Molecular Insight into the Pharmacological Potential of Clerodendrum minahassae Leaf Extract for Type-2 Diabetes Management Using the Network Pharmacology Approach

Abstract

Background and Objectives: The increasing occurrence and prevalence of type-2 diabetes mellitus (T2DM) have led to a growing interest in researching available treatment alternatives. Clerodendrum minahassae, a native plant species of North Sulawesi, has been a focus of ethnopharmacological studies due to its significance contributions to drug development, particularly its potential antidiabetic properties. This study investigated the pharmacological potential of Clerodendrum minahassae (CM) leaf extract for managing type-2 diabetes (T2DM) using a network pharmacology approach. Materials and Methods: Active compounds were extracted from CM leaves, and their interactions with target proteins in T2DM were explored through various in silico analyses. Results: SAR analysis using Way2Drug Pass Online identified 29 bioactive CM leaf extract compounds with promise as T2DM treatments. Additionally, 26 of these met Ro5 criteria for favorable drug-likeness. Most compounds exhibited positive pharmacodynamic and pharmacokinetic profiles, with 22 considered safe, while 7 posed potential toxicity risks when ingested individually. CM leaf extract targeted 60 T2DM-related proteins, potentially affecting T2DM via cytokine regulation, particularly in proteins linked to metabolic processes, cellular response to angiotensin, and the sphingosine-1-phosphate signaling pathway. The network pharmacology analysis identified five genes targeted by CM leaf extract, namely, STAT3, MAPK1, ESR1, PIK3R1, and NFKB1. Among these genes, PIK3R1's interaction with the insulin receptor (INSR) positions it as a crucial candidate gene due to its pivotal role in insulin signal transduction during T2DM development. Conclusions: This research sheds light on the therapeutic potential of CM leaf extract for treating T2DM. This potential is attributed to the diverse array of bioactive compounds present in the extract, which have the capacity to interact with and inhibit proteins participating in the insulin signal transduction pathway crucial for the progression of T2DM. The findings of this study may open up possibilities for future applications of CM leaf extract in the development of novel T2DM treatments.

Keywords: Clerodendrum minahassae; PIK3R1; insulin signal transduction; network pharmacology; type-2 diabetes.

Figure 1

The evaluation of CM leaf extract’s potential for T2DM treatment based on structure–activity relationship (SAR) predictions. The assessment included its impact as an insulin promoter, Nrf2 stimulant, and wound healing agent. Notably, compound C16 (α-amyrin) demonstrated the most consistent effects.

Figure 2

SAR-based prediction of CM leaf extract’s potential for treating T2DM, particularly as an insulin promoter. Seven compounds exhibited Pa values greater than 0.4. Remarkably, compound C16 stood out with the highest predicted value as an insulin promoter (Pa score of 0.75) and as an Nrf2 stimulant (Pa score of 0.46).

Figure 3

Prediction of compound drug-likeness within the CM leaf extract using Lipinski’s Rule of Five (Ro5). The bars outlined in black indicate the seven compounds selected for additional analysis. Out of the 29 bioactive compounds assessed, 26 were found to conform to the Ro5 criteria, indicating their potential as drug-like compounds.

Figure 4

Toxicity prediction based on LD₅₀ and toxicity classes of each compound detected in CM leaf extract. The black-filled labels indicate the seven compounds that are the focus of further analysis. Among the 22 bioactive compounds, the average LD₅₀ and toxicity classes spanned from Class IV to Class VI, indicating that these compounds are predicted to have a low toxicity level and are considered safe.

Figure 5

Venn diagram displaying the shared targets between the CM leaf extract and genes linked to T2DM. The CM leaf extract is suggested to have the potential to impact T2DM by modulating cytokines. This is supported by the fact that a significant number of proteins associated with the immune system are targeted by the extract, and these proteins are also associated with T2DM.

Figure 6

Annotation of gene ontology biological processes for CM leaf extract targets with false discovery rate (FDR) < 0.05. The enrichment score shows CM leaf extract primarily targets metabolic regulation, angiotensin response, and immune pathways. Angiotensin response is closely related to diabetes, while the extract’s main role appears to be in inflammation. Genes associated with angiotensin response, like PRKCD, RELA, NFKB1, and NFE2L2, are also involved in immune responses.

Figure 7

Annotation of the top 25 KEGG pathways targeted by CM leaf extract with FDR < 0.05. The most significant pathway targeted by CM leaf extract is the AGE-RAGE signaling pathway in diabetic complications. This pathway involves various proteins, including CDK4, STAT3, PRKCD, MAPK1, PIK3CD, PRKCA, PIK3R1, F3, RELA, and NFKB1. Some of these proteins are associated with inflammation, while others are involved in the PI3K receptor pathway related to T2DM. RAGE, a receptor for AGEs, plays a crucial role in this pathway, contributing to cellular stress and complications in diabetes.

Figure 8

Protein–protein interaction (PPI) of CM leaf extract targets in T2DM. PIK3R1 is recognized as a prospective target of the extract, demonstrating its ability to interact with the insulin receptor (INSR). The node color intensity indicates the extent of protein interactions. This implies that the CM leaf extract is involved in pathways like PI3K/AKT signaling. Moreover, it affects inflammatory proteins, which are known to influence insulin signaling.