P<0.05 vs Virus control and ^$P<0.05 versus Remdesivir.

Figure 2 The effect of SSV-003 on cellular biomarker CD8+ after treatment. All values are expressed as the mean±SD (n=3, pooled samples of three animals in triplicate). Significant differences between the treated group and Virus control were analyzed by One-way ANOVA, followed by a Dunnett’s multiple comparison test, using GraphPad Prism at 95% confidence levels. *P<0.05 vs Virus control and ^$P<0.05 versus Remdesivir.

Table 6 Percent Increase in Immune Parameters as Compared to the Virus Control Group

	Remdisivir (% Increase)	SSV-003 (500 mg/kg) (% Increase)	SSV- 003 (1,000 mg/kg) (% Increase)	SSV-003 (1,500 mg/kg) (% Increase)
IgG	4.02	8.76	17	16
IgM	5.13	5.13	23.08	20.51
CD4	15.48	37.63	75.76	76.77
CD8	22.19	58.36	64.27	66.11