NCTD-01

C57BL/6 (female, 5-6 weeks old)^[1]

200 mg/kg

p.o.; daily; for 7 days during DSS induction

In the acute colitis model, mice receiving NCTD-01 exhibited reduced body weight loss, showing a protective trend comparable to that of the group receiving 5-Aminosalicylic acid (5-ASA) (HY-15027).
Notably, NCTD-01 improved survival rates relative to the model group.
Histopathological analysis via H&E staining further confirmed these findings.
The model group exhibited extensive mucosal ulceration, destruction of crypt architecture with abscess formation, marked inflammatory cell infiltration in the lamina propria, and near-complete loss of goblet cells.
In contrast, mice receiving NCTD-01 orally at 200 mg/kg showed reduced ulcerated areas in the colorectal mucosa, more regularly arranged crypts, increased goblet cell numbers, and substantially reduced inflammatory cell infiltration.

BALB/c (male, 5-6 weeks old)^[1]

50 mg/kg; 100 mg/kg; 200 mg/kg

p.o.; daily; for 79 days

After 79 days, mice receiving NCTD-01 maintained higher body weights than the model group throughout the experiment.
By the final day, animals in the model group had lost approximately 15% of their initial body weight.
In contrast, NCTD-01 significantly mitigated this loss, compared with the model group, producing weight increases of 5%-15%.
Survival analysis at day 79 revealed clear differences between groups: the model group survival fell to 56%, and the low-dose NCTD-01 group (50 mg/kg) showed 85% survival.
The 5-ASA group had 92% survival.
No mortality occurred in the medium- and high-dose NCTD-01 groups (100 and 200 mg/kg), nor in the NCTD group.
Alleviated this shortening in a dose-dependent manner; the medium- and high-dose groups showed recovery of colorectal length to 8.92 cm (+14.5% vs model) and 9.42 cm (+20.9% vs model), respectively, compared to the model group.
Histopathological grading showed that the model group and the low-dose NCTD-01 group (50 mg/kg) predominantly developed moderately differentiated adenocarcinoma (G2).
In contrast, medium-dose (100 mg/kg) and high-dose (200 mg/kg) NCTD-01 significantly improved histological outcomes.
Notably, 50% of mice in the high-dose group presented with either normal mucosa or high-grade intraepithelial neoplasia, demonstrating significant suppression of malignant progression.
Dose-dependently suppressed this cellular proliferation, with the 200 mg/kg dose exhibiting the most pronounced inhibitory effect.
NCTD-01 significantly downregulated mRNA levels of IL-1β, COX-2, IL-6, and IL-10 in colorectal tissues, with reductions ranging from 40% to 80% compared to the model group.
TGF-β at 50 and 200 mg/kg showed a downward trend.
NCTD-01 induced dose-dependent decreases in serum IL-1β and TNF-α across the 50, 100, and 200 mg/kg groups, with the most pronounced inhibition at 200 mg/kg.
NCTD-01 significantly attenuated Foxp3 expression and reduced CD25 levels (P<0.05 at 50 mg/kg; P<0.001 at 100 and 200 mg/kg).
NCTD-01 downregulated PD-1 at 200 mg/kg.
NCTD-01 attenuated PD-L1 particularly at 50 and 200 mg/kg.
NCTD-01 markedly increased CD8 in the 200 mg/kg group.