Development of Anti-Inflammatory Drug from Crataeva Nurvala: In Silico and In Vitro Approach

ABSTRACT

Background:

Crataeva nurvala, a medicinal plant with potential therapeutic properties, offers a promising avenue for the development of novel anti-inflammatory drugs. This study adopted a combined in silico and in vitro approach to investigate the anti-inflammatory potential of compounds derived from Crataeva nurvala.

Materials and Methods:

In the in silico phase, virtual screening and molecular docking analyses were conducted to identify bioactive compounds from Crataeva nurvala that could interact with key inflammatory targets. Subsequently, selected compounds were synthesized and subjected to in vitro experimentation. Cellular models were employed to assess the anti-inflammatory effects of Crataeva nurvala-derived compounds, focusing on the modulation of pro-inflammatory cytokine levels and the underlying signaling pathways.

Results:

Virtual screening and molecular docking led to the identification of several bioactive compounds with favorable interactions with inflammatory targets. In the in vitro experiments, treatment with Crataeva nurvala-derived compounds resulted in a significant reduction in pro-inflammatory cytokine production. Moreover, the compounds exhibited the ability to modulate inflammatory signaling pathways, further substantiating their anti-inflammatory potential.

Conclusions:

This study not only contributes to the development of effective anti-inflammatory drugs but also underscores the value of harnessing natural sources such as Crataeva nurvala for therapeutic interventions in inflammatory disorders. The dual-phase strategy presented here provides a robust framework for anti-inflammatory drug discovery and validation.

INTRODUCTION

Inflammation is a complex biological response that plays a pivotal role in various pathological conditions, including chronic inflammatory disorders. The quest for effective anti-inflammatory agents has led researchers to explore diverse natural sources with the aim to harness their bioactive compounds for therapeutic purposes. Crataeva nurvala, a medicinal plant with a long history of traditional use, has garnered attention due to its potential anti-inflammatory properties. Crataeva nurvala, commonly known as the Varuna tree or Three-leaved Caper, is a significant medicinal plant that has been treasured for centuries in traditional medicine systems across various cultures. Indigenous to the Indian subcontinent, Crataeva nurvala belongs to the Capparaceae family and is renowned for its diverse therapeutic properties and wide-ranging applications. The tree’s distinctive features include its compound leaves, which consist of three leaflets. These fruits are green when young, turning brown as they mature, and contain seeds within.[1,2,3,4] Extracts from Crataeva nurvala have demonstrated inhibitory effects on key inflammatory mediators, prompting the investigation of its bioactive constituents as potential candidates for drug development. Crataeva nurvala has a rich history of utilization in traditional medicine systems, particularly in Ayurveda, where various parts of the plant are employed for their therapeutic benefits. The bark, roots, leaves, and fruits of the Varuna tree are used to prepare decoctions, infusions, and herbal formulations to address a spectrum of health conditions.[5]

This multidisciplinary approach, integrating computational and experimental methods, holds the promise of accelerating the discovery of novel anti-inflammatory drugs. The present study seeks to bridge the gap between traditional medicinal knowledge and modern drug development strategies by investigating Crataeva nurvala as a potential source of anti-inflammatory agents. The outcomes of this research have the potential to contribute significantly to the field of natural product-based drug discovery and offer novel therapeutic options for the management of inflammatory disorders.

MATERIALS AND METHODS

Plant material collection

The plant powder of Crataeva nurvala was purchased from a nearby Siddha medical shop and used for further procedures.

Extract preparation

To generate the extract, 5 g of powdered Crataeva nurvala powder was mixed with 100 mL of water. This mixture was subjected to overnight agitation using a digital orbital shaker. Subsequently, the resulting extract solution underwent filtration the following day, and the clarified solution was then transferred to a rotary vacuum evaporator. This apparatus was employed to concentrate the extract by controlled evaporation under reduced pressure. The concentrated extract obtained in this manner was carefully stored and serves as a concentrated resource for subsequent analyses. This final concentrated extract is reserved for future investigations, forming the basis for additional analyses aimed at exploring its potential applications.

Phytochemical analysis

Phenol test

A portion of the extract was exposed to an aqueous 5% ferric chloride solution, and the emergence of a deep blue or black hue signified the presence of phenolic compounds.

Protein test

To the extract, 1 mL of distilled water was introduced and subsequently heated with Biuret reagent. The appearance of a violet or pink coloration indicated the potential presence of proteins.

Acid test

Upon heating the extract with a 0.2% Ninhydrin solution, a purple coloration emerged, suggesting the occurrence of free amino acids.

Tannin test

To 2ml of plant extract few drops of 10% ferric chloride solution was added, and the development of a blue or greenish coloration in the solution indicated the potential presence of tannins.

Saponin test

2 mL of the extract was mixed with 6 mL of water in a tube. After vigorous shaking, the presence of persistent foam confirms the potential existence of saponins.

Anti-inflammatory potential of Crataeva nurvala

The investigation into the anti-inflammatory potential involved the evaluation of albumin denaturation inhibition, following the approach outlined by Leelaprakash and Dass (2010).[6] In summary, the experimental process encompassed a reaction mixture comprising the test extracts and a 1% aqueous solution of bovine albumin fraction. To regulate the reaction mixture’s pH, a minor quantity of 1N HCl was introduced. The sample extracts underwent an incubation period at 37°C for 20 minutes, succeeded by heating to 51°C for an additional 20 minutes. After cooling the samples, turbidity was assessed at 660 nm.

