Article Figures & Data
Figures
- Fig. 1.
Chemical structure and in vitro activity of CH6824025 on DDR1 phosphorylation. (A) Chemical structure of CH6824025. (B) Concentration-dependent inhibition of DDR1 phosphorylation by CH6824025 in U2OS cells. Each dot represents the mean of triplicate measurement.
- Fig. 2.
PD evaluation of CH6824025 in UUO mice. (A) Western blotting of pDDR1(pY513), DDR1, and GAPDH in the kidney. The normalized peak area of (B) pDDR1(pY513) and (C) DDR1. The actual values are represented as dots and the columns represent mean (n = 3). *P < 0.05, significant difference versus UUO mice treated with vehicle (one-way ANOVA with posthoc Dunnett’s multiple comparisons test). Sham: sham-operated mice. Vehicle: UUO mice treated with vehicle. 50 mg/kg or 200 mg/kg: UUO mice treated with CH6824025 50 mg/kg or 200 mg/kg.
- Fig. 3.
Effect of CH6824025 on fibrosis in UUO mice. (A) Hydroxyproline amounts in the kidney. (B) Sirius Red positive area of kidney specimen. (C) The ratio of tubular dilation in the kidney. The actual values are represented as dots, and the columns represent mean (n = 3 [sham], 10 [vehicle or 200 mg/kg]). *P < 0.05, significant difference versus UUO mice treated with vehicle (one-way ANOVA with posthoc Dunnett’s multiple comparisons test). Sirius Red stained kidney of (D) sham mouse, (E) vehicle-treated UUO mouse, and (F) CH6824025 200 mg/kg-treated UUO mouse. Arrowheads indicate accumulation of collagen fibers. H&E stained kidney of (G) Sham mouse, (H) vehicle-treated UUO mouse, and (I) CH6824025 200 mg/kg-treated UUO mouse. Arrowheads indicate dilation of tubule. Sham: sham-operated mice. Vehicle: UUO mice treated with vehicle. 200 mg/kg: UUO mice treated with CH6824025 200 mg/kg.
- Fig. 4.
Effect of CH6824025 on the mRNA expression of fibrotic and proinflammatory markers in UUO mice. Relative (A) COL1A1, (B) COL3A1, (C) α-SMA, (D) TGF-β1, (E) TIMP2, (F) CCL2, (G) tumor necrosis factor-α, and (H) CD3E mRNA expression in the kidney. Expression levels are given as ratios to Mrpl19. The actual values are represented as dots, and the columns represent mean (n = 3 [sham], 10 [vehicle or 200 mg/kg]). *P < 0.05, significant difference versus UUO mice treated with vehicle (one-way ANOVA with posthoc Dunnett’s multiple comparisons test). Sham: sham-operated mice. Vehicle: UUO mice treated with vehicle. 200 mg/kg: UUO mice treated with CH6824025 200 mg/kg.
- Fig. 5.
Effect of CH6824025 on F4/80+ macrophages in UUO mice. (A) F4/80 positive area of kidney specimen. The actual values are represented as dots, and the columns represent mean (n = 3 [sham], 10 [vehicle or 200 mg/kg]). *P < 0.05, significant difference versus UUO mice treated with vehicle (one-way ANOVA with posthoc Dunnett’s multiple comparisons test). F4/80-stained kidney of (B) sham mouse, (C) vehicle-treated UUO mouse, and (D) CH6824025 200 mg/kg-treated UUO mouse. Sham: sham-operated mice. Vehicle: UUO mice treated with vehicle. 200 mg/kg: UUO mice treated with CH6824025 200 mg/kg.
- Fig. 6.
10x Visium ST analysis of UUO mouse kidneys treated with CH6824025. DDR1 mRNA expression in the kidney of (A) sham mouse, (B) vehicle-treated UUO mouse, and (C) CH6824025 200 mg/kg-treated UUO mouse. Selected spots as DDR1 mRNA-positive spots in the kidney of (D) sham mouse, (E) vehicle-treated UUO mouse, and (F) CH6824025 200 mg/kg-treated UUO mouse. Red or blue spots indicate DDR1 positive or negative spots, respectively.
- Fig. 7.
Ingenuity pathway analysis of DDR1 positive spots in the kidney of UUO mice treated with CH6824025. (A) Canonical pathway analysis that shows the results of the top 10 pathways. The bar chart is colored based on the z-score. (B) Analysis of disease and functions. The heatmap is colored based on the z-score. The color reflects the direction of change in the biological process. Orange color represents a trend to increase; blue represents a trend to decrease in UUO mice treated with CH6824025 compared with Vehicle-treated UUO mice. Gray indicates that no activity prediction can currently be made. White indicates (1) pathways with z-scores at or very close to 0; or (2) those that are ineligible for analysis because there are fewer than four analysis-ready molecules in the dataset associated with the pathway. The size of each rectangle indicates the P-value significance. The yellow rectangle highlights “cellular movement.”
Tables
- TABLE 1
Concentration of CH6824025 in plasma of individual animals at 4 and 24 h after administration
CH6824025 Time (h) Concentration (ng/ml) Mean SD 50 mg/kg oral administration 4 4260 3900 4150 3720 4010 240 24 164 168 126 123 145 24 200 mg/kg oral administration 4 8040 6060 6690 6630 6860 840 24 3620 4060 3380 3100 3540 410
Data Supplement
- Supplemental Figure -
Supplemental Figure 1. Relative DDR1 mRNA expression in the kidney at day 1 and 7 after the surgery.
- Supplemental Figure -
Supplemental Figure 2. Histopathological images before and after classification of HALO AI v3.4.
- Supplemental Figure -
Supplemental Figure 3. Analysis of DDR1 expression spots in 10x Visium ST analysis.
- Supplemental Table -
Supplemental Table 1. The list of the top 50 upregulated genes and the top 50 downregulated genes in CH6824025 group compared to Vehicle.
- Supplemental Figure -
