# Clerodane diterpenoid glycosides from the tuberous roots of Paratinospora sagittata: targeted isolation, structure characterization and immunomodulatory properties

## TL;DR

This paper reports the discovery of eight new clerodane diterpenoid glycosides from a plant and their potential to boost immune responses.

## Contribution

The study introduces eight new clerodane diterpenoid glycosides with immunomodulatory properties.

## Key findings

- Eight new clerodane diterpenoid glycosides (tinospinosides F–M) were isolated and structurally characterized.
- Compounds 6 and 7 significantly increased NO production in macrophages.
- Compound 6 enhanced immune cytokine release and activated the NF-κB signaling pathway.

## Abstract

Guided by the MS/MS-based molecular networking, eight previously undescribed clerodane diterpenoid glycosides, designated tinospinosides F–M (1−8), along with 12 known analogues (9−20), were isolated from the tuberous roots of Paratinospora sagittata. Structural elucidation of the undescribed compounds was achieved through comprehensive spectroscopic analyses (NMR, HRESIMS), with their absolute configurations confirmed via single-crystal X-ray diffraction, TD-DFT/ECD computational analyses, and chemical degradation. Immunomodulation evaluation on all the isolates revealed that compounds 6 and 7 exerted significant promoting effect toward NO production in RAW264.7 macrophages. Further study demonstrated that 6 could enhance the release of immune cytokines (e.g., TNF-α) and upregulate the protein expression of iNOS and COX-2, which was potentially mediated through the activation of NF-κB signaling pathway.

The online version contains supplementary material available at 10.1007/s13659-025-00555-2.