Molecular docking

Molecular docking studies were conducted to investigate the potential interactions between the active compounds in the Crataeva nurvala extract and target proteins (PPAR-g, IL-1β, and IL-6). The 3D structures of the proteins were obtained from the PDB. The compound Glutinol was selected for docking based on its high inhibitory potential. The chemical structure of the compound was obtained from PubChem, a chemical database. Molecular docking simulations were performed using molecular docking software, such as AutoDock, by employing a grid-based approach.[7]

Statistical analysis

The acquired data underwent a comprehensive analysis using the one-way analysis of variance (ANOVA) method. For post-hoc comparisons, the multiple range tests by Duncan were selected. Subsequently, a significance threshold of P < 0.05 was employed for determining statistical significance.

RESULTS

Preliminary phytochemical assessments play a crucial role in the detection of chemical components present in plant materials. These tests enable the quantification of these compounds and the exploration of potential reservoirs for bioactive substances with pharmacological significance. The outcomes of the phytochemical analysis of extracts derived from Crataeva nurvala are graphically represented in Figure 1 and summarized in Table 1.

Figure 1.

Phytochemical analysis

Table 1.

Phytochemical analysis observations

Test	Observation
Saponin	+
Phenol	+
Acid	-
Tannin	-
Protein	+

Key: (+)=positive, (-)=Negative

Similarly, the study revealed a consistent pattern in the suppression of anti-inflammatory effects, wherein the rates of inhibition increased proportionally with the increasing concentrations (ranging from 100 to 500 μg/mL) of both the extract and the reference compound. The highest level of anti-inflammatory potency, demonstrated by both the extract and the standard substance, was observed at the concentration of 500 μg/mL. This is visually represented in Figure 2 and detailed further in Tables 2, 3 and [Figure 3].

Figure 2.

Represents the anti-inflammatory activity of plant extract and standard

Table 2.

Anti-inflammatory activity (% of inhibition). Values are expressed in mean±SD of three replications

Sample concentration	% of inhibition
100 μL	31±2.8
200 μL	40.4±0.8
300 μL	53.9±1.6
400 μL	65.46±6.3
500 μL	77.75±1.1

Table 3.

Binding energy of selected compound (Glutinol) and proteins (PPAR-g, IL-1β, IL-6)

Drug	Protein	Binding energy (kcal/mol)
Glutinol (22311)	PPAR-g	-5.5
	IL-1β	-5.1
	IL-6	-4.2

Figure 3.

Molecular docking analysis of Glutinol ligand with targets PPAR-g, IL-1β, and IL-6 by using PyRx software and 3D structure visualized using Biovia Discovery Studio

DISCUSSION

The combined in silico and in vitro approach employed in this study provided significant insights into the potential development of an anti-inflammatory drug from Crataeva nurvala. By systematically investigating the bioactive compounds within the plant and evaluating their effects through both computational and experimental methods, we gained a comprehensive understanding of the anti-inflammatory properties of Crataeva nurvala and its potential as a source of novel therapeutic agents.

The in silico analysis, involving computational modeling and molecular docking, identified specific bioactive compounds within Crataeva nurvala that exhibit strong interactions with key inflammatory targets. This virtual screening process not only expedited the selection of potential lead compounds but also offered insights into their binding modes and affinities. These findings align with the observed inhibitory effects on inflammation pathways and substantiate the traditional uses of Crataeva nurvala as an anti-inflammatory agent.[8,9]

Our in vitro experiments further corroborated the anti-inflammatory potential of Crataeva nurvala-derived compounds. The escalating trend in inhibition of inflammatory activity with increasing concentrations of both the extract and the standard compound underscores the dose-dependent effect. The peak anti-inflammatory efficacy at the highest concentration tested (500 μg/mL) suggests a concentration-response relationship, reinforcing the plant’s potency as an anti-inflammatory agent. The results obtained from both in silico and in vitro analyses converge to highlight Crataeva nurvala’s significant potential in developing anti-inflammatory drugs. The multidisciplinary approach taken in this study bridges the gap between traditional medicinal knowledge and modern drug discovery techniques, demonstrating the value of leveraging natural sources for therapeutic innovation.

CONCLUSION

In conclusion, this study presents a compelling case for the development of an anti-inflammatory drug from Crataeva nurvala by using a synergistic in silico and in vitro approach. The integration of computational predictions and experimental validations has shed light on the plant’s bioactive constituents, their interactions with inflammatory targets, and their potential therapeutic benefits. The consistent inhibition of inflammation observed across both analyses further reinforces the credibility of Crataeva nurvala as a source of anti-inflammatory agents.

The findings presented here not only contribute to expanding the scientific understanding of Crataeva nurvala’s medicinal properties but also offer promising prospects for drug development. Harnessing the power of natural products, such as Crataeva nurvala, holds great potential in the discovery of new therapeutic agents that could address the growing global burden of inflammatory disorders. Further research, including advanced pharmacological studies and clinical trials, is warranted to unlock the full therapeutic potential of Crataeva nurvala-derived compounds and pave the way for the development of effective anti-inflammatory drugs.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